Comprehensive characterization of human adipose tissue-derived stem cells expanded in vitro

Adipose tissue seems to be a rich and safe source of mesenchymal stem cells (MSCs). The present study was aimed to investigate the biological and morphological characteristics of human adipose tissue-derived stem cells (ATSCs). Light and transmission electron microscopy were used. Course of proliferation was analyzed by growth curve. Expression of surface antigens was assessed by flow cytometry. Chondrogenic potential was assessed by immunohistochemistry. Obtained results showed morphology typical of fibroblastoid cells. TEM analysis proved ultrastructural morphology similar to MSCs from other sources. ATSCs reflected their proteosynthetic and metabolic activity. Each cell had irregular shape of nucleus with noticeable nucleoli. Abundant cisterns of rough endoplasmic reticulum were present in their cytoplasm. Karyotype mapping showed normal count of human chromosomes (46,XX). The growth curve revealed high capability for proliferation and population doubling time was 27.36 hours. ATSCs were positive for CD13, CD29, CD44, CD73, CD90, CD105 and CD106, but did not express CD14, CD34, CD45 and HLA-DR. It was also proved that ATSCs underwent chondrogenic differentiation in vitro. On the basis of obtained results it should be emphasized that ATSCs are typical MSCs and after further investigations they may be used in tissue engineering and regenerative medicine.     

Laboratory-based surveillance of Shigella spp. from human clinical cases in Colombia, 1997-2018

IntroductionShigellosis is endemic in low-and middle-income countries, causing approximately 125 million episodes of diarrhea and leading to approximately 160 .000 deaths annually one-third of which is associated with children.ObjectiveTo describe the characteristics and antimicrobial resistance profiles of Shigella species recovered in Colombia from 1997 to 2018.Materials and methodsWe received isolates from laboratories in 29 Colombian departments. We serotyped with specific antiserum and determined antimicrobial resistance and minimal inhibitory concentrations for ten antibiotics with Kirby-Bauer tests following the Clinical and Laboratory Standards Institute recommendations.ResultsWe analyzed 5,251 isolates of Shigella spp., most of them obtained from stools (96.4%); 2,511 (47.8%) were from children under five years of age. The two most common species were S. sonnei (55.1%) and S. flexneri (41.7%). The highest resistance rate was that of tetracycline (88.1%) followed by trimethoprim-sulfamethoxazole (79.3%) and ampicillin (65.5%); 50.8% of isolates were resistant to chloramphenicol, 43.6% to amoxicillin/clavulanic acid, and less than 1% to cefotaxime, ceftazidime, gentamicin, and ciprofloxacin. In S. sonnei, the most common resistance profile corresponded to trimethoprim-sulfamethoxazole (92%) whereas in S. flexneri the most common antibiotic profiles were multidrug resistance.ConclusionsIn Colombia, children under five years are affected by all Shigella species. These findings should guide funders and public health officials to make evidence-based decisions for protection and prevention measures. The antimicrobial resistance characteristics found in this study underline the importance of combating the dissemination of the most frequently isolated species, S. sonnei and S. flexneri.     

Patterns of blubber fat deposition and evaluation of body condition in growing southern right whale calves (Eubalaena australis)

Marine mammals rely on blubber mainly for energy storage, buoyancy, and streamlining. Mysticetes are born with a relatively thin fat layer that grows rapidly during nursing. However, little information on blubber deposition patterns is available for baleen whale calves. We measured blubber thickness at nine body locations in 350 southern right whale (Eubalaena australis) newborn to 4–6-month-old calves that died on the Península Valdés (Argentina) calving ground from 2003 to 2019, to document changes in blubber thickness with growth. Additionally, we looked for differences in blubber thickness and lipid content of the outer/superficial blubber in calves that died in years with high (2003, 2005, 2007–2013) and low calf mortality (2004, 2006, 2014–2019) to test whether the former were suffering from gross nutritional stress. Blubber thickness increased at all body locations with calf length. Along the cranio-caudal axis, blubber increased in the dorsal and ventral planes, but decreased laterally towards the peduncle, possibly to improve streamlining. We found no difference in blubber thickness and lipid content between high and low mortality years, suggesting that individuals were not undernourished. This is the first study to describe progressive increases in calf blubber during growth and contributes knowledge to right whale health and ontogeny.     

Correction: Amphibian phylogenetic diversity in the face of future climate change: not so good news for the Chilean biodiversity hotspot

Biodiversity and Conservation -     

Do brassinosteroids alleviate water stress in seedlings of Handroanthus serratifolius?

Limited water availability in reforestation areas can compromise plant growth and development, especially for plants at early stages. In this context, the exogenous application of growth regulators, such as brassinosteroids (BRs), can be used to mitigate the negative effects of water restriction. The aim of this study was to evaluate the effect of the application of BRs (90% brassinolide + a brassinosteroid) on initial growth, gas exchange, leaf water potential and leaf anatomy in yellow Ipê (Handroanthus serratifolius), under water restriction conditions. Throughout the experimental period, plants were maintained in two different water regimes: WR1—plants maintained at 100% of the pot capacity; WR2—plants rehydrated with 50% of the evapotranspiration from the previous day. BRs were applied on three occasions during the experimental period, in five concentrations: C1, 0 g/L (application of distilled water); C2, 0.25 g/L; C3, 0.5 g/L; C4, 0.75 g/L; and C5, 1.00 g/L. The dry mass, root:shoot ratio, water status and leaf anatomy traits were evaluated at the end of the experimental period and were analysed in two-level factorial scheme (2WR × 5BRs concentrations). The plant height, number of leaves, gas exchange and leaf chlorophyll content were evaluated four times during the experimental period and were analysed in a split-split-plot design (two WR × five BRs concentrations × four evaluation times). The differences between the means were evaluated by analysis of variance (ANOVA). Pairs of means were separated using the standard error of the difference between the means (SED) and the Fisher's protected least significant difference test (LSD) at p < .05. In addition, all variables were subjected to regression analysis, being the variables evaluated over time, analysed through a two-stage modelling approach. The results obtained indicate that the water deficit led to reductions in growth and gas exchange parameters, regardless of the exogenous application of the regulator; therefore, higher concentrations (C4 and C5) were the most harmful for the maintenance of metabolic and photosynthetic activity. The exogenous application of BRs in H. serratifolius plants attenuated the effects of water limitation on the leaf water potential, but was not able to mitigate for the negative effects in growth, gas exchange and leaf anatomy.     

CD8(+) T-cell responses to adeno-associated virus capsid in humans

Hepatic adeno-associated virus (AAV)-serotype 2 mediated gene transfer results in transgene product expression that is sustained in experimental animals but not in human subjects. We hypothesize that this is caused by rejection of transduced hepatocytes by AAV capsid-specific memory CD8(+) T cells reactivated by AAV vectors. Here we show that healthy subjects carry AAV capsid-specific CD8(+) T cells and that AAV-mediated gene transfer results in their expansion. No such expansion occurs in mice after AAV-mediated gene transfer. In addition, we show that AAV-2 induced human T cells proliferate upon exposure to alternate AAV serotypes, indicating that other serotypes are unlikely to evade capsid-specific immune responses.     

Functional characterization of the Aspergillus fumigatus calcineurin

Aspergillus fumigatus is an aggressive opportunistic pathogen of humans as well as a major allergen. Environmental sensing and retrieving essential nutrients from the environment are general metabolic traits associated with the growth of this saprophytic fungus. Two important mediators of calcium signals in eukaryotic cells are the Ca(2+)-binding protein calmodulin and the Ca(2+)/calmodulin-dependent phosphatase calcineurin. Calcineurin is a heterodimer that consists of a catalytic subunit A and a Ca(2+)/calmodulin binding unit. We deleted the A. fumigatus calA gene, which encodes the calcineurin A catalytic subunit, and demonstrated that this gene is not essential in this fungus. The DeltacalA mutant strain has severe defects in growth extension, branching and conidial architecture. Furthermore, the A. fumigatus DeltacalA mutant strain has decreased fitness in a low dose murine infection and cannot grow in fetal bovine serum (FBS). After potassium phosphate was added to liquid FBS, the DeltacalA mutant strain could grow with the characteristic phenotype of the DeltacalA mutation. When A. fumigatus calcineurin is inhibited by tacrolimus in a phosphate depleted medium, there is a reduction in the inorganic phosphate transport and six putative phosphate transporter genes have altered mRNA levels. However, there is no effect on the acid phosphatase activity. These results suggest that calcineurin is involved in the regulation of the PHO pathway in A. fumigatus. Our work on calcineurin opens new venues for the research on sensing and nutrient acquisition in A. fumigatus.     

Distribution of nuclear receptors for steroid hormones in the human brain: a preliminary study

BACKGROUND: Expression of the nuclear steroid hormone receptors (SHR) within certain parts of the human brain has been described by many authors. However, a comprehensive analysis of SHR expression in the human brain still has not been performed. AIM: To investigate the expression of SHR in different anatomical areas of the brain, especially within the neocortex. METHOD: Immunohistochemical expression of estrogen receptors (ER), progesterone receptors (PR) and androgen receptors (AR) in different regions of the human brain was examined. RESULTS: Nuclear expression of the AR was found in the mamillary body, praecentral gyrus and hippocampus of males. The same expression in analysed structures of female was not found. The expression of ER and PR was not observed. CONCLUSIONS: The analysis revealed unexpected localization of SHR within the brain cortex, which could be the first step to the explanation of SHR action in brain as an interrelationship to function and behaviour. These results indicate on the possibility of SHR detection in post-mortal brain.     

The hypervirulent Mycobacterium tuberculosis strain HN878 induces a potent TH1 response followed by rapid down-regulation

The HN878 strain of Mycobacterium tuberculosis is regarded as "hypervirulent" due to its rapid growth and reduced survival of infected mice when compared with other clinical isolates. This property has been ascribed due to an early increase in type I IFNs and a failure to generate TH1-mediated immunity, induced by a response to an unusual cell wall phenolic glycolipid expressed by the HN878 isolate. We show, however, that although type I IFN does play an inhibitory role, this response was most apparent during the chronic disease stage and was common to all M. tuberculosis strains tested. In addition, we further demonstrate that the HN878 infection was associated with a potent TH1 response, characterized by the emergence of both CD4 and CD8 T cell subsets secreting IFN-gamma. However, where HN878 differed to the other strains tested was a subsequent reduction in TH1 immunity, which was temporally associated with the rapid emergence of a CD4+CD25+FoxP3+CD223+IL-10+ regulatory T cell population. This association may explain the paradoxical initial emergence of a TH1 response in these mice but their relatively short time of survival.