Improving the stability of insulin through effective chemical modifications: A Comprehensive review.

Insulin, an essential peptide hormone, conjointly regulates blood glucose levels by its receptor and it is used as vital drug to treat diabetes. This therapeutic hormone may undergo different chemical modifications during industrial processes, pharmaceutical formulation, and through its endogenous storage in the pancreatic ß-cells. Insulin is highly sensitive to environmental stresses and readily undergoes structural changes, being also able to unfold and aggregate in physiological conditions. Even; small changes altering the structural integrity of insulin may have significant impacts on its biological efficacy to its physiological and pharmacological activities. Insulin analogs have been engineered to achieve modified properties, such as improved stability, solubility, and pharmacokinetics, while preserving the molecular pharmacology of insulin. The casually or purposively strategies of chemical modifications of insulin occurred to improve its therapeutic and pharmaceutical properties. Knowing the effects of chemical modification, formation of aggregates, and nanoparticles on protein can be a new look at the production of protein analogues drugs and its application in living system. The project focused on effects of chemical modifications and nanoparticles on the structure, stability, aggregation and their results in effective drug delivery system, biological activity, and pharmacological properties of insulin. The future challenge in biotechnology and pharmacokinetic arises from the complexity of biopharmaceuticals, which are often molecular structures that require formulation and delivery strategies to ensure their efficacy and safety.

Duration of the diabetic state altered platelet indices from baseline values in a streptozotocin-induced rat model for type 1 diabetes mellitus.

BACKGROUND: Changes in platelet indices in naturally occurring type I diabetes mellitus (T1DM) have been described in several studies. In this study, platelet indices such as platelet count (PLT), plateletcrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW), and MPV to PLT ratio were investigated according to diabetic duration after streptozotocin (STZ)-induced T1DM, as well as for their correlation with glucose. METHODS: Forty healthy adult Wistar rats were randomly assigned to four experimental groups of ten (5 rats of each sex), including the control group, the 7, 14, and 28 days diabetic groups (D7, D14, and D28, respectively). RESULTS: In diabetic groups, plasma glucose was significantly higher than in control (P < 0.01). D7, D14, and D28 groups presented significantly lower PLT than the control (P < 0. 01). A significant decrease in PCT was observed in D14 and D28 females (P < 0.05). Mean platelet volume was significantly higher in the D28 group than in to control. D28 females also showed a significant difference in PLT, MPV, and the MPV-to-PLT ratio compared with D7 females (P < 0.05). A comparison between D28 females and males showed a significant difference in PDW (P < 0.05). Both females and males showed a significant correlation between glucose and PLT, PCT, MPV, and the MPV-to-PLT ratio. CONCLUSIONS: Platelet indices change significantly with the duration of diabetes compared with the baseline values, and female and male rats did not have significant differences in platelet indices in any period except the 28 days.

Assessment of the utility of platelet indices to diagnose clinical benign prostatic hyperplasia in dogs.

Introduction: Platelet indices changes in severely ill people and in dogs with inflammation are compatible findings. This study aimed to compare platelet indices between dogs with clinical benign prostatic hyperplasia (BPH) and healthy controls. Additionally, to determine whether there is a correlation between the relative prostatic size (S rel) and the platelet indices in BPH dogs. Methods: Thirty-five adult intact male dogs of different breeds were allocated to the experimental groups: dogs with clinical BPH (groups A; n = 24; median age of 6 years; the median weight of 8.50 kg) and healthy dogs (group B; n = 11; median age 5.50 years; the median weight of 7.00 kg) based on physical examination, clinical signs, and S rel detected by ultrasonographic findings. The individual prostatic volume (IPV) was divided by the expected prostatic volume (EPV) to determine the relative prostatic size in dogs over 4 years old. Platelet indices were compared between the two groups, and a correlation between S rel and these indices was calculated. Results: The median S rel of dogs in group A was significantly higher (P = 0.001), and the mean plateletcrit (PCT) was significantly lower (P = 0.003) compared with those in group B. S rel showed a significant negative correlation with PLT and PCT (r = -0.388; P = 0.02 and r = -0.402; P = 0.01). Receiver operating characteristic (ROC) analysis showed PLT and PCT thresholds for estimating S rel > 1 with 75% and 87.5% sensitivity and 71.82 and 63.64% specificity. Discussion: The findings of this study support the use of platelet indices like PLT and PCT to detect clinical BPH in dogs. However, more research is needed to confirm their utility in conjunction with other previously described diagnostic factors.

Evaluation of platelet indices and mean platelet volume to platelet count ratio in experimentally varicocele-induced adolescent and adult rats.

The present study investigated the sperm quality parameters and platelet indices in adolescent and adult varicocele rats. Twenty male Wistar rats were distributed into four groups (n = 5): I-sham adolescent group, II-sham adult group, III-varicocele adolescent group, and IV-varicocele adult group. Rats were sacrificed on the 61st day of the experiment. Sperm count (×106  ml-1 ), sperm motility and viability (%) were significantly (p < 0.05) different between varicocele and sham groups in adolescent and adult rats. Varicocele adults had significant (p < 0.05) lower platelet count (PLT) than sham adolescents and adults. The mean platelet volume (MPV), the MPV/PLT ratio, plateletcrit (PCT) and platelet distribution width (PDW) differ significantly (p < 0.05) between varicocele and sham groups of adolescent and adult rats. It is the first time the ratio MPV/PLT was evaluated in association with other platelet indices in adolescent and adult varicocele rats. Based on the results, it would appear these indicators can become a promising tool in varicocele diagnosis.

Comparison of Cell-Free Fetal DNA Plasma Content Used to Sex Determination Between Three Trimesters of Pregnancy in Torkaman Pregnant Mare.

This investigation aimed to compare the cell-free fetal DNA (cffDNA) plasma present in three trimesters of pregnancy in Torkaman pregnant mare. Peripheral blood samples of 32 pregnant mares in three trimesters of pregnancy were collected in tubes containing ethylenediaminetetraacetic acid at three time points. Circulating cffDNA was extracted from 3 mL of maternal plasma. Using outer and inner primers, a conventional polymerase chain reaction was performed for the sex-determining region Y (SRY) gene present in the Y chromosome. Of the total 32 Torkaman pregnant mares, 24 were carrying male fetuses and eight were carrying female fetuses. In total, the accuracy of the test was 48.75%, 68.75%, and 75% in the first, second, and third trimesters of pregnancy, respectively. The sensitivities were 25%, 58.32%, and 66.66%, respectively, whereas their specificities were 100% in all trimesters. In conclusion, the SRY gene can permit the detection of equine fetal sex with good accuracy through cffDNA analysis in maternal plasma just in the third trimester of pregnancy, although specificity in all duration of pregnancy was 100%.

Comparative evaluation of urinary dipstick and pH-meter for cattle urine pH measurement.

Urinary dipsticks are often used for the diagnosis of kidney, liver, metabolic, and urogenital diseases in cattle clinical practice for the simplicity of use and ease of access. The aims of the present study were to evaluate the agreement between urinary dipsticks and pH-meter for the urine pH in dairy cattle, and also to compare the urine pH before and after centrifugation from each method. The agreement between urine dipstick and pH-meter methods before and after centrifuge were calculated by Passing-Bablok regression analysis demonstrated constant differences (intercept) at 0.60 and -1.01 and positive proportional differences (slope) at 0.94 and 1.13; respectively. Total bias estimated by Bland-Altman plot analysis before (0.20) and after (0.14) centrifuge were lower than the acceptable bias in urine samples. The regression analysis of this study emphasized that the urinary dipstick can be used to determine the cattle urine.

Effect of crocin on biochemical parameters, oxidative/antioxidative profiles, sperm characteristics and testicular histopathology in streptozotocin-induced diabetic rats.

OBJECTIVE: Chronic hyperglycemia and overproduction of reactive oxygen species (ROS) are strong predictors of the development of reproductive complications of diabetes. The present study was conducted to determine the effects of crocin on biochemical parameters, oxidative stress, and sperm characteristics as well as testes histopathology in diabetic rats. MATERIALS AND METHODS: Twenty-four rats were divided into the four groups as follows: control, untreated diabetic and two crocin (40 and 60 mg/kg/day)-treated diabetic groups. Diabetes was induced by injection of a single dose of streptozotocin (STZ, 60 mg/kg). Administration of crocin (intraperitoneally) was started three days after STZ injection and was continued until the 28th day. At the end of the experiment, rats were anesthetized after weighing. Blood samples and epididymal sperm were subsequently collected to measure biochemical parameters (glucose and lipid profile), total oxidant and antioxidant status (TOS and TAS, respectively), oxidative stress index (OSI), and sperm characteristics (count, motility, and viability); also, testes were dissected out for histopathology examination. RESULTS: Our result indicated that blood glucose, cholesterol, triglyceride, LDL cholesterol levels, as well as TOS, and OSI decreased, but body weight, sperm counts, motility and viability, as well as TAS and HDL levels increased significantly in the crocin-treated diabetic rats (P˂0.05). In testis sections from diabetic rats treated with crocin (40 and 60 mg/kg), seminiferous tubules exhibited normal shape and restoration of testis architecture was observed. CONCLUSION: Administration of crocin in the present study, ameliorated blood glucose, lipid abnormalities, oxidative stress, sperm characteristics and testis damage in STZ-diabetic rats.

Multicentric lymphoma in a Rottweiler dog with bilateral ocular involvement: A case report.

A 10-year-old, male Rottweilerdog was presented to Small Animal Hospital of Tehran University with a history of lethargy, anorexia, weight loss, vomiting, polyuria, polydipsia and blindness. The dog showed symptoms of depression, high body temperature (39.2 ˚C), tachypnea (40 breaths min-1) and cachexia. In ophthalmic examination, bilateral hyphema (hemmorrahge in anterior chamber of the eye) and blindness were detected. The pulse of the animal was normal (90 beats min-1) and obvious general lymphadenopathy was determined. Lymph nodes were firm, freely movable, and painless on palpation. Initial differential diagnosis of lymphoma included lymphoma, metastatic neoplasia, lymphoid hyperplasia, and lymphadenitis. In hematology and cytology tests, this case was suspected to lymphoma. Immunohistochemical staining of neoplastic lymph node revealed that nearly 20.00 - 25.00% of neoplastic cells were strongly positive for anti CD3, whereas they were negative for both CD20 and CD79a. According to the microscopic and immunohistochemical findings, a diagnosis of T cell lymphoma was made. The present report is the first case of multicentric lymphoma with ocular metastasis in a dog in Iran.

Diagnostic evaluation of milk lactate dehydrogenase and alkaline phosphatase activities by receiver operating characteristic analysis curve in early lactation of ewes with subclinical mastitis.

The aim of the present study was to investigate the diagnostic evaluation of milk lactate dehydrogenase (mLDH) and alkaline phosphatase (mALP) activities by receiver operating characteristic (ROC) analysis curve in early lactation of ewes with subclinical mastitis (SCM) and determine the correlation between number of somatic cell count (SCC) and mLDH and mALP activities. A total of 196 udder half milk samples were collected within the first 6 weeks of lambing. The SCM was determined by positive milk bacterial culture and positive California mastitis test (CMT); SCC was determined by fossomatic method and enzyme activities were determined spectrophotometrically. The mLDH and mALP of SCM cases were positively correlated with SCC values. Values of mLDH, mALP and SCC were significantly higher in SCM than non-SCM udder halves. The optimum cut-off points of mLDH and mALP activities for SCM diagnosis were determined at 203.61 (U L-1) and 329.84 (U L-1), respectively. In conclusion, SCC has positive correlation with mALP and mLDH activities in SCM ewes and mLDH and mALP activities could be considered as reliable indicators for intra-mammary inflammation diagnosis.

Evaluation of the protective effect of pentoxifylline on carrageenan-induced chronic non-bacterial prostatitis in rats.

Chronic non-bacterial prostatitis (CNP) is the most common type of prostatitis and oxidative stress (OS) was shown to be highly elevated in prostatitis patients. This study aimed to investigate the protective effect of pentoxifylline (PTX) on CNP induced by carrageenan in rats. Male adult Wistar rats (n = 30) were divided into control, CNP and three treatment groups (n = 6) including CNP + cernilton and CNP + PTX groups. CNP was induced by single intraprostatic injection of 1% carrageenan (100 µl). Rats in treatment groups received orally cernilton 100 mg/kg and PTX at 50 and 100 mg/kg 1 week after CNP induction for 21 days. Prostatic index (PI), prostatic specific antigen (PSA), tumor-necrosis factor alpha (TNF-α), serum lipid peroxidation (MDA), blood urea nitrogen, creatinine and histopathological changes were compared between groups. There were significant increase of PI, serum levels of PSA, TNF-α and MDA in CNP group at 29 day. In treatment groups, significant reduction in PI, serum levels of PSA, TNF-α, MDA and creatinine was observed especially in rats treated with dose of 50 mg/kg of PTX. In CNP group, histopathological changes of the prostate such as leucocyte infiltration, large involutions and projection into the lumen and reducing the volume of the lumen were observed as well. Whereas PTX, especially at dose of 50 mg/kg, could improve the above-mentioned changes remarkably in CNP treated rats. For the first time, our findings indicated that PTX improved CNP induced by carrageenan in rats.

Study of Hypoglycemic, Hypocholesterolemic and Antioxidant Activities of Iranian Mentha Spicata Leaves Aqueous Extract in Diabetic Rats.

This study was conducted to evaluate the hypoglycemic, hypocholesterolemic and antioxidant properties of Mentha spicata (Labiateae) leaves aqueous extract (MSLA) in alloxan-induced diabetic rats. In this study, hyperglycemia was induced in male rats by intraperitoneal injection of alloxan monohydrate (150 mg/kg). The aqueous extract of M. spicata was orally administered at a dose of 300 mg/kg body weight to diabetic rats for 21 days and the effects were compared with glibenclamid (2 mg/kg). Fasing blood sugar (FBS), body weight, lipid profile and serum malondialdehyde (MDA) were monitored at 0, 7, 14 and 21 days after induction of diabetes. Total phenol contents (TP) and reducing power (RP) were also evaluated. TP and RP of aqueous extract were 2.763 ± 0.39 mg Galic acid/gr and 0.026 ± 0.001 EC50 mg/mL, respectively. The LD50 of the extract was found to be ˃ 1500 mg/kg. The administration of M. spicata aqueous extract produced a significant reduction (P˂0.01) in FBS, total cholesterol, triglyceride, low density lipoprotein-cholesterol and MDA (101.83 ± 4.33, 95.66 ± 4.75, 89.83 ± 5.26, 26.20 ± 5.10 mg/dl and 1.53 ± 0.61 µmol/l, respectively) in diabetic rats. These effects were comparable with the effects of standard antidiabetic drug (glibenclamide). The results of the investigation indicated that M. spicata leaf aqueous extract possess hypoglycemic, hypocholesterolemic and antioxidant properties in diabetic rats. Therefore, this study suggest a promising use of it for treatment of diabetes.

Follicle stimulating hormone increases spermatogonial stem cell colonization during in vitro co-culture.

The complex process of spermatogenesis is regulated by various factors. Studies on spermatogonial stem cells (SCCs) have provided very important tool to improve herd genetic and different field. 0.2 to 0.3 percent of total cells of seminiferous tubules is consist of spermatogonial stem cells. To investigate and biomanipulation of these cells, proliferation and viability rate of cells should be increased in vitro, at first. Follicle stimulating hormone (FSH) has been suggested to play a determinant role in the survival of germ cells in addition to increasing spermatogonial proliferation. In this study, the in vitro effects of FSH on spermatogonial cell colony formation were investigated. Sertoli and spermatogonial cells were isolated from 3-5 months old calves. The identity of the Sertoli cells and spermatogonial stem cells were confirmed through immunocytochemistry and colony morphology, respectively. Co-cultured Sertoli and spermatogonial cells were treated with FSH in different dose of 10, 20 and 40 IU mL(-1) FSH, before colony assay. Results indicated that, FSH increased in vitro colonization of spermatogonial cells in comparison with control group. In conclusion, using FSH provided proper bovine spermatogonial stem cell culture medium for in vitro study of these cells.