Benzo[a]pyrene exposure causes exonal switch resulting in reduced surface CD5 expression in an AHR-dependent manner.

The function of CD5 protein in T cells is well documented, but regulation of its surface-level expression has yet to be fully understood. However, variation in its surface expression is associated with various immunopathological conditions and haematological malignancies. Briefly, expression of an alternate exon E1B of a human endogenous retroviruses (HERV) origin directly downregulates the conventional transcript variant (E1A), as its expression leads to the retention of the resultant protein at the intracellular level (cCD5). A separate promoter governs the expression of E1B and may be influenced by different transcription factors. Hence, we performed in silico transcription factor binding site (TFBS) analysis of the 3 kb upstream region from TSS of exon E1B and found five putative DREs (Dioxin Response elements) with good similarity scores. Further, we observed the upregulation in E1B expression after the exposure of BaP (a dioxin) and the reduction of E1A expression and their respective protein, i.e. sCD5 and cCD5. The binding of AHR at the predicted DRE sites was confirmed by ChIP qPCR and AHR specific inhibitor and gene silencing studies suggested the involvement of AHR in exonal switch. This study indicates that the polycyclic aromatic hydrocarbon decreases the sCD5 expression by upregulating alternative exon expression, which may adversely affect the overall T cell functions.

Vaccine development: Current trends and technologies.

Despite the effectiveness of vaccination in reducing or eradicating diseases caused by pathogens, there remain certain diseases and emerging infections for which developing effective vaccines is inherently challenging. Additionally, developing vaccines for individuals with compromised immune systems or underlying medical conditions presents significant difficulties. As well as traditional vaccine different methods such as inactivated or live attenuated vaccines, viral vector vaccines, and subunit vaccines, emerging non-viral vaccine technologies, including viral-like particle and nanoparticle vaccines, DNA/RNA vaccines, and rational vaccine design, offer new strategies to address the existing challenges in vaccine development. These advancements have also greatly enhanced our understanding of vaccine immunology, which will guide future vaccine development for a broad range of diseases, including rapidly emerging infectious diseases like COVID-19 and diseases that have historically proven resistant to vaccination. This review provides a comprehensive assessment of emerging non-viral vaccine production methods and their application in addressing the fundamental and current challenges in vaccine development.

HIF-1α regulates the expression of the non-conventional isoform of the cd5 gene in T cells under the hypoxic condition: A potential mechanism for CD5neg/low phenotype of infiltrating cells in solid tumors.

CD5, a T-cell receptor (TCR) negative regulator, is reduced on the surface of CD8+ lymphocytes in the tumor microenvironment (TME). Reduced surface CD5 expression (sCD5) occurs due to the preferential transcription of HERV-E derived exon E1B, i.e., anon-conventional formofthe cd5gene instead of its conventional exon E1A. A tumor employs several mechanisms to evade anti-tumor response, and hypoxia is one such mechanism that prevails in the TME and modulates the infiltrated T lymphocytes. We identified hypoxia response elements (HREs) upstream of E1B. We showed binding of HIF-1α onto these HREs and increased E1B mRNA expression in hypoxic T cells. This results in decreased sCD5 expression and increased cytoplasmic accumulation in T cells. We also validated our study in a solid tumor, i.e., colorectal cancer (CRC) patient samples. This hypoxia-driven mechanism reduces the surface CD5 expression on infiltrated T-cells in solid tumors.

Loss of immune regulation in aged T-cells: A metabolic review to show lack of ability to control responses within the self.

The progressive decline of the anatomical architecture and loss of functional integrity of an individual is aging. Accumulation of degenerative cellular and molecular changes in the aging cells increases the fragility at the cellular and molecular levels. It pushes towards age-associated diseases like Alzheimer's disease, hypertension, cancer, cardiovascular diseases, etc. The impaired T cell function in aging is a leading contributor to increased susceptibility to pathogens, minimized vaccine response, and skewed inflammation. Recent studies about the role of T cells in the remodelling of the immune system have provided ways to examine and explore aging puzzles and their correlation with T cell functions. Here we review the metabolic aspect of T cell function and its possible restoration. IL-7 and mTOR mediated pathways and their association with reactivation of effector T cell function could help understanding the dark side of the compromised adaptive immune system, particularly T cell response, in aging. Understanding these crucial fundamentals could help design and target new molecules to prevent loss of T cell functionality in aging.

Human endogenous retrovirus regulates the initiation and progression of cancers (Review).

The expression of genes is altered in various diseases and is responsible for the disease's initiation, progression and pathology. Several other genes, predominantly inactivated, may become activated in a given condition and contribute to the initiation and progression of the disease. Similarly, human endogenous viruses (HERVs) are an incomplete, non-productive and inactive viral sequence present in the heterochromatin of the human genome, and are often referred to as junk DNA. HERVs were inserted into the host genome millions of years ago. However, they were silenced due to multiple mutations and recombination that occurred over time. However, their expression is increased in cancers due to either epigenetic or transcriptional dysregulation. Some of the HERVs having intact open reading frames have been reported to express virus-like particles, functional peptides and proteins involved in tumorigenesis. To summarize, there is involvement of different HERVs in the initiation and progression of several cancers. The present review aims to provide concise information on HERV and its involvement in the initiation and progression of multiple types of cancer.

Jurkat T Cells are Immunophenotypically Distinct from T-Cell Acute Lymphoblastic Leukemia Cells Due to High-Level Surface Expression of CD5.

Human leukemic T cells show decreased surface CD5 (sCD5) and increased cytoplasmic CD5 (cCD5). When we examined their expressions in the Jurkat T cells, it showed increased sCD5 and decreased cCD5, which is in sharp contrast with the pattern of CD5 expression observed for human leukemic T cells. Furthermore, this opposite pattern was due to the absence of an exonal switch between E1A and E1B. This study suggests that Jurkat cell does not retain all characteristics of T-ALL cells; thus, we should carefully interpret the data obtained using Jurkat T cell as a model cell line of T-ALL.

Retinoic acid restores the levels of cellular cholesterol in Leishmania donovani infected macrophages by increasing npc1 and npc2 expressions.

Visceral leishmaniasis (VL) is a fatal form among all forms of leishmaniasis and is caused by visceralization of the Leishmania donovani (Ld) parasite to the critical organs. Mild to severe malnutrition is common in VL patients and the deficiency of retinoic acid (RA), an important micronutrient, results in a compromised state of immune response in macrophages (mφ) leading to the increased parasite load. In the continuation of our earlier work, we observed loss of cellular cholesterol in infected mφ in the absence of RA i.e., upon inhibition of RALDH pathway. Moreover, the Leishmania utilizes host cholesterol for the establishment of infection and causes a decrease in the expressions of Niemann-Pick C2 (npc2) and Niemann-Pick C1 (npc1) genes involved in the uptake of extracellular cholesterol. This results in reduced levels of cellular cholesterol in infected mφ. Intrigued by this, as the first sign of our hypothesis, we investigated the presence of RA Response Element (RARE) sequences in the upstream of npc1 and npc2 genes. To functionally confirm this, we measured their expressions and the levels of cellular cholesterol in Ld infected mφ in the absence (i.e., using an inhibitor of RALDH pathway) and presence of RA. We found restoration of the levels of cellular cholesterol in infected mφ under the supplementation of RA resulting in the decreased parasite load. Hence, the supplementation of RA with the standard therapy and/or preventive use of RA could be potentially an advancement in the treatment and cure of VL patients.

Indian Morinda species: A review.

Morinda is a largest genus of Rubiaceae family, and its 11 species are found in India. In India, plant species are known by several common names as great morinda, Indian mulberry, noni, beach mulberry and cheese fruit. Various Morinda products (capsules, tablets, skin products and fruit juices) are available in the market, used by people for treatment of several health complaints. A diversity of phytochemicals including iridoids, flavonoids, flavonoid glycosides, anthraquinones, coumarins, lignanas, noniosides, phenolics and triterpenoids have been reported from Morinda species. Morinda species are used in the treatment of inflammation, cancer, diabetes, psyquiatric disorders, and bacterial and viral infections. The noni fruit juice (Morinda citrifolia) and its products are used clinically in the treatment of cancer, hypertension and cervical spondylosis affecting patients. M. citrifolia fuit juice, with different doses, is used in the maintaining blood pressure and reducing of superoxides, HDL and LDL levels. Similarly, oligosaccharide capsules and tablets of root extract of M. officinalis are recommended as medicine for the treatment of kidney problems and sexual dysfunctions of patients. The toxicological studies revealed that higher doses of fruit juice (4,000 or 5,000 mg/kg) of M. citrifolia for 2 or more months cause toxic effects on liver and kidneys. M. officinalis root extracts (ethanolic and aqueous) are found fully safe during treatment of diseases. A large number of reviews are available on M. citrifolia but very few studies are conducted on other Indian Morinda species. This review reports the comprehensive knowledge on state-wise distribution, botany, ethnomedicinal uses, phytochemistry, pharmacological activities, clinical applications and toxicological evaluations of 11 species of Morinda found in India.

Parasitic load determination by differential expressions of 5-lipoxygenase and PGE2 synthases in visceral leishmaniasis.

Infection with L. donovani affects mainly visceral organs. Importantly, the parasitic load differs in different visceral organs; therefore there is a need to understand the organ specific immune regulation, particularly in the spleen and liver. Comparative studies between these organs in Leishmania infected hamster (Mesocricetus auratus) are lacking. Our study highlights the importance of eicosanoids in the organ specific pathology of visceral leishmaniasis. Among other immune cells, macrophages (mφ) which harbor Leishmania parasite are major producers of eicosanoids. In this study, we intend to explore linkage between organ specific immune response and eicosanoids. We suggest that eicosanoids (early immune modulators) and their organ specific expressions, possibly tune the outcome of mφ differently at different sites. We have observed that liver showed better containment of parasitic load than spleen, where we have found higher expression of 5-lipoxygenase (5-LO) enzyme along with IL-12 and iNOS. However, in spleen, enzymes of the PGE2 pathway i.e. PGE2 synthases (cytosolic and microsomal) along with IL-10 were predominantly higher. To further corroborate our findings, in vitro assays were carried out using purified eicosanoids (LTB4 and PGE2) and the inhibitors of these pathways. Findings establish that the 5-lipoxygenase pathway (i.e. LTB4) is anti-parasitic and its inhibition increases the parasitic load (qPCR based kDNA detection). On the contrary, PGES pathway (i.e. PGE2) supports establishment of infection in mφ. Taken together, 5-LO pathway plays a protective role in liver during L. donovani infection. However, the PGES pathway favors the parasite growth, particularly in the spleen at a later stage.

Lymphoepithelioma-like carcinoma of cervix: Cytological Features on Conventional Cervical Smear.

Lymphoepithelioma-like carcinoma (LELC) is a rare neoplasm of the cervix. The importance of distinguishing this undifferentiated carcinoma with a predominant lymphocytic infiltrate lies in the fact that despite being poorly differentiated they have a better prognosis. The diagnosis however becomes more challenging when the pathologist is provided with a small cervical biopsy or a Papanicolaou smear. While the reports describing histology and their relation to Epstein-Barr virus (EBV) are many, there are only few case reports describing the cytology of these tumors. We describe the cytological features of LELC of cervix on conventional smear and correlate it with the histopathological findings of the same. A 67-year-old multiparous Hindu woman presented to the gynecology outpatient department with the history of postmenopausal bleeding for the past six months. The cytological examination of the cervical smear (Papanicolaou stain) was done followed by cervical and endometrial biopsy. Based on Papanicolaou smear and biopsy suggestive of a poorly differentiated carcinoma a radical hysterectomy with pelvic lymphadenectomy was performed. Hysterectomy specimen showed the morphology of LELC and was then correlated with the cervical smears retrospectively. On review of cytological smears it was seen that the tumor cell clusters had an abundant lymphoid background, which was overlooked earlier. Immunohistochemistry for EBV was negative. We conclude that the presence of undifferentiated tumor cell clusters with ill-defined cell borders and large number of lymphoid cells in the background suggest the diagnosis of LELC on cervical cytology. Diagn. Cytopathol. 2017;45:239-242. © 2016 Wiley Periodicals, Inc.

Seroprevalence & changing trends of transfusion-transmitted infections amongst blood donors in a Regional Blood Transfusion Centre in north India.

Background & objectives: Transfusion-transmitted infections (TTIs) are the major problem associated with blood transfusion. Accurate estimates of risk of TTIs are essential for monitoring the safety of blood supply. The present study was undertaken to determine the percentage of voluntary donors (VDs) and replacement donors (RDs) and also, to estimate and compare the seroprevalence and changing trends of TTIs amongst VDs and RDs in a regional blood transfusion centre in north India. Methods: This retrospective study was based on the records of all voluntary and replacement donations which were collected from January 2008 to December 2014 in a Regional Blood Transfusion Centre placed in a tertiary care hospital in Delhi, India. Results: Of the total 220,482 donations, 163,540 (74.17%) were voluntary and 56,942 (25.83%) were replacement donation. The overall seroprevalence of human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), syphilis and malaria were 0.32, 1.61, 0.73, 1.62 and 0.06 per cent, respectively. Furthermore, the TTIs were more frequently encountered in RDs in comparison to VDs. Interpretation & conclusions: The increase in public awareness regarding voluntary blood donation, meticulous donor screening, counselling and use of highly sensitive tests can help in reducing the risk of TTIs.

Flavonoids from Heliotropium subulatum exudate and their evaluation for antioxidant, antineoplastic and cytotoxic activities II.

The flavonoids are the largest group of phenolic compounds isolated from a wide range of higher plants. These compounds work as antimicrobials, anti-insect agents and protect plants from other types of biotic and abiotic stresses. Various researchers have suggested that flavonoids possessed antioxidant, antineoplastic and cytotoxic activities. The main objective of this study was to test dichloromethane fraction of resinous exudate of Heliotropium subulatum for their antioxidant, antineoplastic and cytotoxic activities, as well as to search new antioxidant and antineoplastic agents for pharmaceutical formulations. Five flavonoids were isolated from resinous exudate of this plant species and screened for their in vitro and in vivo antioxidant models (DPPH radical scavenging, reducing power, superoxide anion scavenging, metal chelating scavenging systems, catalase and lipid peroxidation), antineoplastic (Sarcoma 180), and cytotoxic (Chinese hamster V79 cells) activities. Tricetin demonstrated maximum antioxidant activity against both in vitro and in vivo experimental systems while galangin exhibited maximum inhibition (78.35%) at a dose of 10 µg/kg/day against Sarcoma 180. Similarly, it was found that galangin also showed highest activity (21.1 ± 0.15%) at a concentration of 70 µg/ml to Chinese hamster V79 cells. The observed results suggest that tricetin has a potential to scavenge free radicals in both in vitro and in vivo models while the galangin could be considered as antitumor and cytotoxic agent.

Siderophores; iron scavengers: the novel & promising targets for pathogen specific antifungal therapy.

INTRODUCTION: The recent emergence of resistance, toxicity paradigm and limited efficacy of conventional antifungal drugs necessitate the identification of de novo targets in fungal metabolism. One of the most critical physiological processes during in vivo pathogenesis is maintenance of iron homeostasis. The most life threatening opportunistic human fungal pathogens like Aspergillus, Candida and Cryptococcus exploit the siderophore mediated iron uptake mechanism either for survival, virulence, propagation or resistance to oxidative stress envisaged in vivo during infection. Areas covered: In this review, we will highlight the metabolic pathways; specifically siderophore biosynthesis, uptake and utilisation, triggered in the fungal pathogens in iron starving conditions and the various putative targets viable in these pathways to be recruited as novel therapeutic antidotes either via biosynthetic enzymes catalytic site inhibitors or as drug conjugates through trojan horse approach and further role in the development of fungal specific reliable diagnostic markers. Expert opinion: Siderophores are the weapons released by a pathogen to conquer the battle for iron acquisition. Hence, the fungal siderophore biosynthetic pathways along with their uptake and utilisation mechanisms represent an ideal target for pathogen specific, host friendly therapeutic strategy which would block the proliferation of parasite without causing any harm to the mammalian host.

Allergic aspergillosis and the antigens of Aspergillus fumigatus.

Incidence of fungal infections has increased alarmingly in past few decades. Of the fungal pathogens, the Aspergillus fumigatus has been a major cause of allergic bronchopulmonary aspergillosis (ABPA) which has five main stages--the acute, remission, exacerbation, glucocorticoid dependent and fibrotic stage. The diagnosis of ABPA remains difficult due to its overlapping clinical and radiological features with tuberculosis and cystic fibrosis. From past few decades, the crude fractions of A. fumigatus have been used for immunodiagnosis of ABPA. Most of the detection kits based on crude fractions of A. fumigatus are quite sensitive but have low specificity. Till date 21 known and 25 predicted allergens of A. fumigatus have been identified. Of these allergens, only five recombinants (rAsp f1-f4 and f6) are commercially used for diagnosis of allergic aspergillosis. Remaining allergens of A. fumigatus have been restricted for use in specific diagnosis of ABPA, due to sharing of common antigenic epitopes with other allergens. Complete sequencing of A. fumigatus genome identified 9926 genes and the reports on the proteome of A. fumigatus have shown the presence of large number of their corresponding proteins in the pathogen. The analysis of immunoproteomes developed from crude fractions of A. fumigatus by IgG/IgE reactivity with ABPA patients and animal sera have identified the panel of new antigens. A brief description on the current status of A. fumigatus antigens is provided in this review. The implementation of advance recombinant expression and peptidomic approaches on the A. fumigatus antigens may help in the selection of appropriate molecules for the development of tools for more specific early diagnosis of ABPA, and desensitization therapies for patients of allergic disorders.

Anaplastic large cell lymphoma in a patient with systemic onset juvenile arthritis: case report and review of literature.

The association of malignancy with autoimmune rheumatic diseases has been a subject of investigation. It has been shown that there is increased risk of malignancies, mainly non-Hodgkin lymphoma, in patients with autoimmune disorders. There is scarcity of data about malignancy in juvenile idiopathic arthritis (JIA). We report the occurrence of anaplastic large cell lymphoma in a patient with systemic onset juvenile idiopathic arthritis treated with low dose methotrexate (MTX). A relationship between MTX treatment and the occurrence of lymphoma in autoimmune diseases has been suggested. The hypothesis that MTX has a role in the aetiology of lymphoproliferative disorders is supported by the observation of spontaneous remission of lymphoma in few cases on cessation of MTX therapy. However, systemic onset juvenile idiopathic arthritis patients receiving MTX must be periodically examined for the development of lymphoproliferative disorder especially if the disease is difficult to control or patient develop new symptoms on therapy.

ABO blood group discrepancies among blood donors in Regional Blood Transfusion Centre GTB Hospital, Delhi, India.

BACKGROUND: Regional Blood Transfusion Centre (RBTC), GTB Hospital, Delhi is providing safe and quality blood to one third of Delhi population. A discrepancy exists when reactions in forward grouping do not match with reverse grouping or if the previous and current results do not match. AIM: To analyze ABO blood group discrepancies in an algorithmic manner, and to access the incidence and causes of ABO discrepancies among blood donors. MATERIAL AND METHODS: Retrospective data of blood donors with blood group discrepancies was recorded in Regional Blood Transfusion Centre (East) Delhi, during a period of 3years from January 2010 to May 2013. DiaMed-ID Card Micro Typing System using Gel Cards (Cressier sur Morat, Switzerland) were used for determination of the ABO/Rh blood groups combined with reverse grouping. A detailed serological workup of these cases was studied for recognition and resolution of the blood group discrepancy. RESULTS: Total number of donors during the study period were 104,010 (30,120; 31,117; 32,173 and 10,600 respectively). Blood group discrepancies were found in 51 cases (0.04%). There were 30 (58.8%) cases with low avidity anti-B Antibodies, 10 (19.6%) cases with weaker expression or subgroups of A, 2 (3.9%) cases with weaker expression or subgroups of B, 5(9.8%) cases with unexpected alloantibodies (Anti-N and Anti-M, Anti-Lea) and one(1.9%) case of Bombay blood group. In 3 cases, discrepancy could not be resolved and were referred to reference laboratory for confirmation by molecular analysis. The most frequent cause of discrepancy in forward grouping was found to be weak A or B antigen expressions and in reverse grouping decreased anti-B titers was most common. CONCLUSION: All discrepancies reported on ABO cell and serum grouping must be investigated further, so that correct blood group is reported, minimizing the chances of transfusion reaction. A note of caution should be mentioned on the blood group card to prevent ABO incompatibility in case of transfusion.

Coexistent squamous cell carcinoma of the cervix and genital tuberculosis.

BACKGROUND: Tuberculosis and genital malignancies are two common but separate pathologies that rarely coexist. Despite various theories depicting a cause-effect relationship between the two, their association is rarely reported in genital malignancies. CASE REPORT: We report a case of keratinising squamous cell carcinoma of the cervix associated with pelvic granulomatous lymphadenitis, with post-operative flare up of disease responding to anti-tuberculosis treatment. Since coexistence of genital tuberculosis and genital malignancy has been reported rarely, the literature for these cases is reviewed.

Giant subcutaneous synovial sarcoma: an interesting case.

A 45-year-old lady presented to us with a large subcutaneous swelling on her left side of back of 1 year duration. Local examination revealed a 13×12 cm spherical swelling on the left lower paraspinal region. Magnetic resonance imaging of lumbosacral spine revealed a 13×12 cm mass having solid and cystic components with internal septations in the subcutaneous tissue of left posterior paraspinal area. The mass was hetrogenously hyperintense on T 2 weighted images suggestive of synovial sarcoma. Wide local excision of the lump was carried out followed by closure of the defect with a lateral intercostal artery based rotational flap. The histopathology of the excised tumor revealed synovial sarcoma that stained positive for cytokeratin and epithelial membrane antigen. This case highlights an unusually large subcutaneous synovial sarcoma present in para-spinal area.

Proteomic analysis of global changes in protein expression during exposure of gamma radiation in Bacillus sp. HKG 112 isolated from saline soil.

A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a full-system understanding of the radiation stress protection mechanisms of bacteria in different environments.

Nonhealing ulcer--a rare initial presentation of chronic myeloid leukemia diagnosed on aspiration cytology.

Involvement of skin in hematologic malignancies is rare phenomena as majority of the dermatologic lesions in leukemia patients are nonleukemic. In the absence of preceding history of leukemia, the diagnosis is often a surprise. Chronic nonhealing ulcer on the back of a 65-years-old male prompted a fine needle aspiration from its base. Smears showed an unexpected picture comprising of leukocytes in various stages of maturation against a hemorrhagic background. A cytologic diagnosis of leukemia cutis was given. Subsequent hematologic investigations revealed chronic myeloid leukemia in chronic phase. Cutaneous involvement is a rare initial presentation of leukemia in an otherwise asymptomatic patient and also an uncommon cause of nonhealing ulcer. Aspiration cytology scores over biopsy in not only being a rapid and easy diagnostic tool but also allows better appreciation of cellular details like cytoplasmic granules.