Strain Differences Research Articles

Sex-Related Differences of Left Atrial Strain in Patients With Hypertension Using Cardiac Magnetic Resonance Feature Tracking.

Previous studies demonstrated the impact of sex on left ventricular (LV) strain in patients with essential hypertension. However, little is known about the effect of sex on left atrial (LA) strain in patients with hypertension. This study aimed to explore the sex-related differences of LA strain by using cardiac magnetic resonance feature tracking in patients with hypertension and preserved LV ejection fraction. One hundred and fifty hypertensive patients (100 men and 50 women) and 105 age-matched and sex-matched normotensive controls (70 men and 35 women) were retrospectively enrolled and underwent cardiac magnetic resonance examination. LA strain parameters included LA reservoir strain (εs), conduit strain (εe), pump strain (εa), and their corresponding strain rate (SRs, SRe, and SRa). Men had significantly higher LV mass index, lower εs and εe than women in both patients and controls (all P <0.05). LA strain and strain rate were significantly reduced in hypertensive patients compared with controls, both in men and women (all P <0.05). In men, hypertension and its interaction were associated with increased LV mass index and decreased εs and εe. In multivariable analysis, men, LV ejection fraction, and LA minimum volume index remained independent determinants of εs and εe in all hypertensive patients (all P <0.05). LA strain was significantly impaired in hypertensive patients, and men had more impaired LA strain than women. These findings further emphasize the sex-related differences in the response of LA strain to hypertension in the early stage.

Age dynamics of strain differences in the morphofunctional state of pancreatic beta- and amylin-producing cells in SHR and Wistar rats

Large-scale epidemiological studies have shown that cardiac pathology and progressive atherosclerosis in patients with diabetes mellitus occurred already at the stage of prediabetes. Obesity and insulin resistance affect cardiometabolic health due to pleiotropic effects of insulin. Despite the vast range of research, some aspects remain hidden links in the overall pathogenesis of metabolic and hemodynamic disorders. The aim of the work was to study the morphofunctional state of pancreatic islets (PIs), beta- and amylin-producing cells in male rats of Wistar strain (normotensive) and SHR (with spontaneous development of hypertension) in age dynamics. Materials and methods. The study was carried out using 38 male Wistar rats and SHRs aged 7 and 24 months. Non-invasive blood pressure (BP) detection procedures were done using the BP-2000 Blood Pressure Analysis System. The morphofunctional state of PIs was examined in serial 5-μm thick pancreatic tissue sections. Beta- and amylin-producing cells were detected after histological preprocessing and the use of monoclonal FITC-conjugated antibodies. Image file processing was done via ImageJ software (National Institutes of Health, USA). Levels of glycemia were monitored with a SUPER GLUCOCARD-II glucometer. Results. SHRs were hyperglycemic both at 7 and at 24 months, 8.41 ± 0.15 mmol/l and 8.90 ± 0.14 mmol/l, respectively, with elevated BP, 155 ± 5 / 80 ± 5 mm Hg and 165 ± 5 / 90 ± 5 mm, respectively. Old SHRs developed PI hypertrophy mainly associated with the increased number and percentage of beta-cells, apparently in response to hyperglycemia. Both in the PIs of adult and old SHRs, the number of amylin-producing cells was lower while the content of amylin was higher than those in the age-matched Wistar rats. Conclusions. Male SHRs are characterized by a persistent increase in blood pressure and abnormalities of carbohydrate metabolism already at adult age, one of the manifestations of which is hyperglycemia worsening with age. Chronic hyperglycemia in SHRs due to the higher insulin requirement finds its expression in low content of this hormone in the islets at adult age and decreased its content in beta-cells in old animals.

Plastic strain characteristics on frozen silty clay subjected to intermittent vehicle loads

The plastic strain of frozen soil is a key parameter in evaluating the stability of embankment in cold regions. In practical engineering, the vehicle loading is an intermittent cyclic load due to a period of no-load between adjacent vehicles. At this study, a series of dynamic tests were carried out at −3.0℃, and the effects of intermittent ratio (γ), cyclic stress ratio (CSR), frequency (f) and vibration (N) were considered. The results show that the cumulative plastic strain of frozen silty clay increases nonlinearly with increasing vibrations, and with the increase in intermittent ratio it decreases nonlinearly. There exists a critical intermittent ratio, when the intermittent ratio exceeds the critical one, the cumulative plastic strain with a given vibration will keep constant. As the cyclic stress ratio rises, the cumulative plastic strain also grows in nonlinear. For γ ≠ 0.0, the cumulative plastic strain differences slowly tend to stabilize with a CSR value of over 0.7, and for γ = 0.0 the strain difference obviously rises. With increasing frequencies, the cumulative plastic strain decreases in a nonlinear way, and the cumulative plastic strain difference increases and decreases linearly at γ = 0.0 and γ = 1.0, respectively. Finally, a cumulative plastic strain model, considering γ, CSR, f and N, is established. The reliability of model is also verified by comparing the calculated results with the measured ones. This study can provide a reference for predicting the cumulative plastic strain of frozen silty clay under the intermittent vehicle loads in cold regions.

Animals in Search of Stimulation and Information: A Review of over 10 years of our Research on Spontaneous Exploration in Rats as a Response to Novelty in Low-Stress Paradigm

This article comprehensively reviews our studies that analyzed novelty-related behaviors in rats. We developed and utilized techniques and equipment during these studies to ensure our findings' high ecological validity. We allowed rats to explore a testing apparatus freely, and after a long habituation phase consisting of multiple trials, we introduced non-emotionally arousing changes. The research shows that rats demonstrate enhanced abilities in processing and responding to heightened environmental complexity, as evidenced across various studies. Conversely, when environmental complexity diminishes, rats exhibit reduced exploratory behaviors and decreased cognitive effort despite the adaptive importance of such behaviors. Of particular interest is the observation that rats exhibit greater sensitivity to emerging opportunities in contrast to their limited responsiveness to diminishing ones, unveiling a novel facet of the animal mind that warrants further investigation. The influence of individual experiences before testing sessions on the processing of sensory input in terms of complexity is also determined. Our studies highlight the noteworthy impact of environmental unpredictability versus stability on cognitive development, affecting behaviors like food neophobia and exploration. Furthermore, the social environment during development holds wide-ranging implications for individual characteristics, necessitating continued research and refinement of our understanding in this domain. Moreover, the studies recognize strain and species differences in novelty-related behavior, primarily characterized by quantitative variations that do not overshadow the animals' coping strategies in response to environmental changes. Additionally, curiosity is portrayed as an active approach to seeking and processing environmental affordances, with exploratory behaviors fulfilling this cognitive and motivational need. The authors underscore the significance of ecological validity in test methodologies, particularly in designing environments that authentically invite and encourage pertinent behavioral responses. In summary, this research enhances our insight into rat cognition, underscores the role of curiosity, and underscores the imperative of ecological validity in experimental design, ultimately advancing our comprehension of animal behavior and cognition.

Effect of transverse gradient extrusion process on texture and mechanical properties of Mg–3Al–1Zn alloy sheets

Transverse gradient extrusion (TGE) processes with different die inclined angles of 15°, 30°, 45°and 60° were used to produce Mg–3Al–1Zn sheets, thereby tailoring texture and mechanical properties of Mg alloy sheets. Microstructure, deformation texture and final mechanical properties of the obtained sheets were detected and discussed. Furthermore, the rheological behavior of Mg alloys during extrusion was analyzed by finite element method. As the TGE die inclined angle changed, the flow velocity along the transverse direction (TD) and effective strain difference along extrusion direction (ED) were changed during extrusion, and therefore the microstructure and texture of extruded sheets are altered. With increase of TGE die inclined angles, fine grain size and uniform microstructure were obtained. The base poles of most grains for the extruded sheets gradually deviate from normal direction (ND) to ED at center region and inclined from ED to TD at 1/4 edge region. A large elongation of 33.9 % and a low yield stress of 125 MPa can be obtained in the TGE-45 sheet.

Spacecraft Segment Damage Identification Method Based on Fiber Optic Strain Difference Field Reconstruction and Norm Calculation

Real-time online identification of spacecraft segment damage is of great significance for realizing spacecraft structural health monitoring and life prediction. In this paper, a damage response characteristic field inversion algorithm based on the differential reconstruction of strain response is proposed to solve the problem of not being able to recognize the small damages of spacecraft structure directly by the strain response alone. Four crack damage location identification methods based on vector norm computation are proposed, which realize online identification and precise location of structural damage events without external excitation by means of spacecraft structural working loads only. A spacecraft segment structural damage monitoring system based on fiber optic grating sensors was constructed, and the average error of damage localization based on the curvature vector 2 norm calculation was 2.58 mm, and the root-mean-square error was 1.98 mm. The results show that the method has superior engineering applicability for on-orbit service environments.

Study on Mechanical Properties Subjected to Monotonic and Dynamic Loads of Loessal Soil in Songyuan City

In this paper taking loessal soil of Songyuan in Jilin province as the research object，static and dynamic triaxial tests were conducted to investigate the effects of water content and confining pressure on dynamic characteristics of loessal soil. The test results showed that: (1) Under monotonic loading, the deformation of loessal soil exhibits two situations: strain hardening and strain softening, and the deformation behavior is influenced by water content and confining pressure. The static strength decreases with the increase of water content and increases with the increase of confining pressure. (2) The cumulative plastic strain of loessal soil under dynamic load exhibits three modes: plastic shakedown, plastic creep, and incremental collapse. Existence of critical dynamic stress results in significant differences in the cumulative plastic strain of soil. (3) The cumulative plastic strain of soil samples is influenced by water content, confining pressure, and dynamic stress amplitude, low confining pressure, high water content, and high dynamic stress amplitude are adverse to the plastic stability of loessal soil. Based on experimental results, predict the range of critical dynamic stress. The research results have reference value for the evaluation of dynamic deformation stability of settlement of subgrade constructed in the loessal soil.

Cocaine intake correlates with risk-taking behavior and affects estrous cycling in female Sprague-Dawley rats.

Navigating complex decisions and considering their relative risks and rewards is an important cognitive ability necessary for survival. However, use of and dependence on illicit drugs can result in long-lasting changes to this risk/reward calculus in individuals with substance use disorder. Recent work has shown that chronic exposure to cocaine causes long-lasting increases in risk taking in male and female rats, but there are still significant gaps in our understanding of the relationship between cocaine use and changes in risk taking. For example, it is unclear whether the magnitude of cocaine intake dictates the extent to which risk taking is altered. To address this, male and female Sprague-Dawley rats underwent cocaine (or sucrose) self-administration and, following a period of abstinence, were trained and tested in a rodent model of risky decision making. In this behavioral task, rats made discrete-trial choices between a lever associated with a small food reward (i.e., "safe" option) and a lever associated with a larger food reward accompanied by a variable risk of footshock delivery (i.e., "risky" option). Surprisingly, and in contrast to prior work in Long-Evans rats, there were no effects of cocaine self-administration on choice of the large, risky reward (i.e., risk taking) during abstinence in males or females. There was, however, a significant relationship between cocaine intake and risk taking in female rats, with greater intake associated with greater preference for the large, risky reward. Relative to their sucrose counterparts, female rats in the cocaine group also exhibited irregular estrous cycles, characterized by prolonged estrus and/or diestrus phases. Collectively, these data suggest that there may be strain differences in the effects of cocaine on risk taking and highlight the impact that chronic cocaine exposure has on hormonal cyclicity in females. Future work will focus on understanding the neural mechanisms underlying cocaine's intake-dependent effects on risk taking in females, and whether this is directly related to cocaine-induced alterations in neuroendocrine function.

Stress-Mediated Abnormalities in Regional Myocardial Wall Motion in Young Women with a History of Psychological Trauma.

Psychosocial stress has been associated with the development and progression of atherosclerotic cardiovascular disease (CVD). Previously, we reported subtle differences in global longitudinal strain in somatically healthy women with a psychiatric diagnosis of borderline personality disorder (BPD). This study aimed to investigate the impact of BPD on segmental myocardial wall motion using speckle tracking echocardiography (STE) analysis. A total of 100 women aged between 18 and 38 years were included in this study. Fifty patients meeting the diagnostic criteria for BPD were recruited from the Department of Psychiatry (LWL-University Hospital Bochum) and compared with fifty age-matched healthy control subjects without previous cardiac disease. Laboratory tests and STE were performed with segmental wall motion analysis. The BPD group had a higher prevalence of risk factors for CVD, with smoking and obesity being predominant, when compared with the control group. Other cardiovascular parameters such as blood pressure, glucose, and cholesterol levels were also elevated, even though not to pathological values. Moreover, in the STE analysis, the BPD group consistently exhibited decreased deformation in nine myocardial wall regions compared with the control group, along with a shift toward higher values in the distribution of peak pathological segments. Additionally, significantly higher values of free thyroxine concentration and thyroid's secretory capacity were observed in the BPD group, despite falling within the (high-) normal range. BPD is associated with chronic stress, classical risk factors, and myocardial wall motion abnormalities. Further exploration is warranted to investigate the relationship between high-normal thyroid metabolism, these risk factors, and myocardial function in BPD patients. Long-term follow-up studies would be valuable in confirming the potential for predicting adverse events.

Structural Uncertainty Analysis of High-Temperature Strain Gauge Based on Monte Carlo Stochastic Finite Element Method.

The high-temperature strain gauge is a sensor for strain measurement in high-temperature environments. The measurement results often have a certain divergence, so the uncertainty of the high-temperature strain gauge system is analyzed theoretically. Firstly, in the conducted research, a deterministic finite element analysis of the temperature field of the strain gauge is carried out using MATLAB software. Then, the primary sub-model method is used to model the system; an equivalent thermal load and force are loaded onto the model. The thermal response of the grid wire is calculated by the finite element method (FEM). Thermal-mechanical coupling analysis is carried out by ANSYS, and the MATLAB program is verified. Finally, the stochastic finite element method (SFEM) combined with the Monte Carlo method (MCM) is used to analyze the effects of the physical parameters, geometric parameters, and load uncertainties on the thermal response of the grid wire. The results show that the difference of temperature and strain calculated by ANSYS and MATLAB is 1.34% and 0.64%, respectively. The calculation program is accurate and effective. The primary sub-model method is suitable for the finite element modeling of strain gauge systems, and the number of elements is reduced effectively. The stochastic uncertainty analysis of the thermal response on the grid wire of a high-temperature strain gauge provides a theoretical basis for the dispersion of the measurement results of the strain gauge.

Pulsed-field gel electrophoresis typing and molecular characterization of &lt;em&gt;Listeria monocytogenes&lt;/em&gt; isolates in raw milk samples from Polonnaruwa District, Sri Lanka

Listeria monocytogenes is an important food borne human pathogen associated with severe diseases in humans and animals. Human infection is associated with high mortality rates. Our previous studies done in 2012 on milk and milk products revealed 4b as the dominant serotype followed by 1/2a, 1/2b and 1/2c. Listeria monocytogenes is an important food borne human pathogen associated with severe diseases in humans and animals. Human infection is associated with high mortality rates. Our previous studies done in 2012 on milk and milk products revealed 4b as the dominant serotype followed by 1/2a, 1/2b and 1/2c. Therefore, this study was conducted to determine the sources of contamination and to identify the strain differences (pulsotypes) by pulsed-field gel electrophoresis (PFGE) typing and serotype profiles of L. monocytogenes isolates from farm collected raw milk samples from randomly identified locations in Grama Niladhari divisions of the Polonnaruwa district, Sri Lanka. Eighty isolates obtained from raw milk samples were confirmed as L. monocytogenes by Polymerase Chain Reaction, belonging to serotypes 4b, 1/2a, 1/2b and 1/2c. The strains were identified by PFGE. PFGE analysis digested with AscI and ApaI enzymes revealed different banding patterns. All the pulsotypes were found to be serotype 4b. The sub typing indicated the diversity of the Listeria species. The presence of serotypes 1/2a, 1⁄2b, 1/2c, and 4b in raw milk is a public health concern, as these serotypes are frequently associated with foodborne outbreaks and sporadic cases of human listeriosis. In our study, PFGE analysis allowed discrimination among isolates of the same serogroup. Further PFGE analysis showed heterogeneity among isolates recovered from both same sampling areas and different areas.

Antibodies elicited by Plasmodium falciparum circumsporozoite proteins lacking sequentially deleted C-terminal amino acids reveal mouse strain and epitopes specific differences

Malaria affects ∼ ¼ billion people globally and requires the development of additional tools to aid in elimination efforts. The recently approved RTS,S/AS01 vaccine represents a positive step, however, the moderate efficacy necessitates the development of more efficacious vaccines. PfCSP is a key target antigen for pre-erythrocytic vaccines aimed at preventing Plasmodium falciparum malaria infections. Epitopes within the central repeat region and at the junction of the repeat and N-terminal domain are well documented as major protective B cell epitopes. On the other hand, a majority of antibodies against the epitopes in the C-terminal domain, have been shown to be non-protective against sporozoite challenge. The C-terminal domain, however, contains CD4+ and CD8+ T cell epitopes previously shown to be important for regulating immune responses. The present study was designed to further explore the immunomodulatory potential of the C-terminal domain using DNA vaccines encoding PfCSP with sequential C-terminal truncations following known T cell epitopes. Five DNA vaccines encoding different truncations of PfCSP within the C-terminal domain were administered via intramuscular route and in vivo electroporation for effective immunogenicity. Protection in mice was evaluated by challenge with transgenic P. berghei expressing PfCSP. In Balb/c mice, antibody responses and protective efficacy were both affected progressively with sequential deletion of C-terminal amino acid residues. Similar studies in C57Bl/6 mice revealed that immunizations with plasmids encoding truncated PfCSP showed partial protection from sporozoite challenge with no significant differences in antibody titers observed compared to full-length PfCSP DNA immunized mice. Further analysis revealed murine strain-specific differences in the recognition of specific epitopes.

An end is in sight: a perspective on PCR as an endpoint for Chagas disease treatment trials

Novel therapies for chronic indeterminate Chagas disease (CICD) are needed, but trials are limited by the absence of tests to detect infection and early treatment efficacy. This perspective highlights the shortfalls and strengths of polymerase chain reaction (PCR) as a study endpoint for anti-parasitic drug development. Serologic reversion, the gold standard test of cure, may take decades to occur in adults and therefore is challenging as an endpoint for drug development. Use of PCR as a marker of infection and treatment response has notable limitations due to low parasitemia in CICD, fluctuations in circulating (versus tissue) parasite burden, strain differences, and assay performance. It is, however, rapidly responsive to therapy, and technological advances have improved detection of different strains and may allow for parasite quantification. Until we have more sensitive tests for parasitological clearance, PCR as a measure of treatment failure may be the best available efficacy endpoint to accelerate early development of much-needed novel therapies. Adequately designed clinical studies are needed to correlate PCR clearance with clinical outcomes and to identify novel biomarkers predictive of clinical outcomes in patients with CICD. Public-private partnerships and health authority engagement are paramount to identify feasible trial endpoints and deliver promising new drug candidates for Chagas disease.

Mechanism of the Change in the Intestinal Microbiota of C-Strain Spodoptera frugiperda (Lepidoptera: Noctuidae) after an Interspecific Transference between Rice and Corn.

Spodoptera frugiperda (J.E.Smith) (Lepidoptera: Noctuidae) was first found in 2019 in Yunnan, China, and it was characterized as a corn strain; it was also found on rice strains there, and it damages rice in China, but little is known about the effect of host plant transfer on the intestinal microbiota and the activities of detoxification enzymes in the C-strain (corn strain) S. frugiperda. The intestinal microbiota and the protective enzyme activity of S. frugiperda that were transferred from rice plants were assessed, and the fourth generation of insects transferred from corn were studied; the gene types of S. frugiperda that were transferred from rice plants were tested using mitochondrial Tpi gene sequences. The results showed that the intestinal microbiota in the C-strain S. frugiperda were changed after the host transference, and the diversity and richness of the intestinal bacterial communities of the S. frugiperda feeding on rice were significantly reduced after the transfer of the host from corn. The predominant species of intestinal bacteria of the S. frugiperda on rice transferred from corn were Enterococcus and Enterobacter, with relative abundances of 28.7% and 66.68%; the predominant species of intestinal bacteria of the S. frugiperda that were transferred from rice and feeding on corn were Enterococcus (22.35%) and Erysipelatoclostridium (73.92%); and the predominant species of intestinal bacteria of S. frugiperda feeding on corn was Enterococcus, with a relative abundance of 61.26%. The CAT (catalase) activity of the S. frugiperda transferred from corn onto rice from corn was reduced, the POD (peroxidase) activity was significantly increased after the transfer from corn, and no significant variations were found for the SOD (superoxide dismutase), CarE (carboxylesterase), and GST (glutathione S-transferase) activities of S. frugiperda after the host plant transfer. The results showed that after feeding on rice, the activities of CAT and POD in the in S. frugiperda body changed in order to resist plant secondary metabolites from corn or rice, but there was no significant change in the detoxification enzymes in the body. In summary, switching the host plant between corn and rice induced variations in the intestinal microbiota in C-strain S. frugiperda owing to the strain difference between the C-strain and the R-strain (rice strain), and this was consistent with the results of the activities of detoxification enzymes. The results indicat that changes in intestinal microbiota and physiological enzymes may be important reasons for the adaptive capacity of C-strain S. frugiperda to rice.

Adolescent intermittent alcohol exposure produces strain-specific cross-sensitization to nicotine and other behavioral adaptations in adulthood in C57BL/6J and DBA/2J mice

Adolescent alcohol exposure is associated with lasting behavioral changes in humans and in mice. Prior work from our laboratory and others have demonstrated that C57BL/6J and DBA/2J mice differ in sensitivity to some effects of acute alcohol exposure during adolescence and adulthood. However, it is unknown if these strains differ in cognitive, anxiety-related, and addiction-related long-term consequences of adolescent intermittent alcohol exposure. This study examined the impact of a previously validated adolescent alcohol exposure paradigm (2–3 g/kg, i.p., every other day PND 30–44) in C57BL/6J and DBA/2J male and female mice on adult fear conditioning, anxiety-related behavior (elevated plus maze), and addiction-related phenotypes including nicotine sensitivity (hypothermia and locomotor depression) and alcohol sensitivity (loss of righting reflex; LORR). Both shared and strain-specific long-term consequences of adolescent alcohol exposure were found. Most notably, we found a strain-specific alcohol-induced increase in sensitivity to nicotine's hypothermic effects during adulthood in the DBA/2J strain but not in the C57BL/6J strain. Conversely, both strains demonstrated a robust increased latency to LORR during adulthood after adolescent alcohol exposure. Thus, we observed strain-dependent cross-sensitization to nicotine and strain-independent tolerance to alcohol due to adolescent alcohol exposure. Several strain and sex differences independent of adolescent alcohol treatment were also observed. These include increased sensitivity to nicotine-induced hypothermia in the C57BL/6J strain relative to the DBA/2J strain, in addition to DBA/2J mice showing more anxiety-like behaviors in the elevated plus maze relative to the C57BL/6J strain. Overall, these results suggest that adolescent alcohol exposure results in altered adult sensitivity to nicotine and alcohol with some phenotypes mediated by genetic background.

THU043 Single-cell Transcriptomics Show Sex-specific Responses To A High Fat Diet, Under Thermoneutral Conditions, Pointing To Reduced Pituitary Plasticity

Abstract Disclosure: T.K. Miles: None. A.N. Lagasse: None. A.K. Odle: None. S.S. Lira: None. M.M. Syed: None. N. Akhter: None. M.C. MacNicol: None. A.M. MacNicol: None. G.V. Childs: None. Hyperinsulinemia caused by a high fat diet (HFD) directly suppressed pituitary growth hormone (Gh) and growth hormone releasing hormone receptor (Ghrhr) mRNAs (Luque and Kineman, 2006). To study the broader impact of a HFD, we exposed 3 groups of FVB.129P mice to 60% HFD for 10 or 15 wks at thermoneutrality (TN) (30 C), or 15 wks at room temperature (RT). Pituitary transcript expression was analyzed with single-cell RNA-seq. HFD-fed mice gained &amp;gt;20 g by 12 wks, showing increased serum leptin (1.8-2X), insulin (2-8X) and IL-6 (4-6X), decreased glucose clearance after a glucose challenge and decreased respiratory quotient indicating fat burning. Activity was unaffected by HFD. Rank plot analyses showed reduced amplitude GH pulses in HFD females fed at RT (15 wks), but not in mice fed at TN. Serum levels of other pituitary hormones were unaffected in HFD females. In HFD males, only serum TSH was elevated significantly (1.8X, 15 wks, TN). Analysis of the scRNA-seq data detected separate clusters representing each cell type. Two multihormonal cell clusters evident in control females mostly disappeared following HFD (15 wks, RT). At the threshold cutoff of avg Log2FC±0.5 with adj p&amp;lt;0.05, female somatotrope clusters showed reduced Pomc, Cga, and ribosomal proteins. Female lactotrope clusters exhibited decreased Gh mRNA (avg Log2FC -1.8 or -2.5) and galanin at 10 wks (TN). At 15 wks (TN), lactotrope clusters showed significant changes in 46 transcripts including reductions in Prl, 29 ribosomal proteins, galanin, and 3 transcriptional regulators. Notably, this cluster showed an increase in synaptotagmin-4 (Syt4), which is also elevated in oxytocin neurons following a HFD. Stem cell clusters in females were strongly impacted by HFD after 15 weeks (TN) with 94 reduced transcripts of which 48 were involved in cytoplasmic translation and ribosome biogenesis and &amp;gt;20 were involved with signaling and cell differentiation. In males, whereas changes in most clusters fell below the threshold cutoffs, stem cell clusters had 13 reduced transcripts after 10 or 15 weeks, most of which involved immune system processes including defense responses and antigen processing and presentation. The GH deficiency in mice fed under thermoneutral conditions was not as severe as that reported by Luque and Kineman, perhaps due to differences in strain, sex, and/or the environmental temperature. The reduced expression of multihormonal clusters, Gh by lactotropes and corticotropes, and Cga and Pomc by somatotropes suggests that HFD may reduce plasticity and activity of progenitor cells in the pituitary. The changes in lactotropes and somatotropes also indicate that HFD may compromise ribosome function and cytoplasmic translation. The reduced transcriptome in stem cells in both males and females suggests HFD interference with cell differentiation and pituitary plasticity as well as dysfunctional immune defense responses. Presentation: Thursday, June 15, 2023

Zika virus diversity in mice is maintained during early vertical transmission from placenta to fetus, but reduced in fetal bodies and brains at late stages of infection.

Since emerging in French Polynesia and Brazil in the 2010s, Zika virus (ZIKV) has been associated with fetal congenital disease. Previous studies have compared ancestral and epidemic ZIKV strains to identify strain differences that may contribute to vertical transmission and fetal disease. However, within-host diversity in ZIKV populations during vertical transmission has not been well studied. Here, we used the established anti-interferon treated Rag1-/- mouse model of ZIKV vertical transmission to compare genomic variation within ZIKV populations in matched placentas, fetal bodies, and fetal brains via RNASeq. At early stages of vertical transmission, the ZIKV populations in the matched placentas and fetal bodies were similar. Most ZIKV single nucleotide variants were present in both tissues, indicating little to no restriction in transmission of ZIKV variants from placenta to fetus. In contrast, at later stages of fetal infection there was a sharp reduction in ZIKV diversity in fetal bodies and fetal brains. All fetal brain ZIKV populations were comprised of one of two haplotypes, containing either a single variant or three variants together, as largely homogenous populations. In most cases, the dominant haplotype present in the fetal brain was also the dominant haplotype present in the matched fetal body. However, in two of ten fetal brains the dominant ZIKV haplotype was undetectable or present at low frequencies in the matched placenta and fetal body ZIKV populations, suggesting evidence of a strict selective bottleneck and possible selection for certain variants during neuroinvasion of ZIKV into fetal brains.

THU127 Deletion Of VGLUT2 In MCH Neurons Induces Sex-Specific Changes In Metabolic And Reproductive Function

Abstract Disclosure: B.G. Beekly: None. MCH neurons have a well-established role in metabolism and energy balance. MCH-knockout (“MCH-KO”) mice, which lack the MCH peptide itself, are hyperactive, hypophagic, and lean. However, MCH neurons express numerous genes involved in the synthesis, packaging, and release of the classical neurotransmitters GABA and glutamate, which are the central nervous system’s predominant inhibitory and excitatory neurotransmitters, respectively. These genes are also critical for normal metabolism, as the conditional deletion of glutamatergic signaling from MCH neurons also affects energy balance in ways that both mirror and differ from MCH-KO. When the metabolic phenotypes of MCH neuronal ablation (diphtheria toxin), MCH-KO, and transgenic Pmch-Cre;Slc17a6fl/fl mice (which lack VGLUT2 in MCH neurons) are compared, all three experimental groups exhibit decreased body weight and adiposity, increased locomotor activity, late-onset hypophagia, attenuated weight gain on high-fat diet, and reductions in circulating leptin. These data point to some functional redundancy between MCH and glutamate release from MCH neurons in the context of energy balance. By contrast, specific indicators of glucose sensing and handling differ between groups: MCH neuronal ablation and conditional VGLUT2 deletion in MCH neurons, but not global MCH-KO, result in reduced sucrose preference and improved glucose tolerance. These data suggest a role for MCH peptide in body weight regulation, but also a glutamate-dependent mechanism by which MCH neurons contribute to glucose homeostasis. Given the findings related to MCH peptide action on pancreatic islet cells and its effects on insulin resistance, it appears that MCH and glutamatergic signaling from MCH neurons must work synergistically for optimal regulation of energy balance. Importantly, all research to date on the metabolic effect of VGLUT2 deletion from MCH neurons has been in male mice. This constitutes a significant gap in the field’s understanding of these phenomena, particularly because of documented sex differences in glucose handling and associated metabolic disorders like diabetes. To address these gap, we generated Pmch-Cre;Slc17a6fl/fl mice, which lack functional VGLUT2 in MCH neurons. Body weight was measured from weaning until 20 weeks of age and the mice were subjected to comprehensive metabolic testing including oral glucose tolerance testing. We were able to recapitulate some, but not all, of the published findings in males, a fact which may in large part be due to strain differences in metabolism and glucose handling. Notably, several sexually dimorphic phenotypes were also observed, reaffirming the importance of incorporating biological sex variables into metabolic studies. Presentation: Thursday, June 15, 2023

A South African Perspective on the Microbiological and Chemical Quality of Meat: Plausible Public Health Implications.

Meat comprises proteins, fats, vitamins, and trace elements, essential nutrients for the growth and development of the body. The increased demand for meat necessitates the use of antibiotics in intensive farming to sustain and raise productivity. However, the high water activity, the neutral pH, and the high protein content of meat create a favourable milieu for the growth and the persistence of bacteria. Meat serves as a portal for the spread of foodborne diseases. This occurs because of contamination. This review presents information on animal farming in South Africa, the microbial and chemical contamination of meat, and the consequential effects on public health. In South Africa, the sales of meat can be operated both formally and informally. Meat becomes exposed to contamination with different categories of microbes, originating from varying sources during preparation, processing, packaging, storage, and serving to consumers. Apparently, meat harbours diverse pathogenic microorganisms and antibiotic residues alongside the occurrence of drug resistance in zoonotic pathogens, due to the improper use of antibiotics during farming. Different findings obtained across the country showed variations in prevalence of bacteria and multidrug-resistant bacteria studied, which could be explained by the differences in the manufacturer practices, handling processes from producers to consumers, and the success of the hygienic measures employed during production. Furthermore, variation in the socioeconomic and political factors and differences in bacterial strains, geographical area, time, climatic factors, etc. could be responsible for the discrepancy in the level of antibiotic resistance between the provinces. Bacteria identified in meat including Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, Campylobacter spp., Salmonella spp., etc. are incriminated as pathogenic agents causing serious infections in human and their drug-resistant counterparts can cause prolonged infection plus long hospital stays, increased mortality and morbidity as well as huge socioeconomic burden and even death. Therefore, uncooked meat or improperly cooked meat consumed by the population serves as a risk to human health.

Fear-Conditioning to Unpredictable Threats Reveals Sex and Strain Differences in Rat Fear-Potentiated Startle (FPS).

Fear-potentiated startle (FPS) has been widely used to study fear processing in humans and rodents. Human studies showed higher startle amplitudes and exaggerated fear reactivity to unpredictable vs. predictable threats in individuals suffering from post-traumatic stress disorder (PTSD). Although human FPS studies use both sexes, a surprisingly limited number of rodent FPS studies use females. Here we investigate the effects of signal-threat contingency, signal-threat order and threat predictability on FPS in both sexes. We use a classic fear-conditioning protocol (100% contingency of cue and shock pairings, with forward conditioning such that the cue co-terminates with the shock) and compare it to modified fear-conditioning protocols (70% contingency; backward conditioning; or cue and shock un-paired). Although there are no sex differences in the startle amplitudes when corrected for body weight, females consistently demonstrate higher shock reactivity during fear-conditioning. Both sexes and strains demonstrate comparable levels of cued, non-cued, and contextual fear in the classic FPS and FPS following fear-conditioning with 70% contingency or backward order (cue co-starts with shock). However, in the classic FPS, Sprague-Dawley females show reduced proportion between cued fear and cue-elicited vigilant state than males. Lastly, a prominent sex difference is uncovered following unpredictable fear-conditioning (cue and shock un-paired), with Wistar, but not Sprague-Dawley, females showing significantly higher startle overall during the FPS recall, regardless of trial type, and higher contextual fear than males. This striking sex difference in processing unpredictable threats in rodent FPS might help to understand the mechanisms underlying higher incidence of PTSD in women.