Improving Military Medical Retention Through Growth Counseling: A Cross-sectional Retention Survey of U.S. Army Physician Assistants.

INTRODUCTION: Although compensation is often the only factor considered with military medical retention, retention has been shown to have many other important factors. Maximizing the retention of military medical personnel not only decreases the cost of recruiting them but also maintains the institutional knowledge of military medicine needed to conduct military medical operations. This study used a recent retention survey of U.S. Army physician assistants (PAs) to understand military medical retention needs. MATERIALS AND METHODS: The study analyzed a survey created, implemented, and collected by the Statistical Cell at the U.S. Army Center of Excellence. In September 2020, all 864 active duty PAs were sent a link to the survey and 290 completed the survey, with a participation rate of 34%. The study used cross-tabulation, Pearson's chi-square, and multivariate logistical regression to analyze the results. Institutional review board approval was obtained through the Army Medical Center of Excellence and University of the Incarnate Word. RESULTS: Pearson's chi-square found that 18 questions in the survey were answered significantly different (P < .05) among those who intended to stay for 20 years in the Army. Binomial logistical regression found that each point higher on a 5-point Likert scale of the belief a PA could accomplish their career goals in Army medicine increased probability of likelihood to stay for 20 years by 74% (95% CI [0.57-0.98]). Each point higher on a 5-point Likert scale of perception Defense Health Agency control of Army medicine affected retention increased probability of likelihood to stay by 54% (95% CI [0.39-0.75]). Every decade older of age increased probability of increased likelihood to stay for 20 years by 63% (95% CI [0.40-0.98]). CONCLUSIONS: The belief a service member can reach their career goals in the military is critical for retention. Using Army Techniques Publication 6-22.1 as a guide, leaders can help subordinates meet their career goals, and this may be the most important task a leader can accomplish to improve their unit's retention. Strengths of this study include the high response rate of 33.6% and results demonstrating the importance of growth counseling which is already part of Army doctrine. Weaknesses include the use of secondary cross-sectional data which impair the ability to derive conclusions from the data. Future studies should explore how training programs for military leaders in growth counseling can help subordinates find their career goals in the military and the effect on military retention.

Nuclear localization signal peptides enhance genetic transformation of Dunaliella salina.

BACKGROUND: Dunaliella salina (D. salina) expression system shows a very attractive application prospect, but it currently has a technical bottleneck, namely the low or unstable expression of recombinant proteins. Given the characteristics of cell-penetrating peptides or/and nuclear localization signal (NLS) peptides, this study is the first attempt to improve the transformation rate of foreign gene with trans-activating transcriptional (TAT) protein or/and NLS peptides. METHODS AND RESULTS: Using salt gradient method, exogenous plasmids were transferred into D. salina cells with TAT or TAT/NLS complexes simultaneously. The ß-glucuronidase gene expression was identified by means of histochemical stain and RT-qPCR detection. Through observation with light microscope, TAT-mediating cells exhibit an apparent cytotoxicity even at ratios of 0.5, no significant toxicity was noted in the TAT/plasmid/NLS complex group. It is obvious that with the addition of peptides the toxicity decreases significantly. Histochemical staining showed that the transformants presented blue color under light microscope, but the negative control and blank control are not. Furthermore, based on a TAT/plasmids ratio of 4 with 10 µg NLS peptides mediation, RT-qPCR results demonstrated that the transcripts of target gene were increased by 269 times than that of control group. CONCLUSIONS: This study demonstrated that combination of TAT and NLS peptides can significantly improve the transformation rate and expression level of foreign gene in D. salina system. It offers a promising way for promoting the application and development of D. salina bioreactor.

Human esophageal fibroblast-derived exosomal miR-21 reduced the cisplatin sensitivity to esophageal carcinoma EC9706 cells.

The objective of this study was to investigate the effect of human esophageal fibroblast-derived exosomal miR-21 on cisplatin sensitivity against esophageal squamous EC9706 cells. EC9706 cells were co-cultured indirectly with human esophageal fibroblasts (HEF) or miR-21 mimics transfected-HEF in the transwell system. The exosomes in HEF-culture conditioned medium were extracted by differential ultracentrifugation. EC9706 cells were co-cultured with HEF-derived exosomes directly. The cisplatin sensitivity against EC9706 cells was revealed via half maximal inhibitory concentration (IC50) values using MTT assay. The expressions of miR-21, programmed cell death 4 (PDCD4) mRNA, and gene of phosphate and tension homology deleted on chromosome ten (PTEN) mRNA were determined by qRT-PCR. The changes of the protein level were detected using western blot assay. IC50 values of cisplatin against EC9706 cells were increased after EC9706 cells were co-cultured with either HEF or exosomes derived from miR-21 mimics-transfected HEF. Following the increased level of miR-21, the mRNA expression and protein levels of PTEN and PDCD4 were decreased in EC9706 cells. The cisplatin sensitivity to EC9706 cells was reduced by HEF-derived exosomal miR-21 through targeting PTEN and PDCD4. This study suggested that non-tumor cells in the tumor micro-environment increased the tumor anti-chemotherapy effects through their exosomes.

CRISPR/Cas9-induced ß-carotene hydroxylase mutation in Dunaliella salina CCAP19/18.

Dunaliella salina (D. salina) has been exploited as a novel expression system for the field of genetic engineering. However, owing to the low or inconsistent expression of target proteins, it has been greatly restricted to practical production of recombinant proteins. Since the accurate gene editing function of clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system, ß-carotene hydroxylase gene was chosen as an example to explore D. salina application with the purpose of improving expression level of foreign genes. In this paper, based on pKSE401 backbone, three CRISPR/Cas9 binary vectors were constructed to targeting exon 1 and 3 of the ß-carotene hydroxylase of D. salina CCAP19/18 (Dschyb). D. salina mutants were obtained by salt gradient transformation method, and the expression of Dschyb gene were identified through real-time fluorescent quantitative PCR. Moreover, carotenoids content was analyzed by high-performance liquid chromatography at different time points after high intensity treatment. Compared with wild type strains, the ß-carotene levels of mutants showed a significant increase, nearly up to 1.4 µg/ml, and the levels of zeaxanthin decreased to various degrees in mutants. All the results provide a compelling evidence for targeted gene editing in D. salina. This study gave a first successful gene editing of D. salina which has a very important practical significance for increasing carotene yield and meeting realistic industry demand. Furthermore, it provides an approach to overcome the current obstacles of D. salina, and then gives a strong tool to facilitates the development and application of D. salina system.

Human esophageal fibroblast-derived exosomal miR-21 reduced the cisplatin sensitivity to esophageal carcinoma EC9706 cells

The objective of this study was to investigate the effect of human esophageal fibroblast-derived exosomal miR-21 on cisplatin sensitivity against esophageal squamous EC9706 cells. EC9706 cells were co-cultured indirectly with human esophageal fibroblasts (HEF) or miR-21 mimics transfected-HEF in the transwell system. The exosomes in HEF-culture conditioned medium were extracted by differential ultracentrifugation. EC9706 cells were co-cultured with HEF-derived exosomes directly. The cisplatin sensitivity against EC9706 cells was revealed via half maximal inhibitory concentration (IC50) values using MTT assay. The expressions of miR-21, programmed cell death 4 (PDCD4) mRNA, and gene of phosphate and tension homology deleted on chromosome ten (PTEN) mRNA were determined by qRT-PCR. The changes of the protein level were detected using western blot assay. IC50 values of cisplatin against EC9706 cells were increased after EC9706 cells were co-cultured with either HEF or exosomes derived from miR-21 mimics-transfected HEF. Following the increased level of miR-21, the mRNA expression and protein levels of PTEN and PDCD4 were decreased in EC9706 cells. The cisplatin sensitivity to EC9706 cells was reduced by HEF-derived exosomal miR-21 through targeting PTEN and PDCD4. This study suggested that non-tumor cells in the tumor micro-environment increased the tumor anti-chemotherapy effects through their exosomes.

Child welfare workers and peritraumatic distress: The impact of COVID-19.

Whilst there is broad consensus that COVID-19 has had a pernicious impact on child welfare services, in general, and child welfare workers, specifically, this notion has not been thoroughly examined in the literature. This exploratory study examined COVID-19 related peritraumatic distress among child welfare workers (N = 1996) in one southeastern state in the United States (U.S.). Findings suggest that the study sample was experiencing distress levels above normal ranges; 46.4% of participants were experiencing mild or severe distress. Sexual orientation, self-reported physical and mental health, relationship status, supervision status, and financial stability impacted distress levels experienced by child welfare workers. Overall, data suggest that COVID-19 is impacting child welfare workers and there is a need to conceptualize, implement, and evaluate initiatives aimed at assuaging distress among child welfare workers.

Expression of Matrix Metalloproteinase-2, Tissue Inhibitor of Matrix Metalloproteinase-2 and CD147 in the Traditional Chinese Medicine "Compound T11" for Treatment of Chronic Liver Injury.

OBJECTIVES: To measure the expression of matrix metalloproteinase (MMP)-2, tissue inhibitor of matrix metalloproteinase inhibitor (TIMP)-2, and CD147 in mice with chronic liver injury induced by carbon tetrachloride after treatment with the traditional Chinese medicine (TCM) "Compound T11". METHOD: Sixty male ICR mice were divided randomly into 6 groups of 10: control (C), model (M), low-dose treatment (LT; 50 mg/mL of Compound T11), medium-dose treatment (MT, 100 mg/mL), high-dose treatment (HT, 150 mg/mL), and positive drug treatment (YT, 67.5 mg/mL). Each group was modeled for 7 weeks. Groups M, LT, MT, HT, and YT were injected (s.c.) with 20% carbon tetrachloride diluted with olive oil, and group C was given olive oil in the same way twice a week. After modeling, the treatment groups were administered Compound T11 at the concentrations shown above by oral gavage daily for 2 weeks, while group C was given 0.5% carboxymethyl cellulose sodium. After the final treatment, mice were killed and their liver tissues were excised. Immunohistochemical staining was performed to measure the protein expression of MMP-2, TIMP-2, and CD147, and western blotting was used to measure the protein expression of MMP-2, TIMP-2, CD147, and α-smooth muscle actin (SMA). MMP-2, TIMP-2, and CD147 mRNA expression was determined by quantitative fluorescence real-time PCR. RESULTS: Compound T11 increased the protein expression of MMP-2 and CD147 and decreased the protein expression of TIMP-2 and α-SMA. CONCLUSIONS: Treatment of chronic liver injury by TCM Compound T11 may be associated with changes to the expression of MMP-2 and CD147, and the inhibition of TIMP-2 expression.

Ethanol extract of Rehmannia glutinosa exerts antidepressant-like effects on a rat chronic unpredictable mild stress model by involving monoamines and BDNF.

The dried roots of Rehmannia glutinosa Libosch. (Scrophulariaceae) are of both medicinal and nutritional importance. Our previous study has found that the 80% ethanol extract of R. glutinosa (RGEE) produced antidepressant-like activities in mouse behavioral despair depression models. However, its mechanisms are still unclear. The present study aimed to observe the antidepressant-like mechanisms of RGEE on a rat chronic unpredictable mild stress (CUMS) model by involving monoaminergic neurotransmitters and brain-derived neurotrophic factor (BDNF). CUMS-stressed rats were orally given RGEE daily (150, 300, and 600 mg/kg) or fluoxetine hydrochloride (FH) for 3 weeks after starting the CUMS procedure. Sucrose preference test was carried out to observe depression-like behavior, and serum and brain tissues were used for neurochemical and fluorescent quantitative reverse transcription PCR analysis. Results demonstrated that CUMS induced depression-like behavior, whereas RGEE and FH administration inhibited this symptom. Furthermore, CUMS caused excessively elevated levels of serum corticosterone (CORT), an index of hypothalamic-pituitary-adrenal (HPA) axis hyperactivity, in a manner attenuated by RGEE and FH administration. RGEE administration also further elevated monoamine neurotransmitters and BDNF levels, up-regulated the mRNA expression of BDNF and tropomyosin-related kinase B (TrkB) in hippocampus of rats suffering CUMS. Together, our findings suggest that RGEE can improve CUMS-evoked depression-like behavior, and indicate its mechanisms may partially be associated with restoring HPA axis dysfunctions, enhancing monoamineergic nervous systems, and up-regulating BDNF and TrkB expression.

Exploring member perspectives on participation on child welfare Citizen Review Panels: A national study.

The year 2016 marked the 20th anniversary of the Child Abuse Prevention and Treatment Act (CAPTA) amendments (P.L. 104-235) that mandated Citizen Review Panels (CRPs). CRPs are citizen volunteer groups authorized by United States (U.S.) federal law to examine policies and procedures of state child welfare agencies. Despite the potential of CRPs to positively impact child welfare systems outcomes associated with child abuse and neglect, and the millions of dollars in resources allocated to these groups, there remains a dearth in the literature about CRPs. This exploratory study examined CRP member perceptions from across the United States. Researchers collected scaled survey data to examine member knowledge, engagement and assessment of panel influence and impact, membership composition, and meeting structure. Results revealed that panel members lack some knowledge related to the federal mandate guiding their work, and data suggest the need for panels to more adroitly recruit and retain members representative of the communities in which the panels are formed. After a brief review of background literature, this paper will explicate key results, discuss these findings, and identify salient practice, policy and research implications derived from the study.

Planning a mentorship initiative for foster parents: Does gender matter?

Despite the use of mentoring programs in fields such as business, career training, and youth development, little is known about how mentoring can be used to train and support new foster parents. This paper describes how Concept Mapping was used with current foster parents to develop a conceptual framework suitable to plan a foster parent mentor program. A secondary aim of this study was to explore priority differences in the conceptualization by self-reported gender (foster mothers vs. foster fathers). Participant data was collected via three qualitative brainstorming sessions, and analyzed using non-metric multidimensional scaling and hierarchical cluster analysis. Findings indicate that foster parents participating in this study conceptualized effective mentor programs via a seven cluster solution. Study results also showed no significant differences in cluster ratings by gender. Implications for practice and program planning are identified, as well as areas for future research.

BDNF and COX-2 participate in anti-depressive mechanisms of catalpol in rats undergoing chronic unpredictable mild stress.

Catalpol, a major compound in Rehmannia glutinosa with both medicinal and nutritional values, has been previously confirmed to shorten the duration of immobility in mice exposed to tail suspension and forced swimming tests. This study attempted to examine the anti-depressive mechanisms of catalpol in rats undergoing chronic unpredictable mild stress (CUMS) by involving brain-derived neurotrophic factor (BDNF) and cyclooxygenase-2 (COX-2). CUMS-exposed rats were given catalpol daily (5, 10, and 20mg/kg, ig) or a reference drug, fluoxetine hydrochloride (FH, 10mg/kg, ig), at 5 weeks after starting the CUMS procedure. Sucrose preference test was performed to observe depression-like behavior, and serum and brain tissues were used for neurochemical and fluorescent quantitative reverse transcription PCR analysis. CUMS induced depression-like behavior, whereas catalpol and FH administration attenuated this symptom. Moreover, CUMS caused excessively elevated levels of serum corticosterone, an index of hypothalamic-pituitary-adrenal (HPA) axis hyperactivation, in a manner attenuated by catalpol and FH administration. Catalpol administration also further decreased BDNF activities, downregulated the mRNA expression of BDNF and tropomyosin-related kinase B (TrkB), and reversed the excessive elevation in the activities and mRNA expression levels of COX-2 and prostaglandin E2 (PGE2) in the hippocampus and frontal cortex of rats undergoing CUMS. Results indicate that catalpol can ameliorate CUMS-induced depression-like behavior, and suggest its mechanisms may partially be ascribed to restoring HPA axis dysfunctions, upregulating BDNF expression and its cognate receptor TrkB, and downregulating COX-2 expression, thereby reducing PGE2 levels in the brain.

Development and evaluation of a dot blot assay for rapid determination of invasion-associated gene ibeA directly in fresh bacteria cultures of E. coli.

The ibeA gene, one of the important invasion-associated genes in neonatal meningitis Escherichia coli (NMEC), has been recently detected in avian pathogenic E. coli (APEC). Thus, it is necessary to close monitor the possible contamination of the poultry farms and its products to people. Here, a dot blot method for detecting the ibeA gene in E. coli was developed and validated. For the present study, probe sequence was designed and optimized for the specificity of dot blot. A 342-bp conserved fragment of ibeA gene was selected and labeled with digoxigenin (DIG)-dUTP according to the manufacturer's guidelines, which indicated that this probe hybridizes with ibeA. In our established method, the bacteria culture samples were directly spotted on the membrane, following simple lyses on the membrane. Hence, the extraction of genomic DNA is not required, which reduces the workload and shortens the time. Furthermore, this assay was very sensitive, which could detect as few as 2.5 × 10(3) CFU bacteria. The diagnostic reliability of this dot blot was evaluated on 467 APEC bacteria samples by using PCR analysis. Both methods showed that the result was in complete concordance. The dot blot assay was proved to be a simple, rapid, highly accurate, and cost-effective method to identify invasion-associated genes ibeA, which could be applied for initial screening of a large number of clinical samples or direct detection of bacteria culture.

Effects of ibeA deletion on virulence and biofilm formation of avian pathogenic Escherichia coli.

The ibeA gene is located on a genomic island, GimA, which is involved in the pathogenesis of neonatal meningitis Escherichia coli (NMEC) and avian pathogenic E. coli (APEC). The prevalence of ibeA in the APEC collection in China was investigated, and 20 of 467 strains (4.3%) were positive. In addition, analysis of the association of the E. coli reference (ECOR) groups with positive strains revealed that ibeA was linked to group B2. The ibeA gene in DE205B was analyzed and compared to those of APEC and NMEC, which indicated that the specificity of ibeA was not consistent along pathotypes. The invasion of chicken embryo fibroblast DF-1 cells by APEC DE205B and RS218 was observed, which suggested that DF-1 cells could be a model to study the mechanism of APEC invasion. The inactivation of ibeA in APEC DE205B led to the reduced capacity to invade DF-1 cells, defective virulence in vivo, and decreased biofilm formation compared to the wild-type strain. In addition, strain AAEC189 expressing ibeA exhibited enhanced invasion capacity and biofilm formation. The results of the quantitative real-time reverse transcription-PCR (qRT-PCR) analysis and animal system infection experiments indicated that the loss of ibeA decreased the colonization and proliferation capacities of APEC in the brain during system infection.

A monolayer study on phase behavior and morphology of binary mixtures of sulfatides with DPPC and DPPE.

Sulfatides are important constituents of brain myelin membranes and it is thought to be involved in lateral domain formation in biological membranes. In this work, the interaction of mixed systems of sulfatide with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), two of the major components in biological membranes, was investigated using the monolayer technique at the air-water interface. Based on the regular solution theory, the miscibility of the two binary systems in the mixed monolayer was evaluated in terms of mean surface area per molecule (A(m)), excess molecular area (DeltaA((ex))), surface excess Gibbs energy (DeltaG((ex))), interaction parameter (omega) as well as activity coefficients (f(1) and f(2)) of formed films. Thermodynamic analysis indicates in the two binary systems with negative deviations from the ideal behavior. Accordingly, the values of the Gibbs energy of mixing, sulfatide-DPPC form stable mixtures at X(sul)=0.4 (X(sul) is molar ratio of sulfatide in binary mixture) for all the selected pressures. As for sulfatide/DPPE system, at pi=5 and 30 mN m(-1), the minimum for the Gibbs energy of mixing was found at X(sul)=0.6 and 0.2 respectively. But the minimum appeared at X(sul)=0.4 for other surface pressures. The activity coefficients (f(1) and f(2)) of mixed monolayers were evaluated which show a marked dependence on the mole faction of sulfatide X(sul). AFM images could support the above findings as well as interpretation.

Behavior of sulfatide/cholesterol mixed monolayers at the air/water interface.

The monolayer properties of sulfatide and cholesterol binary system have been investigated with surface pressure-mean molecular area isotherms measurements and atomic force microscopy (AFM). The thermodynamic analysis indicates that the obtained negative deviation of the excess molecular area (DeltaA((ex))) and surface excess Gibbs energy (DeltaG((ex))) from the ideal behavior at various molar ratios, suggesting an attractive interaction between sulfatide and cholesterol in the monolayers as compared with the pure components monolayers. Meanwhile, the compression modulus (C(s)(-1)) vs. surface pressure (pi) and activity coefficients (f(1) and f(2)) of mixed films dependencies for mixed monolayers are drawn at different mole fractions. The AFM images for the mixed sulfatide/cholesterol monolayers deposited on the mica at 15 and 30mNm(-1) show the stronger molecular attractive force to form condensed structure. The behavior of sulfatide is thought to be involved in lateral domain formation in biological membranes. Therefore, the interaction between sulfatide and cholesterol becomes more important in mimicking "lipid rafts" domains.