A Well-Curated Cost-Effective Next-Generation Sequencing Panel Identifies a Diverse Landscape of Pathogenic and Novel Germline Variants in a Bone Marrow Failure Cohort in a Resource-Constraint Setting.

The current study is a 4-year experience in diagnosis and screening of inherited and immune bone marrow failure cases using a targeted sequencing panel. A total of 171 cases underwent targeted next-generation sequencing and were categorized as suspected inherited bone marrow failure syndrome (IBMFS) group (106; 62%) and immune/idiopathic aplastic anemia (IAA) group (65; 38%) based on clinical and laboratory criteria. A total of 110 (64%) were pediatric (aged 0 to 12 years) patients and 61 (36%) were adolescent and adult (aged 13 to 47 years) patients. In suspected IBMFS group, 47 (44%), and in IAA group, 8 (12%) revealed a likely germline pathogenic variation. Whole-exome sequencing performed in 15 of 59 suspected IBMFS group cases was negative on targeted panel, and revealed a clinically important variation in 3 (20%) cases. A total of 11 novel variants were identified. The targeted panel helped establish a diagnosis in 44% (27/61) of unclassified bone marrow failure syndrome cases and led to amendment of clinical diagnosis in 5 (4.7%) cases. Overall, diagnostic yield of this well-curated small panel was comparable to Western studies with larger gene panels. Moreover, this was achievable at a much lower cost, making it suitable for resource-constraint settings. In addition, high frequency (>10%) of cryptic pathogenic IBMFS gene variations in IAA cohort suggests routine incorporation of targeted next-generation sequencing screening in these cases.

Recent updates on green synthesis of lignin nanoparticle and its potential applications in modern biotechnology.

Lignin is a complex of organic polymers that are abundantly present in the plant cell wall which considered of emerging substrates for various kinds of value-added industrial products. Lignin has potential use for the production of green nanomaterials, which exhibit improved or different properties corresponding to their parent polymers. Nano lignin has received significant interest in recent years due to its applications in numerous fields. Lignin, the abundant and limited functionality has challenges for its potential uses. Creating advanced functional lignin-derived material like lignin nanoparticles (LNPs) which significantly alter the biological process has great potential for its applications. In the fields of biotechnology, several lignin extraction processes from various raw materials and diverse synthesis techniques, including acid precipitation, dialysis, solvent shifting/solvent exchange, antisolvent precipitation, homogenization, water-in-oil (W/O) microemulsion, ultra-sonication, interfacial crosslinking, polymerization, and biological pathway can be employed to produce LNPs. The scientific community has recently become more concerned about the transformation of lignin to lignin nanomaterials, including nanoparticles, nanocapsules, nanofibers, nanotubes, and nanofilms. Recent research has shown that lignin nanoparticles (LNPs) are: non-toxic at adequate amounts (both in vitro and in vivo), are economical, and can be biodegradable by bacteria and fungi. In promising studies, LNPs have been investigated for their potential applications in gene delivery systems, drug carriers, biocatalysts, tissue engineering, heavy metal absorbers, encapsulation of molecules, supercapacitors, hybrid nanocomposites, and other applications. This current review addresses the recent advances in the synthesis of LNPs, their advanced application in different areas, future perspectives, and challenges associated with lignin-based nanomaterials.

Gingival hyperplasia: An initial oral manifestation of acute myeloid leukemia.

Various systemic diseases can manifest oral signs and symptoms early, which may be crucial for diagnosis and outlining the treatment plan. This case report highlights the presentation of acute leukemia (a malignancy of white blood cells) in a young female. An 11-year-old girl presented with gingival overgrowth and bleeding from the gingiva, weakness, and recent history of weight loss. A detailed workup consisting of complete blood count, bone marrow examination, flow cytometric immunophenotyping, cytogenetics, and molecular studies were carried out. The investigations confirmed the infiltration of blast cells of myelomonocytic origin, and a confirmatory diagnosis of acute myeloid leukemia (French-American-British classification M5) was made. The patient was put on induction chemotherapy and responded well. She developed febrile neutropenia following chemotherapy, which was managed conservatively. Gingival overgrowth subsided after the chemotherapy, and at the time of discharge, she was asymptomatic and hemodynamically stable. The oral health-care professionals must recognize that gingival overgrowth/enlargement may represent an initial manifestation of an underlying systematic disease.

EVI1 upregulates PTGS1 (COX1) and decreases the action of tyrosine kinase inhibitors (TKIs) in chronic myeloid leukemia cells.

Tyrosine kinase inhibitors (TKIs) are highly effective in treating chronic myelogenous leukemia (CML). However, primary and acquired drug resistance to TKIs have been reported. In this study, we used RNA sequencing followed by RQ-PCR to show that the proto-oncogene EVI1 targets the drug-metabolizing gene PTGS1 in CML. The PTGS1 promoter element had an EVI1 binding site, and CHIP assay confirmed its presence. Data from a publicly available CML microarray dataset and an independent set of CML samples showed a significant positive correlation between EVI1 and PTGS1 expression in CML. Downregulation of EVI1 in K562 cells and subsequent treatment with TKIs resulted in a lower IC50 in the control cells. Furthermore, combined inhibition of BCR-ABL with imatinib and PTGS1 with FR122047 (PTGS1 inhibitor) synergistically reduced the viability of imatinib-resistant K562 cells. We conclude that elevated EVI1 expression contributes to TKIs resistance and that combined inhibition of PTGS1 and BCR-ABL may represent a novel therapeutic approach.

Usefulness of Leucocyte Cell Population Data by Sysmex XN1000 Hematology Analyzer in Rapid Identification of Acute Leukemia.

Leukocyte cell population data (CPD) generated by hematology auto analyzers are reported to be useful in screening of sepsis patients. However, there is a paucity of literature highlighting the utility of CPD in screening of acute leukemias (AL). Leucocyte CPD obtained by Sysmex XN1000 hematology analyzer from 210 cases of ALs [22 acute promyelocytic leukemia (APL), 79 non-APL acute myeloid leukemia (non-APL-AML) and 109 acute lymphoblastic leukemia (ALL)] were compared with 100 healthy and 52 reactive controls. Receiver operator curves were drawn to determine the cut-off values of individual parameters. The regression equations combining the best parameters were then formulated to calculate a cut-off value for discrimination among AL subgroups and controls. Acute leukemias showed significant differences (p < 0.05) in various CPD parameters compared to control subjects. A combination of best CPD parameters discriminated ALs from healthy controls (cut off; 0.443, sensitivity of 94% and specificity of 91%), ALs from reactive controls (cut off; 0.576, sensitivity; 97%, specificity; 92%), APL from non-APL-AML (cut off; 0.174, sensitivity of 91% and specificity of 67%), and AML from ALL (cut off; 1.338, sensitivity; 86.1%, specificity; 75%). The CPD from Sysmex XN 1000 analyzer could be a useful tool in screening and lineage characterization of acute leukemias; particularly at centers where high-end technical expertise is still not available. Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-021-01488-9.

Clinical review of COVID-19 in children and adolescents with cancer: Experience from a tertiary care center in East India.

Children with underlying cancer are often immunocompromised. Data on severity of coronavirus disease 2019 (COVID-19) in children with cancer and its outcomes is emerging. Treatment protocols of specific cancers are decided based on the infrastructure, availability of supportive-care, and logistic issues of the locality. The purpose of the study was clinical analysis of COVID-19 in children and adolescents with cancer. The retrospective observational study was conducted at a tertiary healthcare-center in East India. Children and adolescents (aged 0-19 years) with cancer and under treatment with reverse-transcriptase-polymerase-chain-reaction (RT-PCR) confirmed COVID-19 between 5-July-2020 and 5-December-2020 were studied. Median age of the 68 identified patients was six years. Acute leukemia was the most common (66%) diagnosis. COVID-19 was asymptomatic/mildly symptomatic in 91% and moderate to severe in only 9% of patients. Fever (87%) was the commonest symptom, followed-by cough/coryza (75%). Three patients with acute myeloid leukemia (AML) and severe/critical COVID-19 and associated neutropenic sepsis were required transfer to the intensive-care-unit (ICU) for management. Three (4.4%) patients succumbed with COVID-19. Delay in treatment was observed in 63.2% of patients, and the median duration of delay was 28 days after acquiring COVID-19. Median time to attain negative COVID-19 RT-PCR was 16 days, and eight patients were repeat positives. While pediatric and adolescent cancer patients on active treatment may have a higher risk of mortality from severe COVID-19 than their healthy counterparts, the risk may be much lower than deemed. It is essential to continue cancer therapy in these children. Delay in treatment remains a concern.

Primary gastrointestinal anaplastic large cell lymphoma: A critical reappraisal with a systematic review of the world literature.

Anaplastic large cell lymphoma (ALCL) is a distinct T-cell non-Hodgkin lymphoma involving both nodal and extra-nodal sites with a specific anaplastic lymphoma kinase 1 (ALK-1) gene rearrangement. The commonly involved extranodal sites include skin, bone, soft tissue, lungs, and liver. ALCL primarily involving gastrointestinal (GI) tract is rare. In this manuscript, we describe a case of primary esophageal ALK1 positive-ALCL (null phenotype) in a young female, who presented with fleshy mucosal lesion in the lower third of the esophagus and present a systematic review of 35 cases of GI-ALCL reported in the English literature over the past 28 years (1990-2018) with regard to the clinicopathological characteristics, therapy, and outcome.

Pulmonary anaplastic large-cell lymphoma: A case-based systematic review of world literature.

We describe a case of ALK1 negative (-) pulmonary anaplastic large-cell lymphoma (pALCL) in an adult female with an unfavorable outcome following combination chemotherapy and present a systematic review of 39 such sporadic cases reported over the past 28 years (1990-2018). pALCL occurred in 26 males and 13 females (median age, 43 years [5-81]) and 13/39 (33.33%) were ≤18 years. The lesions were endobronchial in 21 (53.85%) and parenchymal in 18 (46.15%) cases. Twenty-six cases were ALK1-; 13 were ALK1+ (positive); and 27/34 cases had a T cell phenotype (where tested). ALK- cases were characterized by higher age (P = 0.012) at presentation, more B symptoms (P = 0.002), and more parenchymal than endobronchial lesions (P = 0.039). The median survival (N = 29/39) was 60 months; pediatric group had a better survival than adult/elderly group (log-rank, P = 0.026). pALCL is rare and may have a distinct biological behavior.

Bone marrow trephine immunohistochemistry is useful in characterizing malignancy-associated myelonecrosis: A retrospective observational study.

OBJECTIVE: We aim to describe the utility of immunohistochemistry (IHC) in characterizing malignancy-associated myelonecrosis (MN) on bone marrow trephine biopsies (BMBx) as a part of initial workup. MATERIALS AND METHODS: Patten and intensity of antigenic immunoexpression in necrotic tumor cells on BMBx were evaluated in a series of cases using standardized avidin-biotin-complex immunoperoxidase technique after heat-induced epitope retrieval and compared the same with viable tumor cells wherever available. RESULTS: Fifteen out of 2494 (0.6%) cases (median age: 28 years; range: 4 to 66 years) had evidence of MN (extensive in eight, moderate in five, and focal in two) secondary to hematological (N = 9) and solid (N = 6) malignancies. Five (33.3%) had pancytopenia, and eight (53.3%) had difficult and/or hemodiluted aspirate. Antigenic expression for CD10, CD79a, CD3, CD7, and CD20 was retained by necrotic leukemic blasts or lymphoma cells; CD34, TdT, and PAX5 showed heterogeneous expression; and a weak Golgi zone (dot like) CD30 positivity was noted in Reed-Sternberg (RS) or RS-like giant cells. Necrotic epithelial metastases retained pancytokeratin in all and showed variable positivity for prostate-specific antigen, carcinoembryonic antigen, CK20, ER, PR, and GATA3. Necrotic neuroblastomas (N = 2) retained positivity for synaptophysin and chromogranin, whereas retained nuclear positivity for NKX2.2 in necrotic Ewing family of tumor (N = 1) aided in early diagnosis. CONCLUSION: Myelonecrosis may retain tumor antigenicity, and immunohistochemistry using selected panel of antibodies should be tried in such challenging cases for an early presumptive diagnosis and further decision making.

Microbial cellulases - An update towards its surface chemistry, genetic engineering and recovery for its biotechnological potential.

The inherent resistance of lignocellulosic biomass makes it impervious for industrially important enzymes such as cellulases to hydrolyze cellulose. Further, the competitive absorption behavior of lignin and hemicellulose for cellulases, due to their electron-rich surfaces augments the inappropriate utilization of these enzymes. Hence, modification of the surface charge of the cellulases to reduce its non-specific binding to lignin and enhance its affinity for cellulose is an urgent necessity. Further, maintaining the stability of cellulases by the preservation of their secondary structures using immobilization techniques will also play an integral role in its industrial production. In silico approaches for increasing the catalytic activity of cellulase enzymes is also significant along with a range of substrate specificity. In addition, enhanced productivity of cellulases by tailoring the related genes through the process of genetic engineering and higher cellulase recovery after saccharification seems to be promising areas for efficient and large-scale enzyme production concepts.

Loss and Reappearance of A Antigen After Chemotherapy Leading to Blood Group Discrepancy in Acute Myeloid Leukemia: A Case Report.

A male patient aged 11 years diagnosed with acute myeloid leukemia presented with complaints of fever, lethargy, and bleeding manifestations. On ordering red blood cells and platelet transfusion, his blood group was tested. Blood group discrepancy was observed in that forward grouping showed the O Rh D positive blood group and reverse grouping revealed the A Rh D positive. The patient's previous blood group record was O Rh D positive, and he had a transfusion history of O Rh D positive red blood cells and platelets in other hospital. Initial immunohematological workup results, including adsorption and heat elution, were consistent with the O Rh D-positive blood group, but further workups on follow-up after the commencement of chemotherapy showed that his original blood group was A Rh D positive, in which the A antigen expression was previously masked by the underlying disease condition of the patient. Hence, the correlation of laboratory results with clinical details and case history is an essential step in resolving such blood group discrepancies.

Development of co-culture yeast fermentation for efficient production of biobutanol from rice straw: A useful insight in valorization of agro industrial residues.

Escalating environmental concerns and petroleum demands leads into the present study. In this investigation delignification of rice straw was optimized by NaOH and H2SO4 pretreatment using L16 Taguchi orthogonal array. NaOH pretreatment revealed higher delignification as compared to H2SO4 and; further subjected to separate enzymatic hydrolysis and co-fermentation (SHCF) using RSM as the SHCF demonstrated a maximum glucose and xylose yield of 575 and 205 mg/g. Further, butanol concentration of 4.32 g/L was achieved from 20 g/L of sugar loadings by co-culture of Saccharomyces cerevisiae and Pichia sp. at 72 h of incubation time which was 79.25% higher as compared to monocultures of Pichia sp. Scale-up experiments with higher sugar loadings (90 g/L) demonstrated a butanol concentration of 13.3 g/L. The release of amino acids in co-culture and monoculture systems demonstrated that the addition of S. cerevisiae promoted the butanol synthesis pathway which led to higher butanol concentration.

Partial consolidated bioprocessing of pretreated Pennisetum sp. by anaerobic thermophiles for enhanced bioethanol production.

Rapid industrialization and consumption of fossil fuels have led to considerable progress in the production of renewable biofuels like bioethanol. Lignocellulosic biomass such as grasses serves as cheap feedstocks for the production of bioethanol. However, the process involved in lignocellulosic bioethanol production is expensive which restricts its industrial production. The present study thus attempted to investigate a partially consolidated bioprocessing (PCB) approach using two isolated anaerobic thermophiles i.e. Bacillus paranthracis and Bacillus nitratireducens for direct conversion of ultra-sonication assisted sodium hydroxide (UA-NaOH) pretreated Denannath grass to bioethanol in co-culture consortium batch fermentation experiments. The process parameters for the PCB approach were optimized using the Box-Behnken design of Response Surface Methodology (RSM). The parameters that were considered were substrate concentration (5-10 g), incubation time (30-66 h), inoculum volume [1:1 to 3:3 (% v/v) and temperature (50-65 °C). The maximum ethanol concentration of 8.46 mM (0.39 g/L from 7.5 g/L of substrate loading) and ethanol yield (Yp/s) of 0.55 g/g of reducing sugar was obtained at 57.5 °C. In the same conditions the cellulase and xylanase activities were 0.8 U/mL and 11.53 U/mL respectively, while the lactate and acetate concentrations were 0.2 mM (0.009 g/L) and 2.9 mM (0.13 g/L) correspondingly. An increase in the substrate loadings to 250 g/L in a batch fermenter (3 L) resulted in the production of 373.35 mM (17.1 g/L) of ethanol concentration and Yp/s of 0.16 g/g of reducing sugar.

Engineering grass biomass for sustainable and enhanced bioethanol production.

MAIN CONCLUSION: Bioethanol from lignocellulosic biomass is a promising step for the future energy requirements. Grass is a potential lignocellulosic biomass which can be utilised for biorefinery-based bioethanol production. Grass biomass is a suitable feedstock for bioethanol production due to its all the year around production, requirement of less fertile land and noninterference with food system. However, the processes involved, i.e. pretreatment, enzymatic hydrolysis and fermentation for bioethanol production from grass biomass, are both time consuming and costly. Developing the grass biomass in planta for enhanced bioethanol production is a promising step for maximum utilisation of this valuable feedstock and, thus, is the focus of the present review. Modern breeding techniques and transgenic processes are attractive methods which can be utilised for development of the feedstock. However, the outcomes are not always predictable and the time period required for obtaining a robust variety is generation dependent. Sophisticated genome editing technologies such as synthetic genetic circuits (SGC) or clustered regularly interspaced short palindromic repeats (CRISPR) systems are advantageous for induction of desired traits/heritable mutations in a foreseeable genome location in the 1st mutant generation. Although, its application in grass biomass for bioethanol is limited, these sophisticated techniques are anticipated to exhibit more flexibility in engineering the expression pattern for qualitative and qualitative traits. Nevertheless, the fundamentals rendered by the genetics of the transgenic crops will remain the basis of such developments for obtaining biorefinery-based bioethanol concepts from grass biomass. Grasses which are abundant and widespread in nature epitomise attractive lignocellulosic feedstocks for bioethanol production. The complexity offered by the grass cell wall in terms of lignin recalcitrance and its binding to polysaccharides forms a barricade for its commercialization as a biofuel feedstock. Inspired by the possibilities for rewiring the genetic makeup of grass biomass for reduced lignin and lignin-polysaccharide linkages along with increase in carbohydrates, innovative approaches for in planta modifications are forging ahead. In this review, we highlight the progress made in the field of transgenic grasses for bioethanol production and focus our understanding on improvements of simple breeding techniques and post-harvest techniques for development in shortening of lignin-carbohydrate and carbohydrate-carbohydrate linkages. Further, we discuss about the designer lignins which are aimed for qualitable lignins and also emphasise on remodelling of polysaccharides and mixed-linkage glucans for enhancing carbohydrate content and in planta saccharification efficiency. As a final point, we discuss the role of synthetic genetic circuits and CRISPR systems in targeted improvement of cell wall components without compromising the plant growth and health. It is anticipated that this review can provide a rational approach towards a better understanding of application of in planta genetic engineering aspects for designing synthetic genetic circuits which can promote grass feedstocks for biorefinery-based bioethanol concepts.

Enhanced reducing sugar production by saccharification of lignocellulosic biomass, Pennisetum species through cellulase from a newly isolated Aspergillus fumigatus.

A cellulose degrading fungus Aspergillus fumigatus (CWSF-7) isolated from decomposed lignocellulosic waste containing soil was found to produce high titer of cellulases. The optimum activity of CMCase and FPase were 1.9 U/mL and 0.9 U/mL respectively while the highest protein concentration was found to be 1.2 mg/mL. Saccharification of two Pennisetum grass varieties [dennanath (DG) and hybrid napier grass (HNG)] were optimized using partially purified CMCase and FPase in equal concentration, i.e. a ratios of 1:1 and further with addition of commercial xylanase using response surface methodology (RSM). The production of total reducing sugar (TRS) using isolated cellulase were 396.6 and 355.8 (mg/g), whereas further addition of xylanase had higher TRS titers of 478.7 and 483.3 (mg/g) for DG and HNG respectively as evident from HPLC analysis. Further, characterization of the enzyme saccharified DG and HNG by SEM and ATR-FTIR revealed efficient hydrolysis of cellulose and partially hydrolysis of hemicellulose.

Gravitational radiation in quantum gravity.

The effective field theory of quantum gravity generically predicts non-locality to be present in the effective action, which results from the low-energy propagation of gravitons and massless matter. Working to second order in gravitational curvature, we reconsider the effects of quantum gravity on the gravitational radiation emitted from a binary system. In particular, we calculate for the first time the leading order quantum gravitational correction to the classical quadrupole radiation formula which appears at second order in Newton's constant.

Physicochemical characterization, modelling and optimization of ultrasono-assisted acid pretreatment of two Pennisetum sp. using Taguchi and artificial neural networking for enhanced delignification.

Acid as well as ultrasono-assisted acid pretreatment of lignocellulosic biomass of two Pennisetum sp.; Denanath grass (DG) and Hybrid Napier grass (HNG) have been investigated for enhanced delignification and maximum exposure of cellulose for production of bioethanol. Screening of pretreatment with different acids such as H2SO4, HCl, H3PO4 and H2NO3 were optimized for different temperature, soaking time and acid concentrations using Taguchi orthogonal array and the data obtained were statistically validated using artificial neural networking. HCl was found to be the most effective acid for pretreatment of both the Pennisetum sp. The optimized conditions of HCl pretreatment were acid concentration of 1% and 1.5%, soaking time 130 and 50 min and temperature 121 °C and 110 °C which yielded maximum delignification of 33.0% and 33.8% for DG and HNG respectively. Further ultrosono-assisted HCl pretreatment with a power supply of 100 W, temperature of 353 K, and duty cycle of 70% has resulted in significantly higher delignification of 80.4% and 82.1% for both DG and HNG respectively than that of acid pretreatment. Investigation using SEM, FTIR and autofloresence microscopy for both acid and ultrasono-assisted acid pretreatment lignocellulosic biomass revealed conformational changes of pretreated lignocellulosic biomass with decreased lignin content and increased exposure of cellulose, with greater effectiveness in case of ultrasono assisted acid pretreatment condition.

Is there an increased risk of metabolic syndrome among childhood acute lymphoblastic leukemia survivors? A developing country experience.

Data on metabolic syndrome (MS) in survivors of childhood acute lymphoblastic leukemia (ALL) from developing countries are lacking. The purpose of this single-center, uncontrolled, observational study was to assess the frequency of MS in our survivors. The survivors of ALL ≤15 years at diagnosis, who had completed therapy ≥2 years earlier, were enrolled. Anthropometric measurements (weight, height, waist circumference), biochemistry (glucose, insulin, triglycerides, high-density lipoprotein [HDL], thyroid function tests, C-reactive protein [CRP], magnesium), measurement of blood pressure, and Tanner staging were performed. MS was defined by International Diabetes Federation (IDF) and the National Cholesterol Education Program Third Adult Treatment Panel guidelines (NCEP ATP III) criteria, modified by Cook et al. (Arch Pediatr Adolesc Med. 2003;157:821-827) and Ford et al. (Diabetes Care. 2005;28:878-881). The median age of 76 survivors was 11.9 years (interquartile range [IQR]: 9.6-13.5). Twenty-four (32%) survivors were obese or overweight. The prevalence of insulin resistance (17%), hypertension (7%), hypertriglyceridemia (20%), and low HDL (37%) was comparable to the prevalence in children/adolescents in historical population-based studies from India. The prevalence of MS ranged from 1.3% to 5.2%, as per different defining criteria. Cranial radiotherapy, age at diagnosis, sex, or socioeconomic status were not risk factors for MS. The prevalence of MS in survivors of childhood ALL, at a median duration of 3 years from completion of chemotherapy, was comparable to the reference population. The prevalence of being obese or overweight was, however, greater than historical controls.