Quantitative Assessment of Morphological Changes in Lipid Droplets and Lipid-Mito Interactions with Aging in Brown Adipose.

The physical characteristics of brown adipose tissue (BAT) are defined by the presence of multilocular lipid droplets (LD) within the brown adipocytes and a high abundance of iron-containing mitochondria, which give it its characteristic color. Normal mitochondrial function is, in part, regulated by organelle-to-organelle contacts. Particularly, the contact sites that mediate mitochondria-LD interactions are thought to have various physiological roles, such as the synthesis and metabolism of lipids. Aging is associated with mitochondrial dysfunction, and previous studies show that there are changes in mitochondrial structure and proteins that modulate organelle contact sites. However, how mitochondria-LD interactions change with aging has yet to be fully clarified. Therefore, we sought to define age-related changes in LD morphology and mitochondria-lipid interactions in BAT. We examined the three-dimensional morphology of mitochondria and LDs in young (3-month) and aged (2-year) murine BAT using serial block face-scanning electron microscopy and the Amira program for segmentation, analysis, and quantification. Analysis showed reductions in LD volume, area, and perimeter in aged samples compared to young samples. Additionally, we observed changes in LD appearance and type in aged samples compared to young samples. Notably, we found differences in mitochondrial interactions with LDs, which could implicate that these contacts may be important for energetics in aging. Upon further investigation, we also found changes in mitochondrial and cristae structure for mitochondria interacting with LD lipids. Overall, these data define the nature of LD morphology and organelle-organelle contacts during aging and provide insight into LD contact site changes that interconnect biogerontology and mitochondrial functionality, metabolism, and bioactivity in aged BAT.

Ablation of Sam50 is associated with fragmentation and alterations in metabolism in murine and human myotubes.

The sorting and assembly machinery (SAM) Complex is responsible for assembling ß-barrel proteins in the mitochondrial membrane. Comprising three subunits, Sam35, Sam37, and Sam50, the SAM complex connects the inner and outer mitochondrial membranes by interacting with the mitochondrial contact site and cristae organizing system complex. Sam50, in particular, stabilizes the mitochondrial intermembrane space bridging (MIB) complex, which is crucial for protein transport, respiratory chain complex assembly, and regulation of cristae integrity. While the role of Sam50 in mitochondrial structure and metabolism in skeletal muscle remains unclear, this study aims to investigate its impact. Serial block-face-scanning electron microscopy and computer-assisted 3D renderings were employed to compare mitochondrial structure and networking in Sam50-deficient myotubes from mice and humans with wild-type (WT) myotubes. Furthermore, autophagosome 3D structure was assessed in human myotubes. Mitochondrial metabolic phenotypes were assessed using Gas Chromatography-Mass Spectrometry-based metabolomics to explore differential changes in WT and Sam50-deficient myotubes. The results revealed increased mitochondrial fragmentation and autophagosome formation in Sam50-deficient myotubes compared to controls. Metabolomic analysis indicated elevated metabolism of propanoate and several amino acids, including ß-Alanine, phenylalanine, and tyrosine, along with increased amino acid and fatty acid metabolism in Sam50-deficient myotubes. Furthermore, impairment of oxidative capacity was observed upon Sam50 ablation in both murine and human myotubes, as measured with the XF24 Seahorse Analyzer. Collectively, these findings support the critical role of Sam50 in establishing and maintaining mitochondrial integrity, cristae structure, and mitochondrial metabolism. By elucidating the impact of Sam50-deficiency, this study enhances our understanding of mitochondrial function in skeletal muscle.

Ablation of Sam50 is associated with fragmentation and alterations in metabolism in murine and human myotubes.

The Sorting and Assembly Machinery (SAM) Complex is responsible for assembling ß-barrel proteins in the mitochondrial membrane. Comprising three subunits, Sam35, Sam37, and Sam50, the SAM complex connects the inner and outer mitochondrial membranes by interacting with the mitochondrial contact site and cristae organizing system (MICOS) complex. Sam50, in particular, stabilizes the mitochondrial intermembrane space bridging (MIB) complex, which is crucial for protein transport, respiratory chain complex assembly, and regulation of cristae integrity. While the role of Sam50 in mitochondrial structure and metabolism in skeletal muscle remains unclear, this study aims to investigate its impact. Serial block-face-scanning electron microscopy (SBF-SEM) and computer-assisted 3D renderings were employed to compare mitochondrial structure and networking in Sam50-deficient myotubes from mice and humans with wild-type (WT) myotubes. Furthermore, autophagosome 3D structure was assessed in human myotubes. Mitochondrial metabolic phenotypes were assessed using Gas Chromatography-Mass Spectrometry-based metabolomics to explore differential changes in WT and Sam50-deficient myotubes. The results revealed increased mitochondrial fragmentation and autophagosome formation in Sam50-deficient myotubes compared to controls. Metabolomic analysis indicated elevated metabolism of propanoate and several amino acids, including ß-Alanine, phenylalanine, and tyrosine, along with increased amino acid and fatty acid metabolism in Sam50-deficient myotubes. Furthermore, impairment of oxidative capacity was observed upon Sam50 ablation in both murine and human myotubes, as measured with the XF24 Seahorse Analyzer. Collectively, these findings support the critical role of Sam50 in establishing and maintaining mitochondrial integrity, cristae structure, and mitochondrial metabolism. By elucidating the impact of Sam50-deficiency, this study enhances our understanding of mitochondrial function in skeletal muscle.

Phenotypic Correction of Murine Mucopolysaccharidosis Type II by Engraftment of Ex Vivo Lentiviral Vector-Transduced Hematopoietic Stem and Progenitor Cells.

Mucopolysaccharidosis type II (MPS II, Hunter syndrome) is an X-linked recessive lysosomal disease caused by deficiency of iduronate-2-sulfatase (IDS). The absence of IDS results in the accumulation of the glycosaminoglycans (GAGs) heparan sulfate and dermatan sulfate. Currently, the only approved treatment option for MPS II is enzyme replacement therapy (ERT), Elaprase. However, ERT is demanding for the patient and does not ameliorate neurological manifestations of the disease. Using an IDS-deficient mouse model that phenocopies the human disease, we evaluated hematopoietic stem and progenitor cells (HSPCs) transduced with a lentiviral vector (LVV) carrying a codon-optimized human IDS coding sequence regulated by a ubiquitous MNDU3 promoter (MNDU3-IDS). Mice treated with MNDU3-IDS LVV-transduced cells showed supraphysiological levels of IDS enzyme activity in plasma, peripheral blood mononuclear cells, and in most analyzed tissues. These enzyme levels were sufficient to normalize GAG storage in analyzed tissues. Importantly, IDS levels in the brains of MNDU3-IDS-engrafted animals were restored to 10-20% than that of wild-type mice, sufficient to normalize GAG content and prevent emergence of cognitive deficit as evaluated by neurobehavioral testing. These results demonstrate the potential effectiveness of ex vivo MNDU3-IDS LVV-transduced HSPCs for treatment of MPS II.

Intrahepatic cholestasis of pregnancy: dilemma in diagnosis and management.

OBJECTIVE: To determine the obstetrical outcomes of women delivered for the diagnosis of intrahepatic cholestasis of pregnancy (ICP). METHODS: Retrospective study of singleton pregnancies diagnosed with ICP between 1 May 2014 and 31 December 2017. Population was analyzed based on bile acids: normal (<10 µmol/L), mild (10 to 40 µmol/L), moderate-severe (>40 µmol/L), and not obtained. Receiver operating characteristic curves established critical values for aspartate aminotransferase (AST) and alanine aminotransferase (ALT) to predict elevated bile acids. Statistical analyses included χ2 for categorical variables and ANOVA for continuous variables. All tests used a 2-sided α level of significance of .05. RESULTS: Bile acids were normal in 39 (45.9%) women, 30 (35.3%) had mild cholestasis, 10 (11.8%) had moderate-severe cholestasis and not obtained for six (7%) women. Gestational diabetes was more common in mild cholestasis (p = .03). There were no differences in demographics, clinical presentation, obstetric interventions and neonatal outcomes. Bile acids took 5-6 days to result. Rate of labor inductions was high in all groups. Postpartum complications occurred in four women in the normal group and in one woman in the mild cholestasis group. Five (12.8%) neonates in the normal group, six (20%) in the mild group, and one (10%) in the severe group were admitted to the NICU. There was no fetal asphyxia, no 5-minute Apgar score <7, and no perinatal deaths. An AST of 27.5 IU/L (p = .002) with sensitivity of 81% and specificity of 76%, and an ALT of 26.7 IU/L (p = .004) with sensitivity of 78% and specificity of 68% predicted elevated bile acids. Improving the sensitivity of AST and ALT to 95%, the ROC curve identified an AST of 62 IU/L with a specificity, positive and negative predictive values of 32, 58 and 86%, respectively; and an ALT of 106 IU/L with a specificity, positive and negative predictive values of 27, 57 and 83%, respectively. CONCLUSIONS: ICP should not be presumed in patients with pruritus. This practice may lead to early term delivery and associated complications.

Chinook and Coho salmon hybrids linked to habitat and climatic changes on Vancouver Island, British Columbia.

Between 2013 and 2019, 63 presumed Chinook salmon Oncorhynchus tshawytscha sampled primarily in the Strait of Georgia (0.63% of total sample) were identified as potential Chinook-Coho (Oncorhynchus kisutch) hybrids by the presence of anomalous microsatellite genotypes. Their hybrid origin was confirmed by single nucleotide polymorphism amplification of two species-specific amplicons. Mitochondrial DNA indicated that most of these fish resulted from the hybridization of Coho salmon females and Chinook salmon males. Although no diagnostic external features were identified, several individuals displayed an abnormal scale arrangement on the caudal peduncle. One hybrid juvenile examined for meristics exhibited a pyloric caeca count intermediate between published values for Chinook and Coho salmon. Most hybrids originated in the Cowichan River during the 2014 brood year. Their prevalence in the watershed is a naturally occurring event, likely exacerbated by prolonged low water levels which limit habitat and delay Chinook salmon spawning, in addition to the differential abundance of the parental species. This research is the first to document ongoing natural hybridization (Chinook-Coho salmon crosses) and link it to habitat and climatic changes, and includes the identification of eight F1 adults and two juvenile backcross or F2 hybrids. The potential negative impacts of hybridization, particularly in Coho salmon through potential introgression, warrant hybrid identification as an ecosystem monitoring tool within a survey program.

Incomplete reproductive isolation and strong transcriptomic response to hybridization between sympatric sister species of salmon.

Global change is altering ecosystems at an unprecedented rate. The resulting shifts in species ranges and reproductive timing are opening the potential for hybridization between closely related species which could dramatically alter the genetic diversity, adaptive capacity and evolutionary trajectory of interbreeding taxa. Here, we used behavioural breeding experiments, in vitro fertilization experiments, and whole-transcriptome gene expression data to assess the potential for and consequences of hybridization between Chinook and Coho salmon. We show that behavioural and gametic prezygotic barriers between socio-economically valuable Chinook and Coho salmon are incomplete. Postzygotically, we demonstrate a clear transcriptomic response to hybridization among F1 Chinook-Coho offspring. Genes transgressively expressed within hybrids were significantly enriched with genes encoded in the nucleus but localized to the mitochondrion, suggesting a potential role for mito-nuclear incompatibilities as a postzygotic mechanism of hybrid breakdown. Chinook and Coho salmon are expected to continue to respond to climate change with shifts in migration timing and habitat use, potentiating hybridization between these species. The downstream consequences of hybridization on the future of these threatened salmon, and the ecosystems they inhabit, is unknown.

The Impact of the Sex of Handlers and Riders on the Reported Social Confidence, Compliance and Touch Sensitivity of Horses in Their Care.

Current evidence of how human sex-related differences in riders and handlers may influence horse behaviour is limited. The Equine Behaviour Assessment and Research Questionnaire (E-BARQ) was used to collect demographic data on riders and handlers (n = 1420) and behavioural data on their horses. It includes demographic items about the sex of the respondent and how frequently the horse has been ridden or handled by male and female humans. The questionnaire then gathers observations on the horse's behaviour on the ground and under saddle or when driven. Using E-BARQ's battery of 97 questions, the current study showed differences in ridden and non-ridden horse behaviour that were related to the sex of the rider or handler. Data were evaluated using multivariate analysis and revealed that horses handled by male humans were significantly more difficult to catch (t-value = -3.11; p = 0.002) and significantly more defensive when approached (t-value = -2.104; p = 0.035), but significantly less likely to pull on the reins/brace the neck or toss their head (t-value 1.980; p = 0.048) than horses handled more frequently by female humans. The differences found between male and female horse handlers suggest that sex is an important factor to consider when understanding equine behaviour. Our study explored reported differences in confidence, handling and working compliance and touch sensitivity among horses ridden and handled by male and female humans and suggested further research into how these differences are gendered.

Chlamydia Prevalence by Age and Correlates of Infection Among Pregnant Women.

BACKGROUND: There is a paucity of population-based data on chlamydia in pregnancy despite rising rates in US women. Our objectives were to assess chlamydia prevalence by age group and to identify factors associated with infection in pregnant women to inform screening guidelines. METHODS: This cross-sectional study included pregnant women tested for chlamydia who delivered at the University of Alabama at Birmingham between November 1, 2012, and December 31, 2017. The primary outcome was chlamydia prevalence, defined as a positive urogenital chlamydia nucleic acid amplification test result documented in the electronic medical record. Multivariable logistic regression was used to identify factors associated with infection. RESULTS: Among 17,796 women who delivered during the study period, 13,657 (77%) had chlamydia testing performed at the University of Alabama at Birmingham. Chlamydia prevalence (95% confidence interval) was 7.4% (7.0%-7.9%). Age-stratified prevalence rates were 14.6%, 4.3%, and 1.7% for women younger than 25 years, 25 to 29 years, and 30 years or older, respectively. Chlamydia in pregnancy remained strongly associated with age (adjusted odds ratio [95% confidence interval], 7.2 [5.6-9.2] for age <25 years, and 2.3 [1.7-3.0] for ages 25-29 years, when compared with >30 years) after adjustment for race, urban residence, and insurance status. CONCLUSIONS: Among pregnant women living in the southeastern United States, chlamydia was detected in 1 of 14 women who were tested. Chlamydia positivity was highest among women younger than 30 years. Study findings support broad screening for chlamydia in pregnancy.

Age-Related Changes in the Behaviour of Domestic Horses as Reported by Owners.

The broad traits of boldness and independence in domestic horses can affect their usefulness and, indirectly, their welfare. The objective of the current study was to explore associations between attributes that reflect equine boldness and independence with both the age of horses and the age at which they were started under saddle, as well as other variables including breed, colour and primary equestrian discipline. All data were sourced from responses (n = 1940) to the 97-question online Equine Behaviour Assessment and Research Questionnaire (E-BARQ). Twenty E-BARQ items from the dataset were selected to reflect boldness and independence and were tested for univariate significance at p < 0.2. Multivariable modelling of the effect of age on remaining traits was assessed by an ordinal logistic regression, using a cumulative log odds model. This revealed that older horses were bolder (p = 0.012). However, horses started under saddle at an older age were less bold and less independent (p = 0.040 and p = 0.010, respectively). Australian Stock Horses were bolder and more independent (p = 0.014 and p = 0.007, respectively) than crossbreed horses. Horses used for breeding conformation (p = 0.039), working equitation (p = 0.045), eventing (p = 0.044) and traditional working horses (p = 0.034) were bolder than those used for other disciplines. Dressage (p = 0.039) and therapy (p = 0.040) horses were less bold than horses used for other disciplines. Stallions were bolder (p = -0.034) than geldings. Brown (p = 0.049) and chestnut (p = 0.027) horses were less bold than bay horses. Compared to crossbreed horses, Thoroughbreds (p = 0.000) and companion horses (p = 0.017) were less bold whilst heavy horses (p = 0.029) and ponies (p = 0.044) were bolder. Compared to pleasure horses, mounted games horses (p = 0.033) were less independent whereas working equitation horses (p = 0.020) were more independent. Riders with more than eight years' experience reported more independence in their horses (p = 0.015) than those who had ridden their whole lives. The study findings suggest that boldness and independence are separate traits and only boldness was associated with the age of the horse. Factors that relate to desirable boldness and independence are important in ridden horses because they can affect rider safety. Results from this study should improve horse-rider matching and thereby potentially enhance horse welfare.

Associations between Owners' Reports of Unwanted Ridden Behaviour and In-Hand Behaviour in Horses.

An evidence-based understanding of dangerous or unwelcome behaviour in horses would greatly benefit both horses and humans who interact with them. Using owner-reported data from the Equine Behaviour Assessment and Research Questionnaire (E-BARQ), the current study investigated in-hand behaviours associated with dangerous or unwelcome ridden behaviours, notably bolting, rearing and bucking. Respondents (n = 1584) to the ridden horse section of the E-BARQ answered 42 demographic questions, followed by 268 behavioural items. Parallel analysis was conducted to group individual behaviours into rotated components to create independent and dependent indices. Multivariable general linear modelling and ordinal logistic regression were used to identify behaviours associated with bolting, rearing and bucking. Results revealed that safety-from-bolt increased as social confidence with horses (Odds ratio (OR) = 1.06; 95% confidence interval (cf = 1.02-1.09) and other animals (OR = 1.08; cf = 1.03-1.12), compliance in-hand (OR = 1.10; cf = 1.06-1.16) and tolerance of restraint (OR = 1.05; cf = 1.0-1.11) increased; and decreased as loading problems (OR = 0.95; cf = 0.92-0.99) increased. Safety-from-rear increased as tolerance of restraint (OR = 1.07; cf = 1.02-1.12) and social confidence with other animals (OR = 1.05; cf = 1.01-1.09) increased; and decreased as loading problems (OR = 0.94; cf = 0.91-0.98) increased. Safety-from-buck increased as social confidence with horses (b-value = 0.011, p < 0.001) and other animals (b-value = 0.010, p = 0.002), compliance in-hand (b-value = 0.015, p < 0.001), tolerance of restraint (b-value = 0.009, p = 0.027) and tolerance of haltering/bridling (b-value = 0.016, p = 0.010) increased, and it decreased as loading problems increased (b-value = -0.011, p < 0.001). By revealing, for the first time, that specific behaviours on the ground are associated with particular responses in the same horses when ridden, this study advances equitation science considerably. Identification of risk factors for dangerous behaviour while under saddle can improve safety for horses and riders and highlights the importance of effective and humane in-hand training.

SARS-CoV-2 serology and virology trends in donors and recipients of convalescent plasma.

SARS-CoV-2 has infected millions worldwide. The virus is novel, and currently there is no approved treatment. Convalescent plasma may offer a treatment option. We evaluated trends of IgM/IgG antibodies/plasma viral load in donors and recipients of convalescent plasma. 114/139 (82 %) donors had positive IgG antibodies. 46/114 donors tested positive a second time by NP swab. Among those retested, the median IgG declined (p < 0.01) between tests. 25/139 donors with confirmed SARS-CoV-2 were negative for IgG antibodies. This suggests that having had the infection does not necessarily convey immunity, or there is a short duration of immunity associated with a decline in antibodies. Plasma viral load obtained on 35/39 plasma recipients showed 22 (62.9 %) had non-detectable levels on average 14.5 days from positive test versus 6.2 days in those with detectable levels (p < 0.01). There was a relationship between IgG and viral load. IgG was higher in those with non-detectable viral loads. There was no relationship between viral load and blood type (p = 0.87) or death (0.80). Recipients with detectable viral load had lower IgG levels; there was no relationship between viral load, blood type or death.

Negotiating Local and Global Values in a Globalized World: The Envisioned Futures of Thai Adolescents.

This study investigates how adolescents growing up in a rapidly globalizing Thai city psychologically manage local and global values when considering their envisioned futures. Qualitative analysis of interviews conducted with 20 (16-18 years old) adolescents reveals four distinct patterns of local-global value negotiation. Findings indicate that adolescents at times dynamically reshape local values in order to encompass global values; at other times, adolescents conceive of themselves as sitting at the crossroads of value systems that cannot be readily integrated. By revealing how global youth negotiate local and global values, this study pushes forward the scientific understanding of biculturalism in contexts of rapid cultural change.

Equine Responses to Acceleration and Deceleration Cues May Reflect Their Exposure to Multiple Riders.

It is logical to assume that horses with multiple riders encounter variation in application of training cues. When training cues are inconsistent, we expect to see a decrease in trained responses or an increase in conflict behaviours. This study investigated the relationship between the number of people that regularly ride or handle a horse and the horse's response to operant cues. Data on 1819 equids were obtained from the Equine Behavior Assessment and Research Questionnaire (E-BARQ), an online global survey of horse owners and caregivers. Three mutually independent indices (acceleration, deceleration, and responsiveness) were derived from a parallel analysis of E-BARQ items related to acceleration and deceleration cues. These indices were then subjected to multivariable modelling against a range of dependent variables including horse and human demographics, horse management, and the number of riders or handlers. The number of riders or handlers was a significant predictor for two out of three indices. As the number of riders or handlers increased, horses were more difficult to accelerate (regression coefficient = 0.0148 ± 0.0071; p = 0.0366) and less difficult to decelerate (regression coefficient = -0.017 ± 0.008; p = 0.030) than those with fewer riders or handlers. These findings suggest that horses' responses to rein tension cues are more persistent than their responses to leg pressure or whip cues. Alternatively, horses with these responses may be actively selected for multiple rider roles. Longitudinal studies of this sort should reveal how the number of riders or handlers affects horse behaviour and could lead to safer and more humane equestrian practices.

Family intervention improves outcomes for patients with delirium: Systematic review and meta-analysis.

OBJECTIVE: To determine if family caregiver involvement in interventions with patients with delirium improves patient outcomes. METHODS: A search of three databases (Medline-Ovid, CINAHL and Embase) was conducted. Eligibility criteria included adult patients and involvement of family caregivers in any delirium intervention. Data were extracted from each study (determined by PEDro scale) using a customised form. A meta-analysis was undertaken which compared the length of hospital stay and duration of delirium. PROSPERO registration number is CRD42017077650. RESULTS: Five studies involving 505 participants published over a 5-year period were suitable for inclusion. Low-level evidence demonstrated family caregiver involvement may reduce caregiver's anxiety and hospital staff viewed administration of education to family caregivers as efficient. Meta-analysis suggested family interventions reduce length of hospital stay for patients with delirium. It remains unclear if it affects the duration of delirium. CONCLUSION: Family caregivers providing interventions to patients with delirium can improve patient outcomes.

Mitochondrial Ecophysiology: Assessing the Evolutionary Forces That Shape Mitochondrial Variation.

The mitonuclear species concept hypothesizes that incompatibilities between interacting gene products of the nuclear and mitochondrial genomes are a major factor establishing and maintaining species boundaries. However, most of the data available to test this concept come from studies of genetic variation in mitochondrial DNA, and clines in the mitochondrial genome across contact zones can be produced by a variety of forces. Here, we show that using a combination of population genomic analyses of the nuclear and mitochondrial genomes and studies of mitochondrial function can provide insight into the relative roles of neutral processes, adaptive evolution, and mitonuclear incompatibility in establishing and maintaining mitochondrial clines, using Atlantic killifish (Fundulus heteroclitus) as a case study. There is strong evidence for a role of secondary contact following the last glaciation in shaping a steep mitochondrial cline across a contact zone between northern and southern subspecies of killifish, but there is also evidence for a role of adaptive evolution in driving differentiation between the subspecies in a variety of traits from the level of the whole organism to the level of mitochondrial function. In addition, studies are beginning to address the potential for mitonuclear incompatibilities in admixed populations. However, population genomic studies have failed to detect evidence for a strong and pervasive influence of mitonuclear incompatibilities, and we suggest that polygenic selection may be responsible for the complex patterns observed. This case study demonstrates that multiple forces can act together in shaping mitochondrial clines, and illustrates the challenge of disentangling their relative roles.

Divinity Revised: The De- and Re-Contextualization of Adolescent Divinity Reasoning in Globalizing Thailand.

This article explores how globalization reshapes moral development in northern Thailand. Employing a cultural-developmental approach to examine interview data gathered over the course of one year, the article discusses variations in Divinity-based moral reasoning among adolescents residing in variously globalized Thai communities. Quantitative analysis shows that moral reasoning diverges across contexts of globalization, with rural adolescents reasoning more in terms of the Ethic of Divinity than urban adolescents. Qualitative analysis shows how the meaning of Divinity diverges, with the Ethic of Divinity co-occurring alongside the Ethic of Community among rural adolescents, and the Ethic of Divinity co-occurring alongside the Ethic of Autonomy among urban adolescents. Analyses further indicate that rural and urban adolescents invoke distinct Divinity principles altogether. Findings suggest that in urban settings, Divinity is decontextualized from traditional community-driven Thai Buddhism and recontextualized to align with values that are adaptive in a globalized society. This article contributes to, and offers suggestions for, the empirical study of the moral psychology of globalization.

Shifting Practices, Shifting Selves: Negotiations of Local and Global Cultures Among Adolescents in Northern Thailand.

Around the world, adolescents increasingly grow up as members of local and global cultures. Little is known, however, about how precisely adolescents in rapidly globalizing societies blend local and global cultures. Interviews with 40 (16- to 19-year old) Thai adolescents, evenly divided between rural and urban communities, were analyzed alongside participant observation data for the interplay between local and global linguistic and dietary practices. Results revealed that urban adolescents inhabited differentiated selves, alternating between local and global practices based on interactional partner. The activation of each assisted them in navigating-and in some cases, reshaping-hierarchies encountered in everyday relationships. Findings contribute to the developmental science of globalization and point to the utility of interrogating cultural practices as sites of self-negotiation in rapidly changing cultural contexts.

Staurosporine Increases Lentiviral Vector Transduction Efficiency of Human Hematopoietic Stem and Progenitor Cells.

Lentiviral vector (LVV)-mediated transduction of human CD34+ hematopoietic stem and progenitor cells (HSPCs) holds tremendous promise for the treatment of monogenic hematological diseases. This approach requires the generation of a sufficient proportion of gene-modified cells. We identified staurosporine, a serine/threonine kinase inhibitor, as a small molecule that could be added to the transduction process to increase the proportion of genetically modified HSPCs by overcoming a LVV entry barrier. Staurosporine increased vector copy number (VCN) approximately 2-fold when added to mobilized peripheral blood (mPB) CD34+ cells prior to transduction. Limited staurosporine treatment did not affect viability of cells post-transduction, and there was no difference in in vitro colony formation compared to vehicle-treated cells. Xenotransplantation studies identified a statistically significant increase in VCN in engrafted human cells in mouse bone marrow at 4 months post-transplantation compared to vehicle-treated cells. Prostaglandin E2 (PGE2) is known to increase transduction efficiency of HSPCs through a different mechanism. Combining staurosporine and PGE2 resulted in further enhancement of transduction efficiency, particularly in short-term HSPCs. The combinatorial use of small molecules, such as staurosporine and PGE2, to enhance LVV transduction of human CD34+ cells is a promising method to improve transduction efficiency and subsequent potential therapeutic benefit of gene therapy drug products.