Dried blood spot‐based host genome analysis technique targeting pathological associations with hepatitis B: Development and clinical application in the Cambodian population

Posted on by
Dried blood spot-based host genome analysis technique targeting pathological associations with hepatitis B: Development and clinical application in the Cambodian population

Genetic analysis based on the DBS system is a feasible strategy for getting a deeper understanding of HBV epidemiology, especially in countries with inadequate infrastructure. Based on the DBS system, we successfully clarified the associations of HLA-DPB1 alleles with chronic HBV infection in the Cambodian population for the first time.


Abstract

Aim

Reports of patients with hepatitis B have highlighted associations between polymorphisms in the human leukocyte antigen (HLA)-DPB1, CXCL13, and CXCR5 genes and disease pathology. Owing to its potential to contribute to the development of new diagnostic and therapeutic methods, we aimed to establish a reliable host genome analysis technique that can be used in countries with inadequate infrastructure.

Method

We compared multiple commercially available kits for dried blood spot (DBS)-based sample collection to develop a basic DBS-based host genome analysis technique. We then collected blood samples from Cambodian patients with hepatitis  B and performed single-nucleotide polymorphism genotyping and HLA allele typing by the DBS system.

Result

We were able to perform single-nucleotide polymorphism genotyping and HLA allele typing with host DNA samples obtained using a combination of a HemaSpot™ filter paper-based device and a SMITEST® EX-R&D DNA extraction kit. The accuracy of genotyping using samples obtained by this method was not inferior to one using samples obtained by venipuncture. In the Cambodian population, significant associations of HLA-DPB1*04:01 with protection against chronic hepatitis B virus (HBV) infection, and HLA-DPB1*05:01 and HLA-DPB1*13:01 with susceptibility to chronic HBV infection were identified.

Conclusion

Based on the DBS system, we clarified the associations of HLA-DPB1 alleles with chronic HBV infection in the Cambodian population for the first time. Because the DBS is a low-cost, durable, transportable, and easy-to-handle modality, genetic analysis based on the DBS system is a feasible strategy for obtaining a deeper understanding of HBV epidemiology, especially in middle- or low-income countries.

Outstanding Gir oocyte donors: How does individual factor affect in vitro embryo production efficiency?

Posted on by

Abstract

The oocyte donor plays a pivotal role in bovine in vitro embryo production (IVP) success. The individual factor affects blastocyst/oocyte ratio and determine the existence of outstanding performing animals. The aim of this study was to assess the extent of individual factor effect to IVP efficiency, in a population of Gir oocyte donors. Extreme (high or low IVP efficiency based on blastocyst/oocyte ratio) animals were selected out of a population of 250 oocyte donors (1,734 observations) to form high (>0.48, n = 40), average (0.17–0.48, n = 168), and low (<0.17, n = 42) efficiency donor groups. Cumulus-oocyte complex indicators (total number, IVF-grade number, and IVF-grade/total ratio) were lower (p < 0.05) in high efficiency donors. The number of blastocysts per OPU was analyzed for highest performing bull, and an increase (p < 0.05) in high efficiency donors and a decrease (p < 0.05) in low efficiency donors were noticed, compared to average efficiency donors. The number of pregnancies obtained per OPU was affected (p = 0.017) by donor's efficiency (low: 0.60±0.09$$ 0.60\pm 0.09 $$, average: 1.17±0.07$$ 1.17\pm 0.07 $$, high: 2.57±0.26$$ 2.57\pm 0.26 $$), being 4.3-fold higher in high than in low efficiency donors. We conclude that producing embryos from high efficiency blastocyst/oocyte ratio donors increases blastocyst and pregnancy numbers by OPU, being an important indicator for donor selection in IVP programs.

Effects of low‐protein diet and feed restriction on mRNA expression of cationic amino acid transporters in porcine skeletal muscles

Posted on by

Abstract

We investigated the effects of a low-protein diet and feed restriction on the mRNA expression of cationic amino acid transporters (CATs) in the longissimus dorsi (LD), rhomboideus (RH), and biceps femoris (BF) muscles of pigs. Eighteen piglets were divided into three groups: a control (CP21%), low-protein diet (LP, CP16%), and feed-restricted diet (FR, CP21%, 76% feed intake of control pigs) groups. The expression levels of CAT-1 in the LD and BF muscles of LP pigs were higher than that of control pigs, whereas that of FR pigs showed no difference. The CAT-2A expression levels in the RH muscle of FR pigs were higher than that of control pigs. The free lysine concentrations in all muscles of LP and FR pigs were lower than that of control pigs. To examine the factors that affect CATs mRNA expression, we evaluated the effects of lysine, arginine, insulin-like growth factor-I, and dexamethasone on the expression of CATs in C2C12 myotubes. CAT-1 expression levels increased in lysine and/or arginine deprivation. We show that CAT-1 and CAT-2A expression levels in skeletal muscles differ in response to dietary treatments and CAT-1 expression in skeletal muscles appears to increase in response to low free lysine concentrations.

The impact of thawing duration on leptin and adiponectin levels in frozen human milk samples

Posted on by

Abstract

Background

Measurements of hormone concentrations in human milk from understudied populations across the world is of growing scientific interest. Due to limited access to suitable laboratory facilities at many research sites, human milk samples are often shipped frozen to distant laboratories for analysis. Shipping and handling exposes samples to the risk of thawing and degradation as the results of delays, mishandling, or other unforeseen circumstances. Similar degradation risk factors are well known in serum samples. However, the vulnerability of hormone degradation in human milk to thawing during transportation is largely unexplored. Leptin and adiponectin are of particular importance due to their roles in feeding behavior and metabolism in infants, hence our focus on these two hormones. In addition, leptin and adiponectin have been shown to be vulnerable to pasteurization temperature degradation in human milk samples.

Aims

We tested the degradation of human milk metabolic hormones – leptin and adiponectin – in a controlled environment in response to a variety of freeze/thaw conditions that are reflective of potential temperature fluctuations during transportation.

Materials & Methods

Human milk samples were experimentally subjected to thaw durations ranging from 4 to 16 h and assayed for hormonal levels to observe changes from baseline. Samples were also subjected to two additional experimental conditions, re-freezing after an extended period outside freezers or remaining thawed with additional ice packs, to investigate the impact of common transportation conditions.

Results

We found the assayed levels of leptin were not significantly impacted by an extended thaw cycle of 16 h. However, leptin levels were impacted by the extended period of 40 h outside of the freezer. Adiponectin showed a decrease in concentration percentage after the initial 16-h thaw period, but the increased degradation between 0–16 h and 16–40 h was not as severe as that seen in the leptin samples.

Discussion & Conclusion

The results of this experiment can be used to inform hormone measurement consistency given different thawing lengths or freezing conditions during transportation. Additionally, this research informs decisions regarding transportation, storage/handling, and data analysis in human milk research.

Facial cues to physical strength are not always associated with facial masculinity: Comparative study of Europeans and Southern Siberians

Posted on by

Abstract

Objectives

The aim of our study was to compare morphometric patterns of facial sexual dimorphism with strength-face relationship in members of two distinct populations of European and Central Asian origin: Russians and Tuvans.

Methods

Handgrip strength (HGS) measures and facial photographs were collected from Russian (n = 233) and Tuvan (n = 187) men and women. We digitized 70 landmarks and semilandmarks on full-face and 54 landmarks and semilandmarks on profile photos. This was done to capture variation in facial morphology. After that, we performed the shape regressions of landmarks' coordinates upon sex and HGS. Results were visualized in forms of thin-plate deformation grids and geometric morphometric morphs.

Results

In both populations, HGS was associated significantly with male facial shape only. In Russian men, strength-related changes of facial shape were almost completely in direction of increase in male-typicality. This was especially evident for the relative lower facial width, which was higher in men compared to women, as well as in stronger men compared to weaker ones. On the contrary, in Tuvans the lower face was relatively narrower in men than in women. However, the facial shape of strong Tuvan men was also associated with relatively wider lower face. Our results indicate that the effect of strength on facial shape is relatively independent of facial sexual dimorphism.

Conclusions

Our findings clearly demonstrate that physical strength is associated with the shape of the lower part of male faces even in populations with a mismatched direction of lower face sexual dimorphism.

Spatiotemporal dynamics of a palm weevils’ outbreak and susceptibility of peach palm (Bactris gasipaes Kunth) landraces in a germplasm bank in southwestern Colombia

Posted on by
Spatiotemporal dynamics of a palm weevils' outbreak and susceptibility of peach palm (Bactris gasipaes Kunth) landraces in a germplasm bank in southwestern Colombia

This study investigated the population dynamics of the palm weevils Rhynchophorus palmarum and Dynamis borassi in a peach palm plantation in southwestern Colombia. We used pheromone traps and conducted palm censuses to analyse temporal and spatial patterns of the weevils. We found that R. palmarum outnumbered D. borassi during periods of high insect populations and observed synchronized population peaks related to the phenological stage of the peach palm. This study also highlighted the need for complementary control strategies and suggested a breeding programme based on identified genotypes for integrated pest management of palm weevils.


Abstract

The palm weevils Rhynchophorus palmarum and Dynamis borassi are prominent pest species in peach palms and coconut in Colombia. Nevertheless, knowledge of the population dynamics of these weevil species in the neotropical region is still scarce. In this study, we systematically sampled these weevils over 2 years using pheromone traps (1 trap ha−1) in an experimental peach palm plantation under heavy weevil infestation in southwestern Colombia. Additionally, we censused the palms in a portion of the plantation (a peach palm germplasm bank) before and after the weevil monitoring. We analysed the data to elucidate sex-specific temporal and spatial patterns of both R. palmarum and D. borassi in a 19.13 ha area. Furthermore, we sought to detect peach palm landraces within the germplasm bank susceptible and resistant/tolerant to the weevil attack. During periods when insect populations were high, we observed that R. palmarum consistently outnumbered D. borassi. Specifically, we found that R. palmarum reached up to 80 insects trap−1 month−1, while D. borassi only reached around 30 insects trap−1 month−1 during the peak months. However, both species exhibited synchronous population peaks, which were not related to rainfall or temperature but rather to the phenological stage of the peach palm. Furthermore, the weevils had a highly aggregated spatial pattern; insect catchments were more prolific in areas where the landraces Nariño and Risaralda were sown. Moreover, the census evidenced that these two landraces were the most affected. Contrarily, weevil capture was overall low around the Guaviare landrace, this was the only landrace that exhibited an increase in stipes between census years. Our results highlight that complementary control strategies are needed to counteract sudden weevil population outbreaks, as pheromone traps are a good tool for monitoring but not for effective control. Furthermore, more importantly, a peach palm breeding programme based on the identified genotypes should be the basis for the integrated pest management of palm weevils.

Increased formation of neutrophil extracellular traps induced by autophagy and identification of autophagy‐related biomarkers in systemic lupus erythematosus

Posted on by

Abstract

Abnormal death of neutrophils and the subsequent ineffective clearance of cell fragments result in production of autoantigens that can lead to systemic lupus erythematosus (SLE). Excessive formation of neutrophil extracellular traps (NETs) can trigger the synthesis of pro-inflammatory cytokines such as type I interferons, leading to tissue damage and immune dysfunction in SLE patients. In this study, we found that a decrease in neutrophil counts in the peripheral blood was correlated with clinical parameters in SLE patients. Patients with low neutrophil counts had high renal activity index and chronicity index scores. NET formation and neutrophil autophagy in SLE patients were increased. The autophagy inhibitor hydroxychloroquine was shown to restrict NET formation. Using comprehensive bioinformatics analysis, we found that the expression of the autophagy-related gene, hypoxia-inducible factor 1A (HIF1A), was enhanced in peripheral neutrophils and in the renal glomeruli in SLE patients. Targeting HIF1A could be a potential therapeutic approach for SLE.

Mesobiliverdin IXα‐enriched microalgae feed additive eliminates reliance on antibiotic tylosin to promote intestinal health of weaning piglets

Posted on by

Abstract

Weaning is a critical period in raising pigs. Novel animal feed additives that promote gut health and regulate immune function of piglets without antibiotics are needed. In this study, we aimed to test the ability of mesobiliverdin IXα-enriched microalgae (MBV IXα-enriched microalgae) to eliminate reliance on antibiotics to promote intestinal health in piglets. Eighty 28-day-old weaned piglets were randomly allocated to four groups each with four replicate pens and five piglets per pen. The dietary treatments were a basal diet as control (NC), basal diet plus 0.05% tylosin (PC), basal diet plus 0.1% or 0.5% MBV IXα-enriched microalgae as low (MBV-SP1) or high (MBV-SP2) dose respectively. All treated animals showed no significant differences in live weight, average daily gain and feed efficiency compared to control animals. Histological examination showed that MBV-SP1 and particularly MBV-SP2 increased the ratio of villus height to crypt depth in the jejunum and ileum compared to NC (p < 0.05). Similarly, tylosin treatment also increased villi lengths and the ratio of villus height to crypt depth in the jejunum and ileum compared to the NC (p < 0.05). MBV-SP1 and particularly MBV-SP2 reduced the levels of inflammatory cytokines interleukin-6 and tumour necrosis factor-alpha in the small intestine. MBV-SP2 and tylosin similarly reduced the lipid peroxidation marker (TBARS value) in the duodenum and ileum. In conclusion, feed supplementation with MBV IXα-enriched microalgae improved gut health by villus height and production of immunomodulators that correlated with down-regulated secretion of inflammatory cytokines.

Response to water‐deficit following waterlogging varies among ecotypes of subterranean clover (Trifolium subterraneum ssp. yanninicum), a waterlogging‐tolerant annual pasture legume

Posted on by
Response to water-deficit following waterlogging varies among ecotypes of subterranean clover (Trifolium subterraneum ssp. yanninicum), a waterlogging-tolerant annual pasture legume

The annual pasture legume Trifolium subterraneum ssp. yanninicum exhibits high waterlogging tolerance. However, the ability of ssp. yanninicum genotypes to alter phenotype under soil moisture fluctuation (SMF) from waterlogged (WL) to water-deficit (WD), relative to a well-watered (WW) control, has not previously been evaluated. This study evaluated phenotypic plasticity under SMF for shoot and root relative growth rates (RGR) and its effects on other traits, including rhizobia nodulation, root morphology and leaf phyto-oestrogen content, in three contrasting ecotypes of ssp. yanninicum. The scientific strength of this paper arises from the imposition of SMF on a WL tolerant legume and the finding that stable shoot growth throughout SMF was attributable to a highly plastic response with capacity for rapid shoot and root growth to post-WL stress in a drying soil profile.


Abstract

The pasture legume Trifolium subterraneum ssp. yanninicum L. is waterlogging tolerant, but water-deficit (WD) susceptible. The interactive effect of waterlogged (WL) and WD (soil moisture fluctuation [SMF]) results in a severe stress impact. We studied three diverse ecotypes to identify traits associated with adaptation to SMF. Ecotypes were established in a glasshouse with two treatments imposed at 21 days: well-watered (WW–WW; 80% field capacity [FC]) and WL to WD (WL–WD). For WL–WD, pots were WL for 28 days (Harvest I), and then transitioned to WD (drained to 40% FC) and maintained for 10 days (Harvest II). For shoot relative growth rates (RGR) at Harvest I, WL had relatively little impact, although there was a greater reduction for Ecotype A (80% of WW) than Ecotypes B (92%) and C (87%). However, between Harvests I and II, the impact of WL–WD varied among ecotypes with Ecotype A being less affected (75% of WW–WW) than Ecotypes B (57%) and C (63%). For root RGR at Harvest I, WL resulted in a greater reduction for Ecotype A (52% of WW) than Ecotypes B (77%) and C (74%), while for WL–WD between Harvests I and II, Ecotype A showed a large increase (117% of WW–WW) compared to Ecotypes B (95%) and C (87%). In conclusion, the response to WD following WL varied among ecotypes, which demonstrates contrasting adaptation responses to SMF unrelated to WL tolerance. High yield under WL together with capacity for rapid growth post-WL in a drying soil profile could enhance adaptation to SMF.