Histochemistry and transcriptomics of mucins and peritrophic membrane (PM) proteins along the midgut of a beetle with incomplete PM and their complementary function.

The midgut of Zabrotes subfasciatus (Coleoptera) and other insects may have regions lacking a peritrophic membrane (matrix, PM) and covered with a jelly-like material known as peritrophic gel. This work was undertaken to test the hypothesis that the peritrophic gel is a vertebrate-like mucus. By histochemistry we identified mucins along the whole midgut, which contrasts with the known occurrence of PM only at the posterior midgut. We also analyzed the expression of the genes coding for mucus-forming mucins (Mf-mucins), peritrophins, chitin synthases and chitin deacetylases along the midgut and carcass (insect without midgut) by RNA-seq. Mf-mucins were identified as proteins with high O-glycosylation and multiple tandem repeats of Pro/Thr/Ser residues. Peritrophins were separated into PM proteins, cuticular proteins analogous to peritrophins (CPAPs) and ubiquitous-chitin-binding domain-(CBD)-containing proteins (UCBPs). PM proteins have at least 3, CPAP one or 3, and UCBPs have a varied number of CBDs. PM proteins are more expressed at midgut, CPAP at the carcass, and UCBP at both. The results showed that most PM proteins are mainly expressed at the posterior midgut, together with midgut chitin synthase and chitin deacetylase, and in agreement with the presence of PM only at the posterior midgut by visual inspection. The excretion of most midgut chitinase is avoided, suggesting that the shortened PM is functional. Mf-mucins are expressed along the whole midgut, probably forming the extracellular mucus layer observed by histochemistry. Thus, the lack of PM at anterior and middle midgut causes the exposure of a mucus, which may correspond to the previously described peritrophic gel. The putative functional interplay of mucus and PM is discussed. The major role of mucus is proposed to be tissue protection and of PM to enhancing digestive efficiency by allowing enzyme recycling.

Racial and Ethnic Disparities in Stillbirth among Pregnant Women with Obesity.

OBJECTIVE: The aim of this study was to examine the relationship between obesity and risk of stillbirth among pregnant women with obesity in the United States, with a focus on racial and ethnic disparities. STUDY DESIGN: We conducted a retrospective cross-sectional analysis of birth and fetal data from the 2014 to 2019 National Vital Statistics System (N = 14,938,384 total births) to examine associations between maternal body mass index (BMI) and risk of stillbirth. Cox's proportional hazards regression model was used to compute adjusted hazard ratios (HR) as a measure of risk of stillbirth in relation to maternal BMI. RESULTS: The stillbirth rate was 6.70 per 1,000 births among women with prepregnancy obesity, while the stillbirth rate among women with a normal (nonobese) prepregnancy BMI was 3.85 per 1,000 births. The risk of stillbirth was greater among women with obesity compared with women without obesity (HR: 1.39; 95% confidence interval [CI]: 1.37-1.41). Compared with non-Hispanic (NH) Whites, women identifying as NH-other (HR: 1.66; 95% CI: 1.61-1.72) and NH-Black (HR: 1.31; 95% CI: 1.26-1.35) were at higher risk of stillbirth, while Hispanic women had a decreased likelihood of stillbirth (HR: 0.38; 95% CI: 0.37-0.40). CONCLUSION: Obesity is a modifiable risk factor for stillbirth. Public health awareness campaigns and strategies targeting weight management in women of reproductive age and racial/ethnic populations at highest risk for stillbirth, are needed. KEY POINTS: · Stillbirth rates differ by race and ethnicity.. · Risk of stillbirth was greatest among women with obesity.. · Stillbirth rates rise with ascending prepregnancy BMI..

Identification and characterization of the T cell receptor (TCR) repertoire of the cynomolgus macaque (Macaca Fascicularis).

BACKGROUND: Cynomolgus macaque (Macaca fascicularis) is an attractive animal model for the study of human disease and is extensively used in biomedical research. Cynomolgus macaques share behavioral, physiological, and genomic traits with humans and recapitulate human disease manifestations not observed in other animal species. To improve the use of the cynomolgus macaque model to investigate immune responses, we defined and characterized the T cell receptor (TCR) repertoire. RESULT: We identified and analyzed the alpha (TRA), beta (TRB), gamma (TRG), and delta (TRD) TCR loci of the cynomolgus macaque. The expressed repertoire was determined using 22 unique lung samples from Mycobacterium tuberculosis infected cynomolgus macaques by single cell RNA sequencing. Expressed TCR alpha (TRAV) and beta (TRBV) variable region genes were enriched and identified using gene specific primers, which allowed their functional status to be determined. Analysis of the primers used for cynomolgus macaque TCR variable region gene enrichment showed they could also be used to amplify rhesus macaque (M. mulatta) variable region genes. CONCLUSION: The genomic organization of the cynomolgus macaque has great similarity with the rhesus macaque and they shared > 90% sequence similarity with the human TCR repertoire. The identification of the TCR repertoire facilitates analysis of T cell immunity in cynomolgus macaques.

Enriching and Characterizing T Cell Repertoires from 3' Barcoded Single-Cell Whole Transcriptome Amplification Products.

Antigen-specific T cells play an essential role in immunoregulation and many diseases such as cancer. Characterizing the T cell receptor (TCR) sequences that encode T cell specificity is critical for elucidating the antigenic determinants of immunological diseases and designing therapeutic remedies. However, methods of obtaining single-cell TCR sequencing data are labor and cost intensive, typically requiring both cell sorting and full-length single-cell RNA-sequencing (scRNA-seq). New high-throughput 3' cell-barcoding scRNA-seq methods can simplify and scale this process; however, they do not routinely capture TCR sequences during library preparation and sequencing. While 5' cell-barcoding scRNA-seq methods can be used to examine TCR repertoire at single-cell resolution, doing so requires specialized reagents which cannot be applied to samples previously processed using 3' cell-barcoding methods.Here, we outline a method for sequencing TCRα and TCRß transcripts from samples already processed using 3' cell-barcoding scRNA-seq platforms, ensuring TCR recovery at a single-cell resolution. In short, a fraction of the 3' barcoded whole transcriptome amplification (WTA) product typically used to generate a massively parallel 3' scRNA-seq library is enriched for TCR transcripts using biotinylated probes and further amplified using the same universal primer sequence from WTA. Primer extension using TCR V-region primers and targeted PCR amplification using a second universal primer result in a 3' barcoded single-cell CDR3-enriched library that can be sequenced with custom sequencing primers. Coupled with 3' scRNA-seq of the same WTA, this method enables simultaneous analysis of single-cell transcriptomes and TCR sequences which can help interpret inherent heterogeneity among antigen-specific T cells and salient disease biology. The method presented here can also be adapted readily to enrich and sequence other transcripts of interest from both 3' and 5' barcoded scRNA-seq WTA libraries.

Multimodal profiling of lung granulomas in macaques reveals cellular correlates of tuberculosis control.

Mycobacterium tuberculosis lung infection results in a complex multicellular structure: the granuloma. In some granulomas, immune activity promotes bacterial clearance, but in others, bacteria persist and grow. We identified correlates of bacterial control in cynomolgus macaque lung granulomas by co-registering longitudinal positron emission tomography and computed tomography imaging, single-cell RNA sequencing, and measures of bacterial clearance. Bacterial persistence occurred in granulomas enriched for mast, endothelial, fibroblast, and plasma cells, signaling amongst themselves via type 2 immunity and wound-healing pathways. Granulomas that drove bacterial control were characterized by cellular ecosystems enriched for type 1-type 17, stem-like, and cytotoxic T cells engaged in pro-inflammatory signaling networks involving diverse cell populations. Granulomas that arose later in infection displayed functional characteristics of restrictive granulomas and were more capable of killing Mtb. Our results define the complex multicellular ecosystems underlying (lack of) granuloma resolution and highlight host immune targets that can be leveraged to develop new vaccine and therapeutic strategies for TB.

Assessment of anterior positioning splint in conjunction with lateral pterygoid BTX injection to treat TMJ disc displacement with reduction - a preliminary report.

OBJECTIVE: Treatment of temporomandibular disc displacement with reduction is controversial. This study assesses the use of an anterior positioning splint with botulinum toxin in the lateral pterygoid muscle (BTX) for such cases. METHODS: Twelve joints were included; groups I and II received BTX injection while group II also received an anterior positioning splint. Pain scores and clicking status were recorded at regular intervals then a postoperative MRI was done after 4 months. RESULTS: Clinical improvement was noted in both groups. Mean pain scores dropped significantly and clicks in the twelve joints disappeared in 83% of group I and 33% of group II. MRIs showed significant disc position improvement with the higher mean change (1.33 ± 0.76) in group I. Group I showed better improvement of discal position and only one joint regained a click. Patients of group II reported discomfort from the splint which may have caused psychological distress and so worst pain scores. CONCLUSIONS: Group I  showed slightly better results but the cost of BTX injections and the complications of the splint should be kept in mind and the decision of treatment selection made according to each condition.

Costs and Charges for Pediatric Tonsillectomy in New York State.

Objective In this study, we aimed to determine the correlation between costs/charges related to admissions for pediatric tonsillectomy in New York State (NYS) and variables including discharge year, All Patient Refined (APR) severity of illness, length of hospital stay, payment typology, location, race, and institutional factors during 2009-2017. Methods Data were extracted from the Statewide Planning and Research Cooperative System (SPARCS) Hospital Inpatient Discharges database developed by the NYS Department of Health. Statistical analysis was employed to determine multiple linear regression coefficients with the costs and charges set as the dependent variable. Results Costs increased by an estimated $230.73 (p<.001) each year, and charges increased by an estimated $1,231.41 (p<.001) annually. For each categorical increase in severity of illness, costs increased by $1,019.21 (p<.001), and charges increased by $3,088.41 (p<.001). For each day spent in the hospital, costs increased by $3,539.23 (p<.001), and charges increased by $8,908.01 (p<.001). Unspecified managed care had the highest mean costs and charges (p<.001). Bronx County had the highest costs, and Queens County had the highest charges. Queens County demonstrated the largest gap between costs and charges. Conclusion This study revealed that the costs and charges related to admissions for elective tonsillectomy had risen from 2009 to 2017, and these changes were not accounted for by inflation alone. We found that the costs and charges for inpatient pediatric tonsillectomy were significantly correlated with discharge year, APR severity of illness, length of hospital stay, location of the hospital, and primary payer.

Pediatric Drug-Induced Sleep Endoscopy: Technique and Scoring System.

Drug-induced sleep endoscopy (DISE) is an invaluable tool for identifying sites of obstruction for patients with obstructive sleep apnea (OSA). During DISE, the patient is in a state of drug-induced sleep, and a flexible laryngoscope is passed through the nose into the upper airway. Sites of obstruction are visualized and scored to guide surgical management. Currently, there is no universally accepted method of DISE analysis and scoring. This limitation in comparability impedes large-scale analysis between clinicians, institutions, and studies. In this report, we propose a standardized method of scoring and performing DISE in children with OSA. Our DISE scoring system is internally developed, consistent through the study, and addresses all levels of potential upper airway obstruction.

Tissue Adhesives for Hernia Mesh Fixation: A Literature Review.

In an effort to optimize the results during inguinal hernia repair, surgeons need to consider the role of different mesh fixation techniques. The use of tissue adhesives is proposed for mesh fixation, which demonstrate similar or improved outcomes in the realm of postoperative pain, hernia recurrence, cost, and formation of a hematoma or seroma. In this review, mesh fixation using fibrin glue and cyanoacrylate glue is compared to standard suture or tack techniques. The results of this investigation warrant consideration by surgeons seeking to improve patient outcomes.

Intestinal microbes influence development of thymic lymphocytes in early life.

The thymus generates cells of the T cell lineage that seed the lymphatic and blood systems. Transcription factor regulatory networks control the lineage programming and maturation of thymic precursor cells. Whether extrathymic antigenic events, such as the microbial colonization of the mucosal tract also shape the thymic T cell repertoire is unclear. We show here that intestinal microbes influence the thymic homeostasis of PLZF-expressing cells in early life. Impaired thymic development of PLZF+ innate lymphocytes in germ-free (GF) neonatal mice is restored by colonization with a human commensal, Bacteroides fragilis, but not with a polysaccharide A (PSA) deficient isogenic strain. Plasmacytoid dendritic cells influenced by microbes migrate from the colon to the thymus in early life to regulate PLZF+ cell homeostasis. Importantly, perturbations in thymic PLZF+ cells brought about by alterations in early gut microbiota persist into adulthood and are associated with increased susceptibility to experimental colitis. Our studies identify a pathway of communication between intestinal microbes and thymic lymphocytes in the neonatal period that can modulate host susceptibility to immune-mediated diseases later in life.

Testemunhos de Sobreviventes ao Desastre de 2011 em Petrópolis: Abordagem Psicossocial em um Campo Ferido / Survivor's testimonies from 2011 disaster in Petrópolis: Psychosocial approach in a wounded field / Testimonios de supervivientes del desastre de 2011 en Petrópolis: Enfoque psicosocial en un campo herido

Este artigo apresenta uma análise dos testemunhos de sobreviventes ao desastre ocorrido em 2011, no Vale do Cuiabá (Petrópolis, RJ), região serrana do Estado do Rio de Janeiro, a partir de uma abordagem psicossocial. Os testemunhos foram coletados sete anos após a tragédia por meio de entrevistas individuais e grupais e mostram que o desastre continua. Dentre os resultados, evidencia-se que as pessoas entrevistadas se percebem como sobreviventes e não como afetados, atingidos ou vítimas; a vivência do abandono permanece com a ausência do Estado; e a precariedade das políticas públicas praticadas fragmentaram as relações comunitárias, gerando comprometimento da comunicação interpessoal. Destaca-se que o desastre permanece enquanto durar o sofrimento social dos envolvidos e que os profissionais que atuam nesse campo necessitam desenvolver uma escuta qualificada.(AU)

This paper presents an analysis of testimonies taken from survivors of 2011 Vale do Cuiabá (Petrópolis, RJ) disaster, located at the highland region of Rio de Janeiro State, based on a psychosocial approach. Testimonies were collected seven years after the tragedy through individual and group interviews. They reveal that the disaster still goes on for those people. Among the results, it is evident that respondents perceived themselves as survivors and not as affected population or victims; the experience of abandonment remains with the absence of State; and the public policies model has shattered communitarian relationship, compromising interpersonal communication. It stands out that the disaster will endure while social suffering from affected people remains, and there is a necessity of developing a more comprehensive and qualified approach for the professionals who take action on disaster situations.(AU)

Este artículo presenta un análisis de los testimonios de los supervivientes del desastre ocurrido en el 2011, en el "Valle del Cuiabá" (Petrópolis, RJ), en la región montañosa del Estado de Rio de Janeiro, a partir de un enfoque psicosocial. Los testimonios fueron recogidos tras siete años de la tragedia por medio de entrevistas individuales y grupales. Para los participantes, el desastre sigue. En los resultados se hace muy evidente que las personas entrevistadas se perciben como supervivientes y no como afectados o víctimas; la vivencia de abandono permanece con la ausencia del Estado; y la precariedad de las políticas públicas implementadas fragmentó las relaciones comunitarias, generando fragilidad en la comunicación interpersonal. Se destaca que el desastre permanece mientras dure el sufrimiento social de los involucrados y que existe necesidad de desarrollo de la escucha cualificada para los profesionales que actúan en los desastres.(AU)

Prevalence of convergence insufficiency among secondary school students in Khartoum, Sudan.

BACKGROUND: Few studies have examined the prevalence of convergence insufficiency (CI) in a school-based population in Sudan. This study sought to determine the prevalence of CI and its related clinical characteristics among Sudanese secondary school students. MATERIALS AND METHODS: A descriptive cross-sectional survey was conducted in Central Khartoum North, Sudan, in the academic year 2013/2014. A total of 4211 secondary school students, with a mean age of 15.5 ± 2.5 years, underwent complete eye examination, and were screened for symptoms associated with near work. Near and distance heterophoria was measured with the alternate cover test using a prism bar; near point of convergence (NPC) and positive fusional vergence (PFV) at near were determined. RESULTS: Of the 4211 students screened, 329 (7.8%) were diagnosed with CI. Of these, 173 (52.6%) students were male and 156 (47.4%) were female; there was no significant relationship between sex and CI (P > 0.05). Standard schools had a higher prevalence of CI (43%) than geographic schools (36%) and there was a significant association between CI and the type of school (P < 0.05). In most of the students (78.42%), CI was due to both remote NPC and decreased PFV; in 20.36% of the students, CI was due to remote NPC only, and in very few students (1.22%), it was due to decreased PFV only. CONCLUSIONS: These findings suggest that CI is prevalent in the secondary school population in Central Khartoum North, Sudan.

Bone Height Changes of the Mandibular Edentulous Ridge in Screw Retained Versus Telescopic Restorations for Completely Edentulous Patients.

AIM: This study was established to evaluate the amount of bone height changes in the posterior mandibular area of edentulous patients receiving screw-retained prostheses versus removable telescopic implant overdentures. MATERIAL AND METHODS: Every patient received four inter-foraminal implants regarding the mandibular arch and four anterior implants for the maxillary arch, computer guided surgical guides were planned for the insertion of the implants accurately with a flapless technique. Panoramic radiographs were made immediately, six months and twelve months after the prostheses` use proportional area and vertical measurements were applied to determine changes in the bone height of the posterior mandibular edentulous area. RESULTS: After twelve months, a statistically non-significant amount of bone resorption was reported for both groups. CONCLUSION: Up to the limitations of this study both treatment options the screw retained and telescopic overdenture can be used for rehabilitation of completely edentulous patients. These cases must be followed for a longer period to have a definite answer regarding their efficiency in the long run.

The clinical outcome of adjuvant therapy with black seed oil on intractable paediatric seizures: a pilot study.

AIM: To evaluate the effect of black seed oil, as add-on treatment to antiepileptic drugs (AEDs), on seizure frequency and severity as well as oxidative stress in intractable epilepsy patients. METHODS: A prospective, randomised, single-blinded, controlled, crossover pilot study. Five healthy children were included as controls. Thirty intractable epileptic children were randomly assigned to either Group I or II. Group I received placebo for four weeks, followed by a two-week washout period, and subsequently black seed oil for four weeks. Group II received the same intervention but in the reverse order. All patients received AEDs throughout the study period. Prior to allocation, all patients underwent a neurological assessment and evaluation of oxidative stress markers; total antioxidant capacity (TAC) and malondialehyde (MDA). Patients were assessed at Weeks 4 and 10 for oxidative stress markers and seizure frequency and severity. RESULTS: At baseline, both groups (I, II) had significantly lower serum TAC levels relative to healthy controls (p=0.007), while MDA levels were unchanged. After the 4-week period of black seed oil administration, there was no significant difference between the two groups with regards to seizure frequency, severity, or oxidative stress markers (TAC and MDA; p>0.05). Eight patients had >50% reduction in seizure frequency/severity after black seed oil versus placebo. CONCLUSION: Children with intractable epilepsy show evidence of oxidative stress. Administration of 40-80 mg/kg/day of black seed oil as add-on therapy did not alter either oxidative stress markers or seizure frequency or severity in intractable epileptic patients.