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A Method For The Rapid Determination Of Alkaline Phosphatase With 5 Cubic Millimeters Of Serum
... The heparinised blood samples were centrifuged at 300 rpm for 10 minutes and the serum collected for analysis. AST and ALT were analysed according to the method of [30] ; ALP according to [31] ; urea according to [32] and creatinine according to the method of [31] . The organ indices were calculated according to the methods of [32] . ...
... The heparinised blood samples were centrifuged at 300 rpm for 10 minutes and the serum collected for analysis. AST and ALT were analysed according to the method of [30] ; ALP according to [31] ; urea according to [32] and creatinine according to the method of [31] . The organ indices were calculated according to the methods of [32] . ...
... Bioassay was performed with instars of test organisms using fractions. Two-day-old fourth instars larvae of treated S. litura and H. armigera were used to quantify the enzyme activities, such as estimation of acid phosphatase (ACP) and alkaline phosphatases (ALP) [19], estimation of adenosine triphosphatase (ATPase) [20] and estimation of lactate dehydrogenase (LDH). Fresh castor and cotton leaves were sprayed with fractions on both surfaces and let to air dry, whereas control leaves were treated with isopropanol and left to air dry. ...
Our study specifically delves into the synthesis of AgNPs utilizing Tinospora malabarica leaf extract fractions (Tm-LMEFs) and explore their efficacy in combating the larvae have progressed to the synthesized AgNPs (Tm-AgNPS) and the leaf methanol extract fractions (LMEFs), our investigation focused on their impact on the targeted larvae species over a 24-h exposure period. Significantly noteworthy mortality rates were observed across all Spodoptera litura and Helicoverpa armigera post-treatment, indicating pronounced efficacy. Notably, the Tm-AgNPS exhibited higher toxicity compared to the LMEFs these selected polyphagous insect pests. Quantitative analysis yielded the highest mortality (98.99%/90.44% and 55.33%/49.60%) values for the synthesized Tm-AgNPs against S. litura and H. armigera, with no observed mortality in the control group. This study marks the first instance of exploring both Tm-LMEFs, Tm-AgNPs and enzyme activity have toxicity properties against selected agriculture pests, highlighting their promising role in pest management and environmental sustainability. ARTICLE HISTORY
... Seminal plasma samples were analyzed for total protein content (TPC) according to the procedure proposed by Weichselbaum (1946). The activity of acid (AcP) and alkaline phosphatase (AP) was measured according to the method described by Bessey et al. (1946). ...
The aim of this study was to evaluate the quality parameters and selected biochemical markers of canine semen sampled at 24-h intervals over a period of 5 days, preceded by 6 months of sexual abstinence. Full ejaculates were obtained from 6 dogs. Ejaculate volume and total sperm counts in the ejaculate decreased gradually on successive sampling days. The percentage of total motile spermatozoa (TMOT), percentage of progressively motile spermatozoa (PMOT), sperm plasma membrane integrity (SPMI), and sperm mitochondrial membrane potential (MMP) increased on successive days of sampling. In addition, ATP content increased in spermatozoa. Total protein content (TPC) and the activity of aspartate aminotransferase (AAT), alkaline phos-phatase (AP), and acid phosphatase (AcP) decreased in seminal plasma. Repeated ejaculation over a period of 5 days induced changes in the qualitative and quantitative parameters of canine semen. A decrease in the values of some biochemical markers of semen, secreted by the epididymis and the prostate gland, could point to disturbances in the secretory activity of these organs. Canine semen sampled after prolonged sexual abstinence is generally characterized by less desirable quality parameters, and this observation should be taken into consideration when semen is collected for artificial insemination or preservation. Semen quality can be significantly improved by repeating the sampling procedure after 24 hours. One the other hand, repeated sampling on successive days can significantly decrease total sperm counts in the ejaculate. As a result, a sufficient number of semen doses for artificial insemination may not be obtained from a single ejaculate.
... Bioassay was performed with instars of test organisms using fractions. Two-day-old fourth instars larvae of treated S. litura and H. armigera were used to quantify the enzyme activities, such as estimation of acid phosphatase (ACP) and alkaline phosphatases (ALP) [19], estimation of adenosine triphosphatase (ATPase) [20] and estimation of lactate dehydrogenase (LDH). Fresh castor and cotton leaves were sprayed with fractions on both surfaces and let to air dry, whereas control leaves were treated with isopropanol and left to air dry. ...
Our study specifically delves into the synthesis of AgNPs utilizing Tinospora malabarica leaf extract fractions (Tm-LMEFs) and explore their efficacy in combating the larvae have progressed to the synthesized AgNPs (Tm-AgNPS) and the leaf methanol extract fractions (LMEFs), our investigation focused on their impact on the targeted larvae species over a 24-h exposure period. Significantly noteworthy mortality rates were observed across all Spodoptera litura and Helicoverpa armigera post-treatment, indicating pronounced efficacy. Notably, the Tm-AgNPS exhibited higher toxicity compared to the LMEFs these selected polyphagous insect pests. Quantitative analysis yielded the highest mortality (98.99%/90.44% and 55.33%/49.60%) values for the synthesized Tm-AgNPs against S. litura and H. armigera, with no observed mortality in the control group. This study marks the first instance of exploring both Tm-LMEFs, Tm-AgNPs and enzyme activity have toxicity properties against selected agriculture pests, highlighting their promising role in pest management and environmental sustainability.
... The ALP was quantified following the method of Bessey et al. (1946) with slight modifications. A working substrate solution was prepared by dissolving an equal amount of glycine buffer (pH 10.5) and substrate stock solution (7.6 mM p-nitrophenyl phosphate). ...
Blood feeding and digestion are vital physiological activities essential for the survival and reproduction of ticks. Chemical acaricides viz., ivermectin, amitraz and fipronil, are known to act on the central nervous system, resulting in the mortality of ticks. The present study is focused on the effect of these acaricides on the midgut and gut enzymes of Rhipicephalus microplus. The ultra-thin sections of midgut of ivermectin-treated ticks showed irregular basal membrane and ruptured digestive vesicles. Amitraz treatment resulted in a notable decrease in digestive cells with pleats in the basal membrane, while fipronil-exposed ticks exhibited reduced digestive cells, loss of cellular integrity, and disintegration of the basal membrane and muscle layer. The gut tissue homogenate of ivermectin and fipronil treated ticks showed a significant reduction of cathepsin D level, 76.54 ± 3.20 μg/mL and 92.67 ± 3.72 μg/mL, respectively, as compared to the control group (150.0 ± 3.80 μg/mL). The leucine aminopeptidase level (4.27 ± 0.08 units/mL) was significantly decreased in the ivermectin treated ticks compared to other treatment groups. The acid phosphatase activity (29.16 ± 0.67 μmole/min/L) was reduced in the ivermectin treated group whereas, increased activity was observed in the fipronil and amitraz treated groups. All the treatment groups revealed increased alkaline phosphatase levels (17.47–26.72 μmole/min/L). The present finding suggests that in addition to the established mechanism of action of the tested acaricides on the nervous system, the alterations in the cellular profile of digestive cells and enzymes possibly affect the blood digestion process and thus the synthesis of vital proteins which are essential for vitellogenesis, and egg production in ticks.
... Activities of antioxidant [25] catalase, [26] glutathione reductase, [27] glutathione peroxidase, [28] and levels of hydrogen peroxide generation, [29] lipid peroxidation [30] were determined in gill, liver and brain tissues. Activities of marker enzymes namely alkaline phosphatase [31] in gill and liver tissues, and acetylcholinesterase [32] activity in brain tissue were also analyzed. ...
... Levels of hydrogen peroxide generation were assayed by the method of Pick and Keisari (1981), levels of lipid peroxidation were measured by the method of Ohkawa et al., 1979. Alkaline phosphatase was measured as described by Bessey et al., 1946. ...
The aim of the present study was to evaluate the toxicity effect of chlordecone on the reactive oxygen species generation in the Cichlid fish, Etroplus maculatus. Chlordecone (0.35 µg/L) was used as the test dose and the fishes were treated for 24, 48 and 96 h maintaining a control group without adding the test chemical. Body weight of the treated fishes remained unchanged throughout the experiments. However, mucous deposition was significantly increased in time-dependent manner which states the defensive mechanism of the exposed fishes to chlordecone. Antioxidant parameters such as superoxide dismutase, catalase, glutathione reductase and the levels of hydrogen peroxide and lipid peroxidation were evaluated in both gill and liver tissues. Statistical analysis reports that chlordecone induced oxidative stress on gill as well as in liver by significant (p<0.05) reduction in the activities of antioxidant enzymes with concomitant increase in the levels of hydrogen peroxide and lipid peroxidation. The present study also showed a significant reduction in the marker enzyme, alkaline phosphatase in gill and liver and it could be due to decreased state of inter and intracellular membrane transport and possibly this could be also due to the toxicity of chlordecone. Histopathological observations reveal chlordecone-treated gill with destruction of primary and secondary lamellae, upliftment of gill epithelium and reduction in the number of chloride cells. Similarly exposure to chlordecone critically affected the architecture of hepatocytes with enucleated cells, cytoplasmic vacuolization and hepatic necrosis. Thus toxicity of chlordecone by reducing the activities of antioxidant enzymes resulted in oxidative imbalance in the vital organs as gill and liver of the fish, Etroplus maculatus.
... Levels of hydrogen peroxide generation [23] and lipid peroxidation [24] were also evaluated. The activity of alkaline phosphatase [25] was analysed in the gill and liver tissue whereas the activity of acetylcholinesterase [26] was measured in the brain tissue of the fish. ...
... According to Rheinhold and Seligron 29 , Bessey et al. 30 , Zimmerman and Weinstein 31 , and Reitman and Frankel 32 , respectively, the total protein, alkaline phosphatase (ALP), lactate dehydrogenase (LDH), aspartate transaminase (AST), and alanine transaminase (ALT) in each rat's serum were estimated using a UVPC spectrophotometer (Jasco V-730, serial No. A 112,361,798, Japan). ...
Tomato pomace (TP), an antioxidant-rich byproduct, may be suitable for noble applications. The regulation of ROS generation and the anti-inflammatory response can help to prevent ulceration. The purpose of this study was to examine TP for antioxidants, in silico anti-inflammatory properties, and its potential to protect against ulceration and erosion triggered by indomethacin. Tomato pomace extract (TPE) was encapsulated either alone or with probiotics to maximize its potential effect. These microcapsules were investigated in indomethacin-treated rats. TPE demonstrated antioxidant activity as well as high levels of carotenoids (15 mg/g extract) and polyphenols. Because of their binding affinity as well as hydrophobic and hydrogen bond interactions with the active sites of TNF-α and IL-1β inflammatory cytokines, ellagic acid and rutin may be implicated in the anti-inflammatory effect of TPE, according to the docking study. TPE microcapsules, either alone or in combination with probiotics, demonstrated a protective effect against enterocolitis by reducing oxidative stress and inflammation, as evidenced by the decrease in stomach and intestinal MDA, NO, IL-1β, IL-6, and TNF-α levels and the increase in CAT, SOD, and GSH activities. The produced microcapsules are suggested to be promising candidates for protection against gastric ulcers and erosion.
... Where indicated, reactions included 20 U/mL superoxide dismutase and/or 30 U/mL catalase. At time points 50 μl aliquots were removed to 1 mL 1 mM p-nitrophenylphosphate in 1 M Tris, pH 8. AP activity was measured based on its ability to hydrolyze p-nitrophenylphosphate to p-nitrophenol, a chromogenic product that absorbs at 405 nm (Bessey et al., 1946). All reactants were incubated with reduced AP at 37°C, and AP was assayed at different time points at room temperature. ...
The oxidizability of cysteine residues is exploited in redox chemistry and as a source of stabilizing disulfide bonds, but it also raises the possibility that these side chains will be oxidized when they should not be. It has often been suggested that intracellular oxidative stress from hydrogen peroxide or superoxide may result in the oxidation of the cysteine residues of cytoplasmic proteins. That view seemed to be supported by the discovery that one cellular response to hydrogen peroxide is the induction of glutaredoxin 1 and thioredoxin 2. In this study we used model compounds as well as alkaline phosphatase to test this idea. Our results indicate that molecular oxygen, superoxide, and hydrogen peroxide are very poor oxidants of N-acetylcysteine and of the protein thiols of alkaline phosphatase in vitro. Copper could accelerate thiol oxidation, but iron did not. When alkaline phosphatase was engineered to remain in the cytoplasm of live cells, unnaturally high concentrations of hydrogen peroxide were required to oxidize it to its active, disulfide-dependent form, and toxic levels of superoxide had no effect. At the same time, far lower concentrations of these oxidants were sufficient to poison key metalloenzymes. The elimination of glutaredoxin 1 and thioredoxin 2 did not change these results, raising the question of why E. coli induces them during peroxide stress. In fact, when catalase/peroxidase mutants were chronically stressed with hydrogen peroxide, the absence of glutaredoxin 1 and thioredoxin 2 did not impair growth at all, even in a minimal medium over many generations. We conclude that physiological levels of reduced oxygen species are not potent oxidants of typical protein thiols. Glutaredoxin and thioredoxin must either have an alternative purpose or else play a role under culture conditions that differ from the ones we tested.
... The protein concentration was measured spectrophotometrically by light absorption at a wavelength of λ = 546 nm and expressed in g/L. 23 The activity of alkaline phosphatase (AP) was determined using a spectrophotometric method based on the property of AP to hydrolyse the ether bond in β-glycerophosphate, releasing phosphoric acid. The generated phosphate content was determined using a reaction with a molybdenum reagent in the presence of ascorbic acid. ...
Sex-related variances in drug metabolism provide a foundation for refining treatment protocols for prevalent conditions based on the patient's sex. Tailoring treatment strategies based on sex is particularly noteworthy among patients with comorbid illnesses due to the potential for drug interactions and the impact of concurrent diseases on clinical outcomes. Aim of this study was to assess the hepatotropic effects of antiulcer drugs (esomeprazole, clarithromycin and metronidazole-E/C/M) and placenta cryoextract (CEP) within a simulated model of tetrachloromethane (CCl 4)-induced hepatitis combined with underlying ethanol-induced liver cirrhosis (EILC), with a focus on the role of subjects' sex. Methods: Using 112 male and female rats, the research explored the effects of different sex hormone levels. Chronic EILC was induced by administering a 50.0 % CCl 4 oil solution (8 mL/kg) twice a week, combined with a 5.0 % ethanol solution, over 45 days. Total protein (TP) levels and alkaline phosphatase (AP) activity were measured spectrophotometrically. Results: The research findings indicate that the onset of EILC and the administration of E/C/M resulted in a significantly greater 10.8 % (p = 0.03) reduction in TP levels among females compared to males, without altering hormonal status. Introducing CEP led to a noteworthy (p < 0.001) rise in TP levels, by 30.8 % in males and 33.9 % in females, in the context of EILC and E/C/M administration, while maintaining hormonal status. Among male rats, the most elevated AP activity was observed with excess testosterone propionate administration (5.0 [5.0; 5.9] μmol/L), while the lowest level was recorded in rats after testectomy, measuring 3.8 [2.5; 4.7] μmol/L, exhibiting a significant 20.8 % decrease (p < 0.05) compared to male rats without hormonal status changes. In female rats, the study revealed that against the backdrop of EILC and E/C/M administration, the highest AP level was seen in ovariectomised females, reaching 5.8 [5.1; 6.2] μmol/L, reflecting a substantial 9.4 % increase compared to rats without hormonal status changes. Conclusions: The administration of CEP under similar experimental conditions led to the recovery of the liver's protein-synthesising function in both male and female rats. When female sex hormones were introduced to sham-operated female rats, a significant 20.8 % greater reduction in AP levels was observed. Additionally, gonadectomy led to a more pronounced decrease in this enzyme's levels in male rats compared to female rats, indicating the cytoprotective properties of female sex hormones.
Hladkykh FV, Koshurba IV, Komorovsky RR, Chyzh MO, Koshurba YuV, Marchenko MM. Sex differences in the hepatotropic effects of antiulcer drugs and placenta cryoextract in an experimental rat liver injury model. Scripta Medica. 2023; 54 (4): 363–370. DOI: https://doi.org/10.5937/scriptamed54-46076
... The gut's brush border was isolated as recommended by Gisbert et al. [48] and Crane et al. [49]. The evaluation of digestive enzymes and soluble protein [50] was determined by standard methods for the following enzymes: trypsin [51], chymotrypsin [52], protease [53,54], ALP [55], α-amylase [56], and lipase [57]. ...
A 16-week feeding trial was done to examine the impacts of continuous feeding (CF) or pulse-feeding (PF) of multi-strain probiotics on Asian seabass (Lates calcarifer, 30.0 ± 0.1 g) juveniles. In this study, three different multi-strain probiotic mixtures were added to a basal diet, including (I) a mixture of different strains of Lactobacillus plantarum, (II) a mixture of the first probiotic (I) + L. delbrueckii sub bulgaricus, L. rhamnosus and L. acidophilus, and (III) a mixture of the second probiotic (II) + two quorum quenching (QQ) bacteria (Bacillus thuringiensis QQ1 and B. cereus QQ2). CF (every day) or PF (every two weeks) strategies were applied for using the abovementioned probiotics to design seven experimental groups including C (control, without probiotics), CF-I (continuous feeding of fish with the probiotic mixture I), CF-II (continuous feeding of fish with the probiotic mixture II), CF-III (continuous feeding of fish with the probiotic mixture III), PF-I (pulse-feeding of fish with the probiotic mixture I), PF-II (pulse-feeding of fish with the probiotic mixture II), and PF-III (pulse-feeding of fish with the probiotic mixture III). Four hundred and twenty fish were stocked into 21 circular polyethylene tanks with 220 L volume (20 fish/tank). Each dietary treatment had three replicates. Tanks were supplied with seawater (temperature = 30.5 °C, salinity = 45 g L⁻¹) in a flow-throw system. Fish in CF-I, CF-II, and CF-III had higher growth rate (ca. 113–145%) and better feed conversion ratio than fish fed C and PF-I (P < 0.05). Fish in the CF-III group had the highest protease activity. Continuous feeding strategy resulted in a higher amount of glutathione and catalase activities in both the liver and plasma as well as higher superoxide dismutase activity in the liver of fish. Pulse-feeding strategy resulted in lower plasma lactate dehydrogenase and aspartate aminotransferase levels than the CF strategy. Regardless of feeding strategy, different probiotic mixtures significantly enhanced blood hemoglobin and hematocrit levels compared to the control. Continuous feeding with the multi-strain probiotics resulted in a higher survival rate against Vibrio harveyi than the PF method. Continuous feeding induced higher mRNA transcription levels of granulocyte-macrophage colony-forming cells and interleukin 10 genes in the gut of fish than PF strategy. In conclusion, continuous feeding with multi-strain probiotics is better than pulse-feeding on growth, feed utilization, antioxidant capacity, and the gut’s immune-related genes and led to higher resistance of L. calcarifer in challenge with V. harveyi.
... The estimation of acid, alkaline and lactate dehydrogenase were carried out with the adapted methodology Bessey et al. (1946). The substrate buffered was incubated with tissue extract for 30 min. ...
22 Dec 2023): In-vitro and in-silico analysis of methanolic crude extracts of Mountain knotgrass Aerva lanta (L.) against two lepidopteran pests and non-target species, Toxin Reviews, ABSTRACT The present research investigates the toxicological screening of crude methanol extracts Avera lanata (Mx-Al) traditionally used for its medicinal properties, and explored its potential as an insecticidal agent against the lepidopteran pests Spodoptera litura Fab. and Plutella xylostella L. and their non-target impact on Eudrilis eugeniae. Chemical characterization of Mx-Al revealed thirteen phyto-compounds and the peak area percentage is maximum at b-D-mannofuranoside (33.96%) and 1-Tetradecyne (21.46%). The larvicidal toxicity of Mx-AI on S. litura (97.23%) and P. xylostella (96.21%) delivered dose-dependent mortality and was significant at 500 ppm. The minimal dosage (200 ppm) extended the larval and pupal durations and significantly reduced the level of ACP, ALP, and LDH against both pests. The non-target toxicity against earthworm displayed higher toxicity to Cypermethrin (0.1 ppm) as compared to Mx-AI (1500 ppm). Further, the In-silico screening showed that the interpretation has significant implications for developing new and more selective inhibitors for P. xylostella and S. litura. Also, Bee-Tox, pkCSM and insecticide -likeness showed that all derivatives of Mx-Al are mostly nontoxic against Bees, protozoa, and rodents respectively, and obeyed the TICE rule violations. Overall, the present results proved that Mx-Al is target specific and harmless to non-target organisms. ARTICLE HISTORY
... According to Friedewald, Levy, and Fredrickson (1972), Serum HDL-cholestrol, VLDL and LDL were estimated as per the standard methods.Plasma SGOT and SGPT enzyme levels were measured by Reitman and Frankel method(Reitman and Frankel 1957). The serum alkaline phosphatase (ALP) was estimated by p-nitro phenyl phosphate method(Bessey, Lowry, and Brock 1946). Serum creatinine & Serum urea concentrations were determined by Jaffe's and diacetyl monoxime methods respectively(Slot 1965; Wybenga, Giorgio, and Pileggi 1971). ...
Green synthesis is based on plant compounds' ability to reduce metal ions and stabilise them into nanoparticles, and it is gaining a lot of attention in recent years. It has been shown to be a reliable and alternate method for the development of novel Nanoparticles . Andrographis echioides (L.) Nees is a used to synthesize zinc oxide nanoparticles . UV-visible spectroscopy, FTIR, Zeta potential, particle size analysis, XRD, SEM, AFM analysis were adopted to characterise the capped ZnONPs. The intense surface Plasmon resonance bands of ZnONPs at 340 nm confirmed the green synthesis of ZnONPs. The various functional groups of A.echioides involved in the synthesis and crystalline nature of ZnO Nanoparticles were identified using FTIR and X-ray diffraction respectively. The average particle size was found to be 4.0 nm, and the zeta potential value was -11 mV. Administration of ZnONPs significantly reduced the blood glucose level and HbA1C when compared with standard drug glibenclamide in STZ- induced in-vivo models. The antidiabetic potential of ZnONPs was confirmed by numerous biochemical parameters like serum lipid profiles, liver and renal functional markers and regeneration β- cells of islets of Langerhans in STZ- induced diabetic rats. Thus, the ZnONPs synthesized from A. echioides were found to be effective anti-hyperglycemic agent in the management of diabetes.
... The activities of serum AST and ALT were determined according to the method of Reitman and Frankel, (1957). The measurement of serum ALP was based on the method of Bessey et al.(1946) and Perry et al.(1983). Serum creatinine was measured according to the methods described previously (Bartels et al., (1972). ...
Email address algraibawi_57@yahoo.com (Al-Graibawi M. A. A.), afaf_a.rahman@yahoo.com (Al-Graibawi M. A. A.) Abstract The present study was designed to investigate the effects of different doses of tramadol hydrochloride (TH) on histopathological and biochemical changes in male rabbits. Fifteen healthy males local breed rabbits weighting 1.6-1.8 kg were distributed randomly and equally into three groups, housed under laboratory condition at the same room with natural light 1 dark cycle at 23 +3 ºC temperature, and given free access to commercial balanced diet and water ad libitum all over the experimental period. Group T 1 injected intramuscularly(i.m) with tramadol 10 mg/kg b.w for 15 days, while the T 2 group injected i.m with tramadol 15mg/kg b.w for 15 days and the 3rd groups T 3 is control inoculated with distilled water 1/M at the same period. Blood samples were collected for biochemical analysis of creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP). After sacrificed the animals, liver and kidney samples were obtained and fixed in 10% buffered formalin, processed, embedded in paraffin, sectioned, and stained with haematoxylin and eosin (H&E) for histopathological changes. The histopathological examination showed cellular degeneration characterized by vacuolization in the endothelium cells of renal tubules of the kidney and degeneration and pyknotic of the hepatocyte nuclei in the liver of the tramadole administrated rabbits only. The biochemical assay revealed that administration of tramadol for 15 days increased serum creatinine, ALT, AST, and ALP in treated rabbits compared to the control. The maximum increases was recorded in group T2, administrated i.m with 15mg/kg tramadol. In conclusion the tramadol side effects should be avoided during long term therapy especially in large doses.
... Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assayed as described by Bergmeyer et al. (1978) with 0.2 M DL-aspartic acid and 20 mM L-ketoglutarate as the substrate and 0.2 M DL-alanine and 2 mM L ketoglutarate, respectively. Also, alkaline phosphatase (ALP) was assayed according to Bessey et al. (1946). The serum triglyceride (TG) level was analyzed using the TG quantification kit (MAK266, Sigma-Aldrich, St Louis, MO, USA). ...
A long-term feeding trial (90 days) was conducted to investigate the effects of dietary supplementation with lavender oil (LO) on the growth performance, innate immunity, antioxidant status, and histomorphometry of different organs of juvenile European seabass (Dicentrarchus labrax). Four groups in triplicate were fed increasing levels of LO (0, 1, 2, and 3 kg⁻¹ diet, expressed as F0, F1, F2, and F3, respectively). Fish weighing 76.87 ± 0.14 g/fish were stocked in 1000-L tanks at an initial stocking density of 25 fish. The results revealed that growth performance and feed utilization indices were significantly improved in the LO-treated groups compared to the control, with the highest values in favor of the F2 group. According to the polynomial second-order regression, the optimal dietary level of LO averaged 2.15–2.24 g/kg diet. Whole body protein and digestive enzyme activities were increased with increasing LO supplementation levels up to 2 mL kg⁻¹. Fish-fed F2 diet exhibited a significant decrease in serum liver function enzymes. A considerable increase in total protein, albumin, and total immunoglobulin was reported in LO-supplemented groups over the control. Dietary LO significantly modulated antioxidant status by increasing antioxidant enzyme activities and decreasing malondialdehyde levels. Microscopic investigation of the gills, liver, and mid-intestine revealed that LO-treated fish had healthier histological features, with normal gill lamella and hepatocytes, and a positive effect of LO on intestinal villi length and goblet cell count. The LO could be used as a dietary supplement in the diet of seabass at the recommended level of 2 g kg⁻¹ to improve growth, feed conversion, physiological status, and organ histomorphometry.
... Then the separated supernatant was centrifuged (34000 g, 20 min, 4°C), the supernatant was removed, and the remained sediment was dissolved in 1 ml DTT HEPES and kept in -80°C freezer according to Gisbert et al. (2018). Standard protocols were used to assess digestive enzymes, including alkaline phosphatase (ALP, Bessey et al., 1946), aminopeptidase N (APN, Appel, 1974), leucine-alanine peptidase (LAP, Nicholson and Kim, 1975), trypsin (Erlanger et al., 1961), chymotrypsin (Worthington, 1991), amylase (Métais and Bieth, 1968), pepsin (Worthington, 1991) and phospholipase A 2 (Reynolds et al., 1992). Bradford method used to evaluate soluble protein in samples (Bradford, 1976). ...
Phospholipid (PL) is an essential nutrient that has vital effects on growth, stress resistance and early development in marine fish larvae. In this regard, a 30-day feeding experiment was conducted in order to examine the effects of live prey enrichment with graded levels of soy lecithin (SL) on some physiological responses of Acanthopagrus latus larvae. Four experimental emulsions levels of SL were used to enrich Rotifer and Artemia including very low (2%, N), low (4%, L), medium (8%, M) and high (12%, H). Newly hatched larvae were distributed into twelve 250-L cylindrical tanks with an initial density of 15000 larvae in each tank that was supplied with natural seawater (23 ± 1 ºC; 40.0 ± 1.0 g L-1). The green water method was used for larviculture and live prey was given to larvae two times daily. Larvae fed 4% SL containing live prey significantly had higher wet weight gain than other treatments. Air exposure and osmotic activity tests were also performed to detect larval resistance to stress. Larvae fed 8% and 12% SL containing live prey had higher survival compared to the other two groups. The accumulation of arachidonic ARA and docosahexaenoic acids was increased in the larval whole body fed high SL-supplemented live foods. Alkaline phosphatase and aminopeptidase N activities in the guts brush border membrane of larvae in M and H groups were higher than other treatments. The trypsin and chymotrypsin activities in the N group were lower than in other treatments. The highest and lowest amylase activities were in the H and N groups, respectively. The activity of catalase and glutathione reductase in the whole body of the M group was higher than the N treatment and other groups had intermediate values. Total antioxidant capacity in the whole body of larvae in the N group was lower than in the other treatments. In summary, moderate levels of SL (4–8%) are suggested for the enrichment of live foods in A. latus .
... Effect of the permissive sociocultural consumption of alcohol on... Enzyme Profile: Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity values were determined using a colorimetric method (Reitman, & Frankel, 1957). Assay of serum alkaline phosphatase (ALP) activity was by a colorimetric method (Bassey et al., 1946). The commercial kits containing the reagents used for these assays were supplied by Randox Laboratories Limited, Ardmore, United Kingdom. ...
To investigate the health risks associated with the permissive sociocultural
use of alcohol by the vast majority of the inhabitants in the Edo-Delta Region
of Nigeria, one hundred and fifty habitual drinkers of alcohol were randomly
selected from the major tribes in the Edo-Delta area of Nigeria. Tribal breakdown
indicates that 40%, 30%, 23% and 7% were Urhobos, Binis, Ibos and Ukwanis,
respectively. These test subjects were males between the ages of 25 and 50
who had been drinking for varying lengths of time and had an average daily
intake of about 1.2g ethanol/kg body weight. Sixty sex- and age-matched individuals in apparent good health who do not consume alcohol were included as
the control subjects. The results obtained showed the mean (mol/L) plasma
total bilirubin for the control group and drinkers to be 16.00.9 and 17.52.7
respectively, while the mean SEM (mol/L) plasma conjugated bilirubin for the
control group and alcohol drinkers was 10.20.3 and 7.252.6 respectively. The
mean SEM (IU/L) plasma alkaline phosphatase (ALP), aspartate aminotranserase
(AST), and alanine aminotransferase (ALT) activities for the alcohol drinkers
were 84.55.2; 53.44.1, and 50.05.0, but those for the control subjects were
58.43.2; 9.11.5, and 11.01.7. The differences demonstrated in the enzymes’
activity values were significant (P<0.05). The data suggest a significant measure of liver dysfunction among alcohol drinkers who are yet to manifest any
recognisable physical symptoms of alcohol toxicity. Thus, a permissive attitude
to alcohol consumption is associated with a high risk of health disturbances. An enlightenment campaign should be initiated and the government should establish rehabilitation programmes and centres.
Keywords: alcohol - aspartate aminotransferase - alanine aminotransferase -
alkaline phosphatase - liver
... ALT and AST levels were measured using a calorimetric method as previously reported 44,45 . Alkaline phosphatase (ALP) was determined using a previously described method 46 . Uric acid and creatinine were measured using a calorimetry method 47,48 . ...
Lactic acid bacteria (LAB) are of major concern due to their health benefits. Fermented food products comprise variable LAB demonstrating probiotic properties. Discovering and evaluating new probiotics in fermented food products poses a global economic and health importance. Therefore, the present work aimed to investigate and evaluate the probiotic potentials of LAB strains isolated from Egyptian fermented food. In this study, we isolated and functionally characterized 100 bacterial strains isolated from different Egyptian fermented food sources as probiotics. Only four LAB strains amongst the isolated LAB showed probiotic attributes and are considered to be safe for their implementation as feed or dietary supplements. Additionally, they were shown to exert antimicrobial activities against pathogenic bacteria and anticancer effects against the colon cancer cell line Caco-2. The Enterococcus massiliensis IS06 strain was exclusively reported in this study as a probiotic strain with high antimicrobial, antioxidant, and anti-colon cancer activity. Hitherto, few studies have focused on elucidating the impact of probiotic supplementation in vivo. Therefore, in the current study, the safety of the four strains was tested in vivo through the supplementation of rats with potential probiotic strains for 21 days. The results revealed that probiotic bacterial supplementation in rats did not adversely affect the general health of rats. The Lactiplantibacillus plantarum IS07 strain significantly increased the growth performance of rats. Furthermore, the four strains exhibited increased levels of antioxidants such as superoxide dismutase and glutathione in vivo. Consistently, all strains also showed high antioxidant activity of the superoxide dismutase enzyme in vitro. Overall, these findings demonstrated that these isolated potential probiotics harbor desirable characteristics and can be applied widely as feed additives for animals or as dietary supplements for humans to exert their health benefits and combat serious diseases.
... Once unfrozen, the mucosa of the digestive tract was collected by scrapping the intestine and was homogenized in cold distilled water with a polytron at maximum speed for 30 s (Crane et al., 1979) to purify brush border membranes (BBM). The BBM enzymes alkaline phosphatase (AP), leucine aminopeptidase (AN), γ-glutamyl transpeptidase (GGT), trypsin, and catalase were assayed according to Bessey et al., (1946), Meister et al., (1981, Holm et al., (1988), Maroux et al., (1973) and Lück, (1965), respectively. Total soluble proteins were determined according to Bradford, (1976) using bovine serum albumin as a standard (Sigma Chemical Co., St. Louis, USA). ...
Morphometric indices of body condition are assumed to reflect an animal’s health and ultimately its fitness, but their physiological significance remains a matter of debate. These indices are indeed usually considered as proxies of energy reserves, ignoring other physiological processes involved in animal health such as nutritional, immune and hormonal states. Given the wide variety of ecological processes investigated through morphometric body condition indices in marine sciences, there is a clear need to determine whether morphometric indices reflect primarily individuals’ energy reserves or their broader integrated physiological status. To address this issue, we used morphometric and physiological data (cortisol level, oxidative stress, digestive enzymes activity, and both fatty acids percentage and total amount) collected in three stocks of the European sardine (Sardina pilchardus) presenting contrasted patterns of growth and body condition. We found that morphometric body condition indices are indeed mainly and consistently linked to a proxy of the amount of lipid reserves (i.e., fatty acids amount), but also significantly to quality of lipid reserves (i.e., fatty acids percentage) and fish chronic stress (scale cortisol levels). We found no significant relationship between morphometric indices of body condition and both oxidative stress proxies and variables measuring digestive enzymes activity. Our study confirmed that morphometric body condition indices primarily reflect the variance in individuals’ lipid reserves and to a lesser extent the actual composition of these reserves (linked to differences in fish diet) and scale cortisol levels (indicating fish metabolism and/or their chronic stress levels). Therefore, some non-energetic aspects should be considered when studying individuals’ responses to environmental changes and other key physiological processes (oxidative stress proxies, activity of digestive enzymes) should be investigated directly to support scientific-based decision-making in the context of climate change.
... Amylase activity was determined using starch as a substrate, dissolved in a pH 7.4 sodium phosphate buffer [27]. Alkaline phosphatase activity was assayed using 5 mM p-nitrophenyl-phosphate (pNPP) as a substrate in a 30 mM carbonate buffer at pH 9.8 [28]. Aminopeptidase activity was assayed using 0.1 M Lleucine p-nitroanilide as a substrate in an 80 mM solution of phosphate buffer at pH 7.0 [29]. ...
Water temperature variations affect fish growth and health, often leading to huge losses in fish production, especially during the cold season. To alleviate this constraint, fish farmers can use a water heating system driven by solar energy during daytime. This action will cause a water temperature drop during the night period, making it important to understand the physiological response of fish exposed to the resulting day/night temperature oscillations. To investigate this scenario, gilthead seabream juveniles (96.3 ± 1.0 g) were exposed to different thermal regimes for 67 days: Tconstant and Tdaily cycles. The latter group was exposed to daily water temperature oscillations between ~19 and 13 °C compared with a constant temperature of ~19 °C for the other experimental group. Temperature fluctuations compromised fish growth efficiency and reduced the proportion of fatty acids in several tissues, with implications for the whole proximate composition. Moreover, temperature oscillations influenced several blood parameters. These results favor the usage of a constant water temperature of ~19 °C for optimal gilthead seabream juvenile production instead of a day/night water temperature oscillating regime. Nevertheless, the type of energy used to warm the water will depend on the operational conditions and/or business strategy of fish farmers.
... Purified BBM from the intestinal segment homogenate were obtained as described above. Activity of alkaline phosphatase (AP), an enzyme of the BBM, was measured using p-nitrophenylphosphate (pNPP) (Bessey et al., 1946). Aminopeptidase N (AN) activity was determined using L-leucyl-β-naphthylamide (Porteous and Clark, 1965). ...
The ontogenesis and specific activities of pancreatic and intestinal enzymes were investigated in sharpsnout sea bream, Diplodus puntazzo, during larval development until the end of weaning on day 50. The green-water technique was carried out for larval rearing in triplicate. Trypsin was first detected as early as hatching and sharply increased related to age and exogenous feeding until day 25, but a sharp decrease was observed towards the end of the experiment. Amylase was determined 2 days after hatching (DAH) and sharply increased to 10 DAH. Afterwards, slight decreases were found between 10 and 20 DAH and then slow alterations were continued until end of the experiment. Lipase was measured for the first time on day 4, and then slight increase was found to 25 DAH. After this date, slow variations were maintained until end of the experiment. Pepsin was firstly assayed 32 DAH related with stomach formation and sharply increased to 40 DAH. Then it was fluctuated until end of the experiment. Enzymes of brush border membranes, alkaline phosphatase and aminopeptidase N, showed similar pattern on specific activities during the first 10 days. Thereafter, while specific activity of alkaline phosphatase slightly decreased to 15 DAH and fluctuated until 20 DAH, aminopeptidase N activity slowly declined to 20 DAH. Afterwards, activity of alkaline phosphatase and aminopeptidase N were sharply increased to 30 DAH, showing maturation of the intestinal digestive process and also these activities continued to slight increase until end of the experiment. The specific activity of cytosolic peptidase, leucine-alanine peptidase sharply increased to on day 8, then suddenly declined to 12 DAH and further decreased until 20 DAH. After this date, in contrast to enzymes of brush border membranes, it sharply decreased to 25 DAH and continued to gradually decline until the end of the experiment. These converse expressions were indicative of a maturation of enterocytes and the transition to an adult mode of digestion.
... Regarding intestinal enzymes, alkaline phosphatase (E.C. 3.1.3.1) was quantified using 4-nitrophenyl phosphate (PNPP) as substrate. One unit (U) was defined as 1 µmol PNPP hydrolyzed per min −1 mL −1 of enzyme extract at 407 nm [76]. Aminopeptidase N (EC 3.4.11.2) activity was determined according to Maroux et al. [77] using sodium phosphate buffer 80 mM (pH 7.0) and L-leucine p-nitroanilide as a substrate in DMSO. ...
Microplastics (MPs) are emergent pollutants in freshwater environments and may impact aquatic organisms, including those of nutritional value. The specific activities of digestive and antioxidant enzymes can be used as good bioindicators of the potential effects of MPs on fish in case of waterborne MP contamination. In this study, we used fluorescent polystyrene microplastics (PS-MPs) to analyze the alterations in enzyme activities in larvae of Coregonus peled Gmelin (peled or Northern whitefish), one of the most valuable commercial fish species of Siberia. Our results indicate that peled larvae can ingest 2 µm PS microspheres in a waterborne exposure model. A positive correlation (rs = 0.956; p < 0.01) was found between MP concentration in water and the number of PS microspheres in fish guts, with no significant differences between 24 h and 6-day exposure groups. The ingestion of MPs caused alterations in digestive enzyme activity and antioxidant responses at the whole-body level. The presence of PS-MPs significantly stimulated (p < 0.05) the specific activity of α-Amylase and non-specific esterases in peled larvae after 24 h. However, a pronounced positive effect (p < 0.05) of MPs on the activity of pancreatic trypsine and bile salt-activated lipase was only found after 6 days of exposure compared to after 24 h. Intestinal membrane enzyme aminopeptidase N was also stimulated in the presence of PS-MPs after 6-day exposure. We also observed a significant increase in the specific activity of catalase in peled larvae after 6 days of exposure, which indicates the MP-induced modulation of oxidative stress. Taken together, these results highlight the potential impact of environmental MPs on northern commercial fish, their importance for estimating fish stocks, and the sustainability of freshwater ecosystems.
... For homogenization, the manufacturer's instructions for the commercial kit were followed using the specific buffer for each enzyme and centrifuging the mixture for 30 min at 3000× g, 4 • C. Before analysis, the supernatants were kept at −80 • C. Measurements of protease: protease, amylase, and lipase activities were carried out as described in [61][62][63]. Alkaline phosphatase was measured as described and modified by Bassey et al. [64] and Wright et al. [65], respectively. ...
Olive leaves are an immense source of antioxidant and antimicrobial bioactive constituents. This study investigated the effects of dietary incorporation of olive leaf extract (OLE) on the growth performance, hematobiochemical parameters, immune response, antioxidant defense, histopathological changes, and some growth- and immune-related genes in the common carp (Cyprinus carpio). A total of 180 fish were allocated into four groups with triplicate each. The control group received the basal diet without OLE, while the other three groups were fed a basal diet with the OLE at 0.1, 0.2, and 0.3%, respectively. The feeding study lasted for 8 weeks, then fish were challenged with Aeromonas hydrophila. The results revealed that the group supplied with the 0.1% OLE significantly exhibited a higher final body weight (FBW), weight gain (WG%), and specific growth rate (SGR) with a decreased feed conversion ratio (FCR) compared to the other groups (p < 0.05). An increase in immune response was also observed in the fish from this group, with higher lysosome activity, immunoglobulin (IgM), and respiratory burst than nonsupplemented fish, both before and after the A. hydrophila challenge (p < 0.05). Similarly, the supplementation of the 0.1% OLE also promoted the C. carpio's digestive capacity pre- and post-challenge, presenting the highest activity of protease and alkaline phosphatase (p < 0.05). In addition, this dose of the OLE enhanced fish antioxidant capacity through an increase in the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) and decreased hepatic lipid peroxidation end products (malondialdehyde—MDA), when compared to the control group, both pre- and post-infection (p < 0.05). Concomitantly with the superior immune response and antioxidant capacity, the fish fed the 0.1% OLE revealed the highest survival rate after the challenge with A. hydrophila (p < 0.05). A significant remarkable upregulation of the hepatic sod, nrf2, and protein kinase C transcription levels was detected as a vital approach for the prevention of both oxidative stress and inflammation compared to the infected unsupplied control group (p < 0.05). Interestingly, HPLC and UPLC-ESI-MS/MS analyses recognized that oleuropein is the main constituent (20.4%) with other 45 compounds in addition to tentative identification of two new compounds, namely oleuroside-10-carboxylic acid (I) and demethyl oleuroside-10-carboxylic acid (II). These constituents may be responsible for the OLE exerted potential effects. To conclude, the OLE at a dose range of 0.66–0.83 g/kg w/w can be included in the C. carpio diet to improve the growth, antioxidant capacity, and immune response under normal health conditions along with regulating the infection-associated pro-inflammatory gene expressions, thus enhancing resistance against A. hydrophila.
... activity was assayed using starch dissolved in NaH2PO4 buffer (0.07 M), pH 7.4 [29], while alkaline phosphatase (E.C.3.1.3.1) activity was assayed using pNPP 5 mM (p-nitrophenylphosphate) in a solution of carbonate buffer (30 mM), pH 9.8 [30]. Aminopeptidase N (E.C.3.4.11.2) activity was quantified using L-leucine p-nitroanilide (0.1 M), as substrate, in buffer phosphate (80 mM), pH 7.0, following Maroux et al. [31] pro-tocol. ...
Efforts have been made to find alternatives to fish meal (FM), as the sustainability of aquaculture depends on it. Insect meal (IM) is a potential candidate to partially replace FM, being more sustainable and economically viable. In this experimental trial, three diets were tested with different yellow mealworm incorporation: a control diet with no IM, a diet with an inclusion of 10% IM (Ins10), and a diet with an incorporation of 20% IM (Ins20). The diets were tested on 10.5 g meagre for 47 days. The results showed that an IM inclusion higher than 10% affected both growth (2.6 vs. 2.2) and FCR (1.5 vs. 1.9) of meagre juveniles. However, this reduction in growth did not result from lower protein retention or changes in muscle fibre area or density. Little differences were observed in the activity of pancreatic and intestinal enzymes except for aminopeptidase total activity which was higher in the control and Ins10 compared to Ins20 (3847 vs. 3540 mU/mg protein), suggesting no limitations in protein synthesis. Also, the alkaline phosphatase intestinal maturation index was higher in the control group compared to the IM groups (437 vs. 296). On the contrary, several differences were also found in the proteolytic activity in the hepatic and muscle tissues of meagre juveniles fed the Ins10 diet. The inclusion of IM had no impact on intestine histomorphology but changes were detected in the enterocytes of fish from control and Ins10 which showed hypervacuolization and nucleus misplacement compared to the Ins20 treatment. Nevertheless, a higher percentage of Vibrionaceae was recorded for meagre fed on the Ins20 diet. Since no signs of inflammation were observed in the distal intestine, this suggests IM incorporation could have had an important impact on intestinal health due to its antimicrobial properties. This is supported by an increase in the haematocrit in the treatments where IM was added (20 to 25%). In conclusion, incorporations of IM at percentages up to 10% do not seem to have a negative impact on meagre performance at this age but can enhance the fish immune system and protection against intestinal inflammation.
... The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were determined according to the method described by the International Federation of Clinical Chemistry (1986). Alkaline phosphatase (ALP) was assayed according to the method of (Bassey et al. 1946) modified by (Wright et al. 1972), while lactate dehydrogenase (LDH) activities were determined according to instructions from Randox diagnostic kit. ...
Cannabichromene (CBC) is one of the non-psychoactive cannabinoids found in Cannabis sativa and the second most abundant after cannabidiol. CBC has been shown to produce antinociception and anti-inflammatory effects in rodents; it is a major non-psychotropic phytocannabinoid that inhibits endocannabinoid inactivation and activates the transient receptor potential ankyrin-1 (TRPA1). The use of non-psychoactive cannabinoids as protective, ameliorative, and preventive agents in diverse disease pathophysiology is increasingly gaining global significance. Cannabidiol (CBD) has been extensively studied in this regard; however, CBC may also possess undiscovered intriguing and appreciable therapeutic properties. Therefore, the effects of 7 days of oral daily administration of 5 mg/kg,10 mg/kg, and 20 mg/kg body weight doses of CBC on antioxidant enzyme system, hepatic, and renal function biomarkers were investigated in this study. Twenty-four (24) male Wistar rats were divided into four groups of 6 animals each (1 control and 3 test groups). Twenty-four hours after the last treatment, the animals were anaesthetized using ketamine/xylazine and then euthanized via cervical dislocation. The liver and kidney were excised, while plasma and red blood cells were processed from the whole blood and used for biochemical analysis. Statistical analysis was done using one-way analysis of variance (ANOVA) with the Tukey’s test of homogeneity where appropriate; p < 0.05 was considered statistically significant. The activities of all antioxidant enzymes (catalase, CAT; superoxide dismutase, SOD; and glutathione peroxidase, GPx) were significantly (p < 0.05) increased in the liver, kidney, red blood cell, and plasma at all doses, especially at the 10 mg/kg body weight dose. The levels of non-enzymatic antioxidants (reduced glutathione, GSH; and nitric oxide, NO) were not significantly (p > 0.05) different at all exposed CBC doses. However, the malondialdehyde level (MDA) was increased notably in the kidney and plasma in the 10 mg/kg group. Hepatic function enzymes: aspartate transaminase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) activities were significantly decreased at the 10-mg/kg dose, although renal function markers seemed consistent at all the doses investigated. Histological evaluations of the kidney and liver revealed severe multifocal necrotizing tubule, interstitial nephritis, and degeneration of renal tubules and periportal hepatocellular necrosis with inflammation of the hepatocytes. Our study revealed modulation of the hepato-renal system via CBC exposure, and despite few reports of beneficial applications, we recommend caution in its use.
... The Biolis 24i kit (Agappe Diagnostics Ltd., India) was used to estimate the levels of AST, ALT, and ALP. The standards proposed by the International Federation of Clinical Chemistry (1980) were used for estimating AST and ALT, while the procedure described by Bassey et al. (1946) and modified by Wright et al. (1972) was used to carry out the ALP assay. A colorimetric assay kit was used to assess CK. ...
Heavy metals have an immense impact on aquatic ecosystems, and their toxic effects are transferred to the inhabiting organisms. Experiments were conducted to investigate the health of snow trout Schizothorax esocinus inhabiting Dal Lake. Heavy metals (Cd > Ni > Cu > Cr) were found to accumulate in the major immune organs of the fish (head kidney, liver, spleen, thymus) which led to change in the overall physiology. The head kidney, liver, and spleen of a fish contain high amount of these metals. The least accumulation of these metals was found in the blood, whereas Cd and Ni were completely absent in the integument. Hepatic marker enzymes (aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP)) were normal, while the renal marker enzyme creatine kinase showed marked difference in its value. The cortisol level was normal, while immunoglobulin M showed elevated level representing active immunity. At a cellular level, the histopathology of immune organs showed marked damage. Metallothionein (MT) and glutathione peroxidase (GPX) genes showed variable expression pattern in the immune organs with the head kidney showing the highest expression of both the genes, and blood showed the least. We observed that the aquatic plants (Nelumbo nucifera and Trapa natans) inhabiting the lake played an important role in phytoremediation. An integrated approach involving biochemical, hematological, genotoxic, and histopathological studies can provide a valuable information to understand fish adaptive patterns and monitor water quality.
Graphical Abstract
... Based on the method of Bessey et al. (1946), 10 µl of enzyme solution was added to the buffered substrate (Tris-HCl, 20 mM, pH 8 for ALP and pH 5 for ACP), phosphate buffer (0.02 m, pH 7.2) and incubated for 5 min. Afterward, 100 µl of NaOH (1 M) was added and the absorbance was read at 405 nm. ...
Toxicity and physiological alterations were determined in Pseudococcus viburni nymphs
treated with Artemisia annua methanolic extract. The leaf dipping bioassay showed LC50
values of 0.287% and 0.194% 24 and 48 hours post-exposure. Activities of general este-
rases were significantly higher in the control nymphs than in those which had been treat-
ed except for the 48 h time interval using α-naphtyl acetate. The activity of glutathione
S-transferase using CDNB (1-chloro-2,4-dinitrobenzene) in the control nymphs, was signif-
icantly higher than in the control at both time intervals while no significant difference was
observed after 24 h in addition to the higher enzymatic activity in the treated nymphs after
48 h. All three aminotransferases were significantly more active in the control nymphs ex-
cept for time intervals of 24 h for γ-glutamyl transferase and 48 h for alanine aminotrans-
ferase. Higher activities of lactate dehydrogenase, acid- and alkaline phosphatase were
found in the control nymphs than in treated nymphs for all time intervals. Activities of
the enzymes involved in the antioxidant system including catalase, peroxidase, superoxide
dismutase, ascorbate peroxidase and glucose-6-phosphate dehydrogenase was increased in
the treated nymphs compared to the control. Results of the current study demonstrated
toxic effects of A. annua methanolic extract on P. viburni nymphs causing mortality and
physiological turbulences.
Keywords: antioxidant response, Artemisia annua, intermediary metabolism, methanolic
extract, Pseudococcus viburni
... Finally, the remaining precipitate was suspended in one mL of buffer (0.1 M KCl, 5 mM Tris-Hepes, 1 mM DTT; pH 7.5) and kept in a freezer (− 80 • C) (Crane et al., 1979). The evaluation of digestive enzymes and soluble protein (Bradford, 1976) were determined by standard methods for the following enzymes: trypsin (Bergmeyer, 1974), chymotrypsin (Hummel, 1959), protease (Folin and Ciocalteau, 1929;Anson, 1938), ALP (Bessey et al., 1946), α-amylase (Bernfeld, 1955), and lipase (Tietz and Fiereck, 1966). ...
A feeding trial was done for 60 days to examine the influence of supplementing low-fish meal (FM) diet with a mixture of acidifiers on the performance of Asian seabass (Lates calcarifer) juveniles (initial weight: 54.2 ± 0.5 g, mean ± standard deviation). Dietary FM was replaced (35% and 70%) with a mixture of alternative protein sources, including soybean meal, corn gluten, wheat gluten, and poultry meal then supplemented with two levels (0.5% and 1.0%) of an acidifier mixture (butyric acid, sodium diformate and fulvic acid, 1:1:1 ratio). In this regard, seven isonitrogenous (45%) and isolipidic (15%) diets were formulated including: FM70 (FM-based diet, control), FMR35 (35% FM replacement), FMR35+0.5% (35% FM replacement + 0.5% acidifier mixture), FMR35+1.0% (35% FM replacement + 1.0% acidifier mixture), FMR70 (70% FM replacement), FMR70+0.5% (70% FM replacement + 0.5% acidifier mixture), and FMR35+1.0% (70% FM replacement + 1.0% acidifier mixture). Fish were stocked into twenty-one 2000-L rectangular concrete tanks (53 fish/tank) that were filled with running seawater in a flow-through system (26.5 ± 1.5 °C and 46.0 ± 0.2 ppt). Fish were fed with the diets twice every day up to visual satiation. Before the beginning (day 0), middle (day 30), and after finishing the feeding trial (day 60), fish were individually weighed, and samples were collected from their blood and gut for evaluating hematological, antioxidant and digestive enzymes, respectively. After finishing the feeding trial, fish fed FMR70 had lower weight gain (89.1% lower than FM70) than other groups (P = 0.001) that associated with the lowest feed intake in this group. The trypsin, protease, and α-amylase activities were decreased by increasing FM replacement level, but it enhanced alkaline phosphatase activity (P
... The alkaline medium's sodium p-nitrophenyl phosphate was used as a substrate to quantify ALP activity. (11) . Oxidative stress parameters in the liver tissues: Glutathione (GSH): its concentrations are given as mg per gram of tissue (12) . ...
Introduction: Lambda Cyhalothrin (LCT) is greatly used to manage a wide variety of pests present in farming and in home procedures. Aim: The current work was intended to demonstrate structural and functional alterations within the liver subsequent to long-standing exposure to LCT. Protective effect of Hesperidin and N-acetylcysteine was also investigated. Material and methods: 40 adult male albino rats were used in this experiment, and they were split into four equal groups: control, LCT group, rats were received LCT at a dose 61.2mg/kg b. wt. per day by oral gavage for 28 days. LCT + Hesperidin group, rats were given the same dose of LCT with simultaneous oral dosage of HSP at a dose of 100mg/kg b. wt., LCT + N-Acetyl cysteine group at which rats were received the same dose of LCT as the previous 2 groups with simultaneous oral administration of N-acetyl cysteine 150 mg/kg b. wt. The liver underwent a number of biochemical, histological, and immunohistochemical analysis. Results: LCT induced oxidative stress which leads to liver damage (increases MDA / decreases GSH). LCT caused degeneration of hepatocytes and increases inflammatory cells, this is followed by rise in liver markers (AST and ALT). While concurrent administration of Hesperidin and N-Acetyl cysteine during LCT exposure period preserved the architecture of the liver, prevents its damage, reduced oxidative stress and normalized liver function tests. Conclusion: Administration of N-Acetyl cysteine during exposure to the insecticide LCT has a protective effect on the liver more than Hesperidin.
... The enzyme assay was done based on the method of Bessey (1954). The buffered substrate, p-nitrophenol phosphate in Tris-HCl (20 mM) pH 8 and (pH 5) was separately used to assay alkalineand acid phosphatase, respectively. ...
The current study aimed to determine antioxidant and detoxifying responses of Chilo suppressalis Walker (Lepidoptera: Crambidae) to Beauveria bassiana (Strains BBRR1, BBAL1, BBLN1, BBLN2), Metarhizium anisopliae and Hirsutella subulata. The interactions of insect humoral immune responses with the entered conidia of entomopathogenic fungi in addition to nodule formation and melanization caused the production of several reactive oxygenate species (ROS), such as hydrogen peroxidase (H2O2), hydroperoxides (ROOH), superoxide radicals (O 2−), and hydroxyl radical (OH −). The highest activity of catalase was recorded by BBRR1 and BBAL1, treatment after 48 to 96 h while the larvae treated by BBRR1 showed the highest peroxidase activity. Both ascorbate peroxidase and glucose-6-phosphate dehydrogenase showed the highest activity in the larvae treated by BBRR1 after 48-96 h. The highest concentration of Malondialdehyde (MDA) reported in the larvae treated by BBRR1, BBAL1 and BBLN1, after 48 hours. The highest activity of general esterases was recorded in the larvae treated by BBRR1 after 48-96 hours. Similar results were recorded in the activity of glutathione-S-transferase but the enzyme had also the highest activity in the larvae treated by BBAL1 and BBLN2 after 48 hours. The larvae treated by BBRR1 and BBLN1 showed the highest activity of acid phosphatase (ACP) after 72 and 96 hours while the highest activity of alkaline phosphatase (ALP) was obtained in the larvae only treated by BBRR1 after 48-96 hours. The results clearly revealed that BBRR1 significantly and severely induced antioxidant and detoxifying systems of C. suppressalis larvae implying on virulence and immune induction of BBRR1 against the larvae.
... Earlier several classical methods were developed using p-nitrophenyl phosphate (pNPP) and disodium phenyl phosphate as substrates of ALP [7,8]. With a further understanding of the character and function of ALP, more facile, sensitive, and precise methods have been developed. ...
Alkaline phosphatase (ALP) is a metalloenzyme, the level of which is clinically significant as an abnormality of ALP activity results in several diseases. In the present study, we introduced a MnO2 nanosheet-based assay for ALP detection employing the adsorption and reduction characteristics of G-rich DNA probes and ascorbic acid (AA), respectively. Ascorbic acid 2-phosphate (AAP) was utilized to act as a substrate for ALP which hydrolyzes AAP generating AA. In the absence of ALP, MnO2 nanosheets adsorb the DNA probe destructing the G-quadruplex formation and showing no fluorescence emission. On the contrary, being present in the reaction mixture ALP hydrolyzes AAP yielding AA, then the AA reduce the MnO2 nanosheets into Mn2+, hence, the probe is free to react with a dye, thioflavin T (ThT), and synthesizes ThT/G-quadruplex to spark high fluorescence intensity. Therefore, under optimized conditions (250 nM DNA probe, 8 μM ThT, 96 μg/mL MnO2 nanosheets, and 1 mM AAP) the sensitive and selective measurement of ALP activity can be achieved through the change of fluorescence intensity, with a linear range and a limit of detection of 0.1-5 U/L and 0.045 U/L. Our assay exhibited its potential to assess the ALP inhibitor when in an inhibition assay Na3VO4 inhibited ALP with an IC50 value of 0.137 mM and also was validated in clinical samples.
... Where indicated, reactions included 20 U/mL superoxide dismutase and 30 U/ mL catalase. At time points, 50 μL aliquots were removed to 1 mL 1 mM p-nitrophenylphosphate in 1 M Tris, pH 8. AP activity was measured based on its ability to hydrolyze p-nitrophenylphosphate to p-nitrophenol, a chromogenic product that absorbs at 405 nm (Bessey et al., 1946). ...
Copper avidly binds thiols and is redox-active, and it follows that one element of copper toxicity may be the generation of undesirable disulfide bonds in proteins. In the present study copper oxidized the model thiol N-acetylcysteine in vitro. Alkaline phosphatase (AP) requires disulfide bonds for activity, and copper activated reduced AP both in vitro and when it was expressed in the periplasm of mutants lacking their native disulfide-generating system. However, AP was not activated when it was expressed in the cytoplasm of copper-overloaded cells. Similarly, this copper stress failed to activate OxyR, a transcription factor that responds to the creation of a disulfide bond. The elimination of cellular disulfide-reducing systems did not change these results. Nevertheless, in these cells the cytoplasmic copper concentration was high enough to impair growth and completely inactivate enzymes with solvent-exposed [4Fe-4S] clusters. Experiments with N-acetylcysteine determined that the efficiency of thiol oxidation is limited by the sluggish pace at which oxygen regenerates copper(II) through oxidation of the thiyl radical-Cu(I) complex. We conclude that this slow step makes copper too inefficient a catalyst to create disulfide stress in the thiol-rich cytoplasm, but it can still impact the few thiol-containing proteins in the periplasm. It also ensures that copper accumulates intracellularly in the Cu(I) valence.
The pervasive presence of electromagnetic fields (EMF) generated by modern technologies poses a significant threat to human health, with ionizing radiation, a byproduct of EMF, potentially contributing to cancer development. This study explores the impact of chronic exposure to GSM-EMFs and 900-1800 MHz-induced electromagnetic radiation (EMR) on liver enzymes, and serum electrolytes in mice and potential mitigating effect of exogenous glutathione administration. Thirty-five adult male mice were randomly divided into seven groups and exposed to various modes of mobile phone radiation for five weeks, with or without glutathione administration. Liver enzymes (ALP, ALT, AST) and serum electrolytes (sodium, potassium, bicarbonate) were analyzed. Results showed significant increases in ALP levels in the Silent, Ringtone + GSH, and Silent + GSH groups compared to controls, while ALT and AST levels remained largely unchanged. Serum electrolyte concentrations did not significantly differ across experimental groups Overall, the study underscores the need for continued research to better understand the health implications of EMF exposure and inform public health measures.
Background: Glycine is a conditional non-essential amino acid in human and other mammals. It is abundant in the liver and is known for a wide spectrum of characteristics including the antioxidant, antiinflammatory, immunomodulatory, and cryoprotective effects. The amino acid is a naturally occurring osmolyte compatible with protein surface interactions and has been reported in literature as a potent therapeutic immuno-nutrient for liver diseases such as alcoholic liver disease. Oral glycine administration protects ethanol-induced liver injury, improves serum and tissue lipid profile, and alleviates hepatic injury in various conditions. In recent years, sodium salt of boron (borax) has been reported for its beneficial effects on cellular stress, including the effects on cell survival, immunity, and tissue redox state. Incidentally both glycine and boron prevent apoptosis and promote cell survival under stress. Objective: This study investigates the beneficial effect of borax on liver protection by glycine. Methods: Briefly, liver toxicity was induced in rats by a single intraperitoneal injection of thioacetamide (400 mg/kg b. wt.). Results: Significant changes in oxidative stress and liver function test parameters, the molybdenum Fe-S flavin hydroxylase activity, nitric oxide and tissue histopathology were observed in thioacetamide treated positive control group. The changes were ameliorated both by glycine as well as borax, but the combinatorial treatment yielded a better response indicating the impact of boron supplementation on glycine mediated protection of liver injury in experimental animal model. Conclusions: The study has clinical implications as the hepatotoxicity caused by thioacetamide mimics features of hepatitis C infection in human.
A 60-day study was carried out to determine the effect of dietary selenium nanoparticles (SeNP) on growth, digestive enzymes, and health status of Asian seabass (Lates calcarifer, 46.5 ± 0.2 g) juveniles reared in high saline water (48 ppt). Five levels of SeNP were added to a basal diet (45% protein, 15% lipid), including control (0), 0.5 (SeNP0.5), 1.0 (SeNP1), 2 (SeNP2), and 4 (SeNP4) mg SeNP kg−1 diet. Fish were stocked into fifteen 2,000 L tanks (50 fish tank−1) filled with 1,800 L sand-filtered seawater (26.5 ± 1.5°C, 48.0 ± 0.2 ppt) in a flow-through system. Each dietary treatment was performed in three replicates. The growth rate positively increased in both linear and quadratic trends with increasing dietary SeNP level ( P < 0.05 ). The liver Se concentration increased with increasing SeNP in diet ( P < 0.05 ). Gut total protease, trypsin, chymotrypsin, alkaline phosphatase (ALP), lipase, and α-amylase activities were significantly enhanced in the SeNP4 group compared to the other treatments ( P < 0.05 ). Antioxidant capacity improved in fish-fed SeNP2 and SeNP4 diets regarding catalase and superoxide dismutase activities and the liver glutathione content. Serum lysozyme and hemolytic activities and white blood cells’ respiratory burst activity in the control were lower than in fish-fed SeNP-supplemented diets. Serum total protein, globulin, and globulin/albumin ratio in fish-fed SeNP1, SeNP2, and SeNP4 diets were higher than the other groups ( P < 0.05 ). The interleukin-10 and granulocyte-macrophage colony-stimulating factor genes’ relative transcription levels in the gut of fish-fed SeNP4 were higher than the other groups. Serum cholesterol, triglycerides, ALP, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase significantly decreased in fish with increasing SeNP content in the diet. In conclusion, supplementing the diet with 4 mg kg−1, SeNP was recommended to improve growth and health indices in L. calcarifer juveniles reared in high saline water (48 ppt).
The aim of this study is to determine to what extent the addition of chitinase to black soldier fly (BSF) larval meal enriched or not with long-chain polyunsaturated fatty acids (LC-PUFAs) could improve growth, protein digestion processes and gut microbial composition in Nile tilapia. Two different types of BSF meal were produced, in which larvae were reared on substrates formulated with vegetable ingredients (VGS) or marine fish offal (FOS). The BSF raised on VGS was enriched in α-linolenic acid-ALA while that raised on FOS was enriched in ALA + eicosapentaenoic acid-EPA +docosahexaenoic-acid-DHA. Six BSF-based diets, enriched or not with chitinase, were formulated and compared with a control diet based on fishmeal and fish oil (FMFO). Two doses (D) of chitinase from Aspergillus niger (2 g and 5 g/kg feed) were added to the BSF larval diets (VGD0 and FOD0) to obtain four additional diets: VGD2, VGD5, FOD2 and FOD5. After 53 days of feeding, results showed that the BSF/FOS based diets induced feed utilization, protein efficiency and digestibility, as well as growth comparable to the FMFO control diet, but better than the BSF/VGS based diets. The supplementation of chitinase to BSF/FOS increased in fish intestine the relative abundance of beneficial microbiota such as those of the Bacillaceae family. The results showed that LC-PUFA enriched BSF meal associated with chitinase could be used as an effective alternative to fishmeal in order to improve protein digestion processes, beneficial microbiota and ultimately fish growth rate.
Arachidonic acid (AA) is an essential fatty acid that has vital influences on early development in marine fish larvae. In this regard, a 30-day research was run to examine the effects of livefoods enrichment with graded levels of AA on some physiological responses of Acanthopagrus latus larvae. Four experimental emulsions containing four levels of AA were used to enrich live foods (Rotifer and Artemia), including very low (6%, N), low (12%, L), medium (24%, M) and high (37.8%, H) AA levels. Newly hatched larvae were distributed into twelve 250-L cylindrical tanks with initial density 15000 larvae in each tank that supplied with seawater (23 ± 1 ºC; 40.0 ± 1.0 g L-1). Nannochloropsis oculata was used in a green water system and live foods were offered to larvae two times daily. Larvae in H group had lower total length (7.24 ± 0.6 mm) than other treatments. The highest survival rate after air exposure and osmotic stress challenges was in H group. The level of n-6 polyunsaturated fatty acids, mainly AA enhanced in the larval whole body with increasing AA level in the emulsion. The highest and lowest alkaline phosphatase activities were in H and N groups, respectively. The aminopeptidase N and leucine alanine paptidase activities in N group were lower than the other treatments. The trypsin activity gradually increased with the increment of AA in the emulsions. The highest and lowest chymotrypsin, amylase and pepsin activities were in H and N groups, respectively. The highest and lowest phospholipase A 2 activity was in M and N groups, respectively. The activity of catalase in the whole body of M group was greater than the rest treatments. The highest and lowest superoxide dismutase activities were in M and N groups, respectively. The activity of glutathione reductase in L and M groups were higher than the rest. Total antioxidant capacity in whole body of larvae in N group was lower than the other treatments and H group showed the highest value. The whole body malondialdehyde level in L group was lower than N and H groups and M showed intermediate value. Based on the findings of the present study, moderate levels of AA suggested for the enrichment of livefoods in A. latus by considering n-3 long chain polyunsaturated fatty acids content in the enrichment solution.
Organotin compounds (OTs) belong to persistent organic pollutants (POPs) group and are capable of persisting up to 40 years due to their chemical nature. Tributyltin (TBT) is an anti‐fouling agent and can easily leach out into water threatening the aquatic life. Molluscs are sensitive towards the presence of toxins in their surrounding environment and respond accordingly to overcome stress conditions. Lamellidens marginalis , a freshwater bivalve is popular for its food value and pink pearls. The present study focuses on the effect of Tributyltin chloride (TBTCl), a form of TBT, on Lamellidens marginalis under laboratory conditions. The 96 h median lethal concentration (LC 50 ) was calculated to be 1 mg L ⁻¹ . Bivalves were segregated into three sub‐groups that is, group I; control, group II; experimental (treated with 0.2 mg L ⁻¹ TBTCl that is, 1/5th of LC 50 value), group III; experimental (treated with 0.2 mg L ⁻¹ TBTCl + 0.003% w/v spirulina). A static non‐renewal toxicity assay was performed for 96 h. Estimation of total protein, acid phosphatase (ACP), alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and glycogen was carried out. An overall decrease in the total protein, ACP, ALP, glycogen, and ALT was observed in both the experimental groups (Groups II and III) in comparison to the control group whereas, the concentration of AST was found to increase in hepatopancreas. Integrated biomarker response index (IBR) value was calculated to be 12.60 for control and 10.56 and 11.29 for groups II and III, respectively in hepatopancreas. Similarly, in gill tissue the values were 3.69, 1.37, and 3.43, respectively. After treatment with nutritional supplement spirulina, a higher BRI (biomarker response index) value of 3.3 for group III was observed, suggesting the therapeutic role of spirulina in test organisms. BRI value derived for group II was 2.7 indicating more severity of TBTCl compared to group III in the absence of spirulina.
Cotton leafworm, Spodoptera littoralis (Boisduval), is one of the major destructive pests of ornamental, industrial, and vegetable crops. The efficacy of technical emamectin benzoate (EMB) and fipronil (FPR) was assessed against the 4th larval instar using leaf-dip bioassay method. EMB was more efficient than FPR based on 96 h LC50 values of 0.004 and 0.023 μg/ml, respectively. Joint toxic action of the dual exposure in sequence with time interval 24 h and in mix were evaluated at LC10:LC10, LC25:LC25 and LC50:LC50 after 96 h posttreatment, as well. Their impacts on detoxification enzymes, esterases (ESTs); alkaline phosphatase (ALP); and glutathione S-transferase (GST) as well as acetylcholine esterase (AChE) were also determined. The sequential exposure of EMB after FPR (S1) produced antagonism, potentiation, and potentiation effects, respectively while sequential exposure of FPR after EMB (S2) interacted as addition, potentiation, and potentiation respectively. The rest of binary mixtures (Mix) revealed antagonistic effect regardless of concentration. Orthogonal contrast analysis showed that the highest elevations of AChE, α-EST, β- EST and ALP enzymes were obtained from Mix at LC50:LC50 (181.6%, 288.4, 229.2 and 460.9%, respectively), LC25:LC25 (131.5%, 252.8, 205.60 and 252.0, respectively) and LC10:LC10 (106.6%, 215.6%, 201.8% and 170.0%, respectively). Differently, the greatest elevation of GST activity (157.7%) resulted from S1 at LC50:LC50, while it was significantly lower at LC25:LC25 and LC10:LC10 as well as Mix and S2 at all concentrations than corresponding concentrations of FPR. These findings shed some light on the role of GST in FPR toxicity and clarified the risk of these dual exposures in elevating detoxification enzymes dangerously compared to their individual insecticides. These dual exposures should be carefully handled. Although rotational exposure at low concentrations may enhance performance and mitigate resistance risk, rotational exposure at high concentrations and Mix may indirectly contribute to the evolution of cross-resistance to other insecticides.
The aim is to investigate cell maturation, synthetic activity, and cell death rate following the exposure of human osteoblasts to alternating biophysical simulation methods. Mechanical vibration, alternating and pulsed electromagnetic fields, and pulsed photobiomodulation can be used for such purposes. Several versatile standard experimental concepts have been described previously and are used primarily in this type of research.
Lipid metabolism is one of energy metabolic pathways that produce adenosine triphosphate (ATP). In this pathway, lysosomal acid lipase (LAL) encoded by Lipase A (LIPA), plays an important role in catalyzing lipids to fatty acids (FAs), which drive oxidative phosphorylation (OXPHOS) and generate ATP. Previously, we found that a LIPA single nucleotide polymorphism rs143793106, which decreases the LAL activity, suppressed the cytodifferentiation of human periodontal ligament (HPDL) cells. However, the mechanisms underlying that suppression are still not fully clarified. Thus, we aimed to investigate the mechanisms regulating the cytodifferentiation of HPDL cells by LAL in terms of energy metabolism. We performed the osteogenic induction of HPDL cells with or without Lalistat-2, a LAL inhibitor. To visualize lipid droplet (LD) utilization, we performed confocal microscopy on HPDL cells. We also performed real-time PCR to analyze the gene expression of calcification-related and metabolism-related genes. Furthermore, we measured the ATP production rate from two major energy production pathways, OXPHOS and glycolysis, and OXPHOS-related parameters of HPDL cells during their cytodifferentiation. We found that LDs were utilized during the cytodifferentiation of HPDL cells. Alkaline phosphatase (ALPL), collagen type 1 alpha 1 chain (COL1A1), ATP synthase F1 subunit alpha (ATP5F1A), and carnitine palmitoyltransferase 1A (CPT1A) mRNA expressions were upregulated, whereas lactate dehydrogenase A (LDHA) mRNA expression was downregulated. Additionally, total ATP production rate was significantly increased. In contrast, in the presence of Lalistat-2, LD utilization was inhibited and ALPL, COL1A1, and ATP5F1A mRNA expression was downregulated. Additionally, ATP production rate and spare respiratory capacity of the OXPHOS pathway were decreased in HPDL cells during their cytodifferentiation. Collectively, the defect of LAL in HPDL cells decreased LD utilization and OXPHOS capacity, resulting in reduced energy to sustain the adequate ATP production required for the cytodifferentiation of HPDL cells. Thus, LAL is important for periodontal tissue homeostasis as a regulator of bioenergetic process of HPDL cells.
Atorvastatin calcium (AC), a cholesterol-lowering medication, has limited oral bioavailability (14%) and adverse impacts on the gastrointestinal tract (GIT), liver, and muscle. So, in an effort to improve the poor availability and overcome the hepatotoxicity complications attendant to peroral AC administration, transdermal transfersomal gel (AC-TFG) was developed as a convenient alternative delivery technique. The impact of utilizing an edge activator (EA) and varying the phosphatidylcholine (PC): EA molar ratio on the physico-chemical characteristics of the vesicles was optimized through a Quality by Design (QbD) strategy. The optimal transdermal AC-TFG was tested in an ex-vivo permeation study employing full-thickness rat skin, Franz cell experiments, an in-vivo pharmacokinetics and pharmacodynamics (PK/PD) evaluation, and a comparison to oral AC using poloxamer-induced dyslipidemic Wister rats. The optimized AC-loaded TF nanovesicles predicted by the 23-factorial design strategy had a good correlation with the measured vesicle diameter of 71.72 ± 1.159 nm, encapsulation efficiency of 89.13 ± 0.125%, and cumulative drug release of 88.92 ± 3.78% over 24 hours. Ex-vivo data revealed that AC-TF outperformed a free drug in terms of permeation. The pharmacokinetic parameters of optimized AC-TFG demonstrated 2.5- and 13.3-fold significant improvements in bioavailability in comparison to oral AC suspension (AC-OS) and traditional gel (AC-TG), respectively. The transdermal vesicular technique preserved the antihyperlipidemic activity of AC-OS without increasing hepatic markers. Such enhancement was proven histologically by preventing the hepatocellular harm inflicted by statins. The results showed that the transdermal vesicular system is a safe alternative way to treat dyslipidemia with AC, especially when given over a long period of time.
Vitamin D insufficiency induces calcification disorder of bone or a decrease in bone mineral density, increasing the risk of fracture. Alkaline phosphatase (ALP) activity, a differentiation marker for intestinal epithelial cells, is regulated by vitamin D. It has also been suggested that ALP may prevent metabolic endotoxemia by dephosphorylating lipopolysaccharide. We hypothesized that vitamin D restriction and/or a high-fat diet influences ALP activity in each tissue and serum lipopolysaccharide concentrations and increases the risk of metabolic endotoxemia. Eleven-week-old female rats were divided into 4 groups: basic control diet (Cont.), basic control diet with vitamin D restriction (DR), high-fat diet (HF), and high-fat diet with vitamin D restriction (DRHF) groups. They were acclimated for 28 days. The results of 2-way analysis of variance showed that intestinal ALP activity, which may contribute to an improvement in phosphate/lipid metabolism and longevity, in the high-fat diet groups (HF and DRHF) was higher than in the low-fat diet groups (Cont. and DR). ALP activity in the vitamin D-restricted groups (DR and DRHF) was lower than in the vitamin D-sufficient groups (Cont. and HF). Furthermore, serum endotoxin concentrations were significantly higher in the high-fat diet groups (HF and DRHF) than in the low-fat diet groups (Cont. and DR). In the vitamin D-restricted groups (DR and DRHF), serum endotoxin concentrations were also significantly higher than in the vitamin D-sufficient groups (Cont. and HF). These results suggest that vitamin D restriction and/or a high-fat diet increases the risk of metabolic endotoxemia.
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