Effects of carbon monoxide releasing molecule-liberated CO on severe acute pancreatitis in rats

Recent studies have suggested that exogenously administered carbon monoxide (CO) is beneficial for resolution of acute inflammation. Severe acute pancreatitis (SAP) is an inflammatory condition which leads to a systemic inflammatory response syndrome (SIRS). In this study, we investigated the role of CO liberated from carbon monoxide releasing molecule-2 (CORM-2) in rats with SAP. SAP was induced by retrograde infusion of 5% sodium taurocholate into the pancreatobiliary duct. Forty Wistar rats were randomly divided into four groups. Sham group was given normal saline after the sham operation. SAP group was treated with normal saline after the induction of SAP. CORM-2 group was injected with CORM-2 (8 mg/kg, i.v.) after the onset of SAP. iCORM-2 group was given iCORM-2 (an inactive compound used as negative control) after SAP induction. All animals were sacrificed at 12 h after the operation. Eighty rats (n = 20 for each group) were monitored for 7 days to observe their survival rates. In another set of experiments, the former three groups received the same treatment as mentioned above. The last group was given ZnPPIX (HO-1 inhibitor) by peritoneal injection at 1 h before the administration of CORM-2 (n = 10 for each group). Serum levels of amylase, tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and interleukin 10 (IL-10) as well as myeloperoxidase (MPO) activity in pancreatic tissue were determined. Histological score, mRNA expression of these cytokines, heme oxygenase-1 (HO-1) expression, HO activity, and nuclear factor κB (NF-κB)-binding activity in the pancreas were also evaluated. Our results showed that compared with SAP group, CORM-2 treatment significantly reduced the serum levels of amylase, TNF-α, and IL-1β, suppressed pancreatic tissue mRNA expression of TNF-α and IL-1β, and decreased MPO activity in the pancreas. In contrast with the pro-inflammatory cytokines, the serum level and pancreatic tissue mRNA expression of IL-10 were markedly increased by the injection of CORM-2. The severity of pancreatic histology and survival rate were also significantly improved by the administration of CORM-2. Treatment with CORM-2 was associated with an increase in HO-1 expression at 12 h after SAP induction. Pretreatment with ZnPPIX had no effect on the production and mRNA expression of these cytokines at 12 h after the development of SAP with the treatment of CORM-2 as compared to CORM-2 group. Furthermore, CORM-2 treatment inhibited the activation of NF-κB in the pancreas. These results indicate that CORM-2-liberated CO exerts protective effects on SAP in rats, and the beneficial effects may be due to the suppression of NF-κB activation and subsequent regulation of NF-κB-dependent expression of cytokines.     

Chitosan cross-linked with poly(ethylene glycol)dialdehyde via reductive amination as effective controlled release carriers for oral protein drug delivery

The covalently cross-linked chitosan-poly(ethylene glycol)₁₅₄₀ derivatives have been developed as a controlled release system with potential for the delivery of protein drug. The swelling characteristics of the hydrogels based on these derivatives as the function of different PEG content and the release profiles of a model protein (bovine serum albumin, BSA) from the hydrogels were evaluated in simulated gastric fluid with or without enzyme in order to simulate the gastrointestinal tract conditions. The derivatives cross-linked with difunctional PEG₁₅₄₀-dialdehyde via reductive amination can swell in alkaline pH and remain insoluble in acidic medium. The cumulative release amount of BSA was relatively low in the initial 2 h and increased significantly at pH 7.4 with intestinal lysozyme for additional 12 h. The results proved that the release-and-hold behavior of the cross-linked CS–PEG₁₅₄₀H-CS hydrogel provided a swell and intestinal enzyme controlled release carrier system, which is suitable for oral protein drug delivery.     

Synthesis and biological evaluation of nimesulide based new class of triazole derivatives as potential PDE4B inhibitors against cancer cells

A new class of 1,2,3-triazol derivatives derived from nimesulide was designed as potential inhibitors of PDE4B. Synthesis of these compounds was carried out via a multi-step sequence consisting of copper-catalyzed azide–alkyne cycloaddition (CuAAC) as a key step in aqueous media. The required azide was prepared via the reaction of aryl amine (obtained from nimesulide) with α-chloroacetyl chloride followed by displacing the α-chloro group by an azide. Some of the synthesized compounds showed encouraging PDE4B inhibitory properties in vitro that is >50% inhibition at 30 μM that were supported by the docking studies of these compounds at the active site of PDE4B enzyme (dock scores ～ ?28.6 for a representative compound). Two of these PDE4 inhibitors showed promising cytotoxic properties against HCT-15 human colon cancer cells in vitro with IC₅₀ ～ 21–22 μg/mL.     

Confirmatory analysis of firocoxib in bovine milk by rapid resolution liquid chromatography tandem mass spectrometry

A rapid method has been developed to analyse for firocoxib (FIRO) residue in bovine milk. Milk samples were extracted with acetonitrile and sample extracts were purified on Evolute? ABN solid phase extraction cartridges. Aliquots were analysed by rapid resolution liquid chromatography tandem mass spectrometry (RRLC–MS/MS). The method was validated in bovine milk, according to the criteria defined in Commission Decision 2002/657/EC. The decision limit (CCα) was 1.18 ng/mL and for the detection capability a (CCβ) value of 2.02 ng/mL was obtained. The measurement uncertainty of the method was 27%. Fortifying bovine milk samples (n = 18) in three separate assays, show the accuracy of the method to be between 96 and 105%. The precision of the method, expressed as RSD values for the within-lab reproducibility at the three levels of fortification (5, 7.5 and 10 ng/mL) was less than 11% respectively.     

Effect of propofol on mitochondrial ATP content and ATPase activity in hippocampus of rats with cerebral ischemia-reperfusion injury

ObjectiveStudy on the influence of the cerebral Ischemia-reperfusion Injury (IRI) on mitochondrial adenosine triphosphate (ATP) content and ATPase activity in hippocampus of rats, as well as the protective effect of propofol on IRI in rats.MethodsA total of 40 male SD rats were randomly divided into 5 groups: sham operation group (Group A), ischemia reperfusion control group (Group B) and ischemic reperfusion with propofol pretreatment group (C group). Group C was further divided into three sub groups according to the different doses of propofol: Group C1 (50 mg/kg), Group C2 (100 mg/kg) and Group C3 (150 mg/kg). The rats from Groups B and C were applied for the IRI model preparation by blockage of the blood flow in arteria carotis communis. For the Groups A, arteria carotis communis were separated without blockage of the blood flow. Before preparation of IRI model for rats in Group C, different doses of propofol were intraperitoneally injected into the rats. For rats in Groups A and B, only saline solution with same volume was intraperitoneally injected at the same time. The ultra-structures of mitochondria in hippocampus of rats were observed under transmission electron microscope, and the mitochondrial degeneration rate was counted. The contents of ATP were determined by HPLC and the ATPase activity was characterized by ATPase activity assay kit.Results(1) Mitochondria in the hippocampus from Groups B and C showed different degrees of ultrastructural damage and more significant mitochondrial degeneration than those from Group A. The degree of damage and the rate of degeneration were in the order of B > C1 > C2 > C3 and the difference was statistically significant (P < 0.01). (2) The contents of ATP and the ATPase activity in hippocampus from Groups B and C were significantly lower than those of Group A, while these indices from Group C were significantly higher than those in the B group, and the sequence was C3 > C2 > C1, indicating that the ATP content and ATPase activity were significantly correlated with the dose of propofol, and the difference was statistically significant (P < 0.05).ConclusionIn summary, the contents of ATP and ATPase activity in hippocampus of rats can be decreased by cerebral IRI. The structure and function of the impaired mitochondria in IRI rats could be significantly improved by propofol, and the improvement effect is related to the dose of propofol.     

Developmental competence and gene expression of immature oocytes following liquid helium vitrification in bovine

The objective of this study was to develop an effective ultra-rapid vitrification method and evaluate its effect on maturation, developmental competence and development-related gene expression in bovine immature oocytes. Bovine cumulus oocyte complexes were randomly allocated into three groups: (1) controls, (2) liquid nitrogen vitrification, and (3) liquid helium vitrification. Oocytes were vitrified and then warmed, the percentage of morphologically normal oocytes in liquid helium group (89.0%) was significantly higher (P < 0.05) than that of the liquid nitrogen group (81.1%). When the vitrified–thawed oocytes were matured in vitro for 24 h, the maturation rate in liquid helium group (50.6%) was higher (P < 0.05) than liquid nitrogen group (42.6%). Oocytes of liquid helium vitrification had higher cleavage and blastocyst rates (41.1% and 10.0%) than that of liquid nitrogen vitrification (33.0% and 4.5%; P < 0.05) after in vitro fertilization. Moreover, the expression of GDF9 (growth/differentiation factor-9), BAX (apoptosis factor) and ZAR1 (zygote arrest 1) was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) when the vitrified–thawed oocytes were matured 24 h. The expression of these genes was altered after vitrification. Expression of GDF9 and BAX in the liquid helium vitrification group was not significantly different from that of the control, however there were significant differences between the liquid nitrogen vitrification group and control. In conclusion, it was feasible to use liquid helium for vitrifying bovine immature oocytes. There existed an association between the compromised developmental competence and the altered expression levels of these genes for the vitrified oocytes.     

Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans

EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N′-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. infantis. The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. K_m values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and V_max values ranging from 3.5 × 10⁻³ to 5.09 × 10⁻³, 4.5 × 10⁻³ to 7.75 × 10⁻³ and 1.9 × 10⁻²to 5.2 × 10⁻² mg/mL × min were determined for ribonuclease B, lactoferrin and whey, respectively. In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q–TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans.     

Effect of serum albumin supplementation on in vitro capacitation and fertilization of caprine oocytes

The aim of the investigation was to study the effect of purity and the type of serum albumin on in vitro fertilization (IVF) and cleavage rate of in vitro matured goat oocytes. Ovaries were collected from the local abattoir and transported within 4 h to the laboratory in warm saline (37 °C) containing 100 IU penicillin-G and 100 μg streptomycin sulfate per ml. A total of 2509 cumulus oocyte complexes (COCs) were collected from 1313 ovaries. Oocytes were matured in TCM-199 medium containing FSH (5 μg/ml), LH (5 μg/ml) and estradiol-17β (1 μg/ml), supplemented with 20% fetal bovine serum at 38.5 °C and 5% CO₂ in an incubator under humidified air for 27 h. After 27 h of in vitro maturation (IVM), oocytes were denuded, washed and randomly divided into 4 groups. Group 1 consisted of in vitro matured oocytes (n = 627) co-incubated with sperm in a 50 μl drop of TALP medium containing a 6 mg/ml crystalline bovine serum albumin (BSA) fraction V and 10 μg/ml heparin. Group 2 was comprised of in vitro matured oocytes (n = 470), co-incubated with sperm in a 50 μl drop of TALP medium containing 3 mg/ml crystalline BSA fraction V, 10% estrous goat serum and 10 μg/ml heparin. Group 3 was comprised of in vitro matured oocytes (n = 489) co-incubated with sperm in a 50 μl drop of TALP medium containing a 6 mg/ml fatty acid free BSA and 10 μg/ml heparin. Group 4 consisted of in vitro matured oocytes (n = 422) co-incubated with sperm in a 50 μl drop of TALP medium containing 20% estrous goat serum and 10 μg/ml heparin. After 18 h of co-incubation, the oocyte–sperm mixture was washed in the culture medium 15–20 times and cultured in 50 μl EDM. Cleavage of the in vitro fertilized oocytes were recorded 48 h post-insemination under an inverted phase contrast microscope. The average oocyte recovery rate/ovary and maturation rate was 1.91% and 80.03%, respectively. The cleavage rate in Group 1, Group 2, Group 3 and Group 4 was 1.59%, 8.93%, 11.86% and 35.30%, respectively. It could be concluded that the use of fatty acid free albumin resulted in a significantly higher (P < 0.05) cleavage rate, compared to unmodified albumin, and the supplementation of 20% estrous goat serum in the fertilization medium, significantly (P < 0.05) increased the cleavage rate of in vitro matured goat oocytes, compared to defatted albumin.     

Metabolic effect of short term administration of Hoodia gordonii,an herbal appetite suppressant

Hoodia gordonii (family: Apocynaceae) is used traditionally by the Khoi-San tribes to control hunger. It has become extremely popular and has triggered commercial interest due to its appetite suppressant property. The present study was undertaken to investigate the appetite regulatory mechanism and associated metabolic changes induced by the herb. Effect of organic solvent extract of H. gordonii on food intake and body weight of male Sprague Dawley rats was monitored at three different doses 50, 100 and 150 mg/kg body weight, given orally for five days. Subsequently, the dose of 100 mg/kg body weight was selected for further studies on the regulatory hormones and biochemical variables. Dose-dependent reduction in food intake (12–26%) was observed at a dose of 100 and 150 mg/kg body weight (p < 0.05). Appetite suppression persisted for 6 h and food intake was restored within 24 h after stopping of the treatment. There was an increase in liver glycogen stores, activity of mitochondrial CPT-1 and thyroid hormones in treated animals. The circulating levels of NPY and IGF-1 were decreased with marginal increase in leptin and CCK, in case of treated rats. There was no change in blood glucose and insulin levels were not affected significantly. The hormonal and metabolic changes due to treatment with the H. gordonii extract may be responsible for its anorectic activity.     

Effects of trehalose supplementation on cell viability and oxidative stress variables in frozen-thawed bovine calf testicular tissue

Trehalose is widely used for cryopreservation of various cells and tissues. Until now, the effect of trehalose supplementation on cell viability and antioxidant enzyme activity in frozen-thawed bovine calf testicular tissue remains unexplored. The objective of the present study was to compare the effect of varying doses of trehalose in cryomedia on cell viability and key antioxidant enzymes activities in frozen-thawed bovine calf testicular tissue. Bovine calf testicular tissue samples were collected and cryopreserved in the cryomedias containing varying doses (0, 5, 10, 15, 20 and 25%; v/v) of trehalose, respectively. Cell viability, total antioxidant capacity (T-AOC) activity, catalase (CAT) activity, superoxide dismutase (SOD) activity, glutathione (GSH) content and malondialdehyde (MDA) content were measured and analyzed. The results showed that cell viability, T-AOC activity, SOD activity, CAT activity and GSH content of frozen-thawed bovine calf testicular tissue was decreased compared with that of fresh group (P < 0.05). MDA content in frozen-thawed bovine calf testicular tissue was significantly increased compared with that of fresh group (P < 0.05). The cryomedia added 15% trehalose exhibited the greatest percentage of cell viability and antioxidant enzyme activity (SOD and CAT) among frozen-thawed groups (P < 0.05). Meanwhile, GSH content was the lowest among frozen-thawed groups (P < 0.05). However, there were no significance differences in MDA content among the groups added 10, 15 and 20% trehalose (P > 0.05). In conclusion, the cryomedia added 15% trehalose reduced the oxidative stress and improved the cryoprotective effect of bovine calf testicular tissue. Further studies are required to obtain more concrete results on the determination of antioxidant capacity of trehalose in frozen-thawed bovine calf testicular tissue.     

The clonogenic potential of hematopoietic stem cells and mesenchymal stromal cells in various hematologic diseases: a pilot study

Background aimsMesenchymal stromal cells (MSC) are the most popular cells used in regenerative medicine and biotechnology. The clonogenic potential of these cells is defined by colony-forming unit-fibroblasts (CFU-F). It is well known that there is an interaction between hematopoietic cells and stromal cells in disease formation pathogenesis. Therefore we hypothesized that there should be a quantitative and qualitative relationship between MSC colonies (CFU-F) and hematopoietic stem cell colonies (colony-forming unit-granulocyte-macrophages; CFU-GM) among patients with and without hematologic diseases.MethodsForty-two patients were included in this study. Patients were divided into three groups: group A, patients with hematologic malignancies (n = 20); group B, patients with bone marrow (BM) failure (n = 11); group C, patients without hematologic diseases (n = 11). BM aspirates were plated in different densities for CFU-F culture. The plating density was the same for CFU-GM culture.ResultsCFU-GM colonies grew in 90% of group A cells and all of group B and C cells (P = 0.0001). CFU-F colonies became visible on the ninth day of plating in group A and on the eight day in groups B and C. There was no statistically significant difference between the groups for the duration of CFU-F colony formation (P = 0.12). There were differences in the morphology of the colonies among the groups.ConclusionsThis is the first study that has compared the clonogenic potential of stromal cells and hematopoietic stem cells in the same subjects with and without hematologic diseases. No correlation was shown between the clonogenic potential of stromal cells and hematopoietic cells.     

Occurrence of cestodes and comparative efficacy of Typha angustata and sulphadimidine against cestodes in Columba livia domestica (Domestic Pigeon)

The occurrence of intestinal parasites of Columba livia domestica has been on the increase, leading to high economic and production losses with more fatal cases. This study was designed to investigate the prevalence of cestodes in pigeons and determine the efficacy of Typha angustata extract and sulphadimidine against these cestodes in the domestic pigeon. A total of 30 pigeons were examined. 18 (60%) pigeons were found infected with only one type of cestode species (Raillietina spp.). The difference in prevalence between males and females was statistically significant (χ² = 8.167, p = 0.004). The mean EPG count in group A (treated with T. angustata extract) before treatment and after treatment was 176 ± 4.33 and 155 ± 4.24, respectively. In group B (treated with sulphadimidine), the mean EPG calculated before treatment and after treatment was 184 ± 6.74 and 35 ± 3.53, respectively. The efficacy at day 28 of T. angustata and Sulphadimidine was 11.93% and 80.97%, respectively. It was concluded on the basis of the EPG and efficacy data that T. angustata extract had low efficacy against raillietiniasis, while as sulphadimidine, which is also used before to treat different intestinal parasites, had a good efficacy against raillietiniasis. Further studies are required to know the prevalence of other gastrointestinal parasites in pigeons and efficacy of different medicinal plants against such parasites.     

Babesia bigemina: Advances in continuous in vitro culture using serum-free medium supplemented with insulin,transferrin, selenite,and putrescine

This study reported that Babesia bigemina (Bbig-SF) was continuously cultured in vitro in a serum-free medium supplemented with a mixture of insulin-transferrin-selenite (M-ITS) and putrescine (Pu). Firstly, the effect of five different types of basal culture media supplemented with 40% bovine serum was evaluated regarding the proliferation of the protozoan parasite. Cultures with the advanced DMEM/F12 medium (A-DMEM/F12) showed the highest percentage of parasitized erythrocytes (PPE) at 8.37%. Using A-DMEM/F12, a strain of B. bigemina (Bbig-SF) was adapted for growth in bovine serum-free medium by a sequential reduction of serum and demonstrated a maximum PPE of 7.18% in the absence of serum. The next study was the evaluation of the effect of adding four different concentrations of M-ITS to the serum-free A-DMEM/F12 medium on Bbig-SF; the optimal concentrations of M-ITS were 2000, 1100, and 1.34 mg/L, which yielded a PPE of 7.23%. Next, eight levels of Pu were evaluated on Bbig-SF cultured in serum-free A-DMEM/F12. After the addition of 0.1012 mg/L of Pu, the maximum PPE was 7.61%. When the combination of serum-free A-DMEM/F12 + M-ITS (2000, 1100, and 1.34 mg/L) + Pu (0.1012 mg/L) was evaluated, it yielded a maximum PPE of 14.80%. Finally, the combination of M-ITS + Pu in A-DMEM/F12 without serum and incorporation of a perfusion bioreactor yielded a maximum PPE of 33.45%. We concluded these culturing innovations for B. bigemina in vitro allow the optimization of small- and large-scale proliferation as a source of this protozoan parasite for future studies.     

Astragaloside IV ameliorates renal injury in streptozotocin-induced diabetic rats through inhibiting NF-κB-mediated inflammatory genes expression

Accumulating evidence suggests that inflammatory processes are involved in the development of diabetic nephropathy (DN). However, there are no effective interventions for inflammation in the diabetic kidneys. Here, we tested the hypothesis that Astragaloside IV(AS-IV), a novel saponin purified from Astragalus membranaceus (Fisch) Bge, ameliorates DN in streptozotocin (STZ)-induced diabetic rats through anti-inflammatory mechanisms. Diabetes was induced with STZ (65 mg/kg) by intraperitoneal injection in rats. Two weeks after STZ injection, rats were divided into three groups (n = 8/each group), namely, diabetic rats, diabetic rats treated with AS-IV at 5 and 10 mg kg^?1 d^?1, p.o., for 8 weeks. The normal rats were chosen as nondiabetic control group (n = 8). The rats were sacrificed 10 weeks after induction of diabetes. AS-IV ameliorated albuminuria, renal histopathology and podocyte foot process effacement in diabetic rats. Renal NF-κB activity, as wells as protein and mRNA expression were increased in diabetic kidneys, accompanied by an increase in mRNA expression and protein content of TNF-α, MCP-1 and ICAM-1 in kidney tissues. The α₁-chain type IV collagen mRNA was elevated in the kidneys of diabetic rats. All of these abnormalities were partially restored by AS-IV. AS-IV also decreased the serum levels of TNF-α, MCP-1 and ICAM-1 in diabetic rats. These findings suggest that AS-IV, a novel anti-inflammatory agent, attenuated DN in rats through inhibiting NF-κB mediated inflammatory genes expression.     

Microbiological assay for quantitative determination of polyoxin B

Polyoxin B, an antifungal agro-antibiotic, is used for treating serious plant diseases. Until now there have been no reports on the antibiotic quantification in liquid in spite of its need for fermentation studies. This work reported a microbiological assay, the cylinder-plate method, for the determination of polyoxin B. The results of assay were treated statistically by analysis of variance (ANOVA) and were found linear in the range of 10–500 μg/ml (r² = 0.998), precise (intra-assay: relative standard deviation (R.S.D.) = 0.64; inter-assay: R.S.D. = 1.70) and accurate. This newly established method was applied to determine the antibiotic titer in Streptomyces cacaoi fermentation with comparison to HPLC analysis. The microbiological assay was satisfactory for polyoxin B quantification, and a simple, sensitive, cost-effective and specific agar diffusion bioassay for polyoxin B was thus developed. This work also demonstrated the usefulness of the microbiological assay for quantitative analysis of antibiotics in fermentation.     

Testosterone may increase selective attention to threat in young male macaques

Animal studies indicate that sex hormones have widespread effects on the brain, cognition and emotion, but findings in humans are inconsistent. Well-controlled studies in nonhuman primates are crucial to resolve these discrepancies. In this study, we examined the effects of testosterone (T) on emotion in male rhesus monkeys. Six young adult males were tested on two emotional tasks during three hormonal conditions in a crossover design: when intact at baseline and when pharmacologically hypogonadal with add-back of T or placebo. The emotional tasks were the Approach–Avoidance task, which tested behavioral responses to three categories of objects (familiar, novel, and negative) and a Social Playback task which tested behavioral responses to scenes of unfamiliar conspecifics engaged in three types of social activities (neutral, positive, or negative). Following a 4-week baseline period, monkeys were treated with Depot Lupron, 200 μg/kg before being randomly assigned to one of two treatment groups: Depot Lupron + Testosterone Enanthate (TE, 20 mg/kg) or Depot Lupron + oil vehicle. In each treatment group, monkeys received one injection of Lupron and one injection of TE or one injection of Lupron and one injection of oil at the onset of a 4-week testing period, before crossing over to the alternate treatment for an additional 4 weeks of testing. TE treatment had no effect on behavioral measures in the Approach–Avoidance task. For the Social Playback task, however, TE significantly increased watching time of video clips which depicted fights between unfamiliar conspecifics. The enhancing effect of T on watching time for negative social scenes is consistent with human data suggesting that T decreases aversion or facilitates approach to threatening social stimuli. Further studies are needed to understand the mechanisms by which T may mediate responsiveness to social threat in male primates.     

Application of enhanced stromal vascular fraction and fat grafting mixed with PRP in post-traumatic lower extremity ulcers

BackgroundThe authors presented their experience in regenerative surgery of post-traumatic lower extremity ulcers, evaluating the effects related to the use of Enhanced Stromal Vascular Fraction (e-SVF) and Fat Grafting with Platelet rich Plasma (PRP). The authors compared the results of two control groups.MethodThe analysis involved 20 patients aged between 23 to 62 years affected by post-traumatic lower extremity ulcers. 10 patients managed with e-SVF and 10 patients managed with Fat grafting + PRP in the Plastic and Reconstructive Surgery Department at “Tor Vergata” University Rome. Patients in the first control group (n = 10), were treated only with curettage and application of hyaluronic acid in the bed of ulcers. Patients in the second control group (n = 10), were treated only with PRP.ResultsThe authors showed that wounds treated with e-SVF healed better than those treated with hyaluronic acid. In fact, after 9.7 weeks, patients treated with e-SVF underwent 97.9% ± 1.5% reepithelialisation compared to 87.8% ± 4.4% of the first control group (only hyaluronic acid; p < 0.05). Patients treated with PRP and fat grafting also showed an improvement in reepithelialisation; in fact after 9.7 weeks, they underwent a 97.8% ± 1.5% reepithelialisation compared to 89.1% ± 3.8% of the second control group (only PRP; p < 0.05). As reported e-SVF and PRP mixed with fat grafting were the two treatments evidencing improvement in the healing of patients post-traumatic extremity ulcers.ConclusionsThe results obtained proved the efficacy of these treatments, and the satisfaction of the patients confirmed the quality of the results.     

The impact of nandrolone decanoate administration on ovarian and uterine tissues in rat: Luteinizing hormone profile,histopathological and morphometric assessment

The study had been conducted to evaluate the effects of nandrolone decanoate (abused repeated doses) on female rat’s ovary and uterus during administration and withdrawal. The study included 18 rats that were divided into control group (n = 6) and treated group (n = 12). The treated group was injected intramuscular (IM) with nandrolone decanoate (7 mg/kg body weight) for three consecutive days, for two weeks. The study stated that nandrolone decanoate increases the weights of body, ovary, and uterus. Moreover, it has a tendency of bringing upon modifications in the biochemical, histopathological, and morphological makeup of the female reproductive aspects. In conclusion, nandrolone decanoate has been identified as deleterious element for the female rats, and it is suggested that keen observations must be made on the human abusers to control and manage the possible pathologies.     

2,5-Diaryloxadiazoles and their precursors as novel inhibitors of cathepsins B,H and L

High levels of cathepsins indicated in various pathological conditions like arthritis, cancer progressions, and atherosclerosis explains the need to explore potential inhibitors of these proteases which can be of great therapeutic significance. We, in the present work, report the synthesis of some 2,5-diaryloxadiazoles from N-subsitutedbenzylidenebenzohydrazides. The synthesized compounds were screened for their inhibitory potential on cathepsins B, H and L. Structure Activity Relationship studies show that 2,5-diaryloxadiazoles were less inhibitory than their precursors. 1i and 2k have been found to be most inhibitory to cathepsins B and L. Their K_i values have been calculated as 11.38 × 10⁻⁸ M and 66.4 × 10⁻⁸ M for cathepsin B and 4.2 × 10⁻⁹ M and 47.31 × 10⁻⁹ M for cathepsin L, respectively. However, cathepsin H activity was maximally inhibited by compounds, 1e and 2c with K_i values of 4.4 × 10⁻⁷ M and 5.6 × 10⁻⁷ M, respectively. Enzyme kinetic studies suggest that these compounds are competitive inhibitors to the enzymes. The results have been compared with docking results obtained using iGemDock.     

Structurally and functionally characterized in vitro model of rabbit vocal fold epithelium

In this paper, we describe a method for primary culture of a well differentiated electrically tight rabbit vocal fold epithelial cell multilayer and the measurement of transepithelial electrical resistance (TEER) for the evaluation of epithelial barrier function in vitro. Rabbit larynges were harvested and enzymatically treated to isolate vocal fold epithelial cells and to establish primary culture. Vocal fold epithelial cells were co-cultured with mitomycin C-treated feeder cells on collagen-coated plates. After 10–14 days in primary culture, cells were passaged and cultured until they achieved 70–90% confluence on collagen-coated plates. Epithelial cells were then passaged onto collagen-coated cell culture inserts using 4.5 cm² membrane filters (1.0 μm pore size) with 10% fetal bovine serum or 30 μg/mL bovine pituitary extract to investigate the effects of growth-promoting additives on TEER. Additional experiments were performed to investigate optimal seeding density (1.1, 2.2, 4.4, or 8.9 × 10⁵ cells/cm²), the effect of co-culture with feeder cells, and the effect of passage number on epithelial barrier function. Characterization of in vitro cultures was performed using hematoxylin and eosin staining and immunostaining for vocal fold epithelial cell markers and tight junctions. Results revealed higher TEER in cells supplemented with fetal bovine serum compared to bovine pituitary extract. TEER was highest in cells passaged at a seeding density of 2.2 × 10⁴ cells/cm², and TEER was higher in cells at passage two than passage three. Ultrastructural experiments revealed a well-differentiated epithelial cell multilayer, expressing the epithelial cell markers CK13, CK14 and the tight junction proteins occludin and ZO-1.