Effect of various antioxidants on ovarian tissue vitrification success in sheep.

BACKGROUND: Vitrification is a technique of cryopreservation that has been proposed as a promising alternative method for the preservation of oocytes, embryos and gonadal tissue. OBJECTIVE: To determine the effect of different antioxidants on post-thaw viability, morphology of retrieved oocytes and histology of vitrified ovarian tissue. MATERIALS AND METHODS: Four different antioxidants [i.e., resveratrol (20 uM), ZnSO4 (500 uM), curcumin (25 uM) and quercetin (1 uM)] were evaluated after their addition to the vitrification and warming media for their effects on the viability and morphology of retrieved oocytes and the histology of vitrified ovarian tissue. RESULTS: The number of oocytes retrieved from ovarian tissue from the above mentioned antioxidants and vitrified control were 34, 41, 26, 31 and 46 respectively. Among these the number of viable oocytes were found to be 24 (70.6%), 30 (73.1 %), 20 (76.9%), 26 (83.9%) and 33 (71.7%) and the number of oocytes found morphologically normal were 24 (70.6%), 26 (63.4%), 18 (69.2%), 21 (67.7%) and 34 (73.9%) for the above mentioned different antioxidants and vitrified control, respectively. Non-significant (P. > 0.05) differences were found between different treatment groups. Histomorphological evaluation of the ovarian cortical tissue showed that the percentage of intact follicles was significantly (P < 0.05) higher in the fresh control (84.19±3.9) than in other groups. Non-significant differences were found between resveratrol (50.2±5.5), curcumin (48.7±5.7), quercetin (51.6±4.8) and the vitrified control (42.7±6.1) groups; however, the ZnSO4 supplemented group (23.1±8.54) differed significantly (P < 0.05) from other antioxidant groups but was non-significant (P > 0.05) with the vitrified control group (42.7±6.1). CONCLUSION: The addition of antioxidants resveratrol, curcumin and quercetin at these concentrations tended to non-significantly improve the follicular integrity after vitrification. Doi.org/10.54680/fr24410110212.

Recent advancements in vitrification cryodevices for gamete and gonadal tissue.

Cryopreservation of gametes and gonadal tissue is nowadays primarily accomplished through vitrification. Variables such as cooling rate, viscosity and volume of vitrification solution are critical in gamete vitrification. In addition, sample size and stepwise exposure are also crucial for gonadal tissue vitrification. Recently a class of cryodevices has been developed to reduce the volume of vitrification solution so as to achieve higher cooling rates. Vitrification devices are classified as "open" or "closed" depending on whether the medium comes into direct contact with liquid nitrogen during the process. Examples of the open cryodevices for gamete vitrification are Cryotop, Cryolock, open pulled straw (OPS), etc., and closed devices are Vitrisafe, CryoTip, and high security vitrification kit. Similarly, for tissue vitrification open cryodevices used are needles, cryovials and closed devices used are Cryotissue, ovarian tissue cryosystem, etc. Among all the gamete cryodevices, Cryotop is unique and the best-selling micro-volume storage device. Use of this device has resulted in the highest number of babies born after embryo or oocyte vitrification. Another novel device, Kitasato vitrification system, is a vitrification solution absorber, which is similar to Cryotop but differs in one way, as it possesses a porous membrane that absorbs extra solution from the gamete. This review provides an update on the recent use of cryodevices for gamete and gonadal tissue vitrification. doi.org/10.54680/fr22310110112.

Role of Endoscopic Ultrasound Guided Fine Needle Aspiration in the Diagnosis of Pancreatic Lesions.

To evaluate the diagnostic utility of endoscopic ultrasound guided fine needle aspiration (EUS-FNA) of pancreatic lesions, EUS-FNA was carried out on a total of 28 cases at the Department of Surgery, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from June 2015 to August 2016. Clinical impression was compared with the final cytological diagnosis and the percentage of non diagnostic smears was calculated. The lesions were categorized according to cytological report. Out of 28 cases, 3(10.71%) cases were normal, 6 cases (21.43%) were reported as inconclusive while a definite diagnosis was given in 19 cases (67.85%). The mean patient age was 47.82 years. There were 16(57.1%) males and 12(42.9%) females. The most common site biopsied was pancreatic head in 21cases (75%) followed by tail in 5 cases (17.9%) & body in 2(7.1%) cases. The average number of passes made was two. Endoscopic Ultrasound Guided Fine-Needle Aspiration in the diagnosis of pancreatic lesion is a useful procedure.

Bilateral Tubercular Mastitis - Case Reports.

Breast tuberculosis is a rare form of tuberculosis. Moreover the disease is often overlooked and misdiagnosed as carcinoma or pyogenic abscess. Reports on breast tuberculosis have been few; reported incidence of breast tuberculosis amongst the total number of mammary conditions varies between 0.1 and 4 percent. Bilateral involvement is still more uncommon (3%). Here we report 3 cases of adult female ranging from 27 to 35 years who presented with 1 to 4 months history of firm lumps in both breasts and no axillary lymphadenopathy. Fine needle aspiration cytology (FNAC) of breast lump in all 3 cases were done but in 1 case showed evidence of tubercular mastitis and other 2 cases findings were inconclusive. Subsequent Excisional Biopsy of those 2 cases showed features of tuberculosis. All 3 were prescribed with four drug anti-tubercular treatments continued for 12 months in 2 cases and 9 months in other case depending upon their response. The lumps disappeared and ulcer healed after anti-tubercular treatment.

Penetrating keratoplasty: outcomes from a corneal unit compared to national data.

AIMS: To determine long term graft survival rates and visual results for different indications for penetrating keratoplasty from a single institution over 10 years and compare these to national outcome data. METHODS: Retrospective chart analysis. 784 records were available for review of 1096 consecutive penetrating keratoplasty procedures performed between 1990 and 1999 (72%). Outcomes of graft survival, visual acuity, and astigmatism were analysed and compared to national outcome data supplied by the UK Transplant Service. RESULTS: At 5 year follow up, overall graft survival was 66%. This was subdivided into 98% for keratoconus, 86% for viral keratitis, 85% for Fuchs' dystrophy, 84% for pseudophakic bullous keratopathy, 55% for regrafts, and 57% for other diagnoses. There was a significantly higher graft survival rate for all diagnostic subgroups except Fuchs' dystrophy at 3 years of follow up compared to the national average. Best corrected visual acuity at 5 years was 6/18 or better in 53% of cases. The mean keratometric astigmatism was 3.4 dioptres. CONCLUSION: Penetrating keratoplasty is a safe and effective treatment for selected corneal disorders. Penetrating keratoplasty for viral keratitis may achieve good results with long term antiviral treatment. Patients may achieve better outcomes if their surgery is performed at specialist centres.

Penetrating keratoplasty: indications over a 10 year period.

AIMS: To determine the indications for penetrating keratoplasty (PK) at the Corneoplastic Unit and Eye Bank, UK, a tertiary referral centre, over a 10 year period. METHODS: Records of all patients who underwent PK at our institution between 1990 and 1999 were reviewed retrospectively. Of the 1096 procedures performed in this period, 784 records were available for evaluation (72%). RESULTS: Regrafting was the most common indication, accounting for 40.9% of all cases. Keratoconus was the second most common indication (15%), followed by Fuchs' endothelial dystrophy (9.3%), pseudophakic bullous keratopathy (7.6%), and viral keratitis (5.9%), which included both herpes simplex and herpes zoster and showed a statistically significant decreasing trend using regression analysis (p<0.005). Among the regraft subgroup, viral keratitis accounted for 21.2% as the underlying primary diagnosis. The most common cause for graft failure in the regraft subgroup was endothelial failure (41.8%). CONCLUSION: Regrafting is the leading indication for PK; viral disease-although declining-is the leading primary diagnosis.

CT-guided percutaneous fibrin glue therapy of cerebrospinal fluid leaks in the spine after surgery.

OBJECTIVE: The objective of this study is to assess CT-guided percutaneous injection of fibrin glue for the management of cerebrospinal fluid leaks within the spine. CONCLUSION: Percutaneous CT-guided placement of fibrin glue can provide a treatment option for postoperative cerebrospinal fluid leaks, potentially allowing a major surgical procedure to be avoided. However, the complication of aseptic meningitis may occasionally result from this procedure.

Echo-planar functional MR imaging of epilepsy with concurrent EEG monitoring.

BACKGROUND AND PURPOSE: The role of functional MR (fMR) imaging in the evaluation of patients with epilepsy has not been systematically studied. Our purpose was to identify the fMR correlates of interictal epileptiform discharges. METHODS: Twenty patients with epilepsy and frequent interictal discharges were studied with concurrent EEG monitoring on a 1.5-T echo-planar magnet to acquire blood-oxygenation-level-dependent (BOLD) images in the baseline (OFF) and immediate post-discharge (ON) states. Analysis was performed using subtraction of average ON and OFF data (method I); cross-correlation analysis between the ON and OFF states (method II); and individual spike analysis (ISA), with which signal intensity in the individual ON states was statistically analyzed using a weighted comparison with the mean and variance of the OFF states (method III). Agreement of fMR activation with EEG localization was determined. RESULTS: Eighteen of 20 patients had interictal discharges during the monitoring period. Method I yielded visually detectable sites of BOLD signal differences in only one patient. Method II resulted in two patients with sites of BOLD activation. Method III, ISA, resulted in regions of increased BOLD signal corresponding to the EEG focus in nine of 10 patients. CONCLUSION: fMR studies can often reveal sites of increased BOLD signal that correspond to sites of interictal EEG discharge activity. Because of variable intensity changes associated with discharge activity, ISA resulted in increased sensitivity.

Cervicomedullary astrocytomas of childhood: clinical and imaging follow-up.

UNLABELLED: BACKGROUND. Cervicomedullary astrocytomas are a unique subset of brainstem tumors in children because they have a good prognosis when compared to the pontine subset of brainstem gliomas. Objective. To review the clinical and imaging findings in a series of children with cervicomedullary astrocytomas as to diagnosis and management. MATERIALS AND METHODS: A retrospective review of eleven children (six females, five males, age range: 10 days-18 years; mean = 7 years) with cervicomedullary tumors was done including the clinical presentation, imaging studies (MR: eleven, CT and MR: four), surgical findings, pathological results, and follow-up clinical and imaging findings (range: 0.2-11 years; mean = 5.2 years). RESULTS: Symptoms and signs were delayed and protracted, often occurring over months to years (mean = 2.3 years, range 0.5-7 years). The tumors expanded the dorsal medulla and involved the upper cervical spinal cord (mean maximum tumor diameter = 4.4 cm). Only three patients had hydrocephalus. In three of four cases the tumor was not seen on CT. On MR, the majority of the tumors were T1 hypointense and T2 hyperintense. Treatment consisted of surgery only in six patients, surgery and radiation therapy in four, and surgery, chemotherapy, and radiation in one. There was recurrent local disease in four patients and on follow-up metastatic disease in the brain in one. On follow-up the majority of the patients are alive and stable (mean = 5.2 years, range 0.2-11 years). There has been one death. The majority of tumors were pilocytic astrocytomas. CONCLUSION: Cervicomedullary tumors are a unique subset of brainstem gliomas in childhood that present with a long duration of symptoms and a greater long-term survival than pontine gliomas.

Echoplanar MR imaging for ultrafast detection of brain lesions.

OBJECTIVE: We retrospectively evaluated the use of echo-planar imaging for ultrafast detection of brain lesions. MATERIALS AND METHODS: In our retrospective study, 61 patients were imaged with the following echo-planar sequences: single-shot proton density-weighted, single-shot T2-weighted, single-shot T2-weighted high-resolution, multishot proton density-weighted, and multishot T2-weighted. Lesions revealed in these patients ranged from 0.5 to 12.0 cm (mean, 3.7 cm) and were the result of tumor (n = 16), stroke (n = 21), demyelination (n = 18), and toxoplasmosis (n = 2). Four patients had scans with normal findings. Two neuroradiologists who were unaware of pertinent clinical data reviewed the images. The images were retrospectively compared with conventional spin-echo images for diagnosis, sensitivity of lesion detection, and qualitative criteria: subjective image quality, gray and white matter differentiation, lesion conspicuity, delineation of lesion borders, and artifacts. (Artifacts included those caused by motion, susceptibility, pulsation, and ghosting.) Quantitative criteria, including signal-to-noise and signal difference-to-noise measurements, were also evaluated in 40 lesions. RESULTS: Sensitivity for lesion detection was 97% for single-shot echo-planar T2-weighted MR images and 100% for multishot echo-planar T2-weighted MR images. Single-shot echo-planar proton density-weighted MR images had the highest signal-to-noise ratio (91.2+/-19.3). Echo-planar T2-weighted MR images had the highest signal difference-to-noise (33.8+/-22.9). Echo-planar sequences were superior to spin-echo sequences regarding motion and pulsation artifacts. Spin-echo sequences lacked susceptibility and ghosting artifacts, and were superior in lesion conspicuity and delineation of lesion borders. CONCLUSION: In this study, echo-planar sequences were as sensitive as conventional spin-echo imaging for the diagnostic assessment of lesions. Echo-planar sequences had a strikingly shorter acquisition time and substantially reduced motion and pulsation artifacts. Echo-planar sequences may be a useful diagnostic tool for use in claustrophobic and unstable patients.

Site-specific delivery of iloprost during experimental angioplasty suppresses smooth muscle cell proliferation.

PURPOSE: The authors have previously reported that intramural delivery of iloprost during angioplasty suppresses local platelet aggregation at 1 hour in undiseased porcine arteries. In this study, the authors sought to quantify the effect of such treatment on medial vascular smooth muscle cell proliferation, an event implicated in the development of intimal hyperplasia. MATERIALS AND METHODS: Three Yorkshire pigs underwent percutaneous transluminal angioplasty with hydrogel-coated balloons for a total of 10 iloprost-treated (experimental) and 10 saline-treated (control) arterial sites. The balloons were prepared with previously reported techniques and loaded with 2.25 microg of iloprost for the experimental sites. On the eighth day after angioplasty, these sites were harvested and prepared for immunohistochemical staining. Thin (4 microm) sections of the specimens were stained with use of monoclonal antibody to proliferating cell nuclear antigen (PCNA). Appropriate positive and negative controls were used. Approximately 350-500 vascular smooth muscle cells were randomly counted under high power (100x) by an experienced physician who was blinded to the origin of the specimen. A PCNA index (%) was calculated as follows: [(#PCNA [+] cells)/(#PCNA [+] cells + #PCNA [-] cells)]x 100. A paired t test was used for statistical comparison. RESULTS: The PCNA indices for eight (n = 8) paired large vessels (iliac, carotid, subclavian) were 7.98 (+/- 1.8)%, for the iloprost-treated experimental sites, and 14.58 (+/- 3.8)% for the saline-treated control sites. This difference was statistically significant (P = .003). One large vessel pair was not available for analysis. When the pair of renal arteries of animal 3 were included (n = 9), the PCNA indices were 8.32 (+/- 2.3)% for the experimental sites, and 13.79 (+/- 4.2)% for the control sites. The differences were again significant (P = .01). CONCLUSION: Intraarterial site-specific delivery of iloprost during angioplasty with drug-loaded, hydrogel-coated balloons significantly suppresses medial smooth muscle cells in swine at the expected peak period of proliferation of 7 days after angioplasty.

Clinical and prognostic significance of in vivo differentiation in acute myeloid leukemia.

Bromodeoxyuridine (BrdU) was administered to 86 newly diagnosed patients with standard risk acute myeloid leukemia (AML) prior to starting induction therapy and the labeling index (LI), durations of S-phase (Ts), and the cell cycle (Tc) of myeloblasts were determined. Induction therapy with cytosine arabinoside and daunomycin was subsequently started. Bone marrow biopsies were obtained on days 6 and 17 and weekly thereafter, and were treated with a monoclonal anti-BrdU antibody to determine the fate of cells labeled on day 0 by BrdU. BrdU labeled granulocytes indicating the presence of in vivo differentiation (Diff+) were identified in 48 patients ranging from 1+ (1-10 labeled cells) to 4+ (greater than 31 labeled granulocytes). When compared to 38 differentiation negative (Diff-) patients, Diff+ group had longer Ts (14.5 hr vs. 10.95 hr, P = 0.015) and Tc (59.7 hr vs. 41.7 hr, P = 0.017). Remission duration was significantly longer (no median) for 3-4+ Diff+ as compared to Diff- (median = 220 days) patients (Wilcoxon P = 0.04). We conclude that the detection of in vivo differentiation in AML patients indicates a favorable long-term prognosis either due to the presence of a substantial amount of normal residual hematopoiesis prior to starting induction therapy or due to the ability of leukemic cells to undergo differentiation.

Cell-cycle characteristics - alterable determinants of remission duration in a study of 179 standard risk newly diagnosed patients with acute myeloid-leukemia.

Prognostic factors were related to remission duration among 179 standard risk newly diagnosed acute myeloid leukemia (AML) patients who received identical induction and consolidation therapies. Following a bromodeoxyuridine infusion, labeling indices of bone marrow aspirate/biopsy, durations of S-phase and cell cycle (Tc) were determined. Patients with slowly cycling myeloblasts had longer remissions (Log rank p=0.03) than those with rapidly cycling myeloblasts. Multivariate analysis demonstrated that both WBC and Tc contributed to remission duration (p=0.01 and 0.005 respectively). Patients with slowly proliferating leukemias have longer remissions probably due to slower regrowth of leukemia between chemotherapy courses.

In vivo cell cycle characteristics of pediatric leukemia patients.

Following i.v. bromodeoxyuridine infusion, a double-label technique using in vitro tritiated thymidine was used to determine the labeling index (LI), duration of S-phase (Ts), and cell cycle time (Tc) in pediatric leukemia patients. Eleven patients with acute lymphoblastic leukemia (ALL) and six patients with acute nonlymphoblastic leukemia (ANLL) were studied. Results of cell cycle kinetic studies are given for each group. Although median values for AML and ALL patients are similar to values reported in previous studies, there is a wide range of values among individual patients. The variation among the kinetic properties of blast cells in these patients reflects the heterogeneity of the acute leukemias of childhood. Further studies will be done to determine if these parameters correlate with outcome of therapy for pediatric leukemia patients.

A novel triple label method validates the double label technique of defining cell cycle kinetics in HL-60 cells.

HL-60 cells were sequentially labeled with the thymidine analogues iododeoxyuridine (IUdR) and bromodeoxyuridine (BrdU). The labeling index (LI), the duration of S-phase (Ts) and the total cell cycle time (Tc) were measured immediately. It was therefore possible to predict the next time when the single versus double labeled cells would re-enter the S-phase. In our study, the Tc was calculated to be 20 hours. The third label, tritiated thymidine (3HTdR), was introduced at the predicted time of 20 hours to confirm the validity of the previously calculated Tc. The actual percentage of cells which were labeled by (3HTdR) was very similar to the predicted value. We conclude, therefore, that the calculated cell cycle time correlated well with the actual cell cycle time, at least in a controlled in vitro culture system. This novel triple label method validates our double-label technique developed for cell cycle measurements.

In vivo determination of cell cycle kinetics of non-Hodgkin's lymphomas using iododeoxyuridine and bromodeoxyuridine.

Using sequential infusions of two S-phase-specific drugs, iododeoxyuridine and bromodeoxyuridine, we have developed an in vivo method for determining the labeling index (LI), the S-phase duration (Ts), and total cell cycle times (Tc) of non-Hodgkin's lymphomas. In nine non-Hodgkin's lymphomas studied, the LI ranged from 1.5% in a follicular small cleaved-cell lymphoma to 29.6% in a diffuse large-cell lymphoma. The Ts ranged from 16 hr in a large-cell lymphoma (immunoblastic type) to 117 hr in a follicular small cleaved-cell lymphoma. The Tc varied from 69 hr in a large-cell lymphoma (immunoblastic type) to over 1000 hr in all low-grade lymphomas studied. Immunohistochemical methods using anti-BrdU antibodies were used to detect cell incorporation of the two S-phase-specific drugs. In this manner, cell cycle times could be calculated while the architecture of the tumor specimen was preserved. Difficulties in using this methodology, specifically in the calculation of the growth fraction and total cell cycle times, are pointed out. This in vivo method does, however, allow for Ts calculations independent of growth fraction considerations. Correlations of cell cycle data with various biological and clinical factors await further patient follow-up.

Contribution of in vivo proliferation/differentiation studies toward the development of a combined functional and morphologic system of classification of neoplastic diseases.

Proliferation kinetics of both leukemia and a variety of solid tumors have been assessed after in vivo infusions of the thymidine analogues, iododeoxyuridine (IUdR) and bromodeoxyuridine (BrdU). In acute myeloid leukemia (AML), these data indicate that the pretherapy cell cycle time (Tc) of myeloblasts is a prognostic indicator for remission duration since patients with slowly cycling myeloblasts had more durable remissions. The presence of in vivo differentiation detected from the day 7 biopsy after chemotherapy was also of favorable prognosis as these individuals had statistically significant improvement in their remission duration. The data in solid tumors are not mature enough for determining their clinical significance. Since cell kinetic information is readily available in a prompt fashion using these novel techniques, data can be used to plan therapeutic strategies for patients. This review discusses the state-of-the-art techniques available for cell cycle kinetic studies and the clinical and prognostic utility of data that have been generated thus far.