Menstrual Disorders and Androgen-Related Traits in Young Women with Type 1 Diabetes Mellitus: a Clinical Study

Objective: To investigate possible causes of menstrual disorders and androgen-related traits in young women with type 1 diabetes mellitus (T1DM).Methods: Fifty-three women with T1DM (duration 8.0 ± 5.6 years), 41 women with (polycystic ovary syndrome) PCOS, and 51 controls matched for age (19.4 ± 4.3 years vs. 21.2 ± 2.7 years vs. 20.8 ± 3.1 years; P>.05) and body mass index (BMI) (22.2 ± 2.7 kg/m² vs. 21.9 ± 2.0 kg/m² vs. 21.4 ± 1.9 kg/m²; P>.05) were prospectively recruited.Results: Two women (3.8%) in the T1DM group had not experienced menarche (at 15.5 and 16.6 years); of the rest, 23.5% had oligomenorrhea, 32.1% hirsutism, and 45.3% had acne. The age at menarche was delayed in the T1DM group compared to controls (12.7 ± 1.3 vs. 12.0 ± 1.0 years; P = .004), while no difference was observed with the polycystic ovary syndrome (PCOS) group (12.4 ± 1.2 years). There were no differences in total testosterone (0.43 ± 0.14 ng/mL vs. 0.39 ± 0.14 ng/mL; P>.05), dehydroepiandrosterone sulfate (DHEA-S) (269 ± 112 μg/dL vs. 238 ± 106 μg/dL; P>.05) or Δ4-androstenedione (2.4 ± 1.3 ng/mL vs. 1.9 ± 0.5 ng/mL; P>.05) concentrations between T1DM and controls. However, patients with T1DM had lower sex hormone binding globulin (SHBG) concentrations than controls (61 ± 17 nmol/L vs. 83 ± 18.1 nmol/L; P = .001), which were even lower in the PCOS group (39.5 ± 12.9 nmol/L; P = .001 compared with T1DM). The free androgen index (FAI) was higher in the PCOS group compared with both other groups (T1DM vs. PCOS vs. controls: 2.53 ± 0.54 vs. 7.88 ± 1.21 vs. 1.6 ± 0.68; P<.001). FAI was higher in patients with T1DM compared to controls as well (P = .038). There was no difference in DHEA-S concentrations between T1DM and PCOS patients (269 ± 112 μg/dL vs. 297 ± 100 μg/dL; P>.05).Conclusion: Menstrual disorders and androgen-related traits in young women with T1DM may be attributed to an increase in androgen bioavailability due to decreased SHBG concentrations.     

Clinical Profile of Nonalcoholic Fatty Liver Disease Among Young Patients With Type 1 Diabetes Mellitus Seen at a Diabetes Speciality Center in India

ObjectiveTo estimate the prevalence and clinical profile of nonalcoholic fatty liver disease (NAFLD) among young type 1 diabetes mellitus (T1DM) patients at a tertiary care diabetes center in India.MethodsElectronic medical records of T1DM patients (age at first diagnosis of T1DM ≤ 25 years) registered between January 1992 and May 2013 who had undergone ultrasonography and denied history of any alcohol intake (n = 736) were reviewed. NAFLD was diagnosed if there was any degree of fatty liver. Retinopathy was initially assessed by direct and indirect ophthalmoscopy and later by retinal photography. Nephropathy was diagnosed if urine protein excretion was > 500 mg/day, and neuropathy was diagnosed if a patient’s vibration perception threshold on biothesiometry was ≥ 20 V.ResultsA total of 204/736 (27.7%) T1DM patients had NAFLD. Compared to T1DM subjects without NAFLD those with NAFLD had higher body mass index (BMI) (18.9 ± 4.2 vs. 20.2 ± 4.7 kg/m², P < .001), waist circumference (67.9 ± 13.2 vs. 71.9 ± 13.3 cm, P < .05), systolic blood pressure (110 ± 15 vs. 116 ± 18 mm Hg, P < .001) and diastolic blood pressure (72 ± 9 vs. 74 ± 10 mm Hg, P < .05), while fasting blood glucose (201 ± 101 vs. 183 ± 101 mg/dL, P < .05) and alkaline phosphatase (419 [12.5] vs. 315 [15.8], P < .001) levels were lower in patients with T1DM with NAFLD. Multiple logistic regression analysis showed a significant association between NAFLD and retinopathy (odds ratio [OR]: 2.01, 95% confidence interval [CI]: 1.13-3.43; P = .017, after adjusting for sex, duration of diabetes, overweight/obesity, hypertension, fasting plasma glucose, nephropathy, and nephropathy (OR: 1.89, 95% CI: 1.02-3.50; P = .042), after adjusting for sex and fasting plasma glucose.ConclusionsThis study suggests that NAFLD is also seen among T1DM patients and that it has an independent and significant association with retinopathy and nephropathy. (Endocr Pract. 2014;20:1249-1257)     

Iodine in Raw and Pasteurized Milk of Dairy Cows Fed Different Amounts of Potassium Iodide

Relation between iodine (I) intake by lactating Holstein cows and iodine concentrations in raw and pasteurized milk were investigated. Four treatment groups with eight cows assigned to each treatment were fed a basal diet containing 0.534 mg I/kg alone or supplemented with potassium iodide at 2.5, 5 or 7.5 mg/kg in 7-week period. Iodine concentrations in raw milk increased with each increase in dietary I from 162.2 ng/ml for basal diet to 534.5, 559.8 and 607.5 ng/ml when 2.5, 5 and 7.5 mg/kg was fed as potassium iodide (P < 0.05). This trend was found for blood plasma and urine iodine concentration. Iodine supplementation had no significant effect on thyroidal hormones. high-temperature short-time (HTST) pasteurization process reduced I concentration. The mean iodine content found in the milk prior to heating processing was 466.0 ± 205.0 ng/ml, whereas for the processed milk this level was 349.5 ± 172.8 ng/ml. It was concluded that iodine supplementation above of NRC recommendation (0.5 mg/kg diet DM) resulted in significant increases in iodine concentrations in milk, although the effect of heating in HTST pasteurization process on iodine concentration was not negligible.     

The Association of Decreased Serum Gdnf Level with Hyperglycemia and Depression in Type 2 Diabetes Mellitus

Objective: Comorbidity of diabetes and depression is a critical problem. Decreased glial-derived neurotrophic factor (GDNF) has been demonstrated in depression, but no evidence of a relationship between GDNF and diabetes has been shown. The present studies were designed to investigate the relationship between GDNF and metabolism.Methods: In Study 1, we performed a case-control study in which subjects with type 2 diabetes mellitus (T2DM), prediabetes (p-DM), and normal glucose tolerance (NGT) were included. In Study 2, we performed a cross-sectional study in 296 patients having pre-existing diabetes in whom the levels of serum GDNF, blood glucose, blood lipids, blood pressure, body mass index, scores from the Patient Health Questionnaire (PHQ-9), the EuroQol-5 scale, and the diabetes distress scale were measured, as well as single-nucleotide polymorphisms of GDNF including rs884344, rs3812047, and rs2075680.Results: In Study 1, serum GDNF concentration was significantly lower in the T2DM group than in the NGT group (NGT: 11.706 ± 3.918 pg/mL; p-DM: 10.736 ± 3.722 pg/mL; type 2 diabetes mellitus &lsqb;T2DM group]: 9.884 ± 2.804 pg/mL, P = .008). In Study 2, significantly decreased serum GDNF levels were observed in subjects with poor glycemic control or depression (glycated hemoglobin &lsqb;HbA1c] <7.0% without depression: 11.524 ± 2.903 pg/mL; HbA1c ≥7.0% without depression: 10.625 ± 2.577 pg/mL; HbA1c <7.0% with depression: 10.355 ± 2.432 pg/mL; HbA1c ≥7.0% with depression: 8.824 ± 2.102 pg/mL, P = .008). Double-factor variance analysis showed that glycemic control and depression were independent factors for the GDNF level. Moreover, the serum GDNF level was significantly inversely associated with the fasting plasma glucose, 2 hours postprandial plasma glucose, HbA1c, and PHQ-9 score.Conclusion: Glycemic dysregulation was an independent factor for the GDNF level. These findings suggest that GDNF level might be involved in the pathophysiology of T2DM and depression through various pathways.Abbreviations: BP = blood pressure; CHO = cholesterol; DDS = diabetes distress scale; DM = diabetes mellitus; EQ-5D = the health-related dimensions of the EuroQol-5 scale; FPG = fasting plasma glucose; GDNF = glial-derived neurotrophic factor; HbA1c = glycated hemoglobin; HDL = high-density lipoprotein; LDL = low-density lipoprotein; NGT = normal glucose tolerance; PHQ-9 = Patient Health Questionnaire; p-DM = prediabetes; PPG = postprandial plasma glucose; SNP = single-nucleotide polymorphism; T2DM = type 2 diabetes mellitus; TG = triglyceride     

Characterization and distribution of cholinesterase activity in mouse uterine horns: changes in estrous cycle

1. Both butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) are present in the mouse uterus, BChE being more abundant.2. Their molecular forms were sequentially solubilized by different extraction media obtaining three ChE fractions whose specific activity was different, depending on the stage of the estrous cycle: hydrosoluble (estrous: 75.5 ± 6.6 and diestrous: 47.9 ± 8.7 μU/mg prot); detergent-soluble or amphiphilic (estrous 26.6 ± 2.4 and diestrous 14.7 ± 3.3 mU/mg prot.), and high ionic strength-soluble (estrous: 18.7 ±4.2 and diestrous 12.8 ± 1.2 mU/mg prot.).3. Histochemical procedures demonstrated a different distribution for both ChE activities. AChE was found in nerves next to smooth muscle cells of the circular layer and blood vessels, while BChE was concentrated in the longitudinal stratum surrounding the smooth muscle cells. Under the predominance of progesterone, BChE was also found in the endometrial glands.4. Maximal contractions evoked by the addition of ACh to the isolated organ bath were concentration dependent and greater in estrous than in diestrous. Nevertheless the difference at the two stages of the estrous cycle disappeared when contractions were normalized to smooth muscle cross-sectional area.5. BChE but not AChE inhibition augmented maximal contractions elicited by ACh in longitudinal but not in circular smooth muscle.6. The effect of BChE inhibition on the contractile force developed was greater at lower concentrations of ACh and did not depend on the stage of the estrous cycle.     

Thiamine modulates intestinal morphological structure and microbiota under subacute ruminal acidosis induced by a high-concentrate diet in Saanen goats

Ruminant animals are generally fed with starch-rich grain as the main energy source, and the incidence of metabolic diseases such as subacute ruminal acidosis (SARA) is high due to the intensive farming. Thiamin has been reported to alleviate SARA caused by high-concentrate diets, but the exact mechanism is not well understood. The goal of this study was to examine the role of thiamine in intestinal inflammation and microbiota caused by high-concentrate diets. The SARA model was induced by low neutral detergent fibre/starch ration to study the effects of thiamine on intestinal tissue structure and microbiota. 18 mid-lactation (148 ± 3 d in milk; milk yield = 0.71 ± 0.0300 kg/d) Saanen goats (BW = 36.5 ± 1.99 kg; body condition score = 2.73 ± 0.16, where 1 = emaciated and 6 = obese) in parities 1 or 2 were selected. The goats were randomly divided into three groups with six replicates: (1) control diet (C; concentrate:forage 30:70), (2) high-concentrate diet (H; concentrate:forage 70:30), and (3) high-concentrate diet with 200 mg of thiamine/kg of DM intake (H + T;concentrate:forage 70:30). The experimental period was lasted for 56 d. The small and large intestine, expression of inflammatory factor genes, tight junction protein genes, total antioxidant capacity, and intestinal microbiota were measured. The results showed that SARA was observed in treatment H, whereas rumen fluid pH was improved in treatment H + T. Treatment H + T also significantly repaired the intestinal tissue structure damaged by SARA, improved the total antioxidant capacity of the small intestinal mucosa, reduced mRNA expression of inflammatory factors in the small intestine tissue, and increased the mRNA expression of tight junction genes in small intestine tissue. The high-concentrate diet reduced the diversity of intestinal microbiota. When thiamine is added to the high-concentrate diet, the relative abundance of intestinal Firmicutes and beneficial bacteria represented by Lactobacilli were upregulated, and the relative abundance of Proteus, a marker of intestinal dysbacteriosis, returned to normal. In conclusion, thiamine supplementation could alleviate the damage to the intestinal tissue structure and microbial environment caused by SARA condition in dairy goats fed a high-concentrate diet.     

Dopaminergic Regulation of Striatal Acetylcholine Release: The Critical Role of Acetylcholinesterase Inhibition

Abstract: This study examined the effects of different levels of acetylcholinesterase (AChE) inhibition on dopaminergic regulation of striatal acetylcholine (ACh) release as estimated by in vivo brain microdialysis. Systemic administration of d-amphetamine (2 or 10 mg/kg) increased the striatal output of ACh when the AChE inhibitor neostigmine (0.1 µM) was present in the perfusion fluid. In contrast, when the same experiments were conducted at 0.01 µM neostigmine, d-amphetamine failed to affect (2 mg/kg) or significantly decreased (10 mg/kg) striatal ACh output. The inhibitory action of the D₂ receptor agonist quinpirole (0.2 mg/kg) was significantly greater at 0.01 µM than at 0.1 µM neostigmine. Similarly, there was a nonsignificant trend for the D₂ antagonist raclopride (1 mg/kg) to stimulate ACh release to a greater extent at the low neostigmine concentration. In contrast, the stimulant effects of systemic administration of the D₁ agonist A-77636 (1.46 mg/kg) on striatal ACh release were the same at the two neostigmine concentrations. These results demonstrate that the concentration of an AChE inhibitor in the perfusion solution can quantitatively and even qualitatively influence the manner in which dopaminergic agents regulate ACh overflow in the striatum. On comparing the present results with earlier reports concerning the effects of d-amphetamine on tissue concentrations of ACh, it is tentatively concluded that a low neostigmine concentration is the more physiologically relevant condition. Under such conditions, at moderate doses d-amphetamine does not appear to alter striatal ACh release, with this likely being due to the opposing actions of D₁ and D₂ receptors. Nevertheless, until the endogenous interstitial concentrations of striatal ACh can be measured by other methods, the physiological relevance of ACh microdialysis studies in the striatum will remain uncertain.     

Evaluation of analgesic,antiamnesic and antidiarrheal potentials of Medicago denticulata extract using animal model

The analgesic, antidiarrheal, and neuro-pharmacological potentials of Medicago denticulata leaves extract were screened in animal models. Potential analgesic response was noted (*P < 0.05, ^**P < 0.01, ^***P < 0.001) in formalin, acetic acid and heat-induced pain models in a dose-dependent manner. Maximum activity by means of writhing inhibition was documented for Medicago denticulata at 300 mg/kg that was found to be 71.79% (17.43 ± 1.31). In first phase, the Medicago denticulata at a dose of 150 and 300 mg/kg showed analgesic activity and reduced the pain by 54.18% (18.39 ± 1.67) and 62.90% (14.89 ± 1.56), respectively. In second phase, the Medicago denticulata at a dose of 150 and 300 mg/kg showed analgesic activity and reduced the pain by 69.48% (19.78 ± 1.44) and 70.89% (18.86 ± 1.58), respectively. In hot plate method, the Medicago denticulata at a dose of 150 and 300 mg/kg showed the maximum response of 61.16% (8.47 ± 1.23) and 67.39% (10.09 ± 1.04), respectively at 60 min. Scopolamine significantly reduces spontaneous alteration in Y-maze model for antiamnesic activity. Medicago denticulata significantly increased the discrimination index in a dose-dependent manner using novel object recognition test (NORT) model. Exploration time in sec for the novel object was increased significantly (P < 0.001) by donepezil decreased for familiar one with a discrimination index (DI) of 62.18%. Medicago denticulata significantly increased the discrimination index by 60.86% and 57.24% at 300 and 150 mg/kg b.w, respectively. The lowest DI of 53.80% at 75 mg/kg was observed in comparison to the amnesic group. The Medicago denticulata significant decreased the elevated levels of acetylcholinesterase (AChE) and malondialdehyde (MDA and enhancing level of acetylcholine (ACh), superoxide dismutase (SOD) and catalase (CAT) acting as an antioxidant agent. Medicago denticulata reduced the total number of diarrheal feces to lesser extent at dose-dependent manner. From the study results, it is suggested that the Medicago denticulata extract possess good analgesic and antiamnesic activity however the antidiarrheal effects of plant were negligible. In the current study, the traditional use of the plant as a source of medicine has been validated.     

Diabetes Status and Race are Associated With Cardiovascular Risk Markers in Obese Adolescents

ObjectiveThe objective of this study was to evaluate differences in cardiovascular disease (CVD) risk markers in obese adolescents based on diabetes status and race in order to improve risk-reduction intervention strategies.MethodsThis was a retrospective, cross-sectional study of obese adolescents, age 10 to 21 years, who were evaluated at Children’s of Alabama between 2000 and 2012. Subjects were classified by glycated hemoglobin (HbA_1c) as having normoglycemia, prediabetes, or type 2 diabetes mellitus (T2DM).ResultsThere were a total of 491 African American (AA) or Caucasian American (CA) subjects. Body mass index was not different between HbA_1c and racial groups. Compared to subjects with normoglycemia or prediabetes, subjects with T2DM had higher levels of total cholesterol (TC) (178.6 ± 43.8 mg/dL vs. 161.5 ± 32.5 mg/dL vs. 162.4 ± 30.6 mg/dL; P < .0001) and low-density-lipoprotein cholesterol (107.4 ± 39.2 mg/dL vs. 97.0 ± 31.0 mg/dL vs. 97.5 ± 26.9 mg/dL; P = .0073). Compared with AA subjects, CA subjects had lower high-density-lipoprotein cholesterol (HDL-C) levels (40.4 ± 10.4 mg/dL vs. 44.3 ± 11.9 mg/dL; P = .0005) and higher non-HDL-C levels (129.6 ± 36.2 mg/dL vs. 122.5 ± 37.5 mg/dL; P = .0490). Of the characteristics studied, HbA_1c had the most significant positive association with dyslipidemia and was strongly correlated with both TC (β, 4.21; P < .0001) and non-HDL-C (β, 4.3; P < .0001).ConclusionObese adolescents with T2DM have more abnormal lipoprotein profiles than those with normoglycemia or prediabetes. Obese CA adolescents have more abnormal lipids than obese AA adolescents. HbA_1c was the characteristic most highly associated with abnormal lipoprotein profiles in our subjects. Our results show that CVD risk markers in obese adolescents vary by race and HbA_1c concentration. (Endocr Pract. 2015;21:165-173)     

Inflammatory and lipid regulation by cholinergic activity in epicardial stromal cells from patients who underwent open‐heart surgery

The modulation of acetylcholine (ACh) release by botulinum toxin injection into epicardial fat diminishes atrial fibrillation (AF) recurrence. These results suggest an interaction between autonomic imbalance and epicardial fat as risk factors of AF. Our aim was to study the inflammatory, lipidic and fibroblastic profile of epicardial stroma from patients who underwent open‐heart surgery, their regulation by cholinergic activity and its association with AF. We performed in vitro and ex vivo assays from paired subcutaneous and epicardial stromal cells or explants from 33 patients. Acute ACh effects in inflammation and lipid‐related genes were analysed by qPCR, in intracellular calcium mobilization were performed by Fluo‐4 AM staining and in neutrophil migration by trans‐well assays. Chronic ACh effects on lipid accumulation were visualized by AdipoRed. Plasma protein regulation by parasympathetic denervation was studied in vagotomized rats. Our results showed a higher pro‐inflammatory profile in epicardial regarding subcutaneous stromal cells. Acute ACh treatment up‐regulated monocyte chemoattractant protein 1 levels. Chronic ACh treatment improved lipid accumulation within epicardial stromal cells (60.50% [22.82‐85.13] vs 13.85% [6.17‐23.16], P < .001). Additionally, patients with AF had higher levels of fatty acid‐binding protein 4 (1.54 ± 0.01 vs 1.47 ± 0.01, P = .005). Its plasma levels were pronouncedly declined in vagotomized rats (2.02 ± 0.21 ng/mL vs 0.65 ± 0.23 ng/mL, P < .001). Our findings support the characterization of acute or chronic cholinergic activity on epicardial stroma and its association with AF.     

Tetramethylpyrazine Attenuates Cognitive Impairment Via Suppressing Oxidative Stress,Neuroinflammation, and Apoptosis in Type 2 Diabetic Rats

Cognitive dysfunction is an important complication observed in type 2 diabetes mellitus (T2DM) patients. Tetramethylpyrazine (TMP) is known to exhibit anti-diabetic and neuroprotective properties. Therefore, the present study aimed to investigate the possible therapeutic effects of TMP against type 2 diabetes-associated cognitive impairment in rats. High-fat diet (HFD) followed by a low dose of streptozotocin (35 mg/kg) was used to induce diabetes in Sprague–Dawley rats. TMP (20, 40, and 80 mg/kg) and Pioglitazone (10 mg/kg) were administered for 4 weeks. The Morris water maze (MWM) and novel objective recognition task (NOR) tests were used to assess memory function. Fasting blood glucose (FBG), lipid profile, HOMA-IR, glycosylated hemoglobin (HbA1c), and glucose tolerance were measured. Acetylcholinesterase (AChE) and choline acetytransferase (ChAT) activity, acetylcholine (ACh) levels, oxidative stress, apoptotic (Bcl-2, Bax, caspase-3), and inflammatory markers (TNF-α, IL-1β, and NF-kβ) were assessed. BDNF, p-AKT, and p-CREB levels were also measured. In the present work, we observed that treatment of diabetic rats with TMP alleviated learning and memory deficits, improved insulin sensitivity, and attenuated hyperglycemia and dyslipidemia. Furthermore, treatment with TMP increased BDNF, p-Akt, and p-CREB levels, normalized cholinergic dysfunction, and suppressed oxidative, inflammatory, and apoptotic markers in the hippocampus. Collectively, our results suggest that the TMP may be an effective neuroprotective agent in alleviating type 2 diabetes-associated cognitive deficits.

     

Retinol‐binding Protein 4 (RBP4) Protein Expression Is Increased in Omental Adipose Tissue of Severely Obese Patients

Visceral fat has been linked to insulin resistance and type 2 diabetes mellitus (T2DM); and emerging data links RBP4 gene expression in adipose tissue with insulin resistance. In this study, we examined RBP4 protein expression in omental adipose tissue obtained from 24 severely obese patients undergoing bariatric surgery, and 10 lean controls (4 males/6 females, BMI = 23.2 ± 1.5 kg/m²) undergoing elective abdominal surgeries. Twelve of the obese patients had T2DM (2 males/10 females, BMI: 44.7 ± 1.5 kg/m²) and 12 had normal glucose tolerance (NGT: 4 males/8 females, BMI: 47.6 ± 1.9 kg/m²). Adipose RBP4, glucose transport protein‐4 (GLUT4), and p85 protein expression were determined by western blot. Blood samples from the bariatric patients were analyzed for serum RBP4, total cholesterol, triglycerides, and glucose. Adipose RBP4 protein expression (NGT: 11.0 ± 0.6; T2DM: 11.8 ± 0.7; lean: 8.7 ± 0.8 arbitrary units) was significantly increased in both NGT (P = 0.03) and T2DM (P = 0.005), compared to lean controls. GLUT4 protein was decreased in both NGT (P = 0.02) and T2DM (P = 0.03), and p85 expression was increased in T2DM subjects, compared to NGT (P = 0.03) and lean controls (P = 0.003). Regression analysis showed a strong correlation between adipose RBP4 protein and BMI for all subjects, as well as between adipose RBP4 and fasting glucose levels in T2DM subjects (r = 0.76, P = 0.004). Further, in T2DM, serum RBP4 was correlated with p85 expression (r = 0.68, P = 0.01), and adipose RBP4 protein trended toward an association with p85 protein (r = 0.55, P = 0.06). These data suggest that RBP4 may regulate adiposity, and p85 expression in obese‐T2DM, thus providing a link to impaired insulin signaling and diabetes in severely obese patients.     

Phytase can reduce theneedfor monocalcium phosphate supplementation in soybean and rapeseed meal‐based diets of black sea bream,Acanthopagrus schlegelii (Bleeker, 1854)

A feeding trial was conducted to study the effect of partial replacement of dietary monocalcium phosphate (MCP) with phytase on growth performance, feed utilization and phosphorus discharge in black sea bream, Acanthopagrus schlegelii. In the feeding trial, the control diet (designated as P1.5) was prepared with 1.5% MCP but without phytase, and the three other diets (designated as PP1.0, PP0.5 and PP0, respectively) were supplemented with 1.0%, 0.5% and 0% MCP, respectively, along with 200 mg (400 U) phytase/kg diet in each. Each diet was tested in triplicate tanks and fish were fed twice daily to satiation. After an 8‐week feeding trial in indoor flow‐through cylindrical fibreglass tanks (25 fish per tank, initial body weight: 11.5 ± 0.12 g), fish fed with PP1.0 and PP0.5 had no significant change in weight gain (WG), specific growth rate (SGR), protein efficiency rate (PER) or feed conversion ratio (FCR) compared to the control (p > .05), whereas fish fed with PP0 showed a significantly lower growth performance in the above parameters (p < .05). The addition of phytase did not affect the body composition or muscle composition. The apparent digestibility coefficients (ADCs) of crude protein and phosphorus increased when fish were fed diets in which MCP was replaced by phytase. Phosphorus discharge was also significantly reduced in fish fed diets in which MCP was replaced by phytase (10.2 ± 0.50 to 8.01 ± 0.47 g/kg weight gain). The present study suggests that dietary MCP can be reduced when phytase is added to the black sea bream diet, with a maximum MCP reduction level of up to 1% when phytase is supplemented at 200 mg (400 U)/kg diet. Thus, phytase in the diet of black sea bream is economically and ecologically beneficial.     

Chenopodium album extract ameliorates carbon tetrachloride induced hepatotoxicity in rat model

Major objective of this study was to explore the protective effect of the methanolic extract of Chenopodium album against carbon tetrachloride induced hepatotoxicity in rats. Chenopodium album has locally been used for multiple medicinal proposes. Methanolic extract of Chenopodium album (whole plant) was prepared with Soxhlet extractor and rotatory evaporator. Antioxidant activity of Chenopodium album was determined by DPPH free radical scavenging assay. Thirty Wister (albino) rats (150–200 g) were divided into six groups for the evaluation of hepatoprotective potential of different concentrations of Chenopodium album against carbon tetrachloride (1:1 CCl₄: Olive oil) under the controlled laboratory conditions. Group-I rats were administrated with olive oil (Normal control), Group-II with CCl₄ only, Group-III with Silymarin (positive control), Group-IV with Chenopodium album (100 mg/kg), Group-V with Chenopodium album (200 mg/kg) and Group-VI rats with Chenopodium album (300 mg/kg) for the period of 28 days. Serum was taken to determine the levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, cholesterol, triglyceride, creatinine and urea in the blood. Formalin stored tissues were examined for histopathological analysis. DPPH assay showed that Chenopodium album has the potential for reduction of oxidative stress. Chenopodium album minimized the levels of ALT (70 ± 8.68 U/L, 68.75 ± 8.38 U/L & 73.5 ± 10.28 U/L), AST (219.5 ± 19.16 U/L, 140.75 ± 13.35 U/L & 221.25 ± 13.33 U/L) and ALP (289.5 ± 28.21 U/L, 258 ± 11.12 U/L & 248.25 ± 4.03 U/L) at different concentrations (100 mg/kg, 200 mg/kg, 300 mg/kg respectively). Chenopodium album enhanced triglyceride level (64.75 ± 12.66 mg/dl at 200 mg/kg) as compared to CCl₄ treated group (33.25 ± 1.26 mg/dl). Carbon tetrachloride elevated urea level (43.25 ± 6.6) was decreased by high dose of Chenopodium album (18 ± 8.17). Moreover, Chenopodium album also improved WBC level (9.69 × 10³ /Cu.mr & 10.59 × 10³ /Cu.mr at low and medium doses respectively), RBCs level (6.97 × 10³ /Cu.mr) and hemoglobin level (13.95 G/dL, 13.467 G/dL & 14.05 G/dL at low, medium and high doses). In vivo study of Chenopodium album methanolic extract demonstrates the potential for protection of liver and after pre-clinical studies the plant can be used as a safe alternative of commercially available hepatoprotective medicines.     

Protective effects of aloe-emodin on scopolamine-induced memory impairment in mice and H2O2-induced cytotoxicity in PC12 cells

Aloe-emodin (AE) is one of the most important active components of Rheum officinale Baill. The present study aimed to investigate that AE could attenuate scopolamine-induced cognitive deficits via inhibiting acetylcholinesterase (AChE) activity and modulating oxidative stress. Kunming (KM) mice were received intraperitoneal injection of scopolamine (2 mg/kg) to induce cognitive impairment. Learning and memory performance were assessed in the Morris water maze (MWM). After behavioral testing, the mice were sacrificed and their hippocampi were removed for biochemical assays (superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA), AChE and acetylcholine (ACh)). In vitro, we also performed the AChE activity assay and H₂O₂-induced PC12 cells toxicity assay. After 2 h exposure to 200 μM H₂O₂ in PC12 cells, the cytotoxicity were evaluated by cell viability (MTT), nitric oxide (NO)/lactate dehydrogenase (LDH) release and intracellular reactive oxygen species (ROS) production. Our results confirmed that AE showed significant improvement in cognitive deficit in scopolamine-induced amnesia animal model. Besides, it increased SOD, GPx activities and ACh content, while decreased the level of MDA and AChE activity in AE treated mice. In addition, AE was found to inhibit AChE activity (IC₅₀ = 18.37 μg/ml) in a dose-dependent manner. Furthermore, preincubation of PC12 cells with AE could prevent cytotoxicity induced by H₂O₂ and reduce significantly extracellular release of NO, LDH and intracellular accumulation of ROS. The study indicated that AE could have neuroprotective effects against Alzheimer’s disease (AD) via inhibiting the activity of AChE and modulating oxidative stress.     

Endocrine responses and milk emission characteristics in high yielding Alpine dairy goats under once daily milking management

This study was done to verify if the lack of increase of milk fat content observed in Alpine dairy goats under once daily milking (ODM) compared to twice daily milking (TDM), results from disturbances of animals and/or milk ejection reflex. In this respect, we determined the milk yield and composition in the same 12 multiparous Alpine dairy goats when they were managed first under TDM (period 1: P1) and then under ODM (period 2: P2). Furthermore, oxytocin (OT) and cortisol (CORT) releases and milk emission kinetic of these goats were measured at morning milking 2 and 4 times in P1 and P2, respectively.ODM compared to TDM, caused 18 and 23% reductions, respectively in daily milk yield and milk fat content without milk protein content modification.Although baseline concentration of blood OT was lower under ODM than TDM management (7.0 pg/mL vs 17.8 ± 2.0 pg/mL, respectively), ODM did not modify the total amount of OT released during milking (15,484 pg/mL/32 min vs 18,996 ± 2865.3 pg/mL/32 min, respectively), the peak concentration of OT (57.2 pg/mL vs 73.6 ± 10.5 pg/mL, respectively) or the time to reach it (3.0 min vs 2.0 ± 0.5 min, respectively) by comparison to TDM.ODM compared to TDM, never modified the baseline concentrations of CORT (6.1 ng/mL vs 7.1 ± 1.1 ng/mL, respectively), the total amount of CORT released (11,727 ng/mL/32 min vs 10,073 ± 1522.2 ng/mL/32 min, respectively), the peak concentrations of CORT (16.1 ng/mL vs 15.1 ± 2.1 ng/mL, respectively) and the time to reach it (13.1 min vs 11.0 ± 1.5 min).During ODM, milk flow latency was reduced (−61%) while the mean flow rate was increased (+28%) by comparison to TDM. ODM compared to TDM management, did not modify the maximum flow rate (1.6 L/min vs 1.6 ± 0.2 L/min, respectively) or the time to reach this maximum (103.0 s vs 95.0 ± 10.0 s, respectively). The total milking duration at morning milking was not different (250.0 s vs 221.0 ± 22.0 s, respectively) although the morning milk yield was significantly higher under ODM management (2.63 kg/d vs 1.87 ± 0.1 kg/d, respectively).This results show that milk emission is improved under ODM management in Alpine goats without inhibition or disturbance of neuro-hypophyseal OT release pattern and lack of activation of the hypothalamic–pituitary–adrenal axis and plasma cortisol increase, disproving our initial hypotheses of a disturbance of animals and/or incomplete milk ejection to explain the low fat content of milk under ODM in this breed.     

Developmental competence of mature yak vitrified–warmed oocytes is enhanced by IGF-I via modulation of CIRP during in vitro maturation

The objective of this study was to investigate whether developmental competence of mature vitrified–warmed yak (Bos grunniens) oocytes can be enhanced by supplemented insulin-like growth factor I (IGF-1) during in vitro maturation (IVM), and its relationship with the expression of cold-inducible RNA-binding protein (CIRP). In experiment 1, immature yak oocytes were divided into four groups, and IVM supplemented with 0, 50, 100 and 200 ng/mL IGF-1 was evaluated; the mRNA and protein expression levels of CIRP in mature oocytes in the four groups were evaluated using quantitative real-time PCR and western blotting analyses. In experiment 2, the mature yak oocytes in the four groups were cryopreserved using the Cryotop (CT) method, followed by chemical activation and in vitro culture for two days and eight days to determine cleavage, blastocyst rates, and total cell number in the blastocysts. Mature yak oocytes without vitrification served as a control group. The outcomes were as following: (1) the expression of CIRP in the matured oocytes was up-regulated in the IGF-1 groups and was highest expression was observed in the 100 ng/mL IGF-1 treatment group. (2) In the vitrified–warmed groups, the rates of cleavage and blastocyst were also highest in the 100 ng/mL IGF-1 treatment group (81.04 ± 1.06%% and 32.16 ± 1.01%), which were close to the rates observed in groups without vitrification (83.25 ± 0.85% and 32.54 ± 0.34%). The rates of cleavage and blastocyst in the other vitrified–warmed groups were 70.92 ± 1.32% and 27.33 ± 1.31% (0 ng/mL); 72.73 ± 0.74% and 29.41 ± 0.84% (50 ng/mL); 72.43 ± 0.61% and 27.61 ± 0.59% (200 ng/mL), respectively. There was no significant difference in the total cell number per blastocysts between the vitrified–warmed groups and group without vitrification. Thus, we conclude that the enhancement in developmental competence of mature yak vitrified–warmed oocytes after the addition of IGF-1 during IVM might result from the regulation of CIRP expression in mature yak oocytes prior to vitrification.     

Plasma Levels of Zinc,Copper, and Ceruloplasmin in Patients after Undergoing Laparoscopic Adjustable Gastric Banding

Laparoscopic adjustable gastric banding (LAGB) causes significant weight loss in morbidly obese adults. However, its consequences on nutritional status still remain unclear. There are a few studies determining the nutritional status after LAGB and none have focused on the serum levels of zinc (Zn), copper (Cu), and ceruloplasmin (CP). We aimed to investigate the effects of LAGB surgery on plasma Zn, Cu, and CP levels. Thirty patients with LAGB with morbid obesity were included. Blood samples were collected preooperatively and in the postoperative third month to determine plasma Zn, Cu, and CP levels. The mean preoperative and postoperative body mass indexes (BMI) were 44.9 ± 7.4 kg/m² and 44.1 ± 6.5 kg/m², respectively. The mean weight loss was 12.9 ± 3.3 kg at the postoperative third month. The postoperative Zn (500 ± 130 ng/ml), Cu (280 ± 80 ng/ml), and CP (23.9 ± 8.8 mg/dl) values were statistically significantly lower than the preooperative Zn (740 ± 230 ng/ml), Cu (370 ± 80 ng/ml) and CP (33.3 ± 15.7 mg/dl) levels (p < 0.05). Decreases in the plasma levels of Zn, Cu, and CP were seen postoperatively following LAGB surgery. The nutritional status of LAGB-applied patients should be monitored and mineral supplementation may be considered.     

Axial BMD in Diabetic and Nondiabetic Southeast Asians with HIP Fractures: Do Race and Body Mass Index Matter?

Objective: Obesity is less prevalent in Asian subjects with type 2 diabetes mellitus (T2DM) in contrast to Caucasians. Whether higher axial bone mineral density (BMD) often reported in T2DM is independent of body mass index (BMI) has not been clearly shown. BMD characterization in T2DM patients with hip fractures has also not been performed. We compared the BMD of Asian diabetic and nondiabetic patients with new hip fractures and explored how BMD was influenced by BMI.Methods: We included 255 diabetic and 148 nondiabetic patients. BMD adjusted for age; BMI; race; sex; renal function; and use of statins, proton pump inhibitors, steroids, anticonvulsants, and calcium and/or vitamin D supplements were compared between the groups. We were particularly interested in the BMD comparison between underweight diabetics and nondiabetics with hip fractures.Results: The presence of T2DM was associated with higher BMD (g/cm²) at the femoral neck (0.527 ± 0.103 vs. 0.491 ± 0.102, P<.01) and lumbar spine [LS] (0.798 ± 0.147 vs. 0.723 ± 0.156, P<.01). This association persisted after adjustment for multiple confounding variables including BMI. The age-, BMI-, and sex-adjusted LS BMD was higher in underweight (BMI <18.5 kg/m²) diabetics compared to similar weight nondiabetics (0.733 ± 0.126 vs. 0.649 ± 0.131 g/cm², P = .014).Conclusion: T2DM is independently associated with higher axial BMD in patients with new hip fractures. The finding of higher BMD even in underweight diabetics with hip fractures compared to their nondiabetic counterparts suggests that higher BMD in subjects with T2DM is not due to higher BMI.Abbreviations:BMD = bone mineral densityBMI = body mass indexCV = coefficient of variationDXA = dual-energy X-ray absorptiometryHbA1c = glycated hemoglobinIGF-1 = insulin growth factor-1LS = lumbar spine25(OH)D = 25-hydroxyvitamin DT2DM = type 2 diabetes mellitus     

Effect of polyphenols extracted from tamarind (Tamarindus indica L.) seed coat on pathophysiological changes and red blood cell glutathione peroxidase activity in heat-stressed broilers

The purpose of this study was to determine the effect of polyphenols extracted from the tamarind seed coat (PETSC) on glutathione peroxidase (GPx) activity, red blood cell parameters and bilirubin in heat-stressed broilers. One hundred forty-seven broilers, 18-days old were divided into two groups. In group 1, broilers were maintained at an environmental temperature of 26?±?2 °C throughout the experimental period. In group 2, the broilers were maintained at 38?±?2 °C (cyclic temperature: 26?±?2 °C; ?38?±?2 °C; and ?26?±?2 °C, and broilers were maintained at 38?±?2 °C for 6 h/ day) and received PETSC at a concentration of 0, 100, 200, 300, 400 or 500 mg/kg in their diet ad libitum. Parameters were investigated on days 1, 7, 14 and 21 of the experimental period. Results showed that GPx activity of heat-stressed broilers that received 100 mg/kg of PETSC in their diet was lower (P?<?0.05) than that in broilers fed the other concentrations. The mean total red blood cell count and hemoglobin concentration of heat-stressed broilers that received 100 mg/kg PETSC was higher (P?<?0.05) than those in broilers in group 1 and those fed the other concentrations. The mean bilirubin level in the excreta of heat-stressed broilers that received 100 mg/kg of PETSC was lower (P?<?0.05) than that in broilers that received 0, 300, 400 and 500 mg/kg of PETSC. This showed that PETSC could reduce GPx activity and bilirubin in feces, and increase red blood cell parameters in heat-stressed broilers.