Molecular monitoring of treatment efficacy in human visceral leishmaniasis.

BACKGROUND: Focused efforts of the visceral leishmaniasis elimination program have led to a drastic decline in cases, and the present challenge is disease monitoring, which this study aimed to assess. METHODS: A Leishmania kinetoplastid-targeted qPCR quantified parasite load at disease presentation, and following treatment completion (n=49); an additional 80 cases were monitored after completion of treatment. RESULTS: The parasite load at disease presentation was 13 461.00 (2560.00-37764.00)/µg gDNA, which upon completion of treatment reduced in 47 of 49 cases to 1(1-1)/µg gDNA, p<0.0001. In 80 cases that presented >2 months post-treatment, their parasite burden similarly decreased to 1(1-1)/µg gDNA except in 6 of 80 cases, which were qPCR positive. CONCLUSION: In 129 cases of visceral leishmaniasis, qPCR by quantification of parasite burden proved effective for monitoring treatment.

Bone mineral density, vertebral fractures and trabecular bone score in primary ovarian insufficiency.

PURPOSE: Bone health in primary ovarian insufficiency (POI) is under-investigated. We assessed patients with spontaneous POI for vertebral fractures (VFs) and related parameters of bone health. METHODS: 70 cases with spontaneous POI (age 32.5 ± 7.0 years) and an equal number of controls were assessed for BMD, TBS, and VFs. BMD at the lumbar-spine (L1-L4), left hip, non-dominant forearm, and TBS (iNsight software) were measured on a dual-energy X-ray absorptiometry (DXA) machine. VFs were assessed by Genant's classification. Serum FSH, LH, estradiol, T4, TSH, iPTH, serum 25(OH)D, total calcium, and inorganic phosphorus were measured. RESULTS: BMD at the lumbar-spine, hip and forearm was reduced by 11.5%, 11.4% and 9.1% in POI as compared to controls (P < 0.001). Degraded or partially degraded microarchitecture on TBS was observed in 66.7% of patients and 38.2% of controls (P = 0.001). 15.7% of the POI patients had VFs, compared to 4.3% of controls (P = 0.045). Age, duration of amenorrhea and duration of HRT use were the significant predictors of TBS (P < 0.01). Serum 25(OH)D was the significant determinant of VFs. TBS abnormalities were higher in patients with POI and VFs. BMD was not significantly different in patients with and without VFs. CONCLUSION: Thus, lumbar-spine osteoporosis, impaired TBS and VFs were present in 35.7%, 66.7% and 15.7% of patients with spontaneous POI in their early third decade. This indicates need for rigorous investigations for impaired bone health in these young patients and management with HRT, vitamin-D, and possible need for bisphosphonate therapy.

Vertebral fractures, trabecular bone score and their determinants in chronic hypoparathyroidism.

PURPOSE: Patients with hypoparathyroidism are at risk of vertebral fractures (VFs) despite high bone mineral density (BMD). We investigated this paradox by assessing trabecular bone score (TBS) and hip structural analysis (HSA) in non-surgical chronic hypoparathyroidism (cHypoPT) with and without VFs. METHODS: 152 cHypoPT patients (age 40.2 ± 13.4 years, M: F = 81:71) with a median follow-up of 8 (2-13) years were assessed for BMD, VFs, TBS, and HSA and compared with 152 healthy controls. VFs at T7-L4 were assessed by Genant's method. Average serum total calcium and phosphorus during follow-up were assessed. RESULTS: The lumbar spine and hip BMD were higher by 25.4 and 13.4% in cHypoPT than controls (P < 0.001). Paradoxically, VFs (30.9 vs.7.9%), including multiple (12.5 vs. 2.6%) were higher in cHypoPT (P < 0.001). Though overall average TBS (1.411 ± 0.091) was normal in cHypoPT, 25.4% of the females had subnormal TBS, more in post than pre-menopausal women (52.3 vs. 14%, P = 0.002) and as compared to males (6.1%, P = 0.001). TBS correlated with menopausal status and follow-up serum calcium-phosphorus product. For every gm/cm2 rise in BMD, TBS increase was only 0.227 in cHypoPT compared to 0.513 in controls. Frequency of VFs increased with declining TBS (P = 0.004). HSA was comparable between cHypoPT with and without VFs. 23.4% of cHypoPT with VFs had subnormal TBS. CONCLUSION: 31% of cHypoPT patients had VFs. TBS indicated degraded bone microarchitecture in 50% of the post-menopausal cHypoPT women. However, TBS has limitations to detect abnormal bone microarchitecture in cHypoPT as only one-fourth of patients with VFs showed low TBS.

Response of modified microclimates on growth, yield, and incidence of rhizome rot disease of ginger in Assam, India.

Field experiments were conducted at Biswanath, Assam, India (26° 42' N and 93° 15' E), during 2016, 2017, and 2018, to evaluate the effect of microclimates on growth, yield, and disease incidence in the ginger crop. The ginger variety Nadia was grown under six microclimates, viz., under shade net for the entire crop season (T1), under shade net from planting to mid-October (T2), with pigeon pea (T3), with maize (T4), with okra (T5), and as a sole crop (T6) in three replicated RBD. Photosynthetically active radiation (PAR), net radiation (Rn), temperature above the ginger canopy, soil temperature, and soil moisture were measured during the critical crop growth period under different microclimates. Recording of rhizome rot disease incidence was done periodically and genomic analysis of pathogen was carried out. PAR recorded above the ginger canopy under T6 was 1688.1 µ mol s-1 m-2, which was attenuated up to 80.1% in other microclimates. The Rn load of the ginger canopy was maximum (446.4 W m-2) under T6, which reduced to below 50 W m-2 under both T3 and T4. Both air temperatures above the ginger canopy and soil temperatures under T3 and T4 were reduced by 3.3 °C and 4.6 °C, respectively, as compared to T6. The pathogen causing the disease in the experimental site was identified as Fusarium oxysporum. Considerable increase in soil and air temperature and soil moisture favored disease incidence (90.3%) under shade net (T1 and T2) treatments, while opposite reason causing significant reduction in disease incidence (16.1%) was observed under T3 and T4. More yield of ginger recorded in treatments T3 (6.21 t ha-1) or T4 (6.48 t ha-1) was attributed to better crop growth and diminutive disease incidence, while the crop was almost damaged due to severe disease incidence under shade net (T1 and T2) treatments.

Bioremediation of heavy metals from wastewater: a current perspective on microalgae-based future.

Heavy metals-containing drinking water and wastewater are posing a severe threat to the environment, and living beings on land, air and water. Different conventional, advanced nanomaterials-based and biological method has been employed for the treatment of heavy metals. Among the biological methods, microalgae are an important group of micro-organisms that have numerous environmental applications and can remediate heavy metals from wastewater. Also, it has numerous advantages over conventional remediation processes. Microalgae cells can uptake the heavy metal via different physiological and biological methods and are utilized as a nutrient source to regulate its metabolic process for the production of biomass. Furthermore, the enhancement in heavy metal removal efficiency can be improved using different strategies such as immobilization of algal cells, development of algal consortia and designing of microalgae-based nanocomposite materials. Also, it can significantly contribute towards environmental sustainability and future. Thus, the review provides a critical overview of heavy metals and their existence along with their negative effects on humans. This review provides insight on recent advanced nanomaterial approaches for the removal of heavy metals, overviews of microalgae-based heavy metal uptake mechanisms and their potential for the amputation of different heavy metals. Furthermore, the special focus is on recent strategies that enhance heavy metal removal efficiency and contribute towards sustainability for the development of a microalgae-based future.

Mechanical properties of cell- and microgel bead-laden oxidized alginate-gelatin hydrogels.

3D-printing technologies, such as biofabrication, capitalize on the homogeneous distribution and growth of cells inside biomaterial hydrogels, ultimately aiming to allow for cell differentiation, matrix remodeling, and functional tissue analogues. However, commonly, only the mechanical properties of the bioinks or matrix materials are assessed, while the detailed influence of cells on the resulting mechanical properties of hydrogels remains insufficiently understood. Here, we investigate the properties of hydrogels containing cells and spherical PAAm microgel beads through multi-modal complex mechanical analyses in the small- and large-strain regimes. We evaluate the individual contributions of different filler concentrations and a non-fibrous oxidized alginate-gelatin hydrogel matrix on the overall mechanical behavior in compression, tension, and shear. Through material modeling, we quantify parameters that describe the highly nonlinear mechanical response of soft composite materials. Our results show that the stiffness significantly drops for cell- and bead concentrations exceeding four million per milliliter hydrogel. In addition, hydrogels with high cell concentrations (≥6 mio ml-1) show more pronounced material nonlinearity for larger strains and faster stress relaxation. Our findings highlight cell concentration as a crucial parameter influencing the final hydrogel mechanics, with implications for microgel bead drug carrier-laden hydrogels, biofabrication, and tissue engineering.

Growth and digestive enzyme activities of rohu labeo rohita fed diets containing macrophytes and almond oil-cake.

The impact of plant-based diets on the digestive physiology of rohu Labeo rohita fingerlings (10.66 ±â€¯0.53 g) was evaluated. A diet with all protein supplied by fishmeal was included as a control (F). Four test diets containing 300 g/kg protein were formulated using the following plant ingredients and fishmeal in a 1:1 blend: almond oil-cake Terminalia catappa (FTC), duckweed Lemna minor (FLM), water fern Salvania molesta (FSM) and combination of these three ingredients (FTCLMSM). The final body weight and specific growth rate were significantly higher in rohu fed diet FLM compared to the other treatments. Significantly lower feed conversion ratio in rohu fed diet FLM showed that diet was utilized efficiently in this feeding regime compared to the other diets. The composition of diets also influenced the digestive enzyme activities of the fish. Thus, amylase, trypsin and chymotrypsin activities were significantly higher in rohu fed diet FLM compared to the rohu fed the other diets. Protease activity was significantly higher in rohu fed diets FTC and F and lipase activity was significantly higher in rohu fed diet FTC compared to the rohu fed the other diets. The inclusion of raw duckweed in feed replaced 300 g/kg of dietary fishmeal without affecting growth.

Polyacrylamide Bead Sensors for in vivo Quantification of Cell-Scale Stress in Zebrafish Development.

Mechanical stress exerted and experienced by cells during tissue morphogenesis and organ formation plays an important role in embryonic development. While techniques to quantify mechanical stresses in vitro are available, few methods exist for studying stresses in living organisms. Here, we describe and characterize cell-like polyacrylamide (PAAm) bead sensors with well-defined elastic properties and size for in vivo quantification of cell-scale stresses. The beads were injected into developing zebrafish embryos and their deformations were computationally analyzed to delineate spatio-temporal local acting stresses. With this computational analysis-based cell-scale stress sensing (COMPAX) we are able to detect pulsatile pressure propagation in the developing neural rod potentially originating from polarized midline cell divisions and continuous tissue flow. COMPAX is expected to provide novel spatio-temporal insight into developmental processes at the local tissue level and to facilitate quantitative investigation and a better understanding of morphogenetic processes.

Non-genetic factors affecting pre-weaning growth and morphometric traits in Assam Hill goat.

AIM: This study aimed to determine the genetic and non-genetic factors affecting pre-weaning body weight (BW) and morphometry in Assam Hill goat along with the genetic parameters. MATERIALS AND METHODS: The detailed information in respect of BW and body measurements of 960 animals at birth and 3 months of age belonging to three different populations of Assam Hill goat maintained at field units, namely, Batabari, Nahira, and Tetelia under "All India Coordinated Research Project on Goat Improvement" were utilized in the present study. The data were analyzed using least squares technique. RESULTS: The least squares means for BW, height at withers (HW), heart girth (HG), and body length (BL) were 1.166±0.008 kg, 26.198±0.070 cm, 26.695±0.096 cm, and 29.482±0.119 cm at birth and 4.590±0.083 kg, 36.850±0.105 cm, 40.741±0.115 cm, and 39.703±0.108 cm at 3 months of age, respectively. Location had a significant effect on BW, HW, and BL at both birth and 3 months and on HG at 3 months of age. Season of birth exerted significant effect only on BL at birth, whereas the significant effect of sex was observed on HG and BL at 3 months of age. The heritability estimates for BW and body measurements were moderate indicating the scope of selection. The phenotypic and genetic correlations among BWs and body measurements at birth and 3 months of age were positive in direction and high in magnitude. CONCLUSION: On the basis of the present findings, it could be concluded that the weaning weight of kids can be considered for the selection of parent stock to increase productivity and eventually the economic efficiency. Further, animals with higher body measurements at initial phases of growth will perform better with respect to even BW at later stages of growth.

An IL-10 dominant polarization of monocytes is a feature of Indian Visceral Leishmaniasis.

Leishmania donovani, the causative parasite of Visceral Leishmaniasis (VL), deviously manipulates host monocytes/macrophages to ensure its survival. Although monocytes/macrophages from patients with VL have demonstrated an impaired oxidative burst and antigen presentation, an unanswered yet pertinent question remains as to whether they are deactivated or alternatively activated. The significantly raised plasma levels of IL-4/IL-13 and IL-10 in VL patients suggested a microenvironment conducive for alternative activation of monocytes/macrophages. Accordingly, the classical markers for IL-4-driven monocytes/macrophages [M(IL-4)] were studied namely intramonocytic CD206+ , circulating CCL22 and CCL17, and were unchanged. Furthermore, the mRNA expression of Kruppel-like factor 4 (KLF4), peroxisome proliferator-activated receptors (PPAR)-γ and arginase-I (ARG-I) in peripheral blood mononuclear cells was unaltered. However, markers for IL-10-driven monocytes/macrophages [M(IL-10)], namely soluble CD163, intramonocytic IL-10, and circulating CXCL13 were significantly increased. Monocytes/macrophages of patients with VL demonstrated an increased expression of markers for M(IL-10), along with the absence of markers for M(IL-4). Taken together, in human VL, manipulation of these IL-10 polarized monocytes-macrophages may pave the way for improved therapeutic outcomes.

Standardized microgel beads as elastic cell mechanical probes.

Cell mechanical measurements are gaining increasing interest in biological and biomedical studies. However, there are no standardized calibration particles available that permit the cross-comparison of different measurement techniques operating at different stresses and time-scales. Here we present the rational design, production, and comprehensive characterization of poly-acrylamide (PAAm) microgel beads mimicking size and overall mechanics of biological cells. We produced mono-disperse beads at rates of 20-60 kHz by means of a microfluidic droplet generator, where the pre-gel composition was adjusted to tune the beads' elasticity in the range of cell and tissue relevant mechanical properties. We verified bead homogeneity by optical diffraction tomography and Brillouin microscopy. Consistent elastic behavior of microgel beads at different shear rates was confirmed by AFM-enabled nanoindentation and real-time deformability cytometry (RT-DC). The remaining inherent variability in elastic modulus was rationalized using polymer theory and effectively reduced by sorting based on forward-scattering using conventional flow cytometry. Our results show that PAAm microgel beads can be standardized as mechanical probes, to serve not only for validation and calibration of cell mechanical measurements, but also as cell-scale stress sensors.

Cardiac tamponade in systemic lupus erythematosus.

BACKGROUND: Cardiac tamponade is a rare but life-threatening complication of systemic lupus erythematosus (SLE). AIMS/OBJECTIVES: To describe incidence, risk factors and treatment of cardiac tamponade in a large cohort of Indian patients with SLE. METHODS: This retrospective study was conducted at the Department of Rheumatology, IPGMER, Kolkata, India from May 2014 to December 2016 on admitted patients with SLE. Lupus-related serositis was diagnosed after excluding other causes, such as infection, malignancy or heart failure. RESULTS: Of 409 patients with SLE, pericarditis was diagnosed in 25.4% (104/409) and cardiac tamponade in 5.9% (24/409). Tamponade was the presenting feature of SLE in 50% (12/24). Tamponade occurred in 77.8% (14/18) of large effusions and in 11.63% (10/86) of small-to-moderate effusions. The commonest autoantibody in serum and pericardial fluid was anti-nucleosme antibody. Large pericardial effusion (>20 mm) (Odd's ratio (OR): 93.2, 95% confidence interval (CI): 11.1-782.5, P < 0.001) predicted tamponade. In the subset of patients with small-to-moderate sized pericardial effusion, tamponade was associated with pleuritis (OR: 44.5, 95% CI: 1.6-1243, P = 0.025), anti-nucleosome antibody (OR: 42.9, 95% CI: 1.6-1176, P = 0.026) and size of pericardial effusion (OR: 1.36, 95% CI: 1.04-1.76, P = 0.025). Repeated pericardiocentesis was required in 3 patients and one needed surgical intervention. Immunosuppressives used were: prednisolone with monthly intravenous cyclophosphamide (in 33.33%) and intravenous methylprednisolone with monthly cyclophosphamide (in 50%). CONCLUSIONS: Pleuritis, anti-nucleosome antibody and size of pericardial effusion predicted development of tamponade. High dose immunosuppression (methylprednisolone and IV cyclophosphamide) alleviated need for surgery in majority.

Fatigue-Assisted Grain Growth in Al Alloys.

Stress-assisted grain growth at room temperature is known for materials with nanocrystalline grains. For larger grain sizes, the grain growth usually takes place at higher homologous temperatures even under stress. Here we report, for the first time, significant grain growth at room temperature under fatigue loading in microcrystalline grains (≥10 µm) in Al 7075. We demonstrate that this grain growth at room temperature is similar to non-uniform grain growth due to grain rotation and coalescence rather than the thermally and the stress-assisted driven grain growth. We show that the grain growth is associated with the formation of a strong near-Cu {112}<111> texture component as a result of fatigue-assisted deformation. These changes in microstructural features (viz., grain size, grain orientations and texture) are fundamentally important in understanding the cyclic crack induced deformation behavior and for predicting the fatigue lifetime in structural materials.

Regulation of PI3K signaling in T-cell acute lymphoblastic leukemia: a novel PTEN/Ikaros/miR-26b mechanism reveals a critical targetable role for PIK3CD.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy, and T-ALL patients are prone to early disease relapse and suffer from poor outcomes. The PTEN, PI3K/AKT and Notch pathways are frequently altered in T-ALL. PTEN is a tumor suppressor that inactivates the PI3K pathway. We profiled miRNAs in Pten-deficient mouse T-ALL and identified miR-26b as a potentially dysregulated gene. We validated decreased expression levels of miR-26b in mouse and human T-ALL cells. In addition, expression of exogenous miR-26b reduced proliferation and promoted apoptosis of T-ALL cells in vitro, and hindered progression of T-ALL in vivo. Furthermore, miR-26b inhibited the PI3K/AKT pathway by directly targeting PIK3CD, the gene encoding PI3Kδ, in human T-ALL cell lines. ShRNA for PIK3CD and CAL-101, a PIK3CD inhibitor, reduced the growth and increased apoptosis of T-ALL cells. Finally, we showed that PTEN induced miR-26b expression by regulating the differential expression of Ikaros isoforms that are transcriptional regulators of miR-26b. These results suggest that miR-26b functions as a tumor suppressor in the development of T-ALL. Further characterization of targets and regulators of miR-26b may be promising for the development of novel therapies.

STAT Transcription Factors in T Cell Control of Health and Disease.

The Jak-STAT pathway is one of many pleiotropic signaling pathways that plays an important role in organismal development and in response to changing environmental cues. As a key signaling cascade for cytokines and growth factors, Jak-STAT plays central role in the innate and adaptive immune system. Cytokines control the stability, commitment, and maturation of cytotoxic and helper T cells, parts of the adaptive immune system that mediate immunity to pathogens and are linked to inflammatory diseases. Dysregulation of Jak-STAT protein expression or function leads to autoimmunity, allergic diseases, and cancer. Because of their central role in these responses, Jak and STAT molecules have been targeted to develop therapeutics. This review extensively discusses the mechanism of how Jak-STAT signaling in T cells defines our immune responses in the battle against foreign pathogens.

Frontotemporal correlates of impulsivity and machine learning in retired professional athletes with a history of multiple concussions.

The frontotemporal cortical network is associated with behaviours such as impulsivity and aggression. The health of the uncinate fasciculus (UF) that connects the orbitofrontal cortex (OFC) with the anterior temporal lobe (ATL) may be a crucial determinant of behavioural regulation. Behavioural changes can emerge after repeated concussion and thus we used MRI to examine the UF and connected gray matter as it relates to impulsivity and aggression in retired professional football players who had sustained multiple concussions. Behaviourally, athletes had faster reaction times and an increased error rate on a go/no-go task, and increased aggression and mania compared to controls. MRI revealed that the athletes had (1) cortical thinning of the ATL, (2) negative correlations of OFC thickness with aggression and task errors, indicative of impulsivity, (3) negative correlations of UF axial diffusivity with error rates and aggression, and (4) elevated resting-state functional connectivity between the ATL and OFC. Using machine learning, we found that UF diffusion imaging differentiates athletes from healthy controls with significant classifiers based on UF mean and radial diffusivity showing 79-84 % sensitivity and specificity, and 0.8 areas under the ROC curves. The spatial pattern of classifier weights revealed hot spots at the orbitofrontal and temporal ends of the UF. These data implicate the UF system in the pathological outcomes of repeated concussion as they relate to impulsive behaviour. Furthermore, a support vector machine has potential utility in the general assessment and diagnosis of brain abnormalities following concussion.