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Abstract Background The genetic diversity of the human immunodeficiency virus type 1 (HIV-1) is critical to lay the groundwork for the design of successful drugs or vaccine. In this study we aimed to characterize and define the molecular prevalence of HIV-1 subclade F1 currently circulating in São Paulo, Brazil. Methods A total of 36 samples were selected from 888 adult patients residing in São Paulo who had previously been diagnosed in two independent studies in our laboratory as being infected with subclade F1 based on pol subgenomic fragment sequencing. Proviral DNA was amplified from the purified genomic DNA of all 36 blood samples by 5 fragments overlapping PCR followed by direct sequencing. Sequence data were obtained from the 5 fragments of pure subclade F1 and phylogenetic trees were constructed and compared with previously published sequences. Subclades F1 that exhibited mosaic structure with other subtypes were omitted from any further analysis Results Our methods of fragment amplification and sequencing confirmed that only 5 sequences inferred from pol region as subclade F1 also holds true for the genome as a whole and, thus, estimated the true prevalence at 0.56%. The results also showed a single phylogenetic cluster of the Brazilian subclade F1 along with non-Brazilian South American isolates in both subgenomic and the full-length genomes analysis with an overall intrasubtype nucleotide divergence of 6.9%. The nucleotide differences within the South American and Central African F1 strains, in the C2-C3 env, were 8.5% and 12.3%, respectively. Conclusion All together, our findings showed a surprisingly low prevalence rate of subclade F1 in Brazil and suggest that these isolates originated in Central Africa and subsequently introduced to South America.

Palliative care focuses on improving the quality of life for people with serious illnesses and their loved ones. This article introduces considerations including barriers to care, intersectionality, minority stress, microaggressions, and social safety that may impact the experience and openness of people to receive this care. The authors outline tools to address these challenges including trauma-informed care and how to recognize bias and earn trust. The authors conclude by offering a model for incorporating these assessments and tools with sample scripts to provide patient-centered and holistic palliative care., Competing Interests: Disclosure The authors have no disclosures., (Published by Elsevier Inc.)

Purpose: The purpose of this study was to characterize the histologic contents of the salpingopharyngeal fold. The primary objective was to observe the presence of salpingopharyngeus (SP) muscle fibers, particularly at the section near the muscle origin at the torus tubarius (TT)., Method: Histologic samples of the salpingopharyngeal fold from 10 cadavers (six females and four males) were analyzed. Following a head bisection, a tissue sample measuring 5 mm in length along the course of the salpingopharyngeal fold was collected from one side (i.e., right or left). The tissue sample was taken from the estimated base of the TT to a point 5 mm inferiorly. Slides were prepared using a standard histological approach and basic pathological staining and analyzed via bright-field microscopy., Results: Skeletal muscle fibers were identified in eight of the 10 tissue blocks of the salpingopharyngeal fold, with dense connective tissue identified in the remaining two tissue blocks. Glandular material was also identified in all 10 tissue blocks., Conclusions: Skeletal muscle fibers and/or dense connective tissue can be consistently identified in the section of the salpingopharyngeal fold near the TT. Glandular material is also consistently present in this same region of the salpingopharyngeal fold. These findings are discussed in relation to possible functional roles of the salpingopharyngeal fold contents, including the SP muscle.

Collaborative learning is a teaching method that brings together students to discuss a topic important for a given course or curriculum and solve a related problem or create a product. By doing this, learners create knowledge together and gain 21 st -century skills such as communication, critical thinking, decision making, leadership and conflict management. Universities had to close their campuses and turn their education fully online in 2020 due to the COVID-19 pandemic, which created a forced step in the evolution of the digitalisation of collaborative teaching. How did TU Delft face this challenge? How did the students experience the online version of collaborative learning? How did distant learning affect their motivation? This article presents four student team projects investigating these questions from the collaborative learning perspective. One of the significant findings of these projects is the lack of socio-emotional interactions during online collaborative work. We present a few guidelines on how to enable these interactions when designing online or blended collaborative education., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

Purpose The aim of the study was to update our information regarding the salpingopharyngeus (SP) muscle using cadaveric and in vivo magnetic resonance imaging (MRI) data. Primary objectives were to (a) observe the presence/absence of the muscle and (b) quantify and describe its dimensions and course. Method SP specimens from 19 cadavers (10 women, nine men) were analyzed. Following head bisection, measurements of SP, including width of the cartilaginous attachment (CW) and width of the superior muscle base (SMW), were taken before and after removal of the overlying mucosa. In addition, SP was analyzed in 15 healthy subjects (eight men, seven women) using high-resolution three-dimensional MRI data. CW and SMW measures were replicated in the paraxial MRI view. Results The presence of the salpingopharyngeal fold and muscle was confirmed bilaterally in all cadaveric and living subjects. Following mucosa removal, mean cadaveric CW and SMW measurements were 5.6 and 3.8 mm, respectively. Mean in vivo CW and SMW were 6.1 and 3.7 mm, respectively. Results from the hierarchical regression analyses revealed that, in both cadaveric and living groups, SMW is dependent on the relationship between age and body weight, after controlling for sex. Conclusions The salpingopharyngeal fold and SP muscle are always present bilaterally and can be quantified at the superior origin using both cadaveric and in vivo three-dimensional MRI data. Though both the superior origin and inferior course of SP are highly variable, the size of the SP muscle is dependent on characteristics known to affect muscle fibers, such as the relationship between age and body weight. Given the consistent and quantifiable presence of the SP muscle, its potential role in velopharyngeal function for speech and swallowing is reconsidered. Supplemental Material https://doi.org/10.23641/asha.14347859.

Objectives: To provide detailed descriptions of contraction-induced morphometric changes in the extravelar segments of the levator veli palatini (LVP) muscle using 3-dimensional (3-D) magnetic resonance imaging (MRI)., Design: Three-dimensional MRI data were acquired at rest and during "silent /i/" from 4 singers. During silent /i/, participants voluntarily sustained velar elevation while breathing orally for the entire scan time. Focusing on the extravelar segments, LVP length, angle of the muscle origin, and cross-sectional area (CSA), measurements were obtained and compared between tasks., Results: Three of the 4 participants exhibited the expected patterns of change following concentric contraction of the LVP muscle. Consistent changes from the resting to the contracted state included reductions in LVP length by 13.5% and angle of the muscle origin by 9.8%, as well as increases in CSAs by 22.1%, on average., Conclusions: This study presented high-resolution data of the LVP muscle behavior with the first in vivo 3-D measurements of the contracted LVP muscle, which can be useful for the validation of computational models that aim at describing biomechanical properties of the LVP muscle in future research. The active behavior of the extravelar LVP muscle also provides some insight on optimal LVP muscle geometry to consider during cleft palate repair.

Progressive curricular changes in medical education over the past two decades have resulted in the diaspora of gross anatomy content into integrated curricula while significantly reducing total contact hours. Despite the development of a wide range of alternative teaching modalities, gross dissection remains a critical component of medical education. The challenge posed to modern anatomists is how to maximize and integrate the time spent dissecting under the current curricular changes. In this study, an alternative approach to the dissection of the pelvis and perineum is presented in an effort to improve content delivery and student satisfaction. The approach involves removal of the perineum en bloc from the cadaver followed by excision of the pubic symphysis, removal and examination of the bladder and associated structures, examination and bisection of the midline pelvic organs in situ, and midsagittal hemisection of the pelvis for identification of the neurovasculature. Results indicate that this novel dissecting approach increases the number of structures identified by 46% ± 14% over current dissecting methods. Survey results indicate that students were better able to integrate lecture and laboratory concepts, understand the concepts, and successfully identify more structures using the new approach (P < 0.05). The concept of anatomic efficiency is introduced and proposed as a standard quantitative measure of gross dissection proficiency across programs and institutions. These findings provide evidence that innovative solutions to anatomy education can be found that help to maintain critical content and student satisfaction in a modern medical curriculum., (© 2019 American Association of Anatomists.)

We present a case of rapidly progressing Addison's disease in adrenal crisis with severe hyponatraemia and absence of hyperkalaemia in a 10-year-old girl. She presented with 2 weeks of vomiting, fatigue and weight loss. Her serum electrolytes obtained 1 week prior to presentation were normal, except for mild hyponatraemia at 131 mmol/L, which dropped to 112 mmol/L on admission. She had normal serum potassium, low-serum osmolality, with elevated urine sodium and osmolality, indistinguishable from syndrome of inappropriate antidiuretic hormone (SIADH). Subsequently, Addison's disease was diagnosed on the basis of gingival hyperpigmentation and undetectable cortisol on adrenocorticotropic hormone stimulation test. She rapidly responded to stress dose hydrocortisone, followed by hydrocortisone and fludrocortisone replacement therapy. The absence of hyperkalaemia in the presence of severe hyponatraemia cannot rule out Addison's disease in children. The mechanism of hypo-osmolar hyponatraemia in primary adrenal insufficiency and clinical clues to differentiate it from SIADH are discussed., (2015 BMJ Publishing Group Ltd.)

ERD10 and ERD14 (for early response to dehydration) proteins are members of the dehydrin family that accumulate in response to abiotic environmental stresses, such as high salinity, drought, and low temperature, in Arabidopsis (Arabidopsis thaliana). Whereas these proteins protect cells against the consequences of dehydration, the exact mode(s) of their action remains poorly understood. Here, detailed evidence is provided that ERD10 and ERD14 belong to the family of intrinsically disordered proteins, and it is shown in various assays that they act as chaperones in vitro. ERD10 and ERD14 are able to prevent the heat-induced aggregation and/or inactivation of various substrates, such as lysozyme, alcohol dehydrogenase, firefly luciferase, and citrate synthase. It is also demonstrated that ERD10 and ERD14 bind to acidic phospholipid vesicles without significantly affecting membrane fluidity. Membrane binding is strongly influenced by ionic strength. Our results show that these intrinsically disordered proteins have chaperone activity of rather wide substrate specificity and that they interact with phospholipid vesicles through electrostatic forces. We suggest that these findings provide the rationale for the mechanism of how these proteins avert the adverse effects of dehydration stresses.

Background: The past few years have seen a vast increase in the amount of genomic data available for a growing number of taxa, including sets of full length cDNA clones and cis-regulatory sequences. Large scale cross-species comparisons of protein function and cis-regulatory sequences may help to understand the emergence of specific traits during evolution., Principal Findings: To facilitate such comparisons, we developed a Gateway compatible vector set, which can be used to systematically dissect cis-regulatory sequences, and overexpress wild type or tagged proteins in a variety of chordate systems. It was developed and first characterised in the embryos of the ascidian Ciona intestinalis, in which large scale analyses are easier to perform than in vertebrates, owing to the very efficient embryo electroporation protocol available in this organism. Its use was then extended to fish embryos and cultured mammalian cells., Conclusion: This versatile vector set opens the way to the mid- to large-scale comparative analyses of protein function and cis-regulatory sequences across chordate evolution. A complete user manual is provided as supplemental material.

Background: All vertebrates share a remarkable degree of similarity in their development as well as in the basic functions of their cells. Despite this, attempts at unearthing genome-wide regulatory elements conserved throughout the vertebrate lineage using BLAST-like approaches have thus far detected noncoding conservation in only a few hundred genes, mostly associated with regulation of transcription and development., Results: We used a unique combination of tools to obtain regional global-local alignments of orthologous loci. This approach takes into account shuffling of regulatory regions that are likely to occur over evolutionary distances greater than those separating mammalian genomes. This approach revealed one order of magnitude more vertebrate conserved elements than was previously reported in over 2,000 genes, including a high number of genes found in the membrane and extracellular regions. Our analysis revealed that 72% of the elements identified have undergone shuffling. We tested the ability of the elements identified to enhance transcription in zebrafish embryos and compared their activity with a set of control fragments. We found that more than 80% of the elements tested were able to enhance transcription significantly, prevalently in a tissue-restricted manner corresponding to the expression domain of the neighboring gene., Conclusion: Our work elucidates the importance of shuffling in the detection of cis-regulatory elements. It also elucidates how similarities across the vertebrate lineage, which go well beyond development, can be explained not only within the realm of coding genes but also in that of the sequences that ultimately govern their expression.

Lesions of the CNS induce a complex cascade of tissue reactions. The purpose of this study was to determine the response of the vasculature to partial spinal cord transection. Adult rat spinal cords were lesioned and then examined during acute, subacute, and chronic periods for the presence of endothelial cells and blood vessels at the lesion site. The association of endothelial cells and astrocytes was examined immunohistochemically (RECA-1 and glial fibrillary associated protein, respectively). During the first 48 h following an incision lesion of the dorsal spinal cord, the vasculature was significantly decreased, concurrently with the tissue loss due to primary and secondary degeneration. Subsequently, at 4 days postlesion, vasculature repair processes were evidenced by a significant increase in the number of vessels present at the lesion center. Blood vessels even formed in areas densely packed with macrophages and tissue debris. After 1 week, the number of blood vessels declined in the lesion center and at the place of the forming caverns. These results show significant initial attempts at repair of the vasculature which do not, however, lead to the restoration of a compact tissue and cannot prevent the subsequent formation of caverns.

Spermagglutination by gosling hepatitis virus was specifically inhibited by sera from sick and convalescent goslings exposed to experimental and natural infection, by hyperimmune horse serum and by sera from vaccinated geese. When compared with passive protection of goslings, neutralization of egg infectivity and precipitation techniques, the spermagglutination-inhibition technique was found to be a highly sensitive serological procedure for antibody detection and assay.

Agglutination of bovine spermatozoa by gosling hepatitis virus grown in embryonated eggs of 3 different hosts and from several different passage levels was demonstrated. A good agreement was found between the egg-infective and spermagglutinating titres of the virus. The reaction closely resembles that previously described for Newcastle disease virus, influenza virus and Mycoplasma. Gosling hepatitis virus differs however from the other spermagglutinating agents by its inability to agglutinate erythrocytes. Optimal physico-chemical requirements for the reaction were determined. The spermagglutination technique provides a rapid and simple method for assaying the virus. The method is also convenient and inexpensive owing to the availability of spermatozoa in pelleted form which can be stored frozen for long periods.