Effect of actin polymerization inhibitor during oocyte maturation on parthenogenetic embryo development and ploidy in Capra hircus.

This study was designed to observe the effect of cytochalasin B (CCB) concentrations on ploidy and early development of parthenogenetic embryos in a caprine species. Caprine oocytes were matured in the presence of different concentrations of CCB (5, 10, 15, and 20 µg/ml) and activated by 7% ethanol followed by incubation with 2 mM DMAP. For embryos fertilized in vitro, oocytes were matured in maturation medium without CCB. The cleavage rate and further embryo development were significantly higher (P < 0.05) when oocytes were treated in this way. The percentage of embryos showed higher diploid values in 15 µg/ml CCB (83.66 ± 1.13), followed by 20 (72.22 ± 1.22), 10 (68.57 ± 1.17), and 5 µg/ml (62.00 ± 2.48). These results indicate that CCB with a concentration of 15 µg/ml in maturation medium can be used for the production of diploid parthenogenetic embryos in the caprine species.

Multi-modality parathyroid imaging: A shifting paradigm.

The goal of parathyroid imaging in hyperparathyroidism is not diagnosis, rather it is the localization of the cause of hyperparathyroidism for planning the best therapeutic approach. Hence, the role of imaging to accurately and precisely localize the abnormal parathyroid tissue is more important than ever to facilitate minimally invasive parathyroidectomy over bilateral neck exploration. The common causes include solitary parathyroid adenoma, multiple parathyroid adenomas, parathyroid hyperplasia and parathyroid carcinoma. It is highly imperative for the radiologist to be cautious of the mimics of parathyroid lesions like thyroid nodules and lymph nodes and be able to differentiate them on imaging. The various imaging modalities available include high resolution ultrasound of the neck, nuclear imaging studies, four-dimensional computed tomography (4D CT) and magnetic resonance imaging. Contrast enhanced ultrasound is a novel technique which has been recently added to the armamentarium to differentiate between parathyroid adenomas and its mimics. Through this review article we wish to review the imaging features of parathyroid lesions on various imaging modalities and present an algorithm to guide their radiological differentiation from mimics.

Illness wellness scale: novel grading system for performance status of patients under surgical care.

BACKGROUND: Grading the illness using clinical parameters is essential for the daily progress of inpatients. Existing systems do not incorporate these parameters holistically. The study was designed to internally validate the illness wellness scale, based upon clinical assessment of the patients requiring surgical care, for their risk stratification and uniformity of communication between health care providers. METHODS: Prospective observational study conducted at a tertiary care hospital. An expert panel devised the scale, and it was modified after feedback from 100 health care providers. A total of 210 patients (150 for internal validation and 60 for inter-observer variability) who required care under the department of surgical disciplines were enrolled. This included patients presenting to surgery OPD, admitted to COVID/non-COVID surgical wards and ICUs, aged ≥16 years. RESULTS: The response rate of the final illness wellness scale was 95% with 86% positive feedback and a mean of 1.7 on the Likert scale for ease of use (one being very easy and five being difficult). It showed excellent consistency and minimal inter-observer variability with the intra-class correlation coefficient (ICC) above 0.9. In the internal validation cohort (n = 150), univariate and multivariable analysis of factors affecting mortality revealed that categorical risk stratification, age ≥ 60 years, presence or absence of co-morbidities especially hypertension and chronic kidney disease significantly affect mortality. CONCLUSIONS: The Illness wellness scale is an effective tool for uniformly communicating between health care professionals and is also a strong predictor of risk stratification and mortality in patients requiring surgical care.

Complex chest wall reconstruction after excision of malignant phyllodes tumour.

Phyllodes tumour is a rare breast tumour which is locally invasive, and full thickness chest wall involvement by phyllodes is rarely reported. A woman presented with recurrent phyllodes tumour and multilobulated exophytic mass of size 18.5×5.3 cm in anterior chest wall which was abutting the right 2nd-7th rib, left 3rd-5th rib and costal cartilages. She underwent wide excision of the left chest wall tumour with excision of left 3rd-5th rib, with polypropylene mesh and pedicled lattismus dorsi myocutaneous flap reconstruction. After recovery, 3 weeks later, she underwent wide excision of the phyllodes tumour on the right chest wall with excision of 2nd-7th rib, lateral border of sternum, right crus of the diaphragm and the lower lobe of the right lung. Anterior chest wall was reconstructed using scaffold made from Steinmann pins and polypropylene mesh with greater omentum pedicled flap and split thickness skin graft.

Comparison of Modified Rose Method of Thyroid Palpation Versus Other Methods for the Detection of Retrosternal and Nodular Goitre.

Objectives: This is a diagnostic test research study to evaluate the various existing methods of thyroid examination and their comparison with the novel modified Rose method. It also aims to measure inter-examiner variation in clinical findings based on the level of education and training, as compared to ultrasonography. Methodology: This cross-sectional study was conducted at a tertiary care hospital with 83 patients presenting to surgery OPD with neck swelling. Each patient was examined by one trained Junior Resident and a Surgery Consultant with all the four methods and with ultrasonography. Data was analysed by Stata-14, agreement between the two categorical variables was assessed by Kappa. In case of continuous variable agreement was assessed by Intra class correlation and Bland-Altman plot. Results: Modified Rose method by the consultant has the highest sensitivity (98%) and diagnostic odds (210) as compared to others but its specificity ranges from 46.7-91.1% to diagnose retro-sternal extension of a goiter. It has 93.98% agreement for identification of nodules. It has a high specificity (Consultant - 100%, Resident - 95.5%) with relatively lower sensitivity (Consultant - 94%, Resident - 86.8%) to diagnose solitary thyroid nodule (STN) but the sensitivity and specificity for diagnosing a multinodular goitre (MNG) was high. However, the highest sensitivity to diagnose STN was highest with Crile's method, but specificity was low. Lahey's method was a better clinical method to palpate lymph nodes compared to the other three. Conclusion: Examination in modified Rose's position is a better method of clinical examination of thyroid especially in patients with occult substernal extension. Lahey's method is a better method to examine cervical lymph nodes.

Management of post-traumatic ischaemic ileal stricture using intraoperative indocyanine green fluorescence-guided resection.

Blunt abdominal trauma can affect mesenteric circulation which may lead to bowel strictures. Indocyanine green (ICG) angiography can be used to assess mesenteric blood flow and bowel perfusion as a guide to resect length intraoperatively. But this concept has not been applied to ischaemic bowel strictures. We present a case of ischaemic ileal stricture induced by blunt abdominal trauma which was managed by resection and anastomosis. Intraoperative near-infrared (NIR) ICG angiography was used as a guide to resect the bowel length. This case emphasises that ischaemic bowel strictures should be suspected in patients presenting with intestinal obstruction following trauma. Resection and anastomosis of the affected segment remains the primary treatment modality with excellent outcomes. NIR ICG angiography is a real-time objective and useful resource for assessing bowel perfusion and could be used to determine the length of the segment to be resected in patients with ischaemic bowel stricture.

Clinicopathological and Radiological Features with Long Term Follow Up of Metaplastic Carcinoma Breast in India.

OBJECTIVE: Metaplastic breast carcinoma (MBC) is a heterogeneous group of invasive carcinomas with squamous and/or mesenchymal differentiation. Because of their rare occurrence, the information regarding the clinical behaviour of metaplastic carcinomas is limited. The purpose of our study was to delineate the clinicopathological and radiological features, treatment outcomes, prognostic factors, and survival of patients with MBC. METHODS: Ambispective observational study with prospective recruitment was done from 1st January 2019 to 31st August 2020. Retrospective data included between 1st January 2009 and 31st December 2018. In the retrospective group surgical database of our department was searched and those with MBC diagnosis on post-operative histopathology recruited. In prospective group patients with MBC on core biopsy were followed and those operated were included. The patients followed up at our breast cancer clinic (BCC) and their demographic, clinical, pathological radiological and treatment details noted. RESULTS: Forty patients formed the study population. The mean age of the patents was 42 years. Ipsilateral axillary lymph node metastasis was present in 22.5%. The pathological median tumor size was 5.4 (range 2.1 to 22 cm). The most common differentiation was cartilaginous (35%) followed by squamous (32.5%). The most common mammographic grading was BIRADS 4 (Breast Imaging Reporting and Data system). Magnetic resonance imaging was T2 hyperintense with peripheral rim enhancement and restriction on DWI. The median overall (OS) and disease-free survival (DFS) was 42 and 40 months, respectively. Fifteen patients (37.5%) had disease related mortality. A subgroup analysis revealed that, type of differentiation, histopathology and tumor size > 5cm affected both OS and DFS significantly. CONCLUSION: Metaplastic breast cancer in our setup presents in young patients with aggressive large tumors at a higher stage and diverse histopathology and with comparable overall and disease-free survival. The histological subtype, tumor differentiation and tumor size are prognostic factors.

Hematobiochemical profile in Surti goats during post-partum period.

AIM: The study was undertaken to find out the changes in hematobiochemical profile in post-partum Surti goats. MATERIALS AND METHODS: The present study was conducted on 40 Surti goats out of which 20 goats who had undergone recent parturition acted as treatment group and 20 non-pregnant animals comprised control group. Blood samples were collected from the treatment group on 0, 7, 14, 21, 30, and 45 days post kidding and once from the control group. Blood samples were analyzed for hematological parameters such as hemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC), differential leukocyte count, and biochemical metabolites such as total protein (TP), albumin, globulin, total cholesterol (TC), triglycerides (TG), non-esterified fatty acid (NEFA), glucose, and urea. RESULTS: Hb level was the highest on the 45(th) day and lowest on the day of parturition. Significantly low level of Hb, PCV, and lymphocyte was found on 0 day and significantly high level was found on the 45(th) day. TEC was also low on the 0 day and high on the 45(th) day post kidding, but the difference was non-significant. Mid-sized cells and granulocyte percentage decreased significantly from 0 to 45(th) day post-partum. TP, albumin, and urea were found to be lowest on 0 day and highest on 45(th) day post-partum. Urea concentration increased parallel to TP indicating an increase in urea production with catabolism of protein. The globulin concentration also increased from 0 to 45(th) day post-partum, but the difference was non-significant. TC, TG, and NEFA were the highest on 0 day and lowest on the 45(th) day post-partum. Decrease in TC and TG from 0 to 45(th) day post-partum indicates that the animals utilized the lipids for the supply of energy for milk production. High level of NEFA acts as an indicator of negative energy status but in the present study, the high glucose level on 0 day indicates that the animals were in positive energy status. The glucose concentration decreased up to 14(th) day post-partum and then started increasing from 21(st) day onward. CONCLUSION: Hematobiochemical parameters are indicative of health status of animals. Significantly low values of Hb, PCV, and TLC during first 2 weeks post-partum are indicative of stress. Catabolism of protein occurred during 2 weeks post-partum, as observed from increase in urea concentration. Decrease TGs and high level of NEFA during the transition period are suggestive of utilization of lipids for the supply of energy. High glucose level on 0 day indicates that the animals were in positive energy status.

Effects of fetal bovine serum and estrus buffalo serum on maturation of buffalo (Bubalus bubalis) oocytes in vitro.

AIM: The aim was to assess the effects of fetal bovine serum (FBS) and estrus buffalo serum (EBS) on in vitro maturation rate of oocytes in buffalo. MATERIALS AND METHODS: Maturation rate of oocytes was assessed in two maturation media supplemented with 20% FBS and EBS. Oocytes maturation rate was evaluated on the basis of cumulus cell expansion and extrusion of polar body after 24 h of in vitro culture in CO2 incubator. RESULTS: The average percentage of in vitro matured oocytes in FBS was 83.80%, and EBS was 77.45%, respectively. The results revealed a significant (p<0.05) increase in maturation rate of oocytes in FBS than EBS. CONCLUSION: Buffalo oocytes were better in vitro matured in FBS than EBS.

Effects of ß-mercaptoethanol on in vitro maturation and glutathione level of buffalo oocytes.

AIM: The present study was carried out to evaluate the effect of supplementation of ß-mercaptoethanol (ß-ME) on in vitro maturation rate and glutathione (GSH) level of buffalo oocytes. MATERIALS AND METHODS: Oocytes were recovered from buffalo's ovaries collected from government approved slaughter house (near Kamela darwaza, Surat) of Surat Municipal Corporation. The obtained oocytes were in vitro matured in maturation media supplemented with 0 µM (117 oocytes), 100 µM (46 oocytes) and 200 µM (42 oocytes) concentration of ß-ME. After 24 h of incubation, maturation rate of oocytes and intra-cellular GSH level were determined. RESULTS: The results showed that the presence of ß-ME did not influence (p>0.05) the oocyte maturation rate. However, GSH level increased significantly (p<0.05) in matured oocytes when supplemented with 100 µM and 200 µM ß-ME (6.19±0.10 and 6.37±0.20 pmol/oocyte) as compared to control media (4.68±0.26 pmol/oocyte). CONCLUSION: It was concluded that ß-ME may have a potential to increase the meiotic maturation of in vitro cultured oocytes and protect it from oxidative damage.

Evaluation of physiological and biochemical responses in different seasons in Surti buffaloes.

AIM: This study was conducted to evaluate the impact of hot dry, hot humid and comfortable season on physiological, hematological, biochemical, and oxidative stress parameters in Surti buffaloes. MATERIALS AND METHODS: Ten lactating Surti buffaloes of similar physiological status were selected. Based on the temperature-humidity index (THI), their natural exposure to the environment was categorized as hot dry (THI1), hot humid (THI2) and moderate winter/comfort season (THI3). Blood/serum samples were collected and analyzed for physiological, hematological, biochemical, and oxidative stress parameters. The results were analyzed using standard statistical methods. RESULTS: With increase in THI, significant rise in physiological parameters such as respiration rate (RR), hematological parameters such as red blood cell (RBC), hematocrit, hemoglobin (Hb) and mean cell Hb concentration (MCHC), biochemical parameters such as alanine aminotransferase (ALT), Na, K, creatinine, blood urea nitrogen, Mn, Cu and Zn, hormones such as cortisol and oxidative stress parameters such as glutathione peroxidase (GPx), superoxide dismutase (SOD), lipid peroxide (LPO) and total antioxidant status (TAS) and significant decline in glucose, cholesterol and triiodothyronine (T3) was observed. CONCLUSION: It was concluded that THI is a sensitive indicator of heat stress and is impacted by ambient temperature more than the relative humidity in buffaloes. Higher THI is associated with significantly increased RR, total RBC count, Hb, hematocrit, MCHC, ALT, urea, sodium, creatinine, triiodothyronine, SOD, GPx, LPO and TAS and with significant decrease in glucose, cholesterol and triiodothyronine (T3).

Reprogramming of fetal cells by avian EE for generation of pluripotent stem cell like cells in caprine.

The present work was carried out to study the ability of avian "Extract Egg" (EE) for reprogramming caprine fetal cells. The isolated caprine fetal cells were cultured in stem cell media supplemented with different percentages of either EE or FBS. The results indicated that the supplementation of 2-4% EE formed lesser but larger size stem cell like cell colonies as compared to 6% or 10% EE. The expression of pluripotent genes were comparatively higher in colonies developed in 2% or 4% as compared to 6% or 10% EE. Further, immunocytochemistry revealed that the colonies developed in all percentage of EE expressed pluripotent markers like Oct4, Nanog, TRA-1-60 and TRA-1-81. Our findings indicated that avian EE has the potentiality to reprogram caprine fetal cells into embryonic state which may help in generation of pluripotent stem cells without using viral vector.

Effect of mitotic inducers and retinoic acid blocker on expression of pluripotent genes in ES cells derived from early stage in vitro-produced embryos in buffalo.

So far, it has been difficult to generate embryonic stem (ES) cell from early stage preimplantation embryos of buffalo. These ES cells will be more helpful for efficient embryo cloning and generation of body cells as they are more primitive than inner cell mass (ICM)-derived ES cells. The present study was conducted to find the effect of lipopolysaccharide (LPS), melatonin (N-acetyl-5-methoxytryptamine, a pineal gland product), and citral (3,7-dimethyl-2,6-octadienal and a retinoic acid synthesis blocker) on establishment of primary ES cell colonies, the comparative size of the ES cell colonies, and expression of pluripotent genes during extended period of culture in buffalo. Zona-free eight-cell stage in vitro fertilization (IVF) embryos were cultured in ES cell medium supplemented with none (media I as control), LPS (media II), citral melatonin (media III), or melatonin (media IV). The multiplication of blastomere leading to ES cell colony formation and expression of pluripotent genes were assessed up to day 20 of culture. The primary colony formation, the comparative size of the ES cell colonies, and expression of pluripotent genes in these colonies were better in the medium supplemented with melatonin in all days of culture. Within melatonin supplementation, the colony size was comparatively larger on day 8 and day 12 of culture. Further, with this supplementation, the Oct-4 and Nanog expression was comparatively higher on all days of culture. The results indicated that supplementation of melatonin helped in the formation of better primary ES cell colony as well as in the maintenance of pluripotency. The results also indicated that primary colonies developed on day 8 to day 12 of culture may be better for passaging them for establishment of ES cell line from early stage preimplantation IVF embryos of in buffalo.