A single-cell atlas of the peripheral immune response in patients with influenza A virus infection.

Influenza A virus (IAV) remains a pressing global health concern, yet our understanding of the specific nature and functional roles of certain circulating cell subsets in relation to this viral infection remains unclear. We performed single-cell RNA sequencing (scRNA-seq) on single-cell whole-blood (scWB) isolated from various populations using the Singleron Matrix platform. Our investigation showed a significant upregulation of the IFN-stimulated gene, IFN-α-inducible protein 27 (IFI27), in patients affected by IAV infection and further found that the heightened expression of IFI27 was primarily concentrated in specific immune cell populations, including monocytes and conventional dendritic cells (cDCs). Notably, we identified a specific subset of neutrophils, neutrophil_ISG15, which implicates interferon (IFN) signaling in IAV infection. Our findings provide a comprehensive understanding of the cellular subtypes and molecular characteristics of scWB across different populations with IAV infection.

Pulmonary infection in patients with severe fever with thrombocytopenia syndrome: A multicentre observational study.

Co-infection in patients with severe fever with thrombocytopenia syndrome (SFTS) has been reported, posing a serious threat to survival and treatment. We aimed to systematically investigate the SFTS associated pulmonary infection, particularly invasive pulmonary fungal infection (IPFI). During April 2019 to October 2021, we conducted a multicentre observational study on adult hospitalized patients confirmed with SFTS from three tertiary hospital in central China. Demographic, clinical and laboratory data of patients were collected and re-assessed. A total of 443 patients (51.7% were male sex) were included for analysis with median age of 65-year-old. Among them, 190 (42.9%) patients met the criteria for pulmonary infection. Pulmonary infection was associated with shorter survival time (p < 0.0001 by log-rank test), and adjusted hazard ratio was 1.729 [95% confidence interval, 1.076-2.780] (p = 0.024). Age (odds ratio (OR) 1.040 [1.019-1.062], p < 0.001), time from onset to admission (OR 1.163 [1.070-1.264], p < 0.001), having severe status (OR 3.166 [2.020-4.962], p < 0.001) and symptoms of skin change (OR 2.361 [1.049-5.316], p < 0.001) at admission and receiving intravenous immunoglobin (OR 2.185 [1.337-3.569], p = 0.002) were independent risk factors for the occurrence of pulmonary infection. A total of 70 (15.8%) patients were defined as IPFI. Multivariate analysis showed that time from onset to admission (OR 1.117 [1.016-1.229], p = 0.022), severe status (OR 5.737 [3.054-10.779], p < 0.001), having smoking history (OR 3.178 [1.251-8.070], p = 0.015) and autoimmunity disease (OR 7.855 [1.632-37.796], p = 0.010), receiving intravenous immunoglobin (OR 3.270 [1.424-7.508], p = 0.005) were independent risk factors for the occurrence of IPFI. In SFTS patients with pulmonary infection, white blood count <2.09 × 109 per L (OR 11.064 [3.708-33.012], p < 0.001) and CD3+ CD4+ T cell count <104.0 per µL (OR 10.429 [3.395-32.038], p < 0.001) could independently predict IPFI. This study showed the high prevalence and poor outcomes of pulmonary infection and IPFI in patients with SFTS. These findings highlighted the need for active surveillance of fungal pathogens and early antifungal treatment in patients with SFTS.

Development and validation of a clinical risk score to predict the occurrence of critical illness in hospitalized patients with SFTS.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease with high mortality. Early identification of patients who may advance to critical stages is crucial. This investigation aimed to establish models to predict SFTS before it reaches the critical illness stage. METHODS: Between January 2016 and September 2022, 278 cases have been included in this study. There were 87 demographic and systemic chosen variables. For selecting the predictive variables from the cohort, the LASSO was utilized, and for identifying independent predictors, multivariate logistic regression was performed. Based on these factors, a nomogram was established for critical illness. Concordance index values, decision curve analysis and the area under the curve (AUC) were also examined. RESULTS: Multivariate logistic regression demonstrated the most important differentiating factors as;> 65 years old (P < 0.001, OR 3.388, 95 % CI 1.767-6.696), elevated serum PT (P = 0.011, OR 6.641, 95 % CI 1.584-31.934), elevated serum TT (P = 0.005, OR 3.384, 95 % CI 1.503-8.491), and elevated serum bicarbonate (P = 0.014, OR 0.242, 95 % CI 0.070-0.707). The C-index of the nomogram was 0.812 (95 % CI: 0.754-0.869), representing good discrimination. The model also showed excellent calibration. The AUC of the nomogram established based on four factors, as mentioned earlier, was 0.806. Furthermore, the model had the excellent net benefit, as revealed by the decision curve analysis. CONCLUSION: An accurate risk score system built on manifestations noted in patients with SFTS upon admission to hospital, might be advantageous in managing SFTS.

Impact of Drinking Water Source and Sanitation Facility on Malnutrition Prevalence in Children under Three: A Gender-Disaggregated Analysis Using PDHS 2017-18.

OBJECTIVES: The proposed research studied the determinants of male and female child malnutrition in Pakistan. More specifically, it observed the role of the sanitation facility and drinking water source as important determinants of malnutrition in a gender analysis. METHODS: Novel data of 1010 children under three years of age from PDHS 2017-18 were used. A CIAF (Cumulative Index for Anthropometric Failure) was established to assess malnourishment in the children. Discrete-choice logistic methodology was applied in this empirical research to study the likelihood of malnourishment in children. RESULTS: The logistic regression results depicted that factors such as a child belonging to a deprived area, the status of home wealth, and the education of the mother were common determinants of malnutrition in children. Factors such as a child having diarrhea (OR = 1.55, CI = 0.96-2.50) and the drinking water source (OR = 0.62, CI = 0.37-1.03) were separate prominent predictors of malnutrition in male children whereas the sanitation facility was the main determinant of malnutrition in female children (OR = 0.64, CI = 0.43-0.95). CONCLUSION: This study concludes that important links exist between the drinking water source and male child malnutrition and between sanitation facilities and female child malnutrition.

Role of vitamin C in preventing of COVID-19 infection, progression and severity.

Vitamin C stands as an essential water-soluble vitamin, antioxidant and has been shown to enhance immunity. SARS-CoV-2 has been spreading rapidly across the worldwide, several cellular processes of innate and adaptive immunity are aided by vitamin C, which strengthens the immune system overall. Multiple lines of evidence in the literature associate vitamin C with antioxidant, anti-inflammatory, anticoagulant and immunomodulatory actions. Pneumonia and sepsis patients had poor ascorbic acid status and high oxidative stress, according to many studies. Pneumonia patients who get vitamin C may have less severe symptoms and a longer course of the illness if they do. To standardize plasma levels in sepsis patients, gram measurements of the vitamin must be administered intravenously (IV). This intervention has been shown in a few trials to reduce mortality. COVID-19 management in China and the United States has exhibited remarkable results when using a high percentage of intravenous vitamins C. It's acceptable to include vitamin C in the COVID-19 treatment protocol as a secondary measure based on the current active clinical studies looking at the impact of vitamin C on the management of COVID-19. Patients with hypovitaminosis C or severe respiratory illnesses, such as COVID-19, may benefit from taking vitamin C, due to its good safety profile, simplicity of use, and potential for rapid production scaling. The study's goal was to see whether high dosage intravenous vitamin C had any impact on individuals with severe COVID-19 (HDIVC). Finally we discuss recent research that has been published on the efficacy of vitamin C administration in the treatment of viral infection and life-threatening conditions. The purpose of this manuscript is to summarise existing research on the efficacy of vitamin C as a treatment for COVID-19 and to discuss possible explanations for why it may work in some individuals but not in others.

Occurrence of Diarrheal Disease among Under-Five Children and Associated Sociodemographic and Household Environmental Factors: An Investigation Based on National Family Health Survey-4 in Rural India.

Diarrheal disease is a significant public health problem leading to mortality and morbidity among children aged 0-59 months in rural India. Therefore, the rationale of this study was to identify the sociodemographic and environmental predictors associated with diarrhea among under-five children in rural India. A total of 188,521 living children (0-59 months) were studied from the National Family Health Survey-4, (NFHS-4) 2015-2016. Bivariate and binary logistic regression models were carried out from the available NFHS-4 data for selected sociodemographic and environmental predictors to identify the relationship of occurrence of diarrhea using STATA 13.1. In rural India, children aged 12-23 months, 24-35 months, 36-47 months, and 48-59 months were significantly improbable to suffer diarrheal disease. Children of the female sex, as well as children of scheduled tribes (ST) and other backward classes (OBC), were less likely to experience diarrhea. The disease was more likely to occur among children of scheduled castes (SC); Muslim or other religions; children belonging to central, eastern, and western regions; children with low birth weight; as well as children with improper stool disposal and rudimentary roof materials. In the rural parts of India, sociodemographic and household environmental factors were most influential. Effective community education; improved handwashing practices; pure water supply; and proper waste disposal, including building and utilizing latrines, would help reduce the burden of diarrheal disease in children.

Socio-Economic and Environmental Determinants of Malnutrition in under Three Children: Evidence from PDHS-2018.

OBJECTIVES: This research investigates the association of malnutrition with social and economic factors in general and environmental factors in specific such as sanitation facilities and drinking water sources for Pakistan. METHODS: Authors used the latest data of 1010 Under-Three children from Pakistan Demographic and Health Survey (PDHS) 2017-2018. Cumulative Index of Anthropometric Failure (CIAF) was developed to measure the malnutrition status among children based on z-scores of WHZ, WAZ, and HAZ, respectively. The study has applied the discrete-choice logistic methodology to find the relationship of malnutrition with socio-economic characteristics. The interaction terms of drinking water source and sanitation facility have been measured to see the impact of environmental factors on child malnutrition. RESULTS: The study results depict that the likelihood of malnutrition increases when the child had diarrhea recently and the child belongs to the deprived region such as KPK, Sind, and Baluchistan. However, the chances of child malnutrition drop with (1) an escalation of mothers' education, (2) a rise in wealth status of the household, and (3) the improved water source and sanitation facility in the household. The only water-improved sanitation category of the interaction term is significant in the model which depicts that households having both improved water and improved sanitation facilities had very fewer chances of malnutrition among their children. CONCLUSION: Authors conclude that malnutrition in younger children is associated with improved water as well as sanitation facilities, maternal education, and household wealth in Pakistan.

The AST/ALT Ratio (De Ritis Ratio) Represents an Unfavorable Prognosis in Patients in Early-Stage SFTS: An Observational Cohort Study.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS), a widely prevalent infectious disease caused by severe fever with thrombocytopenia syndrome virus (SFTSV) that carries with it a high mortality rate, has emerged to be a public health concern. This study aimed to investigate the epidemiological and clinical characteristics of patients infected with SFTSV, seeking novel prognostic risk factors for SFTS. METHODS: In this retrospective and cross-sectional study, confirmed SFTS patients from the First Affiliated Hospital of Anhui Medical University were enrolled from September 1, 2019, to December 12, 2020. Cases were analyzed for epidemiological, demographic, clinical, and laboratory data. Logistic regression models were used to assess the association between predictors and outcome variables. A generalized additive mixed model (GAMM) was conducted to analyze the trending shift of aspartate aminotransferase/alanine transaminase-ratio (AST/ALT-ratio) and platelet (PLT) in SFTS patients treated with ribavirin. p values ≤ 0.05 were considered statistically significant. RESULTS: Clinical and laboratory results of 107 hospitalized patients with SFTSV infection were retrospectively described. The mean age at onset of disease was 60.38 ± 11.29 years old and the ratio between male and female was 1:1.2. Fever and thrombocytopenia are hallmark features of SFTS. Furthermore, multiple cases also experienced neurological complications, gastrointestinal/skeletal muscle symptoms together with other non-specific clinical manifestations; laboratory dataset outcomes reported dysregulated levels for routine blood biomarkers, coagulation function, and biochemistry. Overall, 107 patients were segregated into two groups according to patient condition at the clinical endpoint (survivors/non-survivors). SFTS survivors had a higher level of PLT- counts, total protein (TP), and estimated glomerular filtration rate (eGFR), while levels of activated partial thromboplastin time (APTT), thrombin time (TT), D-dimer (D-D), fibrinogen degradation products (FDP), ALT, AST, AST/ALT-ratio, creatinine (Cr), creatine phosphokinase (CK) and procalcitonin (PCT) was higher in non-survivors. Results from univariate Cox regression revealed that elevated levels of FDP, TT, AST/ALT-ratio, PCT, as well as decreased eGFR level and presence of central nervous system symptoms (CNS), were significant predictors for SFTS prognostic, results from multivariate logistic regression analysis in three adjusted models showed AST/ALT-ratio and PCT were independent risk factors for the prognosis of SFTS patients. Kaplan-Meier survival analysis showed that SFTS patients with AST/ALT-ratio >2.683 were associated with a shorter futime (means survival time), therefore indicating an unfavorable prognosis. Treatment with ribavirin could increase PLT count while decreasing AST/ALT-ratio within SFTS patients. CONCLUSION: SFTS is an emerging infectious disease, possibly leading to multiple-organ injury; AST/ALT-ratio was an independent risk factor for the prognosis of SFTS patients. Further investigation should be performed in order to gain more knowledge on this disease and guide clinical management.

Direct enzyme-linked aptamer assay (DELAA) for diagnosis of toxoplasmosis by detection of SAG1 protein in mice and humans.

Toxoplasma gondii is a single-celled parasite commonly found in mammals and birds. Diagnosis of toxoplasmosis largely depends on measurements of the antibody and/or antigen and Toxoplasma DNAs due to the presence of tissue dwelling duplicating tachyzoites, or quiescent cysts in latent infection of the parasite. As a major surface antigen of T.gondii tachyzoites, SAG1 is a key molecule for laboratory diagnosis. However, there are no methods available yet for SAG1 detection using aptamer-based technology. Recombinant SAG1 (r-SAG1) of Toxoplasma WH3 strain (type Chinese 1) was expressed in E.coli and subjected to the synthetic oligonucleotide library for selection of nucleic acid aptamers which target the r-SAG1 antigen, with systematic evolution of ligands by exponential enrichment (SELEX) strategy. The specific aptamers were screened out and used in direct enzyme-linked aptamer assay (DELAA) for detection of native SAG1 (n-SAG1) obtained from tachyzoite lysates, mouse sera of acute infection, and human sera that had been verified for Toxoplasma DNAs by PCR amplification. As results, the soluble r-SAG1 protein was obtained from E.coli lysates by purification and identification with immunoblotting, followed by biotinylation. The selected aptamers were amplified by PCR and DNA sequencing. The results showed that the aptamer-2, with the highest affinity to n-SAG1 in the sera of animals with minimal difference in the four aptamer candidates, has a high specificity and sensitivity when used in detection of n-SAG1 in the sera of humans when compared with the commercial kit of ELISA for T.gondii circulating antigen test. We concluded that a new direct enzyme-linked aptamer assay (DELAA) was developed for the detection of the n-SAG1 protein of T. gondii. With increased sensitivity and specificity, stability, easy and cheap preparation, the aptamer-based technology is considered an efficient method for the diagnosis of active as well as reactivated toxoplasmosis.

The Effects of Financial Stress and Household Socio-Economic Deprivation on the Malnutrition Statuses of Children under Five during the COVID-19 Lockdown in a Marginalized Region of South Punjab, Pakistan.

The lockdown after the COVID-19 pandemic not only caused public health crises and income stress but also put millions at risk of food insecurity and malnutrition across the globe, especially in low and middle-income countries [LMICs]. This study evaluated the effects of financial stress and household socio-economic deprivation on the nutritional status of 1551 children under the age of five during COVID-19 in Pakistan. A self-administered questionnaire was used between November 2020 and April 2021 to collect information on age, height, children's weight, and socio-economic status from 1152 rural households from underdeveloped regions in Punjab, Pakistan. With the help of the proportionate simple random sampling method, this study employed a model (binary logistic regression) to calculate the likelihood of malnourishment. The findings showed that the stunting, underweight, and wasting prevalence rates during COVID-19 were 58.86%, 41.89%, and 8.11%, respectively, in the Bahawalpur region. According to the binary logistic regression analysis, among the household deprivation status (HDS) categories, the risks of childhood malnutrition were lesser in HDS-2 (OR = 0.05, 95% CI: 0. 005-0.879) and HDS-3 (OR = 0.04, 95% CI: 0.008-0.193). Similar to this, within the financial stress index (FSI) categories, the children in homes with medium financial stress had reduced odds of malnutrition (OR = 0.10, 95% CI: 0.018-0.567), and the children in households with low financial stress had reduced risks of malnutrition (OR = 0.006, 95% CI: 0.005-0.061). The proposed research found that stunting and underweight increased by 17.26% and 12.29% during the COVID-19 lockdown in the Bahawalpur region. Additionally, financial stress and socio-economic deprivation strongly affected children's nutritional statuses during the COVID-19 lockdown in the Bahawalpur region of Southern Punjab.

Late-Onset Peripheral T-Cell Lymphoma Not Otherwise Specified in a Liver Transplant Recipient: A Rare Subtype of Posttransplant Lymphoproliferative Disorder.

INTRODUCTION: Posttransplant lymphoproliferative disorder (PTLD) is a rare complication seen in the period after liver transplant. The commonest subtype is B-cell PTLD which is usually associated with Epstein-Barr virus (EBV) infection. T-cell PTLD is rare and the association with EBV is again rarer. CASE: Our patient, a 21-year-old young adult, presented to us with generalized lymphadenopathy, 5 years after liver transplantation. The biopsy of the lymph node was suggestive of peripheral T-cell lymphoma not otherwise specified, which was associated with EBV infection. The Positron emission tomography and computerised tomography (PET-CT) scan showed stage 3 disease. He was treated with standard cyclophosphamide, doxorubicin, etoposide, vincristine, and prednisolone chemotherapy and is currently in remission. CONCLUSION: Peripheral T-cell lymphoma not otherwise specified is a rare subtype of PTLD and its association with EBV is even more rare. A few patients can achieve complete remission with standard chemotherapy.

SARS-CoV-2 microbiome dysbiosis linked disorders and possible probiotics role.

In December 2019, a pneumonia outbreak of unknown etiology was reported which caused panic in Wuhan city of central China, which was later identified as Coronavirus disease (COVID-19) caused by a novel coronavirus, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) by the Chinese Centre for Disease Control and Prevention (CDC) and WHO. To date, the SARS-CoV-2 spread has already become a global pandemic with a considerable death toll. The associated symptoms of the COVID-19 infection varied with increased inflammation as an everyday pathological basis. Among various other symptoms such as fever, cough, lethargy, gastrointestinal (GI) symptoms included diarrhea and IBD with colitis, have been reported. Currently, there is no sole cure for COVID-19, and researchers are actively engaged to search out appropriate treatment and develop a vaccine for its prevention. Antiviral for controlling viral load and corticosteroid therapy for reducing inflammation seems to be inadequate to control the fatality rate. Based on the available related literature, which documented GI symptoms with diarrhea, inflammatory bowel diseases (IBD) with colitis, and increased deaths in the intensive care unit (ICU), conclude that dysbiosis occurs during SARS-COV-2 infection as the gut-lung axis cannot be ignored. As probiotics play a therapeutic role for GI, IBD, colitis, and even in viral infection. So, we assume that the inclusion of studies to investigate gut microbiome and subsequent therapies such as probiotics might help decrease the inflammatory response of viral pathogenesis and respiratory symptoms by strengthening the host immune system, amelioration of gut microbiome, and improvement of gut barrier function.

Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L.

Apoptosis is a process of programmed cell death that is regulated by genes independently. The Bm30kc6 gene is a kind of small molecular lipoprotein about 30 kDa, expressed highly in the late stage of the silkworm hemolymph. Our study showed that overexpression of Bm30kc6 could decrease caspase-3 activation. Meanwhile, activation of caspase-3 increased when Bm30kc6 expression was disturbed by small interfering RNA (siRNA). Cell apoptosis was decreased when Bm30kc6 was overexpressed under UV treatment. The apoptosis rate induced by actinomycin D is similar to the trend by UV. It was inferred that Bm30kc6 has an inhibitory effect on the apoptosis of silkworm cells. The apoptosis-related genes, such as BmFadd, BmDredd, and BmDaxx were increased after overexpression of Bm30kc6 or decreased after interference of siRNA. It was speculated that there was an interactive relationship between Bm30kc6, BmDaxx, BmFadd, and BmDredd in the apoptosis signaling pathways. We investigated the transcription expression of the Bm30kc6 gene in different growth stages and tissues of the silkworm. The results showed that Bm30kc6 reached its peak in the hemolymph during the 6th to 7th days of the 5th instar, or in spinning post 24 h of the silk gland. In the silkworm BmN cells treated with caspase-3/7 inhibitor, the caspase-3 enzyme activity, and the expression levels of Bm30kc6, BmFadd, BmDredd, and BmDaxx were significantly reduced. The expression levels of Bm30kc6 increased sharply when silkworms were treated by molting hormone at Day 3 or 5 of the 5th instar. The results indicated that the expression of the Bm30kc6 gene was affected by the molting hormone and was likely to be its downstream target. In conclusion, the results suggest that the Bm30kc6 gene is involved in the regulation of the apoptotic signaling pathway and plays a role in the apoptotic process.

Burden of care and Suicidal Ideation among Mothers of Children with Autism Spectrum Disorder: Perceived Social Support as a Moderator.

OBJECTIVE: To examine the moderation effect of perceived social support between burden of care and suicidal ideation among mothers of children suffering from Autism Spectrum Disorder.. METHODS: The descriptive cross-sectional study was conducted from September 2017 to February 2018 in the Pakistani cities of Islamabad Lahore, Gujrat, Gujranwala, Jhelum and Sialkot, and comprised of mothers of Autism Spectrum Disorder children aged 7-12 years who were selected conveniently from the special schools in their respective cities. Standardized tools were used to assess the burden of care, perceived social support and suicidal ideation among the participants. SPSS 21 was used for data analysis. RESULTS: Of the 373 mothers approached, 356(95.4%) responded, but the study was completed by 339(95.2%) of them. There was positive relationship (p<0.01) between burden of care and suicidal ideation, and whereas results showed negative relationship among perceived social support, burden of care (p<0.001) and suicidal ideation (p<0.001). The burden of care positively (p<0.001) and perceived social support negatively (p<0.002) affected suicidal ideation. Perceived social support moderated the burden of care-suicidal ideation relationship (p<0.001) among the mothers.. CONCLUSIONS: Social support provided by the significant others lowered the burden of care and suicidal ideation among mothers of children with Autism Spectrum Disorder.

BRAFV600E mutation in hairy cell leukemia: A single-center experience.

BACKGROUND: BRAFV600E mutation has been reported as a unique genetic lesion of hairy cell leukemia (HCL), a subset of which lacks this lesion and shows adverse outcomes. AIMS: To determine the prevalence of BRAFV600E in HCL from our center and derive clinicopathological correlation, if any. MATERIALS AND METHODS: A 9-year retrospective analysis of 46 consecutive cases of HCL diagnosed on morphology and immunophenotyping was done. Stained smears were used as samples for amplification refractory mutation system polymerase-chain reaction using fluorescent primers for mutation detection. RESULTS: BRAFV600E mutation was detected in 41/46 patients (89.1%) while absent in control samples of chronic lymphocytic leukemia. Cases mimicking HCL-variant clinically or immunophenotypically too showed the presence of this mutation. HCL with mutated BRAF presented at a younger age. No statistical difference in blood counts, tumor load, and immunophenotype patterns existed among BRAF mutated and unmutated group. Nine patients (45%) with mutated BRAF had residual disease following treatment with cladribine. CONCLUSION: BRAFV600E mutation analysis has a definitive role in the diagnosis of HCL.

Evaluation of new markers for minimal residual disease monitoring in B-cell precursor acute lymphoblastic leukemia: CD73 and CD86 are the most relevant new markers to increase the efficacy of MRD 2016; 00B: 000-000.

BACKGROUND: Multiparametric flow cytometry (MFC) is a popular technique for minimal residual disease (MRD) analysis. However, its applicability is still limited to 90% of B-cell precursor acute lymphoblastic leukemia (BCPALL) due to two major issues, i.e. a proportion of cases do not express adequate leukemia associated immunophenotype (LAIPs) with currently used markers and drug-induced antigen modulation. Hence, the incorporation of additional reliable markers is required for the further improvement of MFC-based MRD evaluation. We studied the utility of new markers in improvising MFC-based MRD detection in BCPALL. METHODS: Expression-patterns of six new markers, i.e. CD24, CD44, CD72, CD73, CD86, and CD200 were studied in leukemic-blasts from ninety childhood BCPALL patients and in hematogones from 20 uninvolved staging bone marrow (BM) and ten postinduction non-BCPALL BM samples using eight-color MFC. The utility of these new markers in the day 35 postinduction MRD evaluation was determined. RESULTS: Frequencies of LAIPs of CD73, CD86, CD72, CD44, CD200, and CD24 in diagnostic samples were 76.7, 56.7, 55.6, 50, 28.9, and 20%, respectively. Differential expression of all new markers was highly significant (P < 0.01) between early (CD10+ CD19+ CD34+) hematogones, late (CD10+ CD19+ CD34-) hematogones and BCPALL blasts except between early hematogones and BCPALL blasts for CD200 (P = 0.1). In MRD-positive samples, CD73 showed the maximum (83%) frequency of LAIP and CD86 showed the highest (100%) stability of aberrant expression. Inclusion of CD73 and CD86 increased the applicability of MFC-MRD assay to 98.9% MRD samples. CONCLUSION: CD73 and CD86 are the most relevant markers to incorporate in the routine MRD evaluation of BCPALL. © 2016 International Clinical Cytometry Society.

An integrated genomic profile that includes copy number alterations is highly predictive of minimal residual disease status in childhood precursor B-lineage acute lymphoblastic leukemia.

INTRODUCTION: Copy number alterations (CNA) have been described in childhood precursor B-lineage acute lymphoblastic leukemia (B-ALL) which in conjunction with chromosomal abnormalities drive leukemogenesis. There is no consensus on the clinical incorporation of CNA in B-ALL. An integrated genomic classification (IGC) has been proposed which includes CNA and cytogenetics. METHODS: We correlated this IGC with immunophenotypic minimal residual disease (MRD) as well as other standard criteria for 245 patients of B-ALL such as National Cancer Institute (NCI) risk, D+8 prednisolone response, cytogenetics, and ploidy status. RESULTS: MRD was detectable in 81 patients (33.1%). The most common abnormalities were seen in CDKN2A/B (25.7%) followed by PAX5(20%), ETV6(16.7%), IKZF1(15.5%), Rb1(5.3%), BTG (3.3%), EBF1(2.0%), and PAR1(0.8%). On integrating CNA into the IGC, 170 patients (69.4%) were classified into good genomic risk (GEN-GR) whereas 75 (30.6%) belonged to the poor genomic risk (GEN-PR) category. The IGC showed a significant correlation with MRD and NCI risk. The presence of CNA predicted MRD clearance in intermediate cytogenetics group. CONCLUSION: These data seem to indicate that in addition to cytogenetics, CNA should be incorporated into routine clinical testing and risk algorithms for B-ALL. The IGC is of prognostic relevance and offers an additional avenue for prognostication and risk-adapted therapy.

CD19 negative precursor B acute lymphoblastic leukemia (B-ALL)-Immunophenotypic challenges in diagnosis and monitoring: A study of three cases.

BACKGROUND: CD19 is a B-cell specific marker, expressed on all stages of B-lymphocytes including plasma cells. It is widely used in the flow cytometric immunophenotyping (FCI) of B-cell and plasma cell malignancies. The analysis approach of FCI for the diagnosis and monitoring of B-cell acute lymphoblastic leukemia (B-ALL) is totally based on the CD19-based primary gating strategy and it would be challenging to study B-ALL without CD19 expression. Since CD19 negative B-ALL are extremely rare, we report three cases of B-ALL with negative expression of CD19 and discussed its implication in the diagnosis, residual disease monitoring and future targeted therapy. METHODS: Peripheral blood (PB) and bone marrow (BM) samples from three cases suspicious of acute leukemia were studied for morphology, cytochemistry, immunophenotyping and cytogenetics. FCI was performed using a comprehensive six to eight-color multiparametric assay. The cytogenetic studies for standard recurrent genetic translocations were performed by FISH & Karyotyping. RESULTS: The three cases studied were diagnosed as B-ALL based on the expression of CD10, CD20, CD22, CD34, and CD79a by leukemic blasts. CD19 was studied using two different clones (i.e. J3-119 & HIB-19) and found to be severely down regulated in all three cases. There were no significant differentiating features in morphology. Cytogenetic studies were negative for recurrent translocations. CONCLUSION: We report three cases of extremely rare CD19 negative B-ALL. Lack of awareness of negative CD19 expression in B-ALL can leads to incorrect immunophenotypic diagnosis and monitoring of B-ALL, especially in laboratories using limited markers. © 2016 International Clinical Cytometry Society.

Lymphoblastic leukemia with surface light chain restriction: A diagnostic dilemma.

Surface light chain expression is a feature of mature B-cell neoplasms. Light chain restriction in precursor B acute lymphoblastic leukemia is infrequently seen. We report a case of a 28-year-old female with non-FAB L3 morphology blasts and immunophenotypic features showing overlap between a precursor and mature B-cell neoplasm.

Phytoextraction of HG by parsley (Petroselinum crispum) and its growth responses.

The effect of mercury (Hg) on the growth and survival of parsley (Petroselinum crispum) was explored at various treatments. The plants were grown in pots having Hoagland's solution to which various Hg treatments were applied and placed under greenhouse conditions. The treatments were: no metal applied (control) and six doses of Hg as mercuric chloride for 15 days. Linear trend of Hg accumulation was noted in roots, stems, and leaves with increasing Hg treatments. The maximum Hg concentration in root, stem and leaf was 8.92, 8.27, and 7.88 at Hg treatments of 25 mg l(-1), respectively. On the whole, Hg accumulation in different plant parts was in the following order: leaves > stem > roots. Linear trend was also observed for Bioaccumulation Factor (BF) and Translocation Factor (TF) with increasing Hg concentrations in the growth medium. The highest respective BFHg and TFHg values were 9.32 and 2.02 for the Hg treatments of 25 and 50 mg l(-1). In spite of the reduced growth in the presence of Hg, the plant has phytoremediation potential. It is recommended that parsley should not be cultivated in Hg contaminated sites in order to avoid dietary toxicity.