Differentiation of primate primordial germ cell-like cells following transplantation into the adult gonadal niche.

A major challenge in stem cell differentiation is the availability of bioassays to prove cell types generated in vitro are equivalent to cells in vivo. In the mouse, differentiation of primordial germ cell-like cells (PGCLCs) from pluripotent cells was validated by transplantation, leading to the generation of spermatogenesis and to the birth of offspring. Here we report the use of xenotransplantation (monkey to mouse) and homologous transplantation (monkey to monkey) to validate our in vitro protocol for differentiating male rhesus (r) macaque PGCLCs (rPGCLCs) from induced pluripotent stem cells (riPSCs). Specifically, transplantation of aggregates containing rPGCLCs into mouse and nonhuman primate testicles overcomes a major bottleneck in rPGCLC differentiation. These findings suggest that immature rPGCLCs once transplanted into an adult gonadal niche commit to differentiate towards late rPGCs that initiate epigenetic reprogramming but do not complete the conversion into ENO2-positive spermatogonia

A prospective observational study of on-treatment plasma homocysteine levels as a biomarker of toxicity, depression and vitamin supplementation lead-in time pre pemetrexed, in patients with non-small cell lung cancer and malignant mesothelioma

OBJECTIVES: Vitamin supplementation reduces pemetrexed toxicity. Raised plasma homocysteine reflects deficiency in vitamin B12 and folate, and is suppressed by supplementation. This observational study of 112 patients receiving pemetrexed-based chemotherapy assessed homocysteine levels after 3 weeks of vitamin supplementation, hypothesising high levels would correlate with ongoing deficiency, thus increased toxicity. MATERIAL AND METHODS: Primary endpoint was the composite of proportion of patients with treatment delay/ dose reduction/ drug change or hospitalisation during the first six weeks of chemotherapy, comparing those with normal plasma homocysteine (successfully supplemented, SS) and those with high homocysteine (unsuccessfully supplemented, USS). Secondary endpoints included toxicity and analyses for depression. Post-hoc analysis examined correlation between interval of vitamin and folate supplementation and pemetrexed on primary endpoint and grade 3-4 toxicities. RESULTS: Eighty-four patients (84%) were successfully supplemented (SS group). The proportion of patients undergoing a treatment delay/ dose reduction/ drug change or hospitalisation in SS group was 44.0% (95% confidence interval [CI] 33.2%-55.3%) and in USS group was 18.8% (95% CI 4.0%-45.6%) (p = 0.09). Twelve percent of patients gave a past history of depression however 66% of patients had an on study Hospital Anxiety and Depression (HAD) score of >7. Supplementation status was not associated with depression. The median overall survival (OS) was 11.8 months (95% CI 8.6-16.5) in the SS group and 8.8 months (95% CI 6.6-16.2) in the US group (p = 0.5). The number of days (<7 or ≥ 7 days) between vitamin B12 and folate initiation and pemetrexed administration, had no effect on the primary endpoint and grade 3-4 toxicities. CONCLUSION: On-treatment homocysteine levels were not a biomarker of toxicity or depression. Standard vitamin supplementation is adequate in the majority of patients receiving pemetrexed. High HAD score were noted in this population giving an opportunity for mental health intervention. The lead-in time for vitamin supplementation can be short

The Future of Single- to Multi-band Detector Technologies: Review

Using classical optical components such as filters, prisms and gratings to separate the desired wavelengths before they reach the detectors results in complex optical systems composed of heavy components. A simpler system will result by utilizing a single optical system and a detector that responds separately to each wavelength band. Therefore, a continuous endeavors to develop the capability to reliably fabricate detector arrays that respond to multiple wavelength regions. In this article, we will review the state-of-the-art single and multicolor detector technologies over a wide spectral-range, for use in space-based and airborne remote sensing applications. Discussions will be focused on current and the most recently developed focal plane arrays (FPA) in addition to emphasizing future development in UV-to-Far infrared multicolor FPA detectors for next generation space-based instruments to measure water vapor and greenhouse gases. This novel detector component will make instruments designed for these critical measurements more efficient while reducing complexity and associated electronics and weight. Finally, we will discuss the ongoing multicolor detector technology efforts at NASA Langley Research Center, Jet Propulsion Laboratory, Rensselaer Polytechnic Institute, and others

Molecular epidemiology of leptospirosis in western and southern provinces in Sri Lanka

Objective:The present study investigated the molecular epidemiology of leptospirosis in selected provinces in Sri LankaMethods: A prospective study was carried out between January 2013 to 2014. Adult patients with clinically suspected leptospirosis admitted to the medical wards at selected hospital were included. A whole blood sample was collected and DNA was extracted. leptospira IgM was detected using an immunochromatographic assay. PCR was done using fla B primers followed by RFLP using Hind III and Hae III. Nested PCR was performed using rrs primers which amplifies 16S rRNA gene. Amplicons resulting from rrs PCR were purified and subjected to bidirectional DNA sequencing and phylogenic analyses were conducted.Results: Out of the 168 clinically suspected leptospirosis cases 84 (50%) were IgM positive and 13 (7.7 %) were positive with Fla B PCR while 14 were rrs PCR positive. Based on sequence analysis, Leptospira interrogans (11/14, 79%) was the most common cause of disease followed by Leptospira borgpetersenii (2/14,14%) and Leptospira kirschneri (1/14,7%). Renal failure was the most common complication (4/11, 36%) associated with L. interrogans followed by myocarditis(2/11,18%). Leptospira borgpetersenii was associated with myocarditis and liver failure while Leptospira kirschneri was associated with acute renal failure. RFLP Hind 111 digestions could not differentiate between L. interrogans strains Canicola, Icterrohaemorrgiae and Pyrogenes while Hae 111 digestion showed better discrimination.Conclusions: L. interrogans was the predominant circulating strain in western and southern provinces in 2013 in Sri Lanka. The current data will contribute to determining molecular epidemiological diversity of circulating leptospira in Sri Lank

Single Gene Deletions of Orexin, Leptin, Neuropeptide Y, and Ghrelin Do Not Appreciably Alter Food Anticipatory Activity in Mice

Timing activity to match resource availability is a widely conserved ability in nature. Scheduled feeding of a limited amount of food induces increased activity prior to feeding time in animals as diverse as fish and rodents. Typically, food anticipatory activity (FAA) involves temporally restricting unlimited food access (RF) to several hours in the middle of the light cycle, which is a time of day when rodents are not normally active. We compared this model to calorie restriction (CR), giving the mice 60% of their normal daily calorie intake at the same time each day. Measurement of body temperature and home cage behaviors suggests that the RF and CR models are very similar but CR has the advantage of a clearly defined food intake and more stable mean body temperature. Using the CR model, we then attempted to verify the published result that orexin deletion diminishes food anticipatory activity (FAA) but observed little to no diminution in the response to CR and, surprisingly, that orexin KO mice are refractory to body weight loss on a CR diet. Next we tested the orexigenic neuropeptide Y (NPY) and ghrelin and the anorexigenic hormone, leptin, using mouse mutants. NPY deletion did not alter the behavior or physiological response to CR. Leptin deletion impaired FAA in terms of some activity measures, such as walking and rearing, but did not substantially diminish hanging behavior preceding feeding time, suggesting that leptin knockout mice do anticipate daily meal time but do not manifest the full spectrum of activities that typify FAA. Ghrelin knockout mice do not have impaired FAA on a CR diet. Collectively, these results suggest that the individual hormones and neuropepetides tested do not regulate FAA by acting individually but this does not rule out the possibility of their concerted action in mediating FAA

Effects of past and current crop management on soil microbial biomass and activity

As soil biota is influenced by a number of factors, including land use and management techniques, changing management practices could have significant effects on the soil microbial properties and processes. An experiment was conducted to investigate differences in soil microbiological properties caused by long- and short-term management practices. Intact monolith lysimeters (0.2 m2 surface area) were taken from two sites of the same soil type that had been under long-term organic or conventional crop management and were then subjected to the same 2½-year crop rotation (winter barley (Hordeum vulgare L.), maize (Zea mais L.), lupin (Lupinus angustifolius L.) rape (Brassica napus L. ssp. oleifera)) and two fertiliser regimes (following common organic and conventional practices). Soil samples were taken after crop harvest and analysed for microbial biomass C and N, microbial activity (fluorescein diacetate hydrolysis, arginine deaminase activity, dehydrogenase activity) and total C and N. The incorporation of the green manure stimulated growth and activity of the microbial communities in soils of both management histories. Soil microbial properties did not show any differences between organically and conventionally fertilised soils, indicating that crop rotation and plant type had a larger influence on the microbial biomass and enzyme activities than fertilisation. Initial differences in microbial biomass declined, while the effects of farm management history were still evident in enzyme activities and total C and N. Links between enzyme activities and microbial biomass C varied depending on treatment indicating differences in microbial community composition

Recurrent Plasmodium falciparum Malaria Infections in Kenyan Children Diminish T-Cell Immunity to Epstein Barr Virus Lytic but Not Latent Antigens

Plasmodium falciparum malaria (Pf-malaria) and Epstein Barr Virus (EBV) infections coexist in children at risk for endemic Burkitt's lymphoma (eBL); yet studies have only glimpsed the cumulative effect of Pf-malaria on EBV-specific immunity. Using pooled EBV lytic and latent CD8+ T-cell epitope-peptides, IFN-γ ELISPOT responses were surveyed three times among children (10 months to 15 years) in Kenya from 2002–2004. Prevalence ratios (PR) and 95% confidence intervals (CI) were estimated in association with Pf-malaria exposure, defined at the district-level (Kisumu: holoendemic; Nandi: hypoendemic) and the individual-level. We observed a 46% decrease in positive EBV lytic antigen IFN-γ responses among 5–9 year olds residing in Kisumu compared to Nandi (PR: 0.54; 95% CI: 0.30–0.99). Individual-level analysis in Kisumu revealed further impairment of EBV lytic antigen responses among 5–9 year olds consistently infected with Pf-malaria compared to those never infected. There were no observed district- or individual-level differences between Pf-malaria exposure and EBV latent antigen IFN-γ response. The gradual decrease of EBV lytic antigen but not latent antigen IFN-γ responses after primary infection suggests a specific loss in immunological control over the lytic cycle in children residing in malaria holoendemic areas, further refining our understanding of eBL etiology

Baseline results from the UK SIGNIFY study: a whole-body MRI screening study in TP53 mutation carriers and matched controls

In the United Kingdom, current screening guidelines for TP53 germline mutation carriers solely recommends annual breast MRI, despite the wide spectrum of malignancies typically seen in this group. This study sought to investigate the role of one-off non-contrast whole-body MRI (WB MRI) in the screening of asymptomatic TP53 mutation carriers. 44 TP53 mutation carriers and 44 population controls were recruited. Scans were read by radiologists blinded to participant carrier status. The incidence of malignancies diagnosed in TP53 mutation carriers against general population controls was calculated. The incidences of non-malignant relevant disease and irrelevant disease were measured, as well as the number of investigations required to determine relevance of findings. In TP53 mutation carriers, 6 of 44 (13.6, 95% CI 5.2-27.4%) participants were diagnosed with cancer during the study, all of which would be considered life threatening if untreated. Two were found to have two primary cancers. Two participants with cancer had abnormalities on the MRI which were initially thought to be benign (a pericardial cyst and a uterine fibroid) but transpired to be sarcomas. No controls were diagnosed with cancer. Fifteen carriers (34.1, 95% CI 20.5-49.9%) and seven controls (15.9, 95% CI 6.7-30.1%) underwent further investigations following the WB MRI for abnormalities that transpired to be benign (p = 0.049). The cancer detection rate in this group justifies a minimum baseline non-contrast WB MRI in germline TP53 mutation carriers. This should be adopted into national guidelines for management of adult TP53 mutation carriers in addition to the current practice of contrast enhanced breast MRI imaging.This work was supported by The Annabel Evens Memorial Fund. The investigators at The Institute of Cancer Research and The Royal Marsden National Health Service (NHS) Foundation Trust are supported by an NIHR research grant to the Biomedical Research Centre at The Institute of Cancer Research and The Royal Marsden NHS Foundation Trust