Functional characterization of a cystatin A from the bat Myotis davidii.

Myotis davidii cystatin A (MdCSTA), a stefin A-like from the Chinese native bat species M. davidii, was expressed as a recombinant protein and functionally characterized as a strong inhibitor of the cysteine proteases papain, human cathepsins L and B and the tick cathepsin L-like BmCL1. Despite the highly conserved amino acid sequences among stefins A from different vertebrates, MdCSTA presents a Methionine-2 residue at the N-terminal region and the second binding loop (pos 73-79) that differs from human stefin A (HsCSTA) and might be related to the lower inhibition constant (Ki) value presented by this inhibitor in comparison to human stefin A inhibition to cathepsin B. Therefore, to investigate the importance of these variable regions in cathepsin B inhibition, recombinant stefins A MdCSTA and HsCSTA containing mutations at the second amino acid residue and second binding loop were expressed and evaluated in kinetic assays. Enzymatic inhibition assays with cathepsin B revealed that switching the amino acid residues at position 2 and second binding loop region between bat and human CSTAs improved the HsCSTA's and reduced MdCSTA's inhibitory activity. Additionally, molecular docking analysis estimated lower energy values for the complex between MdCSTA-cathepsin B, in comparison to human CSTA-cathepsin B, while the mutants presented intermediate values, suggesting that other regions might contribute to the higher inhibitory activity against cathepsin B by MdCSTA. In conclusion, MdCSTA, the first bat's stefin A-like inhibitor to be functionally characterized, presented a higher inhibitory activity against cathepsin B in comparison to the human inhibitor, which is partially related to the glutamine-rich second binding loop and Met-2. Further structural analysis should be performed to elucidate potential inhibitor effects on cysteine proteinases.

Is Muscle Pain in Temporomandibular Disorder a Hyperactivity Problem? A Clinical, Thermographic, and Electromyographic Analysis.

ABSTRACT: This study aimed to report clinical data on muscle pain in temporomandibular disorders using thermography, algometry, and surface electromyography (EMG). This was done to support or rule out the hypothesis that painful muscles are always related to muscular hyperactivity. This cross-sectional, analytical study included 50 participants who underwent clinical examination and evaluation of anterior temporal muscles and bilateral masseters using thermography, algometry, and EMG. The muscles with greater pain symptoms had hyporadiation and a lower temperature compared with both contralateral muscles and muscles in the asymptomatic group. There was no muscle hyperactivity at rest on EMG analysis; however, there was a greater capacity for muscle recruitment of these symptomatic muscles after a joint decompression test with cotton rolls, suggesting hypoactivity of the affected muscles in the symptomatic patient group. Muscle pain may be related to hypoactivity of the musculature when there is neurological inhibition of the recruitment of muscle fibers, and treatment must consider these neurophysiological alterations. LEVEL OF EVIDENCE: Level IV.

Schistosoma mansoni infection in residents of a riverside community in Eastern Amazon.

Schistosoma mansoni worms cause a waterborne parasitic disease called schistosomiasis. It commonly affects individuals in lack of sanitation structure. In Brazil, Pará state has Belém as one of the worst sanitation-ranking places in 2023, where schistosomiasis transmission was already documented. This study reports the occurrence of schistosomiasis in residents of Ilha das Onças, an island next to Belém. Stool samples were obtained from participants over 2 years old, all residents from Furo do Rio Grande, one of the rivers on the island. The Kato-Katz technique was performed for parasite investigation in the stool samples. Each participant responded to a sociodemographic and clinical questionnaire. The residences were georeferenced for map designing. Three out of 263 participants were S. mansoni positive, all men, ages ranging from 19 to 41 years old, with low parasitic load. Malacological surveys were carried out, but no Biomphalaria snails were found. Risk factors for schistosomiasis establishment are present on the island, and the lack of sanitation makes it a potential risk area. Malacological surveys are highly encouraged as preventive measures, as well as health surveillance for riverside populations, generating data that will help health authorities in the management and planning of preventive control actions.

Understanding microchromosomal organization and evolution in four representative woodpeckers (Picidae, Piciformes) through BAC-FISH analysis.

The genome organization of woodpeckers has several distinctive features e.g., an uncommon accumulation of repetitive sequences, enlarged Z chromosomes, and atypical diploid numbers. Despite the large diversity of species, there is a paucity of detailed cytogenomic studies for this group and we thus aimed to rectify this. Genome organization patterns and hence evolutionary change in the microchromosome formation of four species (Colaptes campestris, Veniliornis spilogaster, Melanerpes candidus, and Picumnus nebulosus) was established through fluorescence in situ hybridization using bacterial artificial chromosomes originally derived from Gallus gallus and Taeniopygia guttata. Findings suggest that P. nebulosus (2n = 110), which was described for the first time, had the most basal karyotype among species of Picidae studied here, and probably arose as a result of fissions of avian ancestral macrochromosomes. We defined a new chromosomal number for V. spilogaster (2n = 88) and demonstrated microchromosomal rearrangements involving C. campestris plus a single, unique hitherto undescribed rearrangement in V. spilogaster. This comprised an inversion after a fusion involving the ancestral microchromosome 12 (homologous to chicken microchromosome 12). We also determined that the low diploid number of M. candidus is related to microchromosome fusions. Woodpeckers thus exhibit significantly rearranged karyotypes compared to the putative ancestral karyotype.

Chromosome mapping of retrotransposon AviRTE in a neotropical bird species: Trogon surrucura (Trogoniformes; Trogonidae).

Avian genomes are characterized as being more compact than other amniotes, with less diversity and density of transposable elements (TEs). In addition, birds usually show bimodal karyotypes, exhibiting a great variation in diploid numbers. Some species present unusually large sex chromosomes, possibly due to the accumulation of repetitive sequences. Avian retrotransposon-like element (AviRTE) is a long interspersed nuclear element (LINE) recently discovered in the genomes of birds and nematodes, and it is still poorly characterized in terms of chromosomal mapping and phylogenetic relationships. In this study, we mapped AviRTE isolated from the Trogon surrucura genome into the T. surrucura (TSU) karyotype. Furthermore, we analyzed the phylogenetic relationships of this LINE in birds and other vertebrates. Our results showed that the distribution pattern of AviRTE is not restricted to heterochromatic regions, with accumulation on the W chromosome of TSU, yet another species with an atypical sex chromosome and TE hybridization. The phylogenetic analysis of AviRTE sequences in birds agreed with the proposed phylogeny of species in most clades, and allowed the detection of this sequence in other species, expanding the distribution of the element.

Enhancing Cosmetic and Functional Improvement of Recalcitrant Nail Lichen Planus With Resin Nail.

Lichen planus (LP) is one of the few conditions that may cause permanent and debilitating nail loss. Recurrence is common despite treatment with first-line therapies including intralesional and systemic corticosteroids. We describe application of a resin nail for recalcitrant LP of the fingernail for improved cosmesis and functionality.

Geospatial analysis and prevalence of Schistosoma mansoni and soil-transmitted helminth infections in an endemic area in Eastern Brazilian Amazon.

OBJECTIVES: This study evaluated the occurrence of Schistosoma mansoni and soil-transmitted helminths in an endemic area in the Eastern Brazilian Amazon, analysing prevalence and spatial distribution. METHODS: The study was conducted in four localities of Primavera Municipality, in Pará state. Data was obtained from the Decit 40/2012 project and the participants were divided into five age range categories for evaluation: children, adolescents, young adults, adults and elderly individuals. For the diagnostic tests, Kato-Katz slides were prepared to detect S. mansoni and soil-transmitted helminths eggs. The spatial distribution map and the Kernel Density Estimation were performed to assess the presence and location of infections. RESULTS: Stool samples revealed the presence of hookworms, S. mansoni, Ascaris lumbricoides and Trichuris trichiura eggs. Mono-, bi- and poly-parasitic infections were observed, with a significant prevalence of hookworm monoparasitism. CONCLUSIONS: The high frequency of children infected with soil-transmitted helminths confirms their significance as an ongoing public health problem in the poorest municipalities of Brazil. The Geographic Information System plays a crucial role in environmental surveillance and in the control of epidemics and endemic diseases, enabling accurate assessment and informed decision-making for their control.

Profile of Facial Trauma Victims of Motorcycle Accidents Treated at a Hospital in Northeastern Brazil: A Prospective Study.

OBJECTIVE: To outline the profile of patients with facial trauma, victims of motorcycle accidents, treated at the Hospital da Restauração, Recife/PE, Brazil, from December 2020 to July 2021. METHODS: The collection was carried out through questionnaire and analysis of medical records. Data were analyzed descriptively, as well as Pearson's chi-square test or Fisher's exact test. RESULTS: Among the patients, 88.6% were male, 47.5% were between 18 and 29 years old, and 59.6% lived in rural areas; 72.15% of the victims used the motorcycle for work and 43.52% were working at the time of the accident. Most victims did not have a national motorcycle license (62%) and were not wearing a helmet at the time of the accident (60%), with 37.6% wearing a full-face helmet, 16.5% an open helmet, and 5.9% an articulated full-face helmet. Soft tissue injuries were the most prevalent. Zygomatic complex fractures were the most common facial fractures and were significantly associated with helmet use and type. CONCLUSIONS: Most victims of motorcycle accidents are young adult men, without a regular driver's license and residing in rural areas. Preventive and educational actions and continuous traffic inspections are necessary to minimize these accidents.

Evolution of bird sex chromosomes: a cytogenomic approach in Palaeognathae species.

BACKGROUND: Different patterns of sex chromosome differentiation are seen in Palaeognathae birds, a lineage that includes the ratites (Struthioniformes, Rheiformes, Apterygiformes, Casuariiformes, and the sister group Tinamiformes). While some Tinamiform species have well-differentiated W chromosomes, both Z and W of all the flightless ratites are still morphologically undifferentiated. Here, we conducted a comprehensive analysis of the ZW differentiation in birds using a combination of cytogenetic, genomic, and bioinformatic approaches. The whole set of satDNAs from the emu (Dromaius novaehollandiae) was described and characterized. Furthermore, we examined the in situ locations of these satDNAs alongside several microsatellite repeats and carried out Comparative Genomic Hybridizations in two related species: the greater rhea (Rhea americana) and the tataupa tinamou (Crypturellus tataupa). RESULTS: From the 24 satDNA families identified (which represent the greatest diversity of satDNAs ever uncovered in any bird species), only three of them were found to accumulate on the emu's sex chromosomes, with no discernible accumulation observed on the W chromosome. The W chromosomes of both the greater rhea and the emu did not exhibit a significant buildup of either C-positive heterochromatin or repetitive DNAs, indicating their large undifferentiation both at morphological and molecular levels. In contrast, the tataupa tinamou has a highly differentiated W chromosome that accumulates several DNA repeats. CONCLUSION: The findings provide new information on the architecture of the avian genome and an inside look at the starting points of sex chromosome differentiation in birds.

Environmental health of water bodies from a Brazilian Amazon Metropolis based on a conventional and metagenomic approach.

AIMS: The present study aimed to use a conventional and metagenomic approach to investigate the microbiological diversity of water bodies in a network of drainage channels and rivers located in the central area of the city of Belém, northern Brazil, which is considered one of the largest cities in the Brazilian Amazon. METHODS AND RESULTS: In eight of the analyzed points, both bacterial and viral microbiological indicators of environmental contamination-physical-chemical and metals-were assessed. The bacterial resistance genes, drug resistance mechanisms, and viral viability in the environment were also assessed. A total of 473 families of bacteria and 83 families of viruses were identified. Based on the analysis of metals, the levels of three metals (Cd, Fe, and Mn) were found to be above the recommended acceptable level by local legislation. The levels of the following three physicochemical parameters were also higher than recommended: biochemical oxygen demand, dissolved oxygen, and turbidity. Sixty-three bacterial resistance genes that conferred resistance to 13 different classes of antimicrobials were identified. Further, five mechanisms of antimicrobial resistance were identified and viral viability in the environment was confirmed. CONCLUSIONS: Intense human actions combined with a lack of public policies and poor environmental education of the population cause environmental degradation, especially in water bodies. Thus, urgent interventions are warranted to restore the quality of this precious and scarce asset worldwide.

Natural rubber latex-based biomaterials for drug delivery and regenerative medicine: Trends and directions.

Natural Rubber Latex (NRL) has shown to be a promising biomaterial for use as a drug delivery system to release various bioactive compounds. It is cost-effective, easy to handle, biocompatible, and exhibits pro-angiogenic and pro-healing properties for both soft and hard tissues. NRL releases compounds following burst and sustained release kinetics, exhibiting first-order release kinetics. Moreover, its pore density can be adjusted for tailored kinetics profiles. In addition, biotechnological applications of NRL in amblyopia, smart mattresses, and neovaginoplasty have demonstrated success. This comprehensive review explores NRL's diverse applications in biotechnology and biomedicine, addressing challenges in translating research into clinical practice. Organized into eight sections, the review emphasizes NRL's potential in wound healing, drug delivery, and metallic nanoparticle synthesis. It also addresses the challenges in enhancing NRL's physical properties and discusses its interactions with the human immune system. Furthermore, examines NRL's potential in creating wearable medical devices and biosensors for neurological disorders. To fully explore NRL's potential in addressing important medical conditions, we emphasize throughout this review the importance of interdisciplinary research and collaboration. In conclusion, this review advances our understanding of NRL's role in biomedical and biotechnological applications, offering insights into its diverse applications and promising opportunities for future development.

Effects of in ovo vaccination time on broiler performance parameters under field conditions.

Hatchery performance is often evaluated based on descriptors such as hatchability, 7-d mortality, and cost. In addition to these descriptors, it is useful to include in this analysis aspects of chick quality through post-hatch performance. Realizing the bird's complete genetic potential necessitates meeting various criteria, with effective support for the chick's immune system being among the pivotal factors. To be effective, in ovo vaccination systems must deliver the vaccines to specific sites in the egg, a circumstance that directly depends on when the injection is made. We examined production data to evaluate the impact of in ovo vaccination time on performance parameters of male Ross308AP chicks. A comprehensive survey was conducted examining records from 3,722 broiler flocks produced and raised by the same company under standard nutrition and management conditions. The selected data specifically pertained to flocks that underwent slaughter between 41 and 45 d. In our analysis, 4 different linear models were built, one for each response variable: mean weight (MW), body weight gain (BWG), corrected feeding conversion rate (cFCR), and total mortality (TM). The linear models used in the analyses included as main predictor the timing of in ovo vaccination (440, 444, 448, 452, 456, 458, and 460 h of incubation), and as additional predictors: age of the breeding flock (26-35, 36-55 and 56-66 wks old), slaughter age, identity of the hatchery, and the season at which the data was collected. Our results showed that the timing of in ovo vaccination significantly affected BWG and cFCR, with procedures performed at 460 h of incubation showing the best outcomes. Breeding flock age affected all response variables, with older breeding flocks delivering increased MW, BWG and TM, and middle-aged flocks increased cFCR. Increasing slaughter age reduced BWG while MW, cFCR and TM were all increased. These data emphasize the benefits of performing in ovo vaccination as close as possible to 460 h of incubation to extract the best BWG and cFCR from Ross308AP male broiler.

Machine Learning-Based Virtual Screening of Antibacterial Agents against Methicillin-Susceptible and Resistant Staphylococcus aureus.

The application of computer-aided drug discovery (CADD) approaches has enabled the discovery of new antimicrobial therapeutic agents in the past. The high prevalence of methicillin-resistantStaphylococcus aureus(MRSA) strains promoted this pathogen to a high-priority pathogen for drug development. In this sense, modern CADD techniques can be valuable tools for the search for new antimicrobial agents. We employed a combination of a series of machine learning (ML) techniques to select and evaluate potential compounds with antibacterial activity against methicillin-susceptible S. aureus (MSSA) and MRSA strains. In the present study, we describe the antibacterial activity of six compounds against MSSA and MRSA reference (American Type Culture Collection (ATCC)) strains as well as two clinical strains of MRSA. These compounds showed minimal inhibitory concentrations (MIC) in the range from 12.5 to 200 µM against the different bacterial strains evaluated. Our results constitute relevant proven ML-workflow models to distinctively screen for novel MRSA antibiotics.

Protocols for Building and Producing High Diversity Peptide Phage Display Libraries.

Phage display is an important technology to study protein-protein interaction and protein evolution, with applications in basic science and applied biotechnology, such as drug discovery and the development of targeted therapies. However, in order to be successful during a phage display screening, it is paramount to have good phage libraries. Here, we described detailed procedures to generate peptide phage display libraries with high diversity and billions of transformants.

Understanding the Structural Requirements of Peptide-Protein Interaction and Applications for Peptidomimetic Development.

Protein-protein interaction is at the heart of most biological processes, and small peptides that bind to protein binding sites are resourceful tools to explore and understand the structural requirements for these interactions. In that sense, phage display is a well-suited technology to study protein-protein interactions, as it allows for unbiased screening of billions of peptides in search for those that interact with a protein binding domain. Here, we will illustrate how two distinct but complementary approaches, phage display and nuclear magnetic resonance (NMR), can be utilized to unveil structural details of peptide-protein interaction. Finally, knowledge derived from phage mutagenesis and NMR studies can be streamlined for quick peptidomimetic design and synthesis using the retroinversion approach to validate using in vitro and in vivo assays the therapeutic potential of peptides identified by phage display.

Comparative cytogenetics in Tyrannidae (Aves, Passeriformes): High genetic diversity despite conserved karyotype organization.

Introduction Passeriformes has the greatest species diversity among Neoaves, and the Tyrannidae is the richest in this order with about 600 valid species. The diploid number of this family remains constant, ranging from 2n = 76 to 84, but the chromosomal morphology varies, indicating the occurrence of different chromosomal rearrangements. Cytogenetic studies of the Tyrannidae remain limited, with approximately 20 species having been karyotyped thus far. This study aimed to describe the karyotypes of two species from this family, Myiopagis viridicata and Sirystes sibilator. Methods Skin biopsies were taken from each individual to establish fibroblast cell cultures and to obtain chromosomal preparations using the standard methodology. The chromosomal distribution of constitutive heterochromatin was investigated by C-banding, while the location of simple repetitive sequences (SSRs), 18S rDNA, and telomeric sequences were found through fluorescence in situ hybridization. Results The karyotypes of both species are composed of 2n = 80. The 18S rDNA probes hybridized into two pairs of microchromosomes in M. viridicata, but only a single pair in S. sibilator. Only the telomeric portions of each chromosome in both species were hybridized by the telomere sequence probes. Most of the SSRs were found accumulated in the centromeric and telomeric regions of several macro- and microchromosomes in both species, which likely correspond to the heterochromatin-rich regions. Conclusion Although both species analyzed showed a conserved karyotype organization (2n = 80), our study revealed significant differences in their chromosomal architecture, rDNA distribution, and SSR accumulation. These findings were discussed in the context of the evolution of Tyrannidae karyotypes.