Validation of Bos taurus SNPs for Milk Productivity of Sahiwal Breed (Bos indicus), Pakistan.

The aim of the present study was the validation of the already reported Bos taurus SNPs in the Sahiwal breed. A total of nine SNPs of the casein gene were studied. Out of nine, seven Bos taurus SNPs of casein protein genes were found to be significantly associated with milk productivity traits. The genomic DNA was extracted from the mammary alveolar endothelial cells of a flock of 80 purebred Sahiwal lactating dams available at Khizrabad Farm near Sargodha. New allele-specific primers were designed from the NCBI annotated sequence database of Bos taurus to obtain 100 nt-long PCR products. Each dam was tested separately for all the SNPs investigated. Animals with genotype GG for the SNPs rs43703010, rs10500451, and 110323127, respectively, exhibited high milk yield. Similarly, animals with genotype AA for the SNPs rs11079521, rs43703016, and rs43703017 showed high milk yield consistently. For the SNP rs43703015, animals with genotype CC showed high milk productivity. These above-mentioned SNPs have previously been reported to significantly up-regulate casein protein contents in Bos taurus. Our results indicated SNPs that significantly affect the milk protein contents may also significantly increase per capita milk yield. These finding suggest that the above-mentioned reported SNPs can also be used as genetic markers of milk productivity in Sahiwal cattle.

Fortification of Extender with Basella rubra Fruit Extract Enhances the Cryosurvival of Ram Semen.

This study aimed to evaluate the impact of Basella rubra fruit extract (BR-FE) on cryopreserved ram sperm's motility, velocity, and membrane integrity. Thirty ejaculates collected from 3 fertile rams (10 from each) were diluted with semen dilution extender (SDE) in a ratio (1:2) and centrifuged to remove 50% supernatant. The remaining sample was mixed with semen cryopreservation extender (SCE) in 1:4 ratio. Then 1.2 mL of SCE diluted sample was divided in four aliquots (0.3 mL each) that were further extended with [(1) control group (0.7 mL of SCE), (2) BR-FE-0.6% group (0.7 mL of SCE supplemented with 0.6% BR-FE), (3) BR-FE-0.8% group (0.7 mL of SCE supplemented with 0.8% BR-FE), and (4) BR-FE-1.6% group (0.7 mL SCE supplemented with 1.6% BR-FE)]. All extended samples were cooled gradually from 25°C to 4°C in half an hour. The 0.1 mL sample from all aliquots was analyzed for precryopreservation sperm parameters and the remaining sample was loaded in 0.5 mL plastic semen straws, cooled gradually to -20°C, and then dipped in liquid nitrogen. After 24 hours of cryopreservation, the straws were thawed for postcryopreservation sperm evaluations. The results (analysis of variance based) showed significantly enhanced percentage of post-thaw sperm membrane integrity, progressive motility, and velocity in BR-FE-0.6% group at both pre- and postcryopreservation stages as compared with all other groups. However, analysis of covariance revealed concentration-dependent cryoprotective effect of BR-FE with maximum percentage of sperm membrane integrity in the 1.6% group. According to these results, BR-FE supplementation adds enormous sperm protective potential to ram sperm cryopreservation medium.

In ovo exposure of F-ions and organo-fluoride insecticide (Bifenthrin) cause developmental anomalies of eye in chick embryos.

Objective: The developmental abnormalities of the in-ovo exposure of Fluoride ions (F-ions) and Bifenthrin (BF) on the embryonic chick eye were investigated. Materials and methods: 165 fresh fertilized eggs of zero day and 40-50 g weight were divided into three groups (55 eggs each) on the basis of inter-vitelline treatment of eggs on zero day of study: 1) Control group (CG); 0.1 ml of 5 % DMSO aqueous solution 2),3) Fluoride group (FG), and Bifenthrin group (BFG); 0.01 mg/kg F-ions (from NaF) and 0.01 mg/kg BF in 0.1 ml of 5 % DMSO aqueous solution respectively. After incubation for 14 days at 37 ± 0.5 °C embryos were externalized. Eyes of each embryo were removed for micro-anatomical, micrometric and histopathological studies. Results: The histological sections have shown denser and enlarged marginal mitotic region of the developing eye lenses in FG and BFG. In vertical sections of the eye lenses the nuclei of the crystalline cells in FG and BFG show a highly depressed bow shaped arrangement. Moreover, the nuclei of the core crystalline cells of the lens were apparently smaller in FG and BFG than CG. Out of the six anatomical layers of the retina the nuclear and the plexiform layers were highly enlarged in FG and BFG, similarly the three corneal cell layers (endothelial, parenchymal and epithelial) were enlarged in FG and BFG than CG. The morphometric, histometric and micrometric estimations also show significant variations in FG and BFG than CG. Conclusion: The results indicate subtle developmental anomalies of the eyes attributable to the F-ions and BF exposure indicating their developmental neuro-optic disruption potentials. Results further revealed higher toxicity of BF as compared to F-ions.

Evaluation of the anti-inflammatory, analgesic, anti-pyretic and anti-ulcerogenic potentials of synthetic indole derivatives.

A large number of new synthetic compounds are synthesized in the field of heterocyclic chemistry having a variety of biological potentials. In the present study, some synthetic indole derivatives are used to check anti-inflammatory, analgesic, antipyretic and gastroprotective activity in albino mice. Albino mice of either sex of reproductive age were used for each study (n = 5). In anti-inflammatory activity, the negative control (NC) and positive control group animals were treated with normal saline and 10 mg/kg of indomethacin respectively. The treated groups received the twenty four different synthetic chemicals, after 30 min of sub cutaneous injection of carrageenan. In analgesic activity, hot-plate method is used and for each group the latency period was recorded at zero moment of the provision of required dose and after 30, 60, 90, 120 and 180 min. In anti-pyretic activity, Pyrexia was induced by using Brewer's yeast method. Before any treatment and then after duration of 18 h, the rectal temperatures were recorded. Among all the chemicals, only those chemicals which show any potential related to above mentioned activities were selected for gastroprotective activity. The gastroprotective activity was performed to check the gastric ulcers by using 300 mg/kg of single oral dose of indomethacin to animals of all groups except NC group. This study helped to screen out the most potent indole derivatives 3a-II and 4a-II from the 24 synthetic indole derivatives which demonstrated the best biological potential (anti-inflammatory, analgesic, antipyretic, and gastroprotection) as compared to the remaining ones. The micrometric and biochemical results also support the histological findings. Out of the twenty-four novel indole amines tested, 3a-II and 4a-II have shown the effective pharmacological capacity and additionally have not shown any overt and systemic toxicity. Thus these two indole amines need further in-depth pharmacokinetic and pharmacodynamics studies before they are recommended for any pre-clinical trial.

Physiology of a Forgotten Electrolyte-Magnesium Disorders.

Magnesium (Mg2+) is the second most common intracellular cation and the fourth most abundant element on earth. However, Mg2+ is a frequently overlooked electrolyte and often not measured in patients. While hypomagnesemia is common in 15% of the general population, hypermagnesemia is typically only found in preeclamptic women after Mg2+ therapy and in patients with ESRD. Mild to moderate hypomagnesemia has been associated with hypertension, metabolic syndrome, type 2 diabetes mellitus, CKD, and cancer. Nutritional Mg2+ intake and enteral Mg2+ absorption are important for Mg2+ homeostasis, but the kidneys are the key regulators of Mg2+ homeostasis by limiting urinary excretion to less than 4% while the gastrointestinal tract loses over 50% of the Mg2+ intake in the feces. Here, we review the physiological relevance of Mg2+, the current knowledge of Mg2+ absorption in the kidneys and the gut, the different causes of hypomagnesemia, and a diagnostic approach on how to assess Mg2+ status. We highlight the latest discoveries of monogenetic conditions causing hypomagnesemia, which have enhanced our understanding of tubular Mg2+ absorption. We will also discuss external and iatrogenic causes of hypomagnesemia and advances in the treatment of hypomagnesemia.

Histo-anatomical mutilations of developing chick brain induced by in-ovo fluoride and bifenthrin exposure.

Comparative developing brain histo-anatomical pathologies of Fluoride ions and Bifenthrin in-ovo exposures were explored in the golden black variety of domestic chick. Three exposure groups were -the Vehicle control group (Vg); Fluoride (F) group and the Bifenthrin (Bn) group each with forty fertilized eggs and received their respective group treatment at zero day of incubation. Embryos were extracted, dissected from head region and the embryonic whole brains were recovered after 14 days of incubation. The embryonic brains were preserved in bouin fixative for 24 h for further studies. The morphological results show the atrophied and hypertrophied embryonic brain in F and Bn groups respectively as compared to Vg group. The toxicological signs of encephalic anatomy and histology of F and Bn exposure were the enlarged third ventricles, optocoeles and arachnoid mater, encephalic spongiosis and decreased neuroglial density. The morphometric data showed significant decrease (p ≤ 0.05) in mean weight and density of whole brain in F and Bn groups compared Vg. The mean length and width of whole brain in F were significantly lower than that of the Bn and Vg. whereas, the mean breadth of third ventricle in Bn remained significantly lower than F and Vg groups. On the other hand, the mean breadth of optocoele and fourth ventricle in F and Bn groups remained significantly higher than Vg. Conversely the mean optic lobe wall thickness in F remained significantly lower than Bn and Vg. Additionally, the mean neuronal density in diencephalon, optic lobe and cerebellum in F group and Bn group remained significantly (p ≤ 0.05) lower than Vg. Results show that low dose in-ovo fluoride or bifenthrin exposure may cause neuro-developmental abnormalities in the developing chick embryos indicating that the Fluoride-ions and Bifenthrin harbor strong developmental neurotoxic capacity.

Mitigating properties of vitamin E and olive oil on the teratomorphogenic impacts of lambda-cyhalothrin in developing chick embryos.

Embryonic toxicity of lambda-cyhalothrin (LCH) and the mitigating properties of extra virgin olive oil (EVO) and vitamin E were investigated in golden black variety of chick embryos. Fifty fertilized eggs distributed in 5 groups received their respective group treatments at zero days of incubation were recovered after 14 days of incubation and fixed in fixative for 48 h for further studies. Results showed that treatment with LCH caused embryonic death, growth retardation and developmental abnormalities such as limbic developments, reduced muscular growth, and embryonic cataract. Post treatment with vitamin E and EVO alone significantly improved all these developmental defects. Morphometric readings also showed that embryos treated with combined dose of vitamin E and EVO had more resemblance towards the control group embryo. These findings suggested that the use of vitamin E and EVO together during pregnancy have the potential to curtail these accidental, environmental and work place exposures of LCH.

Basella alba, Grewia asiatica, Solanum nigrum and Ficus carica fruit extracts ameliorate the testicular histopathology induced by CCl4 exposure in albino mice: An experimental study.

Background: Carbon tetrachloride (CCl 4 ) is extensively used in various industries and induces oxidative stress in humans. Unfortunately, it is one of the neglected potent male reproductive toxicants. Objective: The present research reports the testicular histopathology of CCl 4 and ameliorations by four medicinal fruit pulp extracts (FPEs) in mice. Materials and Methods: Sixty male albino mice were divided into six groups (10/group) as per the following: 1. Vehicle control (Vc); 2. CCl 4 (C); 3-6. CCl 4 +Basella alba (CBa), CCl 4 +Solanum nigrum (CSn), CCl 4 +Ficus carica (CFc) and CCl 4 +Grewia asiatica (CGa). Except for the control group, CCl 4 (0.1 mL of 0.2 mL kg - 1 ) was given to the animals in corn oil. The four plant extracts (0.1 mL each) were respectively given to the relevant FPE group animals for the next five consecutive days, while the animals in the Vc and CCl 4 groups received water instead of FPE. Results: The CCl 4 exposure led to various histometric and histological alterations (loss of interstitial tissue and various dislodged tailless spermatids with enlarged heads) that were recovered in all except Solanum nigrum FPE mice post-treatment. The micrometric data of testicular sections also indicated significant decline in the number of spermatogonia, while the cross-sectional area of the sperm heads remained significantly higher in the CSn and C groups. Conclusion: Except for Solanum nigrum,the three FPEs, especially Ficus carica,showed rehabilitative properties against CCl 4 exposure-related modifications in testicular histopathologies.

Concurrent Anti-Glomerular Basement Membrane Antibody Disease and Membranous Nephropathy: A Case Series.

RATIONALE & OBJECTIVE: Anti-glomerular basement membrane (GBM) disease is a rapidly progressive glomerulonephritis which, in some instances, occurs concurrently with other diseases such as antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis. Rarely, it also occurs with membranous nephropathy (MN). We report a series of such patients, characterizing their long-term follow up. STUDY DESIGN: Case series. SETTING & PARTICIPANTS: Twelve patients referred to 1 medical center between 2001 and 2019 with anti-GBM disease and MN whose clinical characteristics and course were described. RESULTS: Patients' ages ranged from 20 to 81 years old, and all presented with severe acute kidney injury requiring dialysis on presentation or shortly thereafter. Only 1 patient had pulmonary findings on presentation. The predominant pathology was crescentic and necrotizing glomerulonephritis with linear staining for immunoglobulin G along the GBM associated with epimembranous electron-dense immune-type deposits. All 11 patients who were tested had significant titers of anti-GBM antibodies, but only 1 of the 5 tested for phospholipase A2 receptor (PLA2R) on biopsy was positive. Eight patients received therapy with cyclophosphamide, prednisone, and plasmapheresis; 2 patients with prednisone and plasmapheresis; and 2 with rituximab-based regimens. Progression to a requirement for kidney replacement therapy occurred in all 12 patients, but 2 patients later recovered kidney function. Recurrence of anti-GBM disease did not occur for any of the patients studied. LIMITATIONS: Incomplete testing for PLA2R in biopsy and serum, limited sample size, and lack of uniform treatment regimen. CONCLUSIONS: In this case series, the presentation of concurrent anti-GBM disease and MN was characterized by rapidly progressive glomerulonephritis and poor kidney outcomes. These findings suggest possible value from earlier diagnosis and the need for identification of more effective treatment regimens.

Jambul (Syzygium cumini) Pulp Extract Enhances Viability, Motility, and In Vitro Fertilizability of Cryopreserved Bovine Semen.

To study the ameliorative capacity of Syzygium cumini fruit Pulp Extract (SPE) on cryopreserved bovine semen, three ejaculates each of three fertile bulls (total nine samples) were collected. Each sample was mixed with semen dilution medium in 1:2 ratios, centrifuged, and 50% of the supernatant was discarded. The remaining sample was mixed with semen cryopreservation extender (SCE) in 1:4 ratios. Three aliquots (0.3 mL) of each extended semen sample were further diluted with (1) 0.7 mL SCE (SCE group), (2) 0.7 mL SCE containing SPE equivalent to 7 ppm total antioxidants (SPE7 group), and (3) 0.7 mL SCE equivalent to 14 ppm antioxidants (SPE14 group). For each aliquot, semen quality was analyzed in situ as well as after cryopreservation. Results show significant improvement in sperm membrane integrity, motility, and fertilizability in SPE7 (63.5% ± 2.04%, 56.22% ± 1.7%, 51.1%, respectively) and SPE14 (68% ± 1.12%, 63.67% ± 1.06%, 57.7%, respectively) than the SCE group (62.33% ± 1.22%, 54.9% ± 1.34%, 48.8%, respectively). Similarly, significant improvement in the mean percent number of progressively motile sperm at the expense of the mean percent twitching, shaking, and whirling spermatozoa, both before and after cryopreservation, were observed in SPE7 (72 ± 1.06, 61.2 ± 1.2) and SPE14 (78.89 ± 1.12, 67 ± 1.32) groups to that of the SCE (59.6 ± 0.97, 52.3 ± 3.5). In addition, the mean percent of progressively motile spermatozoa showing 20 µm/s or above velocity, before and after cryopreservation, were significantly higher in SPE7 (58.9 ± 4.7, 43.6 ± 1.5) and SPE14 (66.9 ± 0.8, 51.3 ± 0.8) groups than that of the SCE (42.3 ± 3.1, 30 ± 2.4). These findings indicate that SPE can significantly improve various quality parameters of the cryopreserved bovine semen.

C3 Glomerulopathy: Pathogenesis and Treatment.

C3 glomerulopathy (C3G) is a rare set of kidney diseases with 2 patterns: C3 glomerulonephritis (C3GN) and dense deposit disease. Pathogenesis of both diseases is due to complement dysregulation in the alternative pathway. Acquired or genetic alterations of the regulatory proteins of the complement pathway result in C3G. Although the disease is characterized by low C3 levels in serum and C3-dominant staining by immunofluorescence on biopsy, other disease entities such as infection-related glomerulonephritis and masked monoclonal deposits can present similarly. Both the C3GN and dense deposit disease variants of C3G are progressive and recur in transplanted kidneys. Although no direct treatment is available, complement blockers are either available or in the clinical trial phase. This review will survey the pathogenesis of C3GN and current treatment options.

Antigens, antibodies, and membranous nephropathy: a decade of progress.

Membranous nephropathy (MN) is either primary or associated with various etiologies, each with unique glomerular antigens. The discovery of the phospholipase A2 receptor (PLA2R) antigen in primary MN revolutionized our understanding of MN and led to major clinical progress. Other recently discovered antigens in MN include the THSD7A antigen and exostosin. Sethi et al. have now identified a new antigen, NELL-1, in primary MN, again decreasing the number of patients whose antigen remains unknown.

Tourette Syndrome: a general pediatrician's 35-year experience at a single center with follow-up in adulthood.

A retrospective analysis of a 35-year single-center experience with pediatric tics and Tourette syndrome was conducted. 482 charts from 1972 to 2007 were reviewed. Follow-up surveys were mailed to last known address and 83 patients responded (17%). Response rate was affected by long interval from last visit; contact information was often incorrect as it was the address of the patient as a child. Males constituted 84%. Mean tic onset was 6.6 years. At first visit, 83% had multiple motor tics and >50% had comorbidities. 44% required only 1 visit and 90% less than 12 visits. Follow-up showed positive clinical and social outcomes in 73/83 survey responses. Of those indicating a poor outcome, mean educational level was lower and attention deficit/hyperactivity disorder and learning disabilities were significantly higher. Access to knowledgeable caregivers was a problem for adult patients. A shortage of specialists may in part be addressed by interested general pediatricians.

Specific evoked motor response for supraclavicular brachial block.

OBJECTIVE: To assess and evaluate the accuracy of individual types of evoked motor responses (EMRs) for prediction of successful surgical anesthesia following peripheral nerve stimulator (PNS) assisted supraclavicular block. METHODS: A prospective study was carried out over a period of one and half years from August 2004 to January 2006, at J. N. Medical College, Aligarh Muslim University, Aligarh, India, in 60 patients who underwent various elective operative procedures on the upper limb. Any of the EMR, such as forearm flexion or extension, carpal flexion, or extension, prono-supination, or finger flexion, at a definite current of 0.25 mA for 2 ms was taken as an end point for prediction of successful block, and a local anesthetic solution (0.375% Bupivacaine, 30 ml) was administered at that level. RESULTS: Complete surgical anesthesia was observed in those cases where EMR included: flexion of only second and third fingers (n=15/15) or flexion of all 4 fingers with thumb opposition (n=14/14) or uncommon flexion of all 4 fingers without movement of any other joint of the upper limb (n=2/2), suggesting a sensitivity of 100%. However, thumb opposition to the tip of the flexed little finger revealed a success rate of only 83% (n=5/6), and other EMRs were followed by high rates of inadequate surgical anesthesia or total failure. CONCLUSION: Specific EMR of flexion of second and third fingers, or all the 4 fingers are reliable predictors of complete surgical anesthesia following PNS assisted single injection supraclavicular nerve block with no incidence of pneumothorax and nerve damage.