Iloperidone and Temozolomide Synergistically Inhibit Growth, Migration and Enhance Apoptosis in Glioblastoma Cells.

Glioblastoma (GBM) is a fatal astrocytic glioma with poor prognosis and treatment resistance. Repurposing potential FDA-approved drugs like anti-psychotics can address the concerns in a timely and cost-effective manner. Epidemiological studies have shown that patients with schizophrenic using anti-psychotics have a low incidence of GBM. Therefore, we aimed to investigate the therapeutic potential of atypical anti-psychotic Iloperidone (ILO) alone and in combination with Temozolomide (TMZ) against GBM. The study assessed the growth inhibitory effect of ILO, TMZ, and their combination (ILO + TMZ) on U-87MG and T-98G cell lines using an MTT assay. The drug interaction coefficient (CDI) was determined, and doses with synergistic effects were used for subsequent experiments, including migratory, invasion, and TUNEL assays. The expressions of DRD2, ß-catenin, Dvl2, Twist, and Slug were assessed by RTq-PCR, whereas the ß-catenin protein expression was also determined by immunocytochemistry. ILO (p < 0.05) and TMZ (p < 0.01) significantly inhibited the growth of U-87MG cells at all tested doses. The combination of 60 µM of both drugs showed synergistic activity with CDI < 1. The inhibition of migration and apoptosis was more pronounced in the case of combination treatment (p < 0.001). Inhibition of the invading cells was also found to be significant in ILO- and combination-treated groups (p < 0.001). ILO and combination treatment also significantly downregulated the expression of DRD2, while TMZ upregulated the expression (p < 0.001). The expressions of ß-catenin (p < 0.001), Dvl2 (p < 0.001), Twist (p < 0.001), and Slug (p < 0.001) were also significantly downregulated in all treatment groups as compared to the vehicle control. The data suggest that ILO possesses strong growth inhibitory activity, possibly due to its effect on DRD2 and ß-catenin expression and has the potential to be repurposed against GBM.

Clinician's approach for spectrum of orbital diseases: An overview.

Purpose: To study the varied clinical presentation and outcome of management of orbital diseases and to ascertain the concordance of FNAC with histopathology. Materials and Methods: This is a prospective interventional study wherein each patient's clinical presentation was noted. Outcome of management was evaluated based on symptomatic relief, regression of signs, noting any intraoperative or postsurgical complication, recurrence, and cosmetic result in terms of surgical scar. Results: Neoplasia was commonest 33/76; (43.42%), followed by infective and inflammatory conditions 21/76; (27.63% cases), thyroid-associated orbitopathy accounting for 12/76 cases (15.78%), whereas cystic lesions and vascular malformations were 4/76 (5.26%) each. Other disorders constituted a mere 2.65%. Proptosis was the commonest mode of presentation. It was seen in seventy out of seventy-six patients (92.1%). In 86.3% of the cases, there was successful surgical outcome. Of 22 surgically managed cases, recurrence was noted in three cases (13.6%). 90% concordance was seen with histopathology in cases where FNAC was done prior to surgery for diagnosing nature of disease. Conclusions: Majority of orbital and adnexal conditions can be provisionally diagnosed based on imaging; however, an adjunct invasive investigation such as FNAC or biopsy may be required. In the present study, in seven cases (9.2%) FNAC was needed, whereas in two cases (2.63%) biopsy was needed. A definitive diagnosis was confirmed by histopathology in 22 surgically managed cases and by excision biopsy in two. 22/76 (28.94%) of the cases needed surgical treatment, and other cases required conservative or palliative approach.

Fidelity in Academic Global Surgery and Research: Incorporating Trustworthiness in the Development of Research Partnerships, Infrastructure, and Policy.

Academic global surgery consists of collaborative partnerships that address surgical inequities through research, training, education, advocacy, and diplomacy. It has been characterized by increased scholastic production through global surgery publications, dedicated global surgery sessions within scientific conferences, global surgery-specific research grants, database development to support global surgery research, global surgery research fellowships, and global surgery-based academic promotion paradigms. The increased emphasis on global surgery research has been accompanied by multiple ethical challenges. This article reviews critical ethical dilemmas presented by global surgery research efforts and proposes interventions on the partnership, infrastructural, and policy levels to enhance fidelity within research partnerships.

Modulation of Apoptotic and Akt/PI3K/mTOR pathways to target Glioblastoma Cells using synthetic compound PGEA-AN.

Objectives: To investigate the anticancer potential of a novel synthetic derivative of a naturally occurring diterpenoid against glioblastoma. METHODS: The in vitro study was conducted at the Ojha Campus of Dow University of Health Sciences, Karachi, from February to December 2021, and comprised U87 cells. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to determine the growth inhibitory effect of 16(R and S) - phenylamino-cleroda3, 13(14) Zdiene- 15, 16 olide and standard drug temozolomide against glioblastoma cells, and half-maximal inhibitory concentration was calculated. Microscopy and immunocytochemistry were used to investigate apoptotic morphology and active caspase-3 and B-cell lymphoma 2 (Bcl-2) expression. Quantitative real time polymerase chain reaction was used to investigate the expression of proliferation markers. Data was analysed using SPSS 21. RESULTS: Both the synthetic derivative and the standard drug significantly inhibited growth of U87 cells (p<0.001) with half-maximal inhibitory concentration of 19µM and 185µM, respectively. Apoptotic morphology and upregulation of active caspase-3 protein expression was observed in cells treated with half-maximal inhibitory concentration doses of both the synthetic derivative (p<0.05) and the standard drug (p<0.001), and Bcl-2 was downregulated in both the synthetic derivative (p<0.01) and the standard drug (p=0.05). However, no significant difference was observed in the expression of proliferation markers (p>0.05). CONCLUSIONS: The synthetic diterpene derivative PGEA-AN showed growth inhibitory actiity against glioblastoma.

Unplanned readmissions, community socioeconomic factors, and their effects on long-term survival after complex thoracic aortic surgery.

OBJECTIVE: We evaluated community socioeconomic factors in patients who had unplanned readmission after undergoing proximal aortic surgery (ascending aorta, aortic root, or arch). METHODS: Unplanned readmissions for any reason within 60 days of the index procedure were reviewed by race, acuity at presentation, and gender. We also evaluated 3 community socioeconomic factors: poverty, household income, and education. Kaplan-Meier survival curves were used to assess long-term survival differences by group (race, acuity, and gender). RESULTS: Among 2339 patients who underwent proximal aortic surgery during the 20-year study period and were discharged alive, our team identified 146 (6.2%) unplanned readmissions. Compared with White patients, Black patients lived in areas characterized by more widespread poverty (20.8% vs 11.1%; P = .0003), lower income ($42,776 vs $65,193; P = .0007), and fewer residents with a high school diploma (73.7% vs 90.1%; P < .0001). Compared with patients whose index operation was elective, patients who had urgent or emergency index procedures lived in areas with lower income ($54,425 vs $64,846; P = .01) and fewer residents with a high school diploma (81.1% vs 89.2%; P = .005). Community socioeconomic factors did not differ by gender. Four- and 6-year survival estimates were 63.1% and 63.1% for Black patients versus 89.1% and 83.0% for White patients (P = .0009). No significant differences by acuity or gender were found. CONCLUSIONS: Among readmitted patients, Black patients and patients who had emergency surgery had less favorable community socioeconomic factors and poorer long-term survival. Earlier and more frequent follow-up in these patients should be considered. Developing off-campus clinics and specific postdischarge measures targeting these patients is important.

The International System for Reporting Serous Fluid Cytopathology: An Institutional Experience on its Implication and Assessment of Risk of Malignancy in Effusion Cytology.

Background: The "international system for reporting serous fluid cytopathology"(TIS) consists of five diagnostic categories: nondiagnostic (ND), negative for malignancy (NFM), atypia of undetermined significance (AUS), suspicious for malignancy (SFM), and malignant (MAL). The study was conducted to reclassify effusion cytology samples according to the newly proposed TIS to calculate the risk of malignancy (ROM) for each category and to conduct performance analysis. Materials and Methods: The study was a retrospective observational study conducted at a tertiary care institution in North India. Clinical data of the cases from June 2013 to July 2021 were retrieved and the cases were reviewed by two cytopathologists. All cases were reclassified according to the proposed TIS system into five categories. Results: A total of 2318 patients were included in the study over a time span of 8 years, and 1614 (69.6%) cases of pleural effusion cytology, 612 (26.4%) cases of peritoneal effusion cytology, and 92 (3.9%) cases of pericardial effusion cytology were included. All effusion cytology smears were recategorized as per TIS guidelines into ND, NFM, AUS, SFM, and MAL with 4 (0.17%), 1756 (75.75%), 12 (0.51%), 57 (2.46%), and 489 (21.11%) cases, respectively, and ROM was 25%, 17.9%, 66.7%, 75.4%, and 96.5%, respectively, for the categories. Conclusion: Besides being a simple, easy, and user-friendly system, TIS has the benefit of risk stratification and ROM for each category. The TIS system proposed a tiered scheme, which places the effusion cytology into well-defined categories, and therefore has lesser chances of false-positive and false-negative cases.

Monotherapy with topical Natamycin to treat Scedosporium keratomycosis: A rare case from Central India.

Keratomycosis is common in Indian subcontinent. Diagnosis of the causal agent and successful management is a challenge for the clinician. Scedosporium is a rare fungus species, and it is relatively rare in causing keratomycosis. We report the case of a 29-year-old male who presented with complaints of redness, watering, and white lesion over his left eye. He sustained an injury in the left eye with vegetative matter. Corneal scraping was sent for potassium hydroxide staining and culture; fungal colony was seen in culture. Colony characters on Sabouraud dextrose agar and lactophenol cotton blue enabled a diagnosis of Scedosporium species. The patient was treated with topical Natamycin 5%, and complete resolution was seen at the end of 4 weeks. This case report highlights good response of keratitis caused by Scedosporium to topical Natamycin therapy.

Recent Advancements in Ion-Pair Receptors.

Over the past two decades, non-covalent chemistry has introduced various promising artificial receptors and revolutionized the host-guest chemistry. These versatile receptors have particularly been entertained in sensing and recognizing of diverse neutral molecules and/or ionic entities (e. g. anions, cations and ion-pair) of particular interest. Notably, supramolecular chemistry had given birth to a plethora of important molecules, explored in the chemical, biological, environmental, and pharmacological world to resolve the critical issues related to the human health while keeping environmental concerns in mind. Amongst the various types of supramolecular monotopic receptors (anions, cations, and neutral molecules), heteroditopic receptors (ion-pair receptors) consisting of distinct binding sites in one system for both cation and anion, have gained much interest from the scientific community in recent past because of their unique binding abilities. Interestingly, these promising artificial receptors have shown potential applications in sensing, recognition, transport and extraction processes besides their uses in salt/waste purification. Bearing the importance of these systems in mind, we intended to report the recent developments in ion-pair chemistry. Herein, we divided the whole document into three main sections; first one describes the introduction and history of the ion-pairs receptors. The second portion highlights the synthesis and applications of ion-pair receptors in sensing, recognition, molecular machines, photoswitching behaviour, extraction and transport properties, whereas the last part of this manuscript provides concluding remarks as well as future prospects of ion-pair receptors. We hope that this manuscript will be helpful to stimulating researchers around the globe to find out the hidden opportunities in this and related areas.

Dough kneading exposure at the kitchen workstation: Influence on handgrip strength and implications for therapy in hand rehabilitation.

BACKGROUND: Dough kneading is a commonly performed activity in the kitchen, which influences hand grip strength. OBJECTIVES: To study the influence of dough kneading exposure on hand grip strength and to evaluate the effect of dough kneading intervention on hand grip strength with the purpose of recommending dough kneading as a therapeutic exercise for improving hand grip strength. METHODS: One hundred and fifty healthy females with varying levels of exposure to dough kneading, stratified as occupational dough kneaders, habitual dough kneaders and non-kneaders, were studied. Hand grip strength of all participants was measured with a standard protocol using the Jamar dynamometer. Hand grip strength of occupational, habitual and non-kneaders was compared. Non-kneaders followed a 6-week intervention of dough kneading and grip strength was recorded post-intervention. RESULT: Comparison of hand grip strength between the three groups revealed significant difference (p value < 0.001). Linear contrast analysis, revealed the least hand grip strength in non-kneaders compared to habitual and occupational dough kneaders, with occupational dough kneaders presenting maximum hand grip strength (p value < 0.001). Significant improvement was demonstrated in hand grip strength post-intervention in non-kneaders (p value < 0.001). CONCLUSION: Findings suggest that exposure to dough kneading has a positive influence on hand grip strength. Hand grip strength of non-kneaders was lowest compared to habitual and occupational kneaders. Kneading intervention improved hand grip strength and hence can be used therapeutically as a safe, low cost exercise in hand rehabilitation.

Electrophysiological and Visual Parameter Changes in Retinitis Pigmentosa Patients undergoing Autologous Platelet-Rich Plasma Therapy: A Randomized Control Trial.

Purpose: To assess the efficacy of autologous platelet-rich plasma (PRP) injections in suprachoroidal space and subtenon space in cases of retinitis pigmentosa, which is a genetic disease, leading to gradual loss of vision. Till date, no treatment is available. Methods: Seventy-eight eyes of 39 patients of retinitis pigmentosa having visual acuity ranging from reading of Early Treatment Diabetic Retinopathy Study (ETDRS) chart from 1 m onward to patients who were not able to read the ETDRS chart but whose visual acuity ranged from finger count close to face to <1 m were included in the study. The left and right eyes of each patient were randomized as the intervention eye and control eye. 0.2 mL of autologous PRP was injected in suprachoroidal space and 0.5 mL of PRP was injected in subtenon space of the intervention eye taking aseptic precautions. Injections were repeated at 15-day intervals up to 3 injections. Results: Majority of patients were in the age group of 18-30 years (20 cases) followed by 31-45 years (13 cases) and more than 45 years (6 cases). Intervention eyes showed a statistically significant improvement in visual acuity and multifocal electroretinography (mfERG). Improvement was noted in amplitude density latency and in ring ratio of mfERG. There was a significant improvement in best-corrected visual acuity (BCVA). However, no improvement in mfERG or BCVA was observed in the control group. Conclusions: Gene therapy may be the ultimate cure for retinitis pigmentosa, but it is unaffordable for many patients due to its high cost. PRP may be recognized as a modality to improve vision and stop further deterioration, especially in cases where functional vision is preserved. Negligible treatment costs and affordability will give power to economically disadvantaged patients.

Modified small-incision cataract surgery for combined extraction - A comparative study of two techniques.

Purpose: To explore straight incision technique in terms of efficacy for intraocular pressure (IOP) lowering by small-incision cataract surgery (SICS) trab versus modified "frown" incision with triangular scleral flap technique. Methods: This study was done at a tertiary health center. It included 44 eyes diagnosed with cataract and coexisting primary glaucoma that underwent SICS with trabeculectomy using modified "frown" incision with triangular scleral flap technique and straight incision in group A (n = 22) and B (n = 22), respectively. Postoperative evaluation was done at first postoperative day, then at the end of first week, third week, and 6 weeks; at the end of third month and finally at the end of sixth months. Data were entered and analyzed via Microsoft Excel sheet and SPSS software using Mann-Whitney U test for averages and Chi-square test for categorical values. Results: Mean preoperative IOP in groups A and B were 38.6 and 29.1 mm Hg respectively, by applanation tonometry. After 6-month follow-up, mean of difference in IOP (preoperative - postoperative) for group A was 20.8 ± 8.3 mm Hg and that for group B was 17.2 ± 13.5 mm Hg. Conclusion: Capacity of IOP reduction of both techniques was found to be comparable and did not show much difference up to the end of 6 months. Mastering technique of group A (modified "frown" incision with triangular scleral flap technique) requires more expertise; the simpler straight incision technique provided in group B may be effectively used by the novice and current era Ophthalmologists to combat glaucoma coexistant with cataract.

Comparative analysis of inter-observer variability between conventional reporting system of salivary gland cytology with Milan system for reporting salivary gland cytopathology.

BACKGROUND: Interpretation of salivary gland cytology often leads to inter-observer variability due to heterogenous and complex nature of these lesions. This creates a dilemma regarding their management by clinicians. Proposal of a universal system of reporting of salivary gland lesions leading to agreement in diagnosis and better understanding among clinicians was the need of the hour. Hence Milan system for reporting salivary gland cytopathology (MSRSGC) was proposed in 2015 by the American society of cytopathology and the International Academy of Cytology. Present study was undertaken to assess inter-observer variability in reporting by conventional system and MSRSGC. MATERIAL AND METHODS: One hundred and seventy-six cases of salivary gland lesions were subjected to fine needle aspiration cytology. Cases were interpreted by two experienced cytopathologists and were reported by both conventional system and MSRSGC. Histopathological correlation was available in 81 cases. RESULTS: Inter-observer variability was noted in six cases reported by conventional system and in two cases by MSRSGC. Moreover three cases out six cases had different management protocols while both cases of Milan system for reporting salivary gland cytology had same management. Thirteen cases diagnosed by Milan system and 17 cases diagnosed by conventional system were discordant with histopathological diagnosis. CONCLUSION: Milan system for reporting salivary gland cytology has an edge over conventional system of reporting as it provides better agreement among cytopathologists and better management guidelines for clinicians with the added advantage of assessment of risk of malignancy.

Tubercular granuloma arising from floor of orbit : A rare occurrence.

INTRODUCTION: Tuberculosis of the orbit is a unilateral entity commonly involving the bony wall, periosteum or soft tissue in left orbit. It is a rarity. CASE: A fifty seven years old female presented with a localised mass near inferior orbital margin. Owing to its small size and superficial location in orbit, there was no soft tissue displacement or motility disturbance. OBSERVATIONS: Dissection revealed a localised firm mass attached to periorbita covering the orbital floor. The mass was excised and on histopathological analysis it was found to be composed of multiple granulomas containing abundant Giant cells. Considering such a picture at cellular level, ELISA for tuberculosis was done. Values were suggestive of recent infection. The patient was put on antitubercular multidrug regimen for six months one week after excision of mass, with a follow up of four years, having no recurrence till date. CONCLUSION: Though orbital tuberculosis is much more common in childhood, it should always be considered in the differential diagnosis of orbital mass in adults also. At times, even in an otherwise healthy individual, tuberculosis might be diagnosed because of an orbital mass. Ophthalmologists should have a high index of suspicion since orbital tuberculosis has been underdiagnosed in the past.

Opuntiol Inhibits Growth and Induces Apoptosis in Human Glioblastoma Cells by Upregulating Active Caspase 3 Expression.

BACKGROUND: Glioblastoma Multiforme (GBM) is a deadly tumor with poor prognosis. Resistance to apoptosis considered as an important factor in treatment failure. Therefore, identification of new compounds that facilitates apoptosis is crucial. Natural Anti-inflammatory compounds have emerged as potential anti-cancer agents and should be explored for their apoptotic activity against GBM. Therefore, the present study aims to evaluate growth inhibitory and apoptotic activity of a natural anti-inflammatory compound "Opuntiol" against GBM cell line U87. METHODS: MTT assay was performed to determine the effect of Temozolomide and Opuntiol on growth inhibition of U87 cell. While, TUNEL assay was used to assess their apoptotic activity. To further assess apoptosis, nuclear condensation and nuclear area factor (NAF) was evaluated through DAPI staining. Whereas, active caspase-3 protein expression determined using immunocytochemistry. RESULTS: Significant growth inhibition was observed in U87 cells treated with Temozolomide (IC50 380 µM) and Opuntiol (IC50 357 µM). Temozolomide (p<0.001) and Opuntiol (p<0.001) significantly improved rate of apoptosis when compared to control group. A significant decrease in NAF was also observed in Temozolomide (p < 0.05) and Opuntiol (p < 0.05) treated cells. There was a significant increase in active caspase-3 expression when observed in Temozolomide (p<0.001) and Opuntiol (p<0.05) treated groups as compared to control. CONCLUSION: In conclusion our findings suggests, Opuntiol repress cell viability and possess strong apoptotic activity against GBM cell line U-87. However, further mechanistic studies will be required to confirm whether it can be develop as a potential drug against GBM.

Study to Evaluate Stress Among Ophthalmic Surgeons with Different Levels of Surgical Experience.

AIM: To assess and quantify the stress in two different groups of ophthalmic surgeons while performing cataract surgery. MATERIALS AND METHODS: This is a prospective observational institutional study. Healthy ophthalmic surgeons (4 trainee surgeons and 4 consultants) and 4 OT assistants without any history of systemic illness were studied while performing uncomplicated and uneventful cataract surgery. Resting state and post-operative (immediately after surgery) systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse rate (PR), and oxygen saturation (SPO2) were measured by a Comen C80 multi-parameter monitor. Blood cortisol levels were measured by chemiluminescent assay (CLIA) and urine catecholamines levels (adrenaline, nor-adrenaline and dopamine) were measured by high performance liquid chromatography (HPLC). For assessment of stress among one group, paired t-test is used and for comparison of stress levels between trainee surgeons and consultants unpaired t-test is used. RESULTS: Consultants performed phacoemulsification and trainee surgeons performed small incision cataract surgery. In both the groups, the post-operative values of SBP, DBP, PR, blood cortisol, and urine catecholamines were significantly higher than the pre-operative values. This difference was statistically significant. There was a significantly higher increase in SBP, DBP, PR, blood cortisol, urine adrenaline, and urine dopamine in trainee surgeons as compared to consultants whereas there was a significantly higher increase in urine nor-adrenaline in consultants as compared to trainee surgeons. CONCLUSION: Our study reveals that every surgeon is under stress irrespective of experience though the level of stress is different among surgeons.

The role of iron repletion in adult iron deficiency anemia and other diseases.

Iron deficiency anemia (IDA) is the most prevalent and treatable form of anemia worldwide. The clinical management of patients with IDA requires a comprehensive understanding of the many etiologies that can lead to iron deficiency including pregnancy, blood loss, renal disease, heavy menstrual bleeding, inflammatory bowel disease, bariatric surgery, or extremely rare genetic disorders. The treatment landscape for many causes of IDA is currently shifting toward more abundant use of intravenous (IV) iron due to its effectiveness and improved formulations that decrease the likelihood of adverse effects. IV iron has found applications beyond treatment of IDA, and there is accruing data about its efficacy in patients with heart failure, restless leg syndrome, fatigue, and prevention of acute mountain sickness. This review provides a framework to diagnose, manage, and treat patients presenting with IDA and discusses other conditions that benefit from iron supplementation.

Supplementation of green tea extract (GTE) in extender improves structural and functional characteristics, total antioxidant capacity and in vivo fertility of buffalo (Bubalus bubalis) bull spermatozoa.

The aim of this study was to elucidate the beneficial effects of green tea extract (GTE) in tris citric acid extender on post thaw structural and functional characteristics, DNA fragmentation (%), total antioxidant capacity (TAC, µM/L), lipid peroxidation (LPO, µM/mL) levels and in vivo fertility of buffalo (Bubalus bubalis) bull spermatozoa. GTE is a acknowledged natural antioxidant, act as a free radical scavenger and protects spermatozoa against oxidative stress. Three mature and donor buffalo bulls were used in this experiment. Two ejaculates were collected per bull on each collection day, followed by initial evaluation for consistency (colour), volume (mL), progressive motility and concentration (x109) and were diluted in five extenders @ 50 x 106/ mL (C = control, no GTE; T1 = treatment 1, GTE 0.1%; T2 = treatment 2, GTE 0.5%; T3 = treatment 3, GTE 0.75% and T4 = treatment 4, GTE1.0%). The experiment was repeated thrice. Data analysis showed that sperm progressive motility (%), plasma membrane integrity (%), supravital plasma membrane integrity (%), viable sperm with intact acrosome (%) and TAC were significantly higher (P < 0.05) in extender supplemented with T4 as compared to control. Furthermore, sperm DNA fragmentation and occurrance of LPO in buffalo bull spermatozoa were significantly lowered in T4 than control. In vitro longevity (%) of spermatozoa was significantly higher in T4 compared to control during 45 and 90 min of incubation at 37 °C. In vivo fertility rate of buffalo bull spermatozoa was significantly higher in T4 compared to control (64.96 vs. 48.40%, P < 0.05). It is concluded that supplementation of tris citric acid extender with 1.0% GTE improved structural and functional characteristics, TAC, in vitro longevity (%) and in vivo fertility, whereas decreased DNA fragmentation and LPO occurrence in buffalo bull spermatozoa after freezing and thawing protocol.

Supplementation of l-tryptophan (an aromatic amino acid) in tris citric acid extender enhances post-thaw progressive motility, plasmalemma, mitochondrial membrane potential, acrosome, and DNA integrities, and in vivo fertility rate of buffalo (Bubalus bubalis) bull spermatozoa.

The aromatic amino acid l-tryptophan is an essential and versatile molecule, acts by transferring an electron to free radicals and protects the plasma membrane from injuries. The aim of the present study was to investigate the effects of l-tryptophan in extender on semen quality parameters, in vitro longevity and in vivo fertility rate of buffalo spermatozoa during cryopreservation. Two ejaculates were collected from each bull (n = 2 ejaculates and n = 4 bulls) with artificial vagina at 42 °C followed by initial evaluation for volume, motility, concentrations and were diluted in five extenders (C = lacking l-tryptophan, D1 = 25 µ M l-tryptophan, D2 = 50 µ M l-tryptophan, D3 = 75 µ M l-tryptophan, and D4 = 100 µ M l-tryptophan) respectively, and cryopreserved. The experiment was repeated four times (n = 4 replicates). At post-dilution, sperm plasma membrane integrity (PMI, %), supravital plasma membrane integrity (SVPMI, %), hypo-resistivity (HR, %) and acrosome integrity (ACR-I, %) were significantly higher (P < 0.05) in extender supplemented with D4 than control. At post-thawing, progressive motility (PM, %), PMI, SVPMI, HR, ACR-I, and DNA-I of buffalo bull spermatozoa were significantly higher in D4 than control. Sperm in vitro longevity (%) assessed in terms of PM, SVPMI, and ACR-1 were significantly higher in D4 than control. Sperm mitochondrial membrane potential (%) was higher in treated groups than the control. The in vivo fertility rate was significantly higher in D4 than control (60.17% vs. 44.17%, P < 0.05). It is concluded that the supplementation of l-tryptophan in tris citric acid extender improves semen quality parameters, in vitro longevity and in vivo fertility rate of buffalo spermatozoa during freezing and thawing process.

Milan System for Reporting Salivary Gland Cytopathology: An Experience with the Implication for Risk of Malignancy.

INTRODUCTION: Fine-needle aspiration cytology (FNAC) is a well-established technique for evaluation of salivary gland lesions, but because of the heterogenicity and morphological overlap between spectrum of lesion, there are a few challenges in its wide use. Recently, "The Milan system for reporting salivary gland cytopathology" (MSRSGC) was introduced, providing guide for diagnosis and management according to the risk of malignancy (ROM) in different categories. The current study was conducted retrospectively to reclassify the salivary gland lesions from previous diagnosis and to evaluate the ROM in different categories. MATERIAL AND METHODS: Clinical data, FNAC specimen, histological, and clinical follow-up of cases were retrieved, cytological features were re-evaluated, and cases were reclassified as follows: Category 1: Non-diagnostic (ND); Category 2: Non-neoplastic (NN); Category 3: Atypia of undetermined significance (AUS); Category 4a: Neoplasm: benign (NB), Category 4b: Neoplasm: salivary gland neoplasm of uncertain malignant potential (SUMP); Category 5: suspicious of malignancy (SM); and Category 6: Malignant (M). RESULT: Total 293 cases were evaluated cytologically, and histological follow-up was available in 172 cases. The distribution of cases into different categories was as follows ND (6.1%), NN (38.2%), AUS (2.7%), NB (33.4%), SUMP (2.0%), SM (2.4%), and M (15%). Overall, ROM reported were 25%, 5%, 20%, 4.4%, 33.3%, 85.7%, and 97.5%, respectively for each category. Overall, sensitivity was 83.33%, specificity was 98.31%, positive predictive value was 95.74%, and negative predictive value was 92.80%. CONCLUSION: MSRSGC is a recently proposed six category scheme, which places salivary gland FNAC into well-defined categories that limit the possibilities of false negative and false positive cases.

Clinical and Cyto-Morphological Characterization of Triple Negative Breast Cancer.

OBJECTIVE: Triple negative breast cancer (TNBC), despite being the uncommon subtype, contributes a major portion to mortality and associated with poor prognosis. The purpose of this study was to evaluate the cytological criteria for the diagnosis of TNBC through fine-needle aspiration cytology (FNAC). MATERIAL AND METHOD: Clinical, cytological, histological, and immunohistochemical (IHC) data of 256 patients were evaluated, and patient were classified as TNBC and non-TNBC phenotype by IHC. All cytological specimens were reviewed for 12 criteria: cellularity, tubule/gland formation, syncytial clusters, large bare nuclei, nuclear atypia, chromatin pattern, cell borders, nucleolus, cytoplasm, lymphocytic infiltrate, calcification, and necrosis. The Fischer's exact test was used to show test association. RESULT: Out of 256 patients, 82 patients were TNBC, and 174 patients were non-TNBC. TNBC phenotype showed statistically significant association to cellularity, tubule/gland formation, syncytial cluster formation, bare nuclei, nuclear atypia, cell borders, lymphocyte infiltration, and necrosis. CONCLUSION: FNAC can be helpful in making diagnosis of TNBC and along with ER, PR, HER2 characterization, helpful in planning treatment strategy, saving time, manpower, and resources in the patient management.