Thymosin ß4 Is an Endogenous Iron Chelator and Molecular Switcher of Ferroptosis.

Thymosin ß4 (Tß4) was extracted forty years agofrom calf thymus. Since then, it has been identified as a G-actin binding protein involved in blood clotting, tissue regeneration, angiogenesis, and anti-inflammatory processes. Tß4 has also been implicated in tumor metastasis and neurodegeneration. However, the precise roles and mechanism(s) of action of Tß4 in these processes remain largely unknown, with the binding of the G-actin protein being insufficient to explain these multi-actions. Here we identify for the first time the important role of Tß4 mechanism in ferroptosis, an iron-dependent form of cell death, which leads to neurodegeneration and somehow protects cancer cells against cell death. Specifically, we demonstrate four iron2+ and iron3+ binding regions along the peptide and show that the presence of Tß4 in cell growing medium inhibits erastin and glutamate-induced ferroptosis in the macrophage cell line. Moreover, Tß4 increases the expression of oxidative stress-related genes, namely BAX, hem oxygenase-1, heat shock protein 70 and thioredoxin reductase 1, which are downregulated during ferroptosis. We state the hypothesis that Tß4 is an endogenous iron chelator and take part in iron homeostasis in the ferroptosis process. We discuss the literature data of parallel involvement of Tß4 and ferroptosis in different human pathologies, mainly cancer and neurodegeneration. Our findings confronted with literature data show that controlled Tß4 release could command on/off switching of ferroptosis and may provide novel therapeutic opportunities in cancer and tissue degeneration pathologies.

Toward strengthening active case finding for ending tuberculosis in India.

Active case finding (ACF) is critical for the timely detection and treatment of diseases like tuberculosis (TB). ACF is crucial in India, where health-seeking behaviour across various populations is low and non-reporting is a barrier to breaking disease transmission cycles and eliminating the disease. As an integral part of India's national TB control programs, there is further scope for improvement in its design. This paper outlines a roadmap for ACF for TB in the country. It is based on current national strategic plans for public health monitoring and TB case identification. It suggests that ACF should be strengthened by developing it as a learning surveillance system involving a change from a siloed public health approach to a more comprehensive surveillance exercise. It defines operational requirements for establishing predictive capabilities and introducing feedback and learning in the system as a precondition. It highlights the importance of establishing proper processes, as well as extending and expanding on existing infrastructural and human resource, and inter-programmatic coordination and collaboration in the health sector.

Genomic surveillance and phylodynamic analyses reveal emergence of novel mutation and co-mutation patterns within SARS-CoV2 variants prevalent in India

Emergence of distinct viral clades has been observed in SARS-CoV2 variants across the world and India. Identification of the genomic diversity and the phylodynamic profiles of the prevalent strains of the country are critical to understand the evolution and spread of the variants. We performed whole-genome sequencing of 54 SARS-CoV2 strains collected from COVID-19 patients in Kolkata, West Bengal during August to October 2020. Phylogeographic and phylodynamic analyses were performed using these 54 and other sequences from India and abroad available in GISAID database. Spatio-temporal evolutionary dynamics of the pathogen across various regions and states of India over three different time periods in the year 2020 were analyzed. We estimated the clade dynamics of the Indian strains and compared the clade specific mutations and the co-mutation patterns across states and union territories of India over the time course. We observed that GR, GH and G (GISAID) or 20B and 20A (Nextstrain) clades were the prevalent clades in India during middle and later half of the year 2020. However, frequent mutations and co-mutations observed within the major clades across time periods do not show much overlap, indicating emergence of newer mutations in the viral population prevailing in the country. Further, we explored the possible association of specific mutations and co-mutations with the infection outcomes manifested within the Indian patients.

Rare Intracranial Recurrent Meningioma with Unique Coexisting Atypical, Papillary and Lipomatous Component: Case Report and Review of Literature.

BACKGROUND: Meningiomas are common central nervous system tumors with a wide range of morphologic variants, the pathogenesis being their complex embryogenesis. Intracranial meningiomas with heterogenous histopathology in the same lesion are common in low-grade meningiomas but less frequent in recurrent and high-grade variants. CASE DESCRIPTION: A 75-year-old male presented elsewhere a year ago with complaints of slurred speech. Magnetic resonance imaging revealed a left frontotemporal, dural-based, extraaxial, solid cystic lesion with doubtful infiltration into the adjacent brain parenchyma. A subtotal excision was done. A diagnosis of meningioma with an aggressive biological behavior was rendered on histology, but the grade was deferred, in view of a single focus of small cell formation and no adjacent brain parenchyma to comment on invasion. The patient presented here with recurrence of his original symptoms. Magnetic resonance imaging of the brain with contrast revealed 2 dural-based, solid cystic enhancing lesions of sizes 29 mm × 25 mm × 24 mm and 25 mm × 16 mm seen at the left frontal region, indenting the adjacent brain parenchyma with diffuse meningeal thickening in the postoperative bed. Microscopy revealed a hypercellular meningeal neoplasm with increased mitosis and a pseudopapillary pattern with lipomatous changes. CONCLUSIONS: To date there are no case reports in current literature with such rare combinations in a recurrent meningioma. This highlights the multipotency of phenotypic transformation of primary meningothelial cells. The presence of papillary features, even if focal, should be quantified in the diagnosis. This is of importance because the most current literature suggests that meningioma harboring a papillary component has an increased risk of recurrence and progression to aggressive behavior.

Clinicopathological Analysis of HIF-1alpha and TERT on Survival Outcome in Glioblastoma Patients: A Prospective, Single Institution Study.

Glioblastoma multiforme is a highly malignant and aggressive primary brain tumor with a dismal prognosis. We studied the association of immunohistochemical expression of hypoxia inducible factor-1 alpha (HIF-1α), telomerase reverse transcriptase (TERT), isocitrate dehydrogenase 1 (IDH1) and tumor protein p53 with overall survival (OS) in glioblastoma patients uniformly treated by standard of care, with adequate follow-up. In 87 patient samples studied, 59 were male and 28 were female. The median age was 55 years. The median follow-up was 27.7 months and the median overall survival was 14.9 months. Nuclear staining of HIF-1α was expressed in all samples and scored as strong in 42 (48%) and weak in 45 (52%). Multivariable Cox regression revealed strong HIF-1α expression as an independent poor prognostic factor (Hazard Ratio 2.12, 95% CI 1.20 - 3.74, P = 0.01). There was a statistically significant difference in OS (9.8 months vs. 16.3 months) between the "HIF-1α - strong and TERT - strong" and the "HIF-1α - weak and TERT - weak" patient subgroups, as evaluated by Kaplan-Meier analysis (P = 0.005). In our study, HIF-1α expression was an independent predictor of OS. The subgroup of patients with strong expression of both HIF-1α and TERT had the poorest prognosis.

Evaluation of a new flow cytometry based method for detection of BCR-ABL1 fusion protein in chronic myeloid leukemia

BACKGROUND: Philadelphia chromosome, a hallmark of chronic myeloid leukemia (CML), plays a key role in disease pathogenesis. It reflects a balanced reciprocal translocation between long arms of chromosomes 9 and 22 involving BCR and ABL1 genes, respectively. An accurate and reliable detection of BCR-ABL fusion gene is necessary for the diagnosis and monitoring of CML. Previously, many technologies, most of which are laborious and time consuming, have been developed to detect BCR-ABL chimeric gene or chromosome. METHODS: A new flow cytometric immunobead assay was used for detection of BCR-ABL fusion proteins and applicability, sensitivity, reliability, efficacy and rapidity of this method was evaluated. RESULTS: From February 2009 to January 2014, a total 648 CML patients were investigated for the status of BCR-ABL1 protein. Among them, 83 patients were enrolled for comparative study of BCR-ABL1 positivity by three routinely used procedures like karyotyping, and quantitative real time PCR (RT-PCR) as well as immunobead flow cytometry assay. BCR-ABL protein analysis was found consistent, more sensitive (17% greater sensitivity) and reliable than the conventional cytogenetics, as flow cytometry showed 95% concordance rate to RT-PCR. CONCLUSION: BCR-ABL fusion protein assay using a new flow cytometric immunobead might be useful in the diagnosis and monitoring CML patients.

Psammomatous melanotic schwannoma as a component of Carney complex.

Melanotic schwannoma is a rare form of pigmented neural tumor commonly arising from the posterior spinal nerves and ganglia. Two variants have been described, psammomatous and nonpsammomatous. 50% of psammomatous tumors are associated with Carney complex. The biologic behavior of the tumor is difficult to predict and slightly over 10% of the tumors follow malignant course. We present a case of psammomatous melanotic schwannoma as part of Carney complex in a 67-year-old male. Clinical examination revealed oral mucosal and abdominal skin pigmentation. Magnetic resonance imaging showed an intradural extramedullary lesion at D8-D12 level. Intraoperative squash smear study showed sheets of spindle cells with abundant intracytoplasmic melanin pigmentation and few psammoma bodies. Based on clinical, radiologic, and histopathological findings with immunohistochemistry correlation a final diagnosis of psammomatous melanotic schwannoma was rendered.

Pleomorphic xanthoastrocytoma with anaplastic features: a case report.

Pleomorphic xanthoastrocytoma has been considered as an astrocytic tumor with relatively favorable prognosis. It corresponds to WHO Grade-II neoplasm. Recently, several patterns with relatively poor prognosis have been recorded and a new concept of "PXA with anaplastic features" has been proposed. The present case is about a 9-year-old girl who presented with symptoms of recurrent headache, seizures and poor academic performance. MRI revealed left fronto-parietal irregular enhancing mass lesion with callosal involvement and right mid-brain arteriovenous malformation. Clinical and radiological examination was suggestive of a high grade glial neoplasm/PNET. A diagnosis of high grade glial neoplasm was rendered on the squash smears submitted for frozen sections based on the presence of spindle cells, admixed with pleomorphic bizarre, giant cells with multilobated nuclei showing few atypical mitosis and abundant eosinophilic cytoplasm. Frontal craniotomy with debulking of the tumor was performed and permanent sections revealed a biphasic glial neoplasm with spindle cells arranged in fascicles admixed with bizarre multinucleated giant cells showing abundant vacuolated and lipidized cytoplasm, nuclear hyperchromasia with intranuclear inclusions. Eosinophilic granular bodies, mitosis of 7/10 HPF, micro vascular proliferation, necrosis and invasion into the underlying brain parenchyma were noted. With these histomorphological findings a diagnosis of pleomorphic xanthoastrocytoma with anaplastic features was rendered.

Primary central nervous system lymphoma: a clinicopathological and cytomorpholgical study from a tertiary care centre in Chennai, India.

BACKGROUND: The aim of this study was to analyze the clinicopathological and immunohistochemical features of primary central nervous system lymphoma (PCNSL) cases occurring in Indian patients and also study the utility of the crush smear preparation in intraoperative diagnosis. MATERIALS AND METHODS: The immune status, clinical, radiological details, immunohistochemical profile, histopathological findings and cytological features in smear preparation of 32 cases of PCNSL were analyzed. Patients with systemic NHL and skull-base lymphomas were excluded. RESULTS: The mean age of our patients was 52 years with a male: female ratio 1:1. A periventricular location was found in 62.5% of patients. None of our PCNSL cases were associated with AIDS. All cases except one were diffuse large B-cell lymphomas. Intraoperative diagnosis using crush smears allowed correct prediction in 93% of cases. CONCLUSIONS: Our study shows that PCNSL is seen predominantly in immunocompetent patients in India .The age of presentation is relatively young as compared to the West. Our study also stresses the utility of crush smear preparation in establishing an intraoperative diagnosis.

Modulation of conjunctival goblet cell function by inflammatory cytokines.

Ocular surface inflammation associated with Sjögren's syndrome is characterized by a loss of secretory function and alteration in numbers of mucin secreting goblet cells. Such changes are a prominent feature of ocular surface inflammatory diseases and are attributed to inflammation; however, the exact effect of the inflammatory cytokines on conjunctival goblet cell function remains largely unknown. In this study, we developed a primary culture of mouse goblet cells from conjunctival tissue and evaluated the effects on their function by inflammatory cytokines detected in the conjunctiva of mouse model of Sjögren's syndrome (Thrombospondin-1 deficient mice). We found that apoptosis of goblet cells was primarily induced by TNF-α and IFN-γ. These two cytokines also inhibited mucin secretion by goblet cells in response to cholinergic stimulation, whereas IL-6 enhanced such secretion. No changes in secretory response were detected in the presence of IL-13 or IL-17. Goblet cells proliferated to varying degrees in response to all the tested cytokines with the greatest response to IL-13 followed by IL-6. Our results therefore reveal that inflammatory cytokines expressed in the conjunctiva during an ocular surface disease directly disrupt conjunctival goblet cell functions, compromising the protective function of tears, thereby contributing to ocular surface damage.

Practical value of MIB-1 index in predicting behavior of astrocytomas.

BACKGROUND: The MIB-1 labeling index (LI) has proved to be useful in assigning grading and prognosis to astrocytomas. The purpose of our study was to analyze the utility of MIB-1 LI in differentiating astrocytomas of varying grades and the possible relationships of MIB-1 LI with clinical parameters like age and sex. We also wanted to study the prognostic role of MIB-1 index in predicting behavior of astrocytomas. MATERIALS AND METHODS: Our study included 145 patients with astrocytic tumors of varying grades. Immunolabeling for all patients was done using MIB-1 antibody. Survival data could be obtained for 64 patients. A Mann-Whitney U test was used to test the difference in MIB-1 LI between different histological grades. The univariate analysis was done by the Kaplan-Meier method, and the multivariate analysis for survival was performed using the Cox proportional hazard model. RESULTS: Significant differences were noted in mean MIB-1 LI of high-grade and low-grade diffuse astrocytomas. MIB-1 LI did not vary significantly with age and sex. Univariate analysis showed favorable prognostic factors for low histopathological grade, young patient age and low MIB-1 LI; however, multivariate analysis showed that only histopathological grade had independent prognostic significance. CONCLUSIONS: Our study proves that MIB-1 LI is not dependent on factors like age and sex and is solely dependent on histological grade. Though the average level of MIB-1 LI varies considerably in the different grades of astrocytomas, considerable overlap can be observed between them. MIB-1 LI is a very useful adjunct to the histopathological diagnosis and can be of great help in situations where the clinical and radiological findings do not correlate with histological diagnosis.

Immunohistochemical profile of breast cancer patients at a tertiary care hospital in South India.

AIMS: 1) To evaluate the estrogen receptor(ER), progesterone receptor (PR) and Her-2 /neu expression in invasive breast carcinomas by immunohistochemistry and 2) to compare the pattern of expression with clinico-pathological parameters like patient's age, tumor size, mitotic index, histological type and grade and lymph node metastasis. METHODS: This is a retrospective study of 321 female invasive breast carcinomas diagnosed in the Department of Histopathology, Apollo Speciality Hospital, Chennai from January 2009 to June 2010. RESULTS: The age of the patients ranged from 24 to 99 years, with a mean of 53.8, and the majority of the tumors were T2 (83.8% in range of 2-5 cms), predominantly histological grade 2 (57.3%), followed by grade 3 (33.3%). ER, PR and Her-2/neu expression was seen in 59, 51 and 27% of cases respectively. Triple-negative breast cancers constituted 25 % of our cases. We also found characteristic associations between hormonal receptor and Her-2/neu expression and various clinico-pathological parameters. CONCLUSIONS: The hormonal receptor expression appears to be lower in the Indian population compared to the West. A significant proportion of tumors in our study with Her2/neu overexpression also showed ER and PR positivity. Triple-negative breast tumors were most commonly grade 3, in women aged more than 50 years.

The role of immunohistochemistry in predicting behavior of astrocytic tumors.

The purpose of this study was to analyze the significance of p53, bcl-2 and EGFR expression in the grading and biological behavior of astrocytic tumors, especially in the Indian population. A total of 117 cases of astrocytomas graded using the WHO grading system published in 2007 were immunolabeled using p53, EGFR and bcl-2 monoclonal antibodies and analyzed with respect to grade and other relevant parameters. The 117 cases included 16 cases of pilocytic astrocytomas and 25, 15 and 61 cases of diffuse fibrillary astrocytomas WHO grade II, anaplastic astrocytomas WHO grade III and glioblastomas (GBM), respectively. Our results showed that p53 alterations is an early event in astrocytic gliomagenesis, but is not significant in the evolution of pilocytic astrocytomas. Bcl-2 expression did not correlate with grade and no statistical correlation was seen with p53 expression. EGFR protein expression correlated with the severity of tumor grade. Of the GBM cases, 47.5% were p53 positive only, 18% were EGFR positive only, 16.5% were negative for both and 18% were positive for both. The mean age in the dual positive category was significantly higher when compared to the others. EGFR and p53 alterations are not mutually exclusive and might act synergistically to promote progression. We also noted a significantly higher p53 expression in females in GBMs. Though most of our findings correlated with those of previous studies, some differences were noted, especially in the pattern of immunoexpression in GBMs, perhaps because of ethnicity.

Desmoplastic infantile ganglioglioma -- a case report.

Desmoplastic infantile ganglioglioma is a very rare supratentorial tumor occurring in the first two years of life. A five-month-old female infant presented with recurrent seizures, large head and loss of acquired milestones. Computerized Tomographic Scan of brain showed a large subarachnoid cyst with a solid intensely contrast enhancing tumor in the right temporoparietal region with severe degree of mass effect. Craniotomy and total excision of the tumor followed subsequently by subduro-peritoneal shunt for the extracerebral fluid collection was done. The child made good recovery. Histopathology revealed features of desmoplastic infantile ganglioglioma, viz., marked desmoplastic component with glial and neuronal elements. Immunohistochemistry showed positive staining for glial fibrillary acidic protein (GFAP) with areas of synaptophysin and chromogranin positivity. Desmoplastic infantile ganglioglioma is a rare tumor of infancy, which has excellent prognosis after total excision. No adjuvant therapy is required. This is the first Indian report of desmoplastic infantile ganglioglioma out of less than fifty cases reported worldwide.