Impact of Covid-19 on gastrointestinal cancer surgery: A National Survey.

PURPOSE: To understand the actual impact of the Covid-19 pandemic and frame the future strategies, we conducted a pan India survey to study the impact on the surgical management of gastrointestinal cancers. METHODS: A national multicentre survey in the form of a questionnaire from 16 tertiary care gastrointestinal oncology centres across India was conducted from January 2019 to June 2021 that was divided into a 15-month pre-Covid era and a similar period of active Covid pandemic era. RESULTS: There was significant disruption of services; 13 (81%) centres worked as dedicated Covid care centres and 43% reported suspension of essential care for more than 6 months. In active Covid phase, there was a 14.5% decrease in registrations and proportion of decrease was highest in the centres from South zone (22%). There was decrease in resections across all organ systems; maximum reduction was noted in hepatic resections (33%) followed by oesophageal and gastric resections (31 and 25% respectively). There was minimal decrease in colorectal resections (5%). A total of 584 (7.1%) patients had either active Covid-19 infection or developed infection in the post-operative period or had recovered from Covid-19 infection. Only 3 (18%) centres reported higher morbidity, while the rest of the centres reported similar or lower morbidity rates when compared to pre-Covid phase; however, 6 (37%) centres reported slightly higher mortality in the active Covid phase. CONCLUSION: Covid-19 pandemic resulted in significant reduction in new cancer registrations and elective gastrointestinal cancer surgeries. Perioperative morbidity remained similar despite 7.1% perioperative Covid 19 exposure.

Exploring the relationship between LST and land cover of Bengaluru by concentric ring approach.

The present study aims at investigating the impact of land cover features in enhancing or mitigating Land Surface Temperature (LST) in a semi-arid tropical metropolitan city of Bengaluru, India. Spatial distribution of LST and land cover types of the area were examined in the circumferential direction, and the contribution of land cover classes on LST was studied over 28 years. Urban growth and LST were modelled using Landsat and MODIS data for the years 1989, 2001, 2005 and 2017 based on the concentric ring approach. The study provides an efficient methodology for modelling and parameterisation of LST and urban growth by fitting an inverse S-curve into urban density (UD) and mean LST data. In addition, multiple linear regression models which could effectively predict the LST distribution based on land cover types were developed for both day and night time. Based on the analysis of remotely sensed data for LST, it is observed that over the years, urban core area has increased circumferentially from 5 to 10 km, and the urban growth has spread towards outskirts beyond 15 km from the city centre. As urban expansion occurs, the area under the study experiences an expansive cooling effect during day time; at night, an expansive heating effect is experienced in accordance with the growth in UD in the suburban area and outskirts. The regression models that were developed have relatively high accuracy with R2 value of more than 0.94 and could explain the relationship between LST and land cover types. The study also revealed that there exists a negative correlation between urban, vegetation, water body and LST during day time while a positive correlation is observed during night. Thus, this study could assist urban planners and policymakers in understanding the scientific basis for urban heating effect and predict LST for the future development for implementing green infrastructure. The proposed methodology could be applied to other urban areas for quantifying the distribution of LST and different land cover types and their interrelationships.

The impact of spatiotemporal patterns of land use land cover and land surface temperature on an urban cool island: a case study of Bengaluru.

In most of the developing countries, man-made developments in the environment have led to the growing demand to contextualize the land use land cover (LULC) changes and land surface temperature (LST) variations. Due to the modification in the surface properties of the cities, a difference in energy balance between the cities and its nonurban surroundings is observed. The aim of this study is to analyze the spatial and temporal patterns of LULC and LST and its interrelationship in Bengaluru urban district, India, during the period from 1989 to 2017 using remote sensing data. Intensity analysis was performed for the interval to analyze the LULC change and identify the driving forces. The impact of LULC change on LST was assessed using hot spot analysis (Getis-Ord Gi* statistics). The results of this study show that (a) dominant LULC change experienced is the increase in urban area (approximately 40%) and the rate of land use change was faster in the time period 1989-2001 than 2001-2017; (b) the major transition witnessed is from barren and agricultural land to urban; (c) over the period of 28 years, LST patterns for different land use classes exhibit an increasing trend with an overall increase of approximately 6 °C and the mean LST of urban area increased by about 8 °C; (d) LST pattern change can be effectively analyzed using hot spot analysis; and (e) as the urban expansion occurs, the cold spots have increased, and it is mainly clustered in the urban area. It confirms the presence of an urban cool island effect in Bengaluru urban district. The findings of this work can be used as a scientific basis for the sustainable development and land use planning of the region in the future.

Localization techniques for guided surgical excision of non-palpable breast lesions.

BACKGROUND: Breast cancer is the most common form of cancer and the second leading cause of death amongst women in Europe. Amongst five invasive cancers per 1000 women detected in screening, 2.7 were < 15 mm in diameter; and others reported that over one third of excised breast lesions were clinically occult. The challenge is to accurately locate small non-palpable lesions intraoperatively for optimal therapeutic outcome. A secondary important goal is to remove the smallest amount possible of healthy glandular tissue for optimal cosmesis. Currently the most widely adopted approach (80% in one survey) in guided breast-conserving surgery for excising non-palpable breast lesions is wire-guided localization (WGL). With the clinical setting shifting towards earlier non-palpable breast lesions being detected through screening, we investigated whether the current standard in assisting surgical excision of these lesions, WGL, yields the best therapeutic outcome for women with breast cancer. OBJECTIVES: To assess the therapeutic outcomes of any new form of guided surgical intervention for non-palpable breast lesions against wire-guided localization, the current gold standard. SEARCH METHODS: We searched the Cochrane Breast Cancer Group's (CBCG) Specialized Register, MEDLINE (via PubMed), the Cochrane Central Register of Controlled Trials (CENTRAL), and the World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) search portal from the earliest available date up to 30 March 2015. We also handsearched recent conference proceedings and sought information from experts in the field. SELECTION CRITERIA: Two review authors, BC and RJ, independently screened by title and abstract the studies we had identified through the search strategy; when this was inconclusive, they examined the full-text article for inclusion. We resolved any discrepancies regarding eligibility by discussion with a third review author, RA. DATA COLLECTION AND ANALYSIS: Three review authors, BC, JW, and RJ, independently extracted data using a standardized data sheet. We performed all analyses using Review Manager (RevMan) or the R meta package, and in accordance with the Cochrane Handbook for Systematic Reviews of Interventions. We reported results via a graphical assessment using forest plots showing the study estimates. We considered and discussed additional subgroup and sensitivity analyses. MAIN RESULTS: We identified 11 randomized controlled trials (RCTs) that met the inclusion criteria of this Cochrane review and included eight trials in the meta-analyses. Six RCTs compared radioguided occult lesion localization (ROLL) versus WGL, and two RCTs compared radioactive iodine ((125)I) seed localization (RSL) versus WGL. Of the three remaining trials, one RCT compared cryo-assisted techniques (CAL) versus WGL, one compared intraoperative ultrasound-guided lumpectomy (IOUS) versus WGL, and one compared modified ROLL technique in combination with methylene dye (RCML) versus WGL. Of the trials we included in the meta-analysis, there were a total of 1273 participants with non-palpable breast lesions (627 participants (WGL); 443 participants (ROLL); and 203 participants (RSL)). The participant population varied considerably between included trials, which included participants with both non-palpable benign and malignant lesions, and varied in defining clear margins. The included trials did not report any long-term outcomes.In general, the outcomes of WGL, ROLL and RSL were comparable.ROLL demonstrated favourable results in successful localization (risk ratio (RR) 0.60, 95% confidence interval (CI) 0.16 to 2.28; 869 participants; six trials), positive excision margins (RR 0.74, 95% CI 0.42 to 1.29; 517 participants; five trials), and re-operation rates (RR 0.51, 95% CI 0.21 to 1.23; 583 participants; four trials) versus WGL, but none were statistically significant. WGL was significantly superior to RSL in successfully localizing non-palpable lesions (RR 3.85, 95% CI 1.21 to 12.19; 402 participants; two trials). However, for successful excision, ROLL and RSL have comparable outcomes versus WGL (ROLL versus WGL: RR 1.00, 95% CI 0.99 to 1.01; 871 participants; six trials; RSL versus WGL: RR 1.00, 95% CI 0.99 to 1.01; 402 participants; two trials). These findings were similar in that RSL demonstrated favourable results over WGL in positive tumour margins (RR 0.67, 95% CI 0.43 to 1.06; 366 participants; two trials), and re-operation rates (RR 0.80, 95% CI 0.48 to 1.32; 305 participants; one trial) but neither reached statistical significance. In contrast, WGL had fewer postoperative complications to both ROLL (RR 1.18, 95% CI 0.71 to 1.98; 642 participants; four trials) and RSL (RR 1.51, 95% CI 0.75 to 3.03; 305 participants; one trial), although this was also not statistically significant.The overall quality of evidence was good. The main risk of bias amongst included studies consisted of incomplete data sets, selective reporting, and allocation concealment. Interpretation and applicability of this meta-analysis was hindered by the mixed indication of diagnostic versus therapeutic purposes when undertaking WGL, ROLL, or RSL, leading to a high level of mixed pathology in numerous trials. Other limitations include underpowered studies, lack of data in standardized format for meta-analysis, lack of complete data amongst the trials, and absence of long-term data. AUTHORS' CONCLUSIONS: Owing to a lack of trials in certain localization techniques, we could only draw conclusions about ROLL and RSL versus WGL. There is no clear evidence to support one guided technique for surgically excising a non-palpable breast lesion over another. Results from this Cochrane review support the continued use of WGL as a safe and tested technique that allows for flexibility in selected cases when faced with extensive microcalcification. ROLL and RSL could be offered to patients as a comparable replacement for WGL as they are equally reliable. Other techniques such as IOUS, RCML, and CAL are of academic interest, but recommendation for routine use in the clinical environment and oncological outcomes require further validation. The results of this Cochrane review also stress the need for more fully powered RCTs to evaluate the best technique according to the comprehensive criteria described, with a more consistent and standardized approach in outcome reporting.

Hypolipidemic effect of oryzanol concentrate and low temperature extracted crude rice bran oil in experimental male wistar rats.

Blends of refined groundnut oil (GNO) and oryzanol concentrate having 3, 5, and 10% oryzanol in the blend, and a rice bran oil (RBO) which had retained all the nutrients such as oryzanol, tocopherols and tocotrienols and the unsaponifiable matter components of crude oil (GWF RBO) were prepared. Weanling rats were fed with diet containing the oil blends/rice bran oil at 10% level for 60 days and then dissected. The lipid profiles in serum, liver were investigated and the cholesterol levels were marginally reduced (7-16% in serum, 10-14.5% in liver) in rats fed oryzanol containing diet. RBO, GWF RBO containing diets showed a reduction of serum cholesterol by 14%, 15% respectively when compared to those fed with GNO. Serum and liver lipid analysis also showed significant change in TG concentration in rats fed blended oils containing oryzanol compared to the rats given GNO. Histology of liver and kidneys did not show changes. These studies indicated that oryzanol has an effect in lowering serum and liver cholesterol and shows antiatherogenic properties when incorporated into groundnut oil.

Progression of early phase diabetic nephropathy in streptozotocin-induced diabetic rats: evaluation of various kidney-related parameters.

Diabetic nephropathy (DN) is one of the serious secondary complications of diabetes, which results in end-stage renal failure. Reports on the progressive nature of early phase DN especially with respect to kidney parameters such as kidney weight, type IV collagen excretion, total kidney and urinary glycosaminoglycans (GAGs) are few. This work was undertaken to determine systematically the progression of early phase DN in relation to various kidney-related parameters for a period of four months. Experimentally-induced diabetic rats were grouped based on fasting blood glucose levels. Various basic and kidney-related parameters such as kidney weight, microalbuminuria, urinary excretion of GAGs and type IV collagen, total kidney GAGs, histopathology, glomerular area and glomerular volume were examined in control and diabetic rats. There was a progressive increase in fasting blood sugar, urine sugar, kidney weight, microalbuminuria, urine glycosaminoglycans, urine type IV collagen, glomerular area and glomerular volume but there was a progressive decrease in kidney glycosaminoglycans. Glomerular sclerotic condition was aggravated with the increase in duration of diabetes from 1 to 4 months. Onset of DN in rats begins subtly after one month of diabetes but gets vitiated and more pronounced at the end of four months.

SNOLL. Sentinel node and occult (impalpable) lesion localization in breast cancer.

AIM: To evaluate the efficacy and safety for use of dual radioisotopes for localization of occult (impalpable) breast lesions and sentinel node biopsy as a combined technique (SNOLL) using a lower dose than previous studies. MATERIALS AND METHODS: One hundred and twenty-seven SNOLL procedures were performed. The impalpable breast lesions were localized with an intra-lesion injection of 0.2 ml of (99m)Tc MAA (1 MBq) with a particle size of 10-90 µm (radio-guided occult lesion localization, or ROLL) 1 to 4 h before surgery. Sentinel node localization was performed using 0.2 mls of (99)Tc nanocolloid (20 MBq) particle size no greater than 80 nm injected subdermally in the periareolar region within the index quadrant, the night before or the morning of surgery. RESULTS: Lesion localization was consistently achieved with a lower dose than that described in other studies without the need to use scintigraphy or additional imaging with radioopaque contrast medium. One hundred percent lesion localization with a negative clearance margin of 94.8% and 100% sentinel node localization was achieved. The use of dual radioisotopes with the lower dose used for ROLL did not compromise the localization of the impalpable lesion or the sentinel nodes. CONCLUSION: The combined use of radioisotopes for lesion and sentinel node removal is feasible and reliable with the lower radioisotope dose suggested compared with previously published studies. This method should be recommended as a standard procedure for SNOLL.

Effectiveness of Yoga Lifestyle on Lipid Metabolism in a Vulnerable Population-A Community Based Multicenter Randomized Controlled Trial.

Background: Dyslipidemia poses a high risk for cardiovascular disease and stroke in Type 2 diabetes (T2DM). There are no studies on the impact of a validated integrated yoga lifestyle protocol on lipid profiles in a high-risk diabetes population. Methods: Here, we report the results of lipid profile values of 11,254 (yoga 5932 and control 5322) adults (20-70 years) of both genders with high risk (≥60 on Indian diabetes risk score) for diabetes from a nationwide rural and urban community-based two group (yoga and conventional management) cluster randomized controlled trial. The yoga group practiced a validated integrated yoga lifestyle protocol (DYP) in nine day camps followed by daily one-hour practice. Biochemical profiling included glycated hemoglobin and lipid profiles before and after three months. Results: There was a significant difference between groups (p < 0.001 ANCOVA) with improved serum total cholesterol, triglycerides, low-density lipoprotein, and high-density lipoprotein in the yoga group compared to the control group. Further, the regulatory effect of yoga was noted with a significant decrease or increase in those with high or low values of lipids, respectively, with marginal or no change in those within the normal range. Conclusion: Yoga lifestyle improves and regulates (lowered if high, increased if low) the blood lipid levels in both genders of prediabetic and diabetic individuals in both rural and urban Indian communities.

An Unusual Cause of Chylothorax after Esophagectomy.

Chylothorax due to inadvertent thoracic duct injury after esophagectomy is a well-known complication and requires careful postoperative management and timely intervention to prevent potential morbidity and mortality. We present a case of high-output chylothorax after esophagectomy where the source of chyle leak was not in the thorax.

Isolation and culture of ovine and bubaline small and large pre-antral follicles: effect of cyclicity and presence of a dominant follicle.

Studies were conducted to examine the effects of the cyclicity and the presence of a dominant follicle (DF) in ovary on the recovery and in vitro growth of pre-antral follicles (PFs) in sheep and buffalo. Small pre-antral follicles (SPFs, 100-250 microm) and large pre-antral follicles (LPFs, 250-450 microm) were isolated from slaughterhouse ovaries in the breeding seasons by a mechanical and enzymatic method. The sheep and buffalo PFs were cultured in vitro for 6 and 15 days, respectively, and examined for their growth, survival and antrum formation rates and growth rates of oocytes in cultured pre-antral follicles. The follicles of the sheep and buffalo were recovered and cultured simultaneously within replicates. The recovery rates (number per ovary) of both SPFs and LPFs were significantly (p < 0.05) higher in cyclic ewes (SPFs: 22.0 +/- 3.3 vs 12.1 +/- 2.6 and LPFs: 16.0 +/- 3.6 vs 9.2 +/- 1.8) and buffaloes (SPFs: 9.2 +/- 1.3 vs 4.1 +/- 1.0 and LPFs: 10.3 +/- 2.7 vs 5.4 +/- 0.7) compared with those recovered from acyclic ones. Presence of a DF in ovary significantly (p < 0.05) reduced the recovery rates of LPFs in ewes (9.06 +/- 2.7 vs 16.4 +/- 3.8) but had no effect in buffalo. Cyclicity of animals or follicular dominance had no effects on in vitro growth, survival and antrum formation rates and growth rates of oocytes in cultured PFs of SPFs and LPFs in both sheep and buffalo. The in vitro growth, survival and antrum formation rates of LPFs and growth rates of oocytes in cultured LPFs were significantly (p < 0.05) higher than those observed in SPFs in both sheep and buffalo. The overall recovery and growth rates of the PFs were lower in buffaloes compared with ewes.

Respiratory performance and behavioral responses of the freshwater fish, Cyprinus carpio (Linnaeus) under sublethal chlorpyrifos exposure.

A short-term definitive test by static renewal bioassay method was conducted to determine the acute toxicity (LC50) of an organophosphate insecticide, chlorpyrifos on the fish, Cyprinus carpio. Carp fingerlings were exposed to different concentrations (0.120 to 0.200 mg/L) of chlorpyrifos for 96 h. The acute toxicity of chlorpyrifos was found to be 0.160 m g/L. One-seventh (1/7th, 0.0224 mg/L) and one-fourteenth (1/14th, 0.0112 mg/L) of the acute toxicity value were selected as sublethal concentrations for subchronic studies. The fish were exposed to both the sublethal concentrations for 1, 7, and 14 days and allowed to recover in toxicant-free medium for 7 days. Behavioral responses and respiratory rate were studied in experimental periods. Fish in toxic media exhibited irregular, erratic, and darting swimming movements, hyperexcitability, and loss of equilibrium and sinking to the bottom. Caudal bending was the chief morphological alterations during the exposure tenures. The behavioral and morphologic changes might be due to the inhibition of acetylcholinesterase (AChE) activity. The carp were found under stress but mortality was insignificant at both the sublethal concentrations. Considerable variation in respiratory rates (-71.85 to 119.52%; -75.65 to 17.58%) were observed in the 1/7th and 1/14th of lethal concentration of chlorpyrifos respectively. The alteration in respiratory rate is due to respiratory distress, this may be a consequence of impaired oxidative metabolism and elevated physiological response during chlorpyrifos exposure. The impairments in fish respiratory physiology and behavioral responses even under recovery periods may be due to slow release of sequestered chlorpyrifos from storage tissues.

Respiratory distress and behavioral changes induced by sodium cyanide in the fresh water TELEOST, Cyprinus carpio (Linnaeus).

An acute toxicity (LC50) test using a static renewal bioassay method was conducted to determine the toxicity of sodium cyanide in freshwater exotic carp, Cyprinus carpio exposed for 96 h to different concentrations of sodium cyanide. The acute toxicity value was found to be 1 mg/L; one third of the LC50 (0.33 mg/L) was selected as the sublethal concentration for subacute studies. Behavioral patterns were observed in lethal (1, 2, 3, and 4 d) and sublethal concentrations (1, 5, 10, and 15 d). Cyprinus carpio in toxic media exhibited irregular, erratic, and darting swimming movements, hyperexcitability, loss of equilibrium, and sinking to the bottom, which might be due to inhibition of cytochrome c oxidase activity and decreased blood pH. The combination of cytotoxic hypoxia with lactate acidosis depresses the central nervous system and myocardium, the most sensitive critical sites for anoxia, resulting in respiratory arrest and death. A decrease in oxygen consumption (-28.36 to -78.28%; -14.22 to -47.25%) was observed at both lethal and sublethal concentrations of sodium cyanide respectively. Fish at the sublethal concentration were found under stress, but that was not fatal.

Thoracoscopic Esophagectomy for a Huge Leiomyosarcoma.

Esophageal leiomyosarcoma is the commonest of all esophageal sarcomas but yet has a very low incidence. These tumors have been resected by the open approach so far. We describe the steps and challenges involved in the thoracoscopic excision of a huge leiomyosarcoma of the esophagus.

Surgery for cystic tumors of pancreas: Report of high-volume, multicenter Indian experience over a decade.

BACKGROUND: Pancreatic cystic neoplasms remain uncommon. Although data are accumulating on the incidence of pancreatic cystic neoplasms in the published literature, Indian data on these tumors are sparse. MATERIAL AND METHODS: We collated data from prospectively maintained databases of patients operated for cystic tumors of the pancreas from 2007 to 2016 at 7 academic centers across India to gain insights into clinical presentation and outcome of the operative treatment of these tumors. Data were compared with large series across the world to understand the regional differences in this pathology. RESULTS: Of the 423 patients, there were 98 (23.2%) serous cystic neoplasms, 128 (30.2%) mucinous neoplasms, 34(8%) intraductal papillary mucinous neoplasms, and 121 (28.6%) solid pseudopapillary epithelial neoplasms managed in these 7 academic centers. Malignancy (adenocarcinoma, malignant intraductal papillary mucinous neoplasms, and mucinous cystadenocarcinoma) was reported in 39 (9.2%) patients. Median age at presentation was 41 years, and the female-to-male ratio was 3.4:1. At presentation, 81% of patients were symptomatic. A total of 66.7% of lesions were located in body and tail region of the pancreas. Median tumor size was 6 cm. Operative resection with curative intent was performed in 405 of these 423 patients. Major morbidity occurred in 12%, and 30-day perioperative mortality was 0.9%. Laparoscopic resections were performed in 18% and spleen-preserving resections were performed in 3% of patients. CONCLUSION: Female preponderance, young age, and a benign nature of most pancreatic cystic neoplasms were observed. Large size of tumors on presentation, fewer intraductal papillary mucinous neoplasm resections, and a much greater incidence of solid pseudopapillary epithelial neoplasms were distinctive of this study. Although the proportion of laparoscopic resections and splenic preservation was less compared with Western centers, the perioperative morbidity and mortality was on par with established standards.

Shall we operate? Preoperative assessment in elderly cancer patients (PACE) can help. A SIOG surgical task force prospective study.

BACKGROUND: A number of elderly cancer patients do not receive standard surgery for solid tumors because they are considered unfit for treatment as a consequence of inaccurate estimation of the operative risk. To tailor treatment to onco-geriatric series, oncologists are now beginning to use a comprehensive geriatric assessment (CGA). This study investigates the value of an extended CGA in assessing the suitability of elderly patients for surgical intervention. PATIENTS AND METHODS: Preoperative assessment of cancer in the elderly (PACE) incorporates validated instruments including the CGA, an assessment of fatigue and performance status and an anaesthesiologist's evaluation of operative risk. An international prospective study was conducted using 460 consecutively recruited elderly cancer patients who received PACE prior to elective surgery. Mortality, post-operative complications (morbidity) and length of hospital stay were recorded up to 30 days after surgery. RESULTS: Poor health in relation to disability (assessed using the instrumental activities of daily living (IADL)), fatigue and performance status (PS) were associated with a 50% increase in the relative risk of post-operative complications. Multivariate analysis identified moderate/severe fatigue, a dependent IADL and an abnormal PS as the most important independent predictors of post-surgical complications. Disability assessed by activities of daily living (ADL), IADL and PS were associated with an extended hospital stay. CONCLUSION: PACE represents a valuable tool in enhancing the decision process concerning the candidacy of elderly cancer patients for surgical intervention and can reduce inappropriate age-related inequity in access to surgical intervention. It is recommended that PACE be used routinely in surgical practice.

Follicular fluid concentrations of glucose, lactate and pyruvate in buffalo and sheep, and their effects on cultured oocytes, granulosa and cumulus cells.

The objective was to determine ovarian follicular fluid concentrations of glucose, lactate, and pyruvate in relation to follicle size in buffalo and sheep. The effect of varying concentrations of these substances on in vitro oocyte maturation, oocyte protein content, and granulosa and cumulus cell growth was also investigated. Follicular fluid was aspirated from various sizes of follicles (from ovaries without a dominant follicle) collected from adult, cycling nonpregnant buffalo (Bubalus bubalis) and sheep (Ovis aries) during the breeding season. Overall, mean (+/-S.E.M.) concentrations (mM) were glucose 2.42+/-0.31 and 1.40+/-0.22, lactate 7.56+/-2.61 and 10.42+/-1.64, and pyruvate 0.02+/-0.01 and 0.002+/-0.00, in buffalo and sheep, respectively. In both species, as follicles became larger, concentrations of glucose significantly increased, lactate significantly decreased, but pyruvate was not affected. Oocyte maturation was higher (P<0.05) in medium containing supra-physiological concentrations of either glucose (5 mM), or pyruvate (10 mM) alone, or physiological concentrations of glucose, lactate and pyruvate in combination, compared to supra-physiological concentrations of lactate (15 mM) alone, or sub- or supra-physiological concentrations of glucose, lactate and pyruvate in combination (both species). The protein content of oocytes was not significantly affected by the concentration of glucose, lactate, and pyruvate in the maturation medium. However, growth of granulosa and cumulus cells was higher (P<0.05) in medium containing supra-physiological concentrations of glucose (5 mM) alone, or pyruvate (10 mM) alone, or physiological, or supra-physiological concentrations of glucose, lactate and pyruvate in combination, compared to supra-physiological concentrations of lactate (15 mM) alone, or sub-physiological concentrations of glucose, lactate and pyruvate in combination (both species). In conclusion, concentrations of glucose, pyruvate and lactate in the medium had cell type-specific effects on oocyte maturation, and on growth of granulosa and cumulus cells. Furthermore, glucose and pyruvate were the principal energy sources for oocytes and follicular somatic cells in buffalo and sheep.

Co-culture of buffalo preantral follicles with different somatic cells.

The effect of co-culture of buffalo preantral follicles (PFs) with different somatic cells, i.e, cumulus, granulosa, ovarian mesenchymal and oviductal epithelial cells was studied. Large PFs (250-450 microm) were isolated by microdissecting the trypsin (1%) digested ovarian cortical slices. Cumulus cells were isolated by repeated pipetting of oocytes, granulosa cells were isolated by aspirating from punctured PFs and ovarian mesenchymal cells were isolated from ovarian cortex by scraping the cortical slices and passing through 20 microm filter. Preantral follicles were cultured in standard culture medium without somatic cells or co-cultured with cumulus cells, granulosa cells, ovarian mesenchymal cells and oviductal epithelial cells for 80 days. The growth rate (microm/day) of the PFs was monitored by measuring follicular diameter on day 0, 30, 60 and 80 days of culture. The viability of PFs was evaluated by trypan blue staining. The results indicated that PFs co-cultured with cumulus, granulosa and ovarian mesenchymal cells had a better development and survivality compared with control and those co-culture with oviductal epithelial cells. Maximum growth and survivality of PFs were achieved when cultured with cumulus cells. It is concluded that inclusion of somatic cells in PF culture media had beneficial effect on the growth of PFs and cumulus cells supported maximum growth and survivality of PFs in vitro of all somatic cells tested.