Zinc Ortho Methyl Carbonodithioate Improved Pre and Post-Synapse Memory Impairment via SIRT1/p-JNK Pathway against Scopolamine in Adult Mice.

Alzheimer's disease (AD) is globally recognized as a prominent cause of dementia for which efficient treatment is still lacking. New candidate compounds that are biologically potent are regularly tested. We, therefore, hypothesized to study the neuroprotective potential of Zinc Ortho Methyl Carbonodithioate (thereafter called ZOMEC) against Scopolamine (SCOP) induced Alzheimer's disease (AD) model using adult albino mice. We post-administered ZOMEC (30 mg/Kg) into two group of mice for three weeks on daily basis that received either 0.9% saline or SCOP (1 mg/Kg) for initial two weeks. The other two groups of mice received 0.9% saline and SCOP (1 mg/Kg) respectively. After memory related behavioral analysis the brain homogenates were evaluated for the antioxidant potential of ZOMEC and multiple protein markers were examined through western blotting. Our results provide enough evidences that ZOMEC decrease oxidative stress by increasing catalase (CAT) and glutathione S transferase (GST) and decreasing the lipid peroxidation (LPO). The SIRT1 and pre and post synaptic marker proteins, synaptophysin (SYP) as well as post synaptic density protein (PSD-95) expression were also enhanced upon ZOMEC treatment. Furthermore, memory impairment was rescued and ZOMEC appreciably abrogated the Aß accumulation, BACE1 expression C and the p-JNK pathway. The inflammatory protein markers, NF-kß and IL-1ß in ZOMEC treated mice were also comparable with control group. The predicted interaction of ZOMEC with SIRT1 was further confirmed by molecular docking. These findings thus provide initial reports on efficacy of ZOMEC in SCOP induced AD model.

Comparative binding analysis of WGX50 and Alpha-M with APP family proteins APLP1 and APLP2 using structural-dynamics and free energy calculation approaches.

A.D. is a common disease among other neurodegenerative disorders primarily developing due to amyloid-ß (Aß) neurotoxicity derived from the amyloid-ß protein precursor (AßPP). The amyloid precursor-like proteins 1 and 2 (APP1 and APLP2) biochemically behave similarly in many aspects to AßPP. We, therefore, proposed to test WGX-50 and Alpha-M for their interaction mechanism with APLP1 and APLP2 because both these drug candidate compounds previously showed inhibition of Aß aggregation. We employed a comparative atomic investigation on Alpha-M and WGX-50 in complex with novel targets, i.e., APLP1 and APLP2, using biophysical and molecular simulation methods. The docking score was -6.83 kcal mol-1 for Alpha-M-APLP1, -8.41 kcal mol-1 for WGX-50-APLP1, -7.02 kcal mol-1 for Alpha-M-APLP2 and -8.25 kcal mol-1 for the WGX-50-APLP2 complex. Our results also elaborate that in the case of their interaction with both APLP1 and APLP2, the WGX-50 complex exhibits better stability than the APLP1/2-Alpha-M complexes during simulation. Furthermore, WGX50 in both APLP1 and APLP2 stabilized the internal flexibility upon binding in contrast to the Alpha-M complexes. The data showed that the BFE for Alpha-M-APLP1 was calculated to be -27.38 ± 0.93 kcal mol-1, for WGX-50-APLP1 -39.65 ± 0.95 kcal mol-1, for Alpha-M-APLP2 -24.80 ± 0.63 kcal mol-1 while for WGX-50-APLP2 the BFE was -57.16 ± 1.03 kcal mol-1 respectively. These results highlight that APLP2-WGX50 has greater binding energies in all four systems. PCA and FEL analysis further revealed variations in the dynamic behavior of these complexes. Overall, our findings demonstrate that WGX50 potentially acts as a more potent inhibitor for APLP1 and APLP2 than Alpha-M and thus shows the diverse pharmacological potential of WGX50. Due to its stable binding interaction, WGX50 might be a suitable candidate drug compound for targeting these precursors under pathological conditions.

Predictive Factors of Outcomes in Acute Subdural Hematoma Evacuation.

OBJECTIVE: This comparative cross-sectional study was conducted in the Departments of Trauma and Neurosurgery, Ayub Teaching Hospital, Abbottabad, Pakistan from September 2021 to February 2022 to study predictive factors of outcomes in acute subdural hematoma evacuation. METHODOLOGY: A total of 101 patients with confirmed diagnosis of acute subdural hematoma (ASDH) who underwent surgical evacuation by consultant neurosurgeon were included in the study. A detailed clinical proforma was designed to document all the clinical and demographic details of these patients at the time of admission. Glasgow Coma Scale outcome score (GOS) was used to assess the outcome of patients after the surgery. Sociodemographic and clinical parameters were associated with outcome of surgery in our study participants. RESULTS: Out of 101 patients, 55 (54.5%) were males and 46 (45.5%) were females. Mean age was 43.66±19.66 years with 7.39 as mean Glasgow Coma Scale (GCS) at presentation. Road traffic accident (RTA) 62 (61.4%) was most frequent mechanism of injury followed by fall from height (19.8%) and history of assault (13.9%). In our study, 59 patients had poor outcomes while 42 had good outcomes. Elder age, low GCS at presentation, and use of oral anticoagulant were associated with poor outcomes while pupillary reaction had no effect on the outcome after application of test of significance. CONCLUSION: More than half of the patients managed with surgical evacuation for acute subdural hematoma as per guidelines at our neurosurgical unit had poor outcomes according to Glasgow Coma Scale. In this study, advancing age (>60 years), low GCS score at presentation, and use of oral anticoagulation therapy emerged as significant risk factors for poor outcomes in participants. Pupillary reaction had no effect on outcomes as per this study but this needs further evaluation in future studies.

Efficacy Of Conservative Management Of Early Post Traumatic Cerebrospinal Fluid Rhinorrhoea.

Background: The incidence of Post-traumatic Cerebrospinal Fluid Rhinorrhoea (PCSFR) has been decreased due to advanced therapeutic measures. The current investigative study has been arranged to assess the efficacy of conservative management of early PCSFR. Method: This cross-sectional study was conducted at departmental of Neurosurgery, Ayub Medical Institute, Abbottabad. Patients with traumatic brain injury having Cerebrospinal fluid rhinorrhoea with either gender having age 5-50 year and presenting within seven days of traumatic brain injury were included. Moreover, those with nasal fractures, penetrating head injuries having fever and neck stiffness were also included in the study. Results: A total number of 120 patients having male dominancy, i.e., 86 (72%) were included in the study with the mean age of 27 years ± 8.741 in which 77 (92%) patients were in the age range of 2nd to 4th decades. The commonest cause was trauma due to Road Traffic Accidents (RTA) having 65 (54%) patients. Conservative treatment was effective in 62 (52%) patients predominantly in the patients of 3rd decade, i.e., 31 (50%), in which the effectiveness in male gender was revealed to be 52.32% (45) and effectiveness in RTA patients was recorded to be 54.83%. Similarly, the Chi-Square value was calculated for the PCSFR patients for four groups of patients (5-20, 21-30, 31-40, 41-50) to be 48.27 having critical value of 7.81 with the p-value of 1.87e-10, which completely rejects the Null-hypothesis for the patients of various ages. Conclusion: Based on the current investigative study, it may be concluded that PCSFR is common in middle age population with slight male dominancy. It may also be inferred that RTA is the leading cause of PCSFR in our set up and majority of the patients shows improvement after conservative management. Moreover, the effectiveness of conservative management of PCSFR could be predominantly observed in the patients of 3rd decade.

Surgical Treatment Options For Chronic Subdural Haematoma.

Subdural hematoma is an encapsulated collection of blood under the dura matter. This commonly encountered neurosurgical disorder is best managed by surgical evacuation; however, contemporary neurosurgery lacks a consensus regarding surgical technique of choice. Due to high incidence of the condition and associated complications, vast amount of literature is available on the subject; including studies comparing efficacy of various available treatment modalities. Herein, literature on surgical techniques employed for management of Chronic Subdural Hematoma (CSDH) has been reviewed to provide an evidence-based review on best surgical practices. Following conclusions can be made on basis of evidence of various levels provided in the studied literature: (1) Twist-drill craniostomy is a relatively safe technique that can be employed under local anaesthesia, thus can be considered as first line treatment in high risk surgical candidates. (2) Single and double burr-hole craniostomies have shown comparable results. (3) Intraoperative irrigation during burr-hole craniostomy doesn't affect outcome. (4) Drain insertion after hematoma evacuation lowers recurrence risk. (5) Position of drain is not significant but early drain removal is associated with higher recurrence rates. (6) Craniotomy is associated with high morbidity and mortality, hence should be reserved for recurrent and large septate hematoma cases. (7) Head elevation in postoperative period reduces recurrence. (8) Embolization of middle meningeal artery (EMMA): A novel treatment modality, is promising but requires further approval in terms of large sample sized multicenter randomized control trials. In conclusion further research is required on the subject to formulate guidelines regarding management of this common neurosurgical emergency.

Extracellular vesicles improve embryo cryotolerance by maintaining the tight junction integrity during blastocoel re-expansion.

Cryopreservation is a process in which the intact living cells, tissues, or embryos are preserved at subzero temperatures for preservation. The cryopreservation process highly impacts the survival and quality of the in vitro-produced (IVP) embryos. Some studies have highlighted the use of oviduct extracellular vesicles (EVs) to improve the cryotolerance of IVP embryos but the mechanism has not been well studied. The present study unravels the role of in vitro cultured bovine oviduct epithelial cells-derived EVs in improving the re-expansion and hatching potential of thawed blastocysts (BLs). The comparison of cryotolerance between synthetic oviduct fluid (SOF) and SOF + EVs-supplemented day-7 cryopreserved BLs revealed that the embryo's ability to re-expand critically depends on the intact paracellular sealing which facilitates increased fluid accumulation during cavity expansion after shrinkage. Our results demonstrated that BLs cultured in the SOF + EVs group had remarkably higher re-expansion (67.5 ± 4.2%) and hatching rate (84.8 ± 1.4%) compared to the SOF group (53.4 ± 3.4% and 63.9 ± 0.9%, respectively). Interestingly, EVs-supplemented BLs exhibited greater influence on the expression of core genes involved in trophectoderm (TE) maintenance, formation of tight junction (TJ) assembly, H2O channel proteins (aquaporins), and Na+/K+ ATPase alpha 1. The EVs improved the fluid flux and allowed the transport of H2O into an actively re-expanded cavity in EVs-cultured cryo-survived BLs relative to control BLs. Our findings explored the function of EVs in restoring the TE integrity, improved the cell junctional contacts and H2O movement which helps the blastocoel re-expansion after thawing the cryopreserved BLs.

Hidden allosteric sites and De-Novo drug design.

INTRODUCTION: Hidden allosteric sites are not visible in apo-crystal structures, but they may be visible in holo-structures when a certain ligand binds and maintains the ligand intended conformation. Several computational and experimental techniques have been used to investigate these hidden sites but identifying them remains a challenge. AREAS COVERED: This review provides a summary of the many theoretical approaches for predicting hidden allosteric sites in disease-related proteins. Furthermore, promising cases have been thoroughly examined to reveal the hidden allosteric site and its modulator. EXPERT OPINION: In the recent past, with the development in scientific techniques and bioinformatics tools, the number of drug targets for complex human diseases has significantly increased but unfortunately most of these targets are undruggable due to several reasons. Alternative strategies such as finding cryptic (hidden) allosteric sites are an attractive approach for exploitation of the discovery of new targets. These hidden sites are difficult to recognize compared to allosteric sites, mainly due to a lack of visibility in the crystal structure. In our opinion, after many years of development, MD simulations are finally becoming successful for obtaining a detailed molecular description of drug-target interaction.

Level Of Pre-Tertiary Care Hospital Management Of Traumatic Spinal Cord Injured Patients; Where Do We Stand?

BACKGROUND: Traumatic spinal cord injury is a debilitating condition that may cause long term disabilities with tremendous socioeconomic impact on affected individuals and their families. Secondary injuries can best prevent or minimized by appropriate pre hospital management and proper referral and transfer. This study was conducted to assess the clinical profile of traumatic spinal cord injuries and level of pre-hospital care provided to patients either at the site of injury or at other healthcare facilities. METHODS: This prospective study was conducted in the Department of Neurosurgery Ayub Teaching Hospital Abbottabad, from January 2012 to January 2017. All patients with suspected spinal injury were included in the study. Age, gender, mode of injuries and the pre-tertiary care provided were recorded. RESULTS: Out of 4464 patients with suspected spinal cord injury, 3685 (82.5%) were male, 779 (17.4%) were female. Age ranged from 10-70 years. 1685 (37.8%) were diagnosed as having spinal injury. Cervical spine was the most common affected level 743 (44.09%), followed by thoracic spine 135 (8.01%). 1441 (85.5%) were incomplete while 224(14.5%) were complete spinal cord injuries. Road traffic accident was the most common mechanism of injury 884 (52.4%). Only 4 (0.23%) patients directly received in our unit were properly transported, 66 (3.91%) were brought after proper spinal immobilization, intravenous line was maintained in 584 (34.66%) patients, 410 (24.3%) patients received some fluid resuscitation, parenteral analgesia was given to 441 (26.17%) patients while urinary catheterization was done in 195 (11.75%) patients. Those received from other healthcare facilities only 4 (0.23%) were brought by properly equipped ambulance, intravenous access was maintained in 438 (25.99%),320 (18.99%) received some fluid resuscitation, urinary catheterization was done in 229(13.59%) while proper parenteral analgesia was given to 988 (58.63%) patients. CONCLUSIONS: There is a complete lack of proper transport and referral of trauma patients in our area which reflects almost non-existent emergency medical (rescue) services, deficient health care facilities.

Bovine Oviduct Epithelial Cell-Derived Culture Media and Exosomes Improve Mitochondrial Health by Restoring Metabolic Flux during Pre-Implantation Development.

Oviduct flushing is enriched by a wide variety of nutrients that guide the 3-4 days journey of pre-implantation embryo through the oviduct as it develops into a competent blastocyst (BL). However, little is known about the specific requirement and role of these nutrients that orchestrate the early stages of embryonic development. In this study, we aimed to characterize the effect of in vitro-derived bovine oviduct epithelial cell (BOECs) secretion that mimics the in vivo oviduct micro-fluid like environment, which allows successful embryonic development. In this study, the addition of an in vitro derived BOECs-condition media (CM) and its isolated exosomes (Exo) significantly enhances the quality and development of BL, while the hatching ability of BLs was found to be high (48.8%) in the BOECs-Exo supplemented group. Surprisingly, BOECs-Exo have a dynamic effect on modulating the embryonic metabolism by restoring the pyruvate flux into TCA-cycle. Our analysis reveals that Exo treatment significantly upregulates the pyruvate dehydrogenase (PDH) and glutamate dehydrogenase (GLUD1) expression, required for metabolic fine-tuning of the TCA-cycle in the developing embryos. Exo treatment increases the influx into TCA-cycle by strongly suppressing the PDH and GLUD1 upstream inhibitors, i.e., PDK4 and SIRT4. Improvement of TCA-cycle function was further accompanied by higher metabolic activity of mitochondria in BOECs-CM and Exo in vitro embryos. Our study uncovered, for the first time, the possible mechanism of BOECs-derived secretion in re-establishing the TCA-cycle flux by the utilization of available nutrients and highlighted the importance of pyruvate in supporting bovine in vitro embryonic development.

Phylogenetic Analysis and Structural Perspectives of RNA-Dependent RNA-Polymerase Inhibition from SARs-CoV-2 with Natural Products.

Most recently, an outbreak of severe pneumonia caused by the infection of SARS-CoV-2, a novel coronavirus first identified in Wuhan, China, imposes serious threats to public health. Upon infecting host cells, coronaviruses assemble a multi-subunit RNA-synthesis complex of viral non-structural proteins (nsp) responsible for the replication and transcription of the viral genome. Therefore, the role and inhibition of nsp12 are indispensable. A cryo-EM structure of RdRp from SARs-CoV-2 was used to identify novel drugs from Northern South African medicinal compounds database (NANPDB) by using computational virtual screening and molecular docking approaches. Considering Remdesivir as the control, 42 compounds were shortlisted to have docking score better than Remdesivir. The top 5 hits were validated by using molecular dynamics simulation approach and free energy calculations possess strong inhibitory properties than the Remdesivir. Thus, this study paved a way for designing novel drugs by decoding the architecture of an important enzyme and its inhibition with compounds from natural resources. This disclosing of necessary knowledge regarding the screening and the identification of top hits could help to design effective therapeutic candidates against the coronaviruses and design robust preventive measurements.

Role of Wnt Signaling During In-Vitro Bovine Blastocyst Development and Maturation in Synergism with PPARδ Signaling.

Wnt/ß-catenin signaling plays vital role in the regulation of cellular proliferation, migration, stem cells cell renewal and genetic stability. This pathway is crucial during the early developmental process; however, the distinct role of Wnt/ß-catenin signaling during pre-implantation period of bovine embryonic development is obscure. Here, we evaluated the critical role of Wnt/ß-catenin pathway in the regulation of bovine blastocyst (BL) development and hatching. 6 bromoindurbin-3'oxime (6-Bio) was used to stimulate the Wnt signaling. Treatment with 6-Bio induced the expression of peroxisome proliferator-activated receptor-delta (PPARδ). Interestingly, the PPARδ co-localized with ß-catenin and form a complex with TCF/LEF transcription factor. This complex potentiated the expression of several Wnt directed genes, which regulate early embryonic development. Inhibition of PPARδ with selective inhibitor 4-chloro-N-(2-{[5-trifluoromethyl]-2-pyridyl]sulfonyl}ethyl)benzamide (Gsk3787) severely perturbed the BL formation and hatching. The addition of Wnt agonist successfully rescued the BL formation and hatching ability. Importantly, the activation of PPARδ expression by Wnt stimulation enhanced cell proliferation and fatty acid oxidation (FAO) metabolism to improve BL development and hatching. In conclusion, our study provides the evidence that Wnt induced PPARδ expression co-localizes with ß-catenin and is a likely candidate of canonical Wnt pathway for the regulation of bovine embryonic development.

An Integrated Systems Biology and Network-Based Approaches to Identify Novel Biomarkers in Breast Cancer Cell Lines Using Gene Expression Data.

Breast cancer is the most common cause of death in women worldwide. Approximately 5%-10% of instances are attributed to mutations acquired from the parents. Therefore, it is highly recommended to design more potential drugs and drug targets to eradicate such complex diseases. Network-based gene expression profiling is a suggested tool for discovering drug targets by incorporating various factors such as disease states, intensities based on gene expression as well as protein-protein interactions. To find prospective biomarkers in breast cancer, we first identified differentially expressed genes (DEGs) statistical methods p-value and false discovery rate were initially used. Of the total 82 DEGs, 67 were upregulated while the remaining 17 were downregulated. Sub-modules and hub genes include VEGFA with the highest degree, followed by 15 CCND1 and CXCL8 with 12-degree score was found. The survival analysis revealed that all the hub genes have important role in the development and progression of breast cancer. Enrichment analysis revealed that most of these genes are involved in signaling pathways and in the extracellular spaces. We also identified transcription factors and kinases, which regulate proteins in the DEGs PPI. Finally, drugs for each hub genes were identified. These results further expanded the knowledge regarding important biomarkers in breast cancer.

Dysregulation of neuron differentiation in an autistic savant with exceptional memory.

Autism spectrum disorder (ASD) is a heterogeneous group of complex neurodevelopmental disorders without a unique or definite underlying pathogenesis. Although savant syndrome is common in ASD, few models are available for studying the molecular and cellular mechanisms of this syndrome. In this study, we generated urinary induced pluripotent stem cells (UiPSCs) from a 13-year-old male autistic savant with exceptional memory. The UiPSC-derived neurons of the autistic savant exhibited upregulated expression levels of ASD genes/learning difficulty-related genes, namely PAX6, TBR1 and FOXP2, accompanied by hypertrophic neural somas, enlarged spines, reduced spine density, and an increased frequency of spontaneous excitatory postsynaptic currents. Although this study involved only a single patient and a single control because of the rarity of such cases, it provides the first autistic savant UiPSC model that elucidates the potential cellular mechanisms underlying the condition.

Status and Magnitude of Grey Wolf Conflict with Pastoral Communities in the Foothills of the Hindu Kush Region of Pakistan.

Pastoralist-wolf conflict over livestock depredation is the main factor affecting conservation of grey wolf worldwide. Very limited research has been carried out to evaluate the pattern and nature of livestock depredation by wolf. This study aims to determine the status and nature of human-wolf conflict across different villages in the Hind Kush region of Pakistan during the period January 2016-December 2016. For this purpose, a total of 110 local male respondents from all walks of life were interviewed using a semi-structured questionnaire. The grey wolf was declared as a common species in the area by 51.3% of the locals with an annual sighting rate of 0.46 each. During the year (2016), a total of 358 livestock were lost to grey wolf predation and disease. Of the total livestock loss, grey wolf was held responsible for a total 101 livestock losses. Goat and sheep were the most vulnerable prey species as they accounted for 80 (79.2%) of the total reported depredations. Out of the total economic loss (USD 46,736, USD 424.87/household), grey wolf was accountable for USD 11,910 (USD 108.27 per household), while disease contributed 34,826 (USD 316.6 per household). High depredation was observed during the summer season 58.42% (n = 59) followed by spring and autumn. Unattended livestock were more prone to grey wolf attack during free grazing in forests. Most of the respondents (75.45%) showed aggressive and negative attitudes towards grey wolf. The herders shared more negative attitude (z = -3.21, p = 0.001) than businessman towards the species. Herders having larger herd size displayed more deleterious behavior towards wolves than those having smaller herd size. Active herding techniques, vaccinating livestock, educating locals about wildlife importance, and initiating compensating schemes for affected families could be helpful to decrease negative perceptions.

Comparison Of Lumbar Drain Insertion And Conservative Management In The Treatment Of Traumatic CSF Rhinorrhoea.

BACKGROUND: Conservative management of traumatic CSF rhinorrhoea is associated with a greater risk of developing meningitis in the presence of active CSF leak. Lumbar drains have been reported to be better than conservative management alone in stopping CSF leaks following traumatic brain injury. METHODS: This randomized controlled trial enrolled 60 patients with CSF rhinorrhoea and divided them into two groups. One group was managed with conservative management plus a lumbar drain (group A) and the other was managed with conservative management alone (Group B). Length of CSF rhinorrhoea in days was estimated in both groups. RESULTS: There was a statistically significant difference in in mean length of CSF rhinorrhoea in both groups. In group A, mean Length of CSF rhinorrhoea was found to be 3.4 days ±1.1 SD, while in group B it was 6.75 days ±1.96 SD (p=0.001). Stratification with respect to gender, age, duration and type of trauma showed similar trend (p<0.05 in all cases). CONCLUSIONS: Patients who underwent lumbar drain insertion plus conservative management demonstrated significantly shorter length of CSF rhinorrhoea when compared to conservative management alone in the treatment of traumatic CSF rhinorrhoea.

Immunoinformatics and structural vaccinology driven prediction of multi-epitope vaccine against Mayaro virus and validation through in-silico expression.

The Mayaro virus (MAYV) belongs to genus "Alphavirus" and family "Togaviridae". MAYV has distribution in the Amazonia, Central and Northeastern regions of Brazil. The abundance of mosquito vector Haemagogus janthinomys has major role in the outbreaks of arthralgia disease in Brazil. Vaccination or immunization is an alternative approach for the protection against this disease. To search the effective candidate for vaccine against Mayaro virus, various immunoinformatics tools were used to predict both the B and T cell epitopes from five structural polyproteins (capsid, E2, 6K, E3and E1). A multi subunit vaccine was designed and the final sequence was modeled for docking with TLR-3. Human b defensin based on previous studies was used as linker. The docked complexes of vaccine-TLR-3 were then subjected to dynamics stability and RMSD and RMSF results suggested that the complexes are stable. Further, to validate our final vaccine construct, in silico cloning was carried out using E. coli as host. The CAI value of 0.96 suggests that the vaccine construct properly expresses in the host. The current findings will be useful for the future experimental validations to ratify the immunogenicity and safety of the supposed structure of vaccine, and ultimately to treat the Mayaro virus, associated infections.

Efficacy Of Phenytoin In Prevention Of Early Posttraumatic Seizures.

BACKGROUND: The use of anti-epileptic drugs for prophylaxis of early post-traumatic seizures after traumatic brain injury has been very promising. The objective of this study was to determine the outcome of phenytoin in prevention of early post-traumatic seizures in moderate to severe traumatic brain injuries and to compare the frequency of seizures in moderate to severe traumatic brain injury, with phenytoin started within 12 hours and after 12 hours of injury. METHODS: This cross-sectional study was conducted at Department of Neurosurgery, Ayub Medical Institute, Abbottabad from April to October, 2015. All the patients with moderate to severe head injury presenting within 48 hours of injury were included in this study in consecutive manner. Patients were started on phenytoin and observed for early post-traumatic seizures. RESULTS: A total of 163 patients were included in this study with a mean age of 24.69±10.186 years. One hundred and twenty-two (74.8%) were males and rest of 41 (25.2%) were females. A total of 26 (16%) patients had early post-traumatic seizures. 9.89% patients in whom phenytoin was started within 12 hours had seizures, while 23.11% patients in whom phenytoin was started after 12 hours of injury had seizures, the difference being statistically significant (p-value .018).. CONCLUSIONS: Frequency of early post-traumatic seizures is high in patients with moderate to severe head injured patients. Anti-epileptics like phenytoin should be started within 12 hours for seizure prophylaxis.

Difference in Developmental Kinetics of Y-Specific Monoclonal Antibody Sorted Male and Female In Vitro Produced Bovine Embryos.

Sex-related growth differences between male and female embryos remain an attractive subject for reproductive biologists. This study aimed to investigate the endogenous factors that play a crucial role in the pace of early development between male and female bovine embryos. Using sex pre-selected semen by Y-specific monoclonal antibodies for the production of bovine embryos, we characterized the critical endogenous factors that are responsible for creating the development differences, especially during the pre-implantation period between male and female embryos. Our results showed that at day seven, (57.8%) Y-sperm sorted in vitro cultured embryos reached the expanded blastocyst (BL) stage, whereas the X-sperm sorted group were only 25%. Y-BLs showed higher mRNA abundance of pluripotency and developmental competency regulators, such as Oct4 and IGF1-R. Interestingly, Y-sperm sorted BLs had a homogeneous mitochondrial distribution pattern, higher mitochondrial membrane potential (∆Ñ°m), efficient OXPHOS (oxidative phosphorylation) system and well-encountered production of ROS (reactive oxygen species) level. Moreover, Y-blastocysts (BLs) showed less utilization of glucose metabolism relative to the X-BLs group. Importantly, both sexes showed differences in the timing of epigenetic events. All these factors directly or indirectly orchestrate the whole embryonic progression and may help in the faster and better quality yield of BL in the Y-sperm sorted group compared to the X counterpart group.

A novel role of metal response element binding transcription factor 2 at the Hox gene cluster in the regulation of H3K27me3 by polycomb repressive complex 2.

Polycomb repressive complex 2 (PRC2) is known to play an important role in the regulation of early embryonic development, differentiation, and cellular proliferation by introducing methyl groups onto lysine 27 of histone H3 (H3K27me3). PRC2 is tightly associated with silencing of Hox gene clusters and their sequential activation, leading to normal development and differentiation. To investigate epigenetic changes induced by PRC2 during differentiation, deposition of PRC2 components and levels of H3K27me3 were extensively examined using mouse F9 cells as a model system. Contrary to positive correlation between PRC2 deposition and H3K27me3 level, down-regulation of PRC2 components by shRNA and inhibition of EZH1/2 resulted in unexpected elevation of H3K27me3 level at the Hox gene cluster despite its global decrease. We found that metal response element binding transcriptional factor 2 (MTF2), one of sub-stoichiometric components of PRC2, was stably bound to Hox genes. Its binding capability was dependent on other core PRC2 components. A high level of H3K27me3 at Hox genes in Suz12-knock out cells was reversed by knockdown of Mtf2.This shows that MTF2 is necessary to consolidate PRC2-mediated histone methylation. Taken together, our results indicate that expression of Hox gene clusters during differentiation is strictly modulated by the activity of PRC2 secured by MTF2.

Vocalizations of adult male Asian koels (Eudynamys scolopacea) in the breeding season.

Defining the vocal repertoire provides a basis for understanding the role of acoustic signals in sexual and social interactions of an animal. The Asian koel (Eudynamys scolopacea) is a migratory bird which spends its summer breeding season in the plains of Pakistan. The bird is typically wary and secretive but produces loud and distinct calls, making it easily detected when unseen. Like the other birds in the wild, presumably Asian koels use their calls for social cohesion and coordination of different behaviors. To date, the description of vocal repertoire of the male Asian koel has been lacking. Presently we analyzed and described for the first time the vocalizations of the adult male Asian koel, recorded in two consecutive breeding seasons. Using 10 call parameters, we categorized the vocalization type into six different categories on the basis of spectrogram and statistical analyses, namely the; "type 1 cooee call", "type 2 cooee call", "type 1 coegh call", "type 2 coegh call", "wurroo call" and "coe call". These names were assigned not on the basis of functional analysis and were therefore onomatopoeic. Stepwise cross validated discriminant function analysis classified the vocalization correctly (100%) into the predicted vocal categories that we initially classified on the basis of spectrographic examination. Our findings enrich the biological knowledge about vocalizations of the adult male Asian koel and provide a foundation for future acoustic monitoring of the species, as well as for comparative studies with vocalizations of other bird species of the cuckoo family. Further studies on the vocalizations of the Asian koel are required to unravel their functions in sexual selection and individual recognition.