Hematopoietic stem cells are regulated by Cripto, as an intermediary of HIF-1α in the hypoxic bone marrow niche

Cripto has been known as an embryonic stem (ES)- or tumor-related soluble/cell membrane protein. In this study, we demonstrated that Cripto has a role as an important regulatory factor for hematopoietic stem cells (HSCs). Recombinant Cripto sustained the reconstitution ability of HSCs in vitro. Flow cytometry analysis uncovered that GRP78, one of the candidate receptors for Cripto, was expressed on a subset of HSCs and could distinguish dormant/myeloid-biased HSCs and active/lymphoid-biased HSCs. Cripto is expressed in hypoxic endosteal niche cells where GRP78(+) HSCs mainly reside. Proteomics analysis revealed that Cripto-GRP78 binding stimulates glycolytic metabolism-related proteins and results in lower mitochondrial potential in HSCs. Furthermore, conditional knockout mice for HIF-1α, a master regulator of hypoxic responses, showed reduced Cripto expression and decreased GRP78(+) HSCs in the endosteal niche area. Thus, Cripto-GRP78 is a novel HSC regulatory signal mainly working in the hypoxic niche.     

A shift in the salt bridge interaction of residues D620 and E621 mediates the constitutive activation of Jak2-H538Q/K539L

Jak2 mutations in the exon 14 and exon 12 regions that cause constitutive activation have been associated with myeloproliferative neoplasms. We have previously shown that a pi stacking interaction between F617 and F595 is important for the constitutive activation of Jak2-V617F (Gnanasambandan et al., Biochemistry 49:9972-9984, 2010). Here, using a combination of molecular dynamics (MD) simulations and in vitro mutagenesis, we studied the molecular mechanism for the constitutive activation of the Jak2 exon 12 mutation, H538Q/K539L. The activation levels of Jak2-H538Q/K539L were found to be similar to that of Jak2-V617F, and Jak2-H538Q/K539L/V617F. Data from MD simulations indicated a shift in the salt bridge interactions of D620 and E621 with K539 in Jak2-WT to R541 in Jak2-H538Q/K539L. When compared to Jak2-WT, K539A mutation resulted in increased activation, while K539D or K539E mutations diminished Jak2 activation by 50 %. In the context of Jak2-H538Q/K539L, R541A mutation reduced its activation by 50 %, while R541D and R541E mutations returned its activation levels to that of Jak2-WT. Collectively, these results indicate that a shift in the salt bridge interaction of D620 and E621 with K539 in Jak2-WT to R541 in Jak2-H538Q/K539L is critical for constitutive activation of this Jak2 exon 12 mutant.     

Fluidization, resolidification, and reorientation of the endothelial cell in response to slow tidal stretches

Mechanical stretch plays an important role in regulating shape and orientation of the vascular endothelial cell. This morphological response to stretch is basic to angiogenesis, neovascularization, and vascular homeostasis, but mechanism remains unclear. To elucidate mechanisms, we used cell mapping rheometry to measure traction forces in primary human umbilical vein endothelial cells subjected to periodic uniaxial stretches. Onset of periodic stretch of 10% strain amplitude caused a fluidization response typified by attenuation of traction forces almost to zero. As periodic stretch continued, the prompt fluidization response was followed by a slow resolidification response typified by recovery of the traction forces, but now aligned along the axis perpendicular to the imposed stretch. Reorientation of the cell body lagged reorientation of the traction forces, however. Together, these observations demonstrate that cellular reorientation in response to periodic stretch is preceded by traction attenuation by means of cytoskeletal fluidization and subsequent traction recovery transverse to the stretch direction by means of cytoskeletal resolidification.     

Mapping QTLs for Plant Phenology and Production Traits Using Indica Rice (Oryza sativa L.) Lines Adapted to Rainfed Environment

Drought is a major abiotic stress limiting rice production and yield stability in rainfed ecosystems. Identifying quantitative trait loci (QTL) for rice yield and yield components under water limited environments will help to develop drought resilient cultivars using marker assisted breeding (MAB) strategy. A total of 232 recombinant inbred lines of IR62266/Norungan were used to map QTLs for plant phenology and production traits under rainfed condition in target population of environments. A total of 79 QTLs for plant phenology and production traits with phenotypic variation ranging from 4.4 to 72.8% were detected under non-stress and drought stress conditions across two locations. Consistent QTLs for phenology and production traits were detected across experiments and water regimes. The QTL region, RM204-RM197-RM217 on chromosome 6 was linked to days to 50% flowering and grain yield per plant under both rainfed and irrigated conditions. The same genomic region, RM585-RM204-RM197 was also linked to harvest index under rainfed condition with positive alleles from Norungan, a local landrace. QTLs for plant production and drought resistance traits co-located near RM585-RM204-RM197-RM217 region on chromosome 6 in several rice genotypes. Thus with further fine mapping, this region may be useful as a candidate QTL for MAB, map-based cloning of genes and functional genomics studies for rainfed rice improvement.     

Histopathological changes in Poecilia latipinna male gonad due to Tribulus terrestris administration

Tribulus terrestris is a traditionally known non-toxic aphrodisiac herb for maleness. It was experimented recently to understand the effect and mechanism on mono sex production in Poecilia latipinna. It would help to develop a new eco-friendly way to masculinize P. latipinna, since males have higher commercial value than females. The different concentration (100, 150, 200, 250 and 300 mg/L) of T. terrestris extract and a control were tested for their effect on sex transformation/reversal in P. latipinna by immersing the newly born young ones in the respective concentrations for 2 mo. The obtained results indicate that a dose dependant masculinization is obtained due to T. terrestris administration, which improved the male proportion. Histological results revealed that the testes of fish treated with T. terrestris extract contained all stages of spermatogenesis, clearly demonstrating that the administration of T. terrestris extract to P. latipinna stimulated spermatogenesis. Thus, it is discernible that 0-d-old hatchlings of P. latipinna exposed to T. terrestris extract orient/reverse their sex more towards maleness besides yielding better growth and spermatogenesis which is a mandate for fancy fish industry.     

Effect of alternate nostril breathing on cardio respiratory parameters and muscle strength among rotating shift workers

Night shifts at work is the most frequent reasons for circadian rhythm disruption and subsequent psychological and physiological disturbances, especially increased risk of cardiovascular and respiratory ailments compared to daytime workers. Alternate nostril breathing for about 15 minutes was known to have effect over cardiac, respiratory parameters and muscle strength. Hence aim is of interest to assess the effects of alternate nostril breathing (ANB) on cardio-respiratory parameters and muscle strength among the rotating shift workers in the tertiary care hospital. This observational study was carried out in the department of Physiology after getting institutional ethical committee clearance. Around 140 rotating night shift workers of both sex of age 25-40 years with normal BMI and 140 non-shift workers age, sex and BMI matched were selected as study and control group respectively. Heart rate, blood Pressure, respiratory rate, peak expiratory flow rate, respiratory endurance, respiratory burst test, muscle strength and fatigue were recorded before and after 15 minutes of ANB. Shift workers were found to have significantly altered systolic (P=0.000) and diastolic (P=0.002) blood pressure and heart rate (P=0.010) compared to non-shift workers. Fatigue is altered significantly (P< 0.05) after ANB between both shift and non- shift workers. ANB can be used as a therapeutic module among the shift workers, to maintain their sound health and to improve their performance in the night duty.     

Distributions of Competing Container Mosquitoes Depend on Detritus Types, Nutrient Ratios, and Food Availability

Coexistence of competitors may result if resources are sufficiently abundant to render competition unimportant, or if species differ in resource requirements. Detritus type has been shown to affect interspecific competitive outcomes between Aedes albopictus (Skuse) and Aedes aegypti (L.) larvae under controlled conditions. We assessed the relationships among spatial distributions of detritus types, nutrients, and aquatic larvae of these species in nature. We collected mosquitoes, water, and detritus from artificial containers across 24 Florida cemeteries that varied in relative abundances of Ae. aegypti and Ae. albopictus.We measured nutrient content of fine particulate organic matter in water samples as total N, P, and C and ratios of these nutrients. We quantified food availability via a bioassay, raising individual Aedes larvae in the laboratory in standard volumes of field-collected, particulate-containing water from each cemetery. Quantities of detritus types collected in standard containers were significant predictors of nutrients and nutrient ratios. Nutrient abundances were significant predictors of relative abundance of Ae. aegypti, and of larval survival and development by both species in the bioassay. Survival and development of larvae reared in particulate-containing water from sites decreased with decreasing relative abundance of Ae. aegypti. These data suggest that N, P, and C availabilities are determined by detritus inputs to containers and that these nutrients in turn determine the feeding environment encountered by larvae, the intensity of interspecific competition among larvae, and subsequent relative abundances of species at sites. Detritus inputs, nutrients, and food availability thus seem to contribute to distributions of Ae. aegypti and Ae. albopictus in cemetery containers throughout Florida.     

Design of Site-Specific Microwave Phased Array Hyperthermia Applicators Using 434 MHz Reduced Cavity-Backed Patch Antenna

Cancers of the neck, breast, and lower extremities are common malignancies diagnosed in India with a higher incidence of advanced-stage disease. Phased array (PA) applicators reported for hyperthermia treatment (HT) of the breast have small focal region and high cross-coupling, and those reported for lower extremities provide regional heating and limited steering. In this study, we present the numerical design of site-specific PA applicators for HT of large solid tumors in the neck, breast, and lower extremities using a miniaturized 434 MHz cavity-backed water-loaded patch antenna. The fabricated antenna has 38 × 36 mm² aperture, more than 90% power coupling, 25 MHz bandwidth, and good agreement between simulated and measured specific absorption rate (SAR) in phantom. The site-specific applicators demonstrated less power reflection (<−17.9 dB) and cross-coupling (<−26.8 dB) for 5 mm inter-ring spacing. SAR indicators for 64 cc tumor at varying locations in simplified layered three-dimensional (3D) tissue models of the neck, breast, and leg showed average power absorption ratio (aPA_ratio) ≥ 3.16, target to hotspot quotient (THQ) ≥ 0.57, 25% iso-SAR coverage (TC₂₅) ≥ 81%, and 50% iso-SAR coverage (TC₅₀) ≥51.8%. Simulation results of site-specific applicators for 3D inhomogeneous patient models showed aPA_ratio ≥ 5.98, THQ ≥ 0.9, TC₅₀ ≥ 86%, and 100% TC₂₅ for all sites. It is concluded that the 434 MHz miniaturized cavity-backed patch antenna can be used to develop high-density PA applicators with 12–24 antennas for HT of large solid tumors (≥4 cm) in the neck, breast, and lower extremities with 3D steering ability and less cross-coupling (≤−26.8 dB). © 2020 Bioelectromagnetics Society     

Synthesis and cytotoxic activity of novel quinazolino-beta-carboline-5-one derivatives

A novel series of quinazolino-beta-carbolinone derivatives was synthesized and evaluated for their in vitro and in vivo anticancer activity. Many compounds have shown good in vitro activity in the range 1-8 microM concentration. Three of the compounds were further tested in nude mice bearing HT-29 colon cancer xenografts.