Characterization of sperm morphology in two species of Neotropical bats: Artibeus planirostris and Sturnira erythromos (Phyllostomidae, Chiroptera).

Sperm morphology is considered the best indicator of male fertility. In Neotropical bats, important aspects of sperm morphology have been scantly studied. The aim of the present study was to characterize and compare the sperm morphology and morphometry of Artibeus planirostris and Sturnira erythromos. A total of 11 specimens were analyzed from the Colección de Mamíferos Lillo: five A. planirostris and six S. erythromos. The fixed epididymis were extracted and macerated in Farmer's solution, followed by the routine cytological procedure with different stains. To carry out the description and morphometric analysis, microphotographs were taken under an optical, epifluorescence and scanning electron microscope. A total of 50 sperm from each individual were measured for morphometric analysis. The percentage of normal/abnormal spermatozoa was estimated and the sperm abnormalities were classified. Both species showed morphologically simple spermatozoa with a spatulate head, a short neck, a helical midpiece and a tail that tapers at the final end, similar to other species of Phyllostomidae. The differences observed were: apex of the head was conical in A. planirostris and was oval in S. erythromos; longer head and midpiece in S. erythomos and longer sperm in A. planirostris. Both species showed a high percentage of sperm with normal appearance: 65% for A. planirostris and 72% for S. erythromos. The main sperm abnormalities were: scattered tails and heads, coiled tails, folded midpieces and presence of cytoplasmic droplets. The present work will improve the understanding of their reproductive biology. RESEARCH HIGHLIGHTS: Morphological descriptions and morphometric analyses of the sperm of Artibeus planirostris and Sturnira erythromos were carried out with optical, epifluorescence and scanning electron microscopy.

Prostatic morphological changes throughout life: Cytochemistry as a tool to reveal tissue aging markers.

The prostate undergoes normal or pathological morphological changes throughout life. An understanding of these changes is fundamental for the comprehension of aging-related pathological processes such as benign prostatic hyperplasia (BPH) and cancer. In the present study, we show some of these morphological changes, as well as histochemical techniques like Weigert's resorcin-fuchsin method, Picrosirius Red, and Gömöri's reticulin for use as tools in the study of prostate tissue under light microscopy. For this purpose, prostates of the Mongolian gerbil (n = 9), an experimental model that develops BPH spontaneously, were analyzed at three life stages: young (1 month old), adult (3 months old), and old (15 months old). The results showed that fibrillar components such as collagen, and reticular and elastic fibers, change throughout life. In young animals, the prostate has cuboidal epithelium surrounded by thin layers of smooth muscle, continuous collagen fibers, winding reticular fibers, and sporadic elastic fibers. With adulthood, the epithelium becomes columnar, encircled by compacted muscle cells among slender collagen fibers, elongated reticular fibers, and linear elastic fibers. In aging individuals, the prostate's epithelium stratifies, surrounded by thick muscle layers among dense collagen fibers, disordered reticular fibers, and elastic fibers in different planes. We also identified a few accumulations of lipid droplets and lipofuscin granules in adult animals and high accumulation in old animals evidenced by Oil red O and Gömöri-Halmi techniques, respectively. The histochemical techniques presented here have been demonstrated to be useful and accessible tools in prostate studies. RESEARCH HIGHLIGHTS: Cytochemical techniques to study prostate morphology. The prostate changes with age.

Antimicrobial Efficacy and Dentin Collagen Damage Caused by Calcium Hypochlorite and Sodium Hypochlorite

Abstract This study aimed to evaluate the antimicrobial activity of calcium hypochlorite (Ca (OCl)2) and sodium hypochlorite (NaOCl) using confocal laser scanning microscopy (CLSM) and dentin organic matrix alteration by picrosirius staining and light microscopy (LM). Samples of human extracted teeth were infected with Enterococcus faecalis by centrifugation of the bacterial suspension and were treated with Ca(OCl)2 or NaOCl at 0.5%, 2.5%, and 6% for 15, 30, and 60 seconds. CLSM and viability staining were used to quantitatively analyze the proportions of dead/live bacteria in the canal lumen and border of the root canal. The data were analyzed by ANOVA and Fisher test. For LM analysis, one hundred bovine teeth were randomly divided into 10 test groups (n=10): G1- Without treatment; G2- 17% EDTA; G3- 6% NaOCl; G4- 6% NaOCl + EDTA; G5- 0.5% Ca(OCl)2; G6- 0.5% Ca(OCl)2 + EDTA; G7- 2.5% Ca(OCl)2; G8- 2.5% Ca(OCl)2 + EDTA; G9- 6% Ca(OCl)2; G10- 6% Ca(OCl)2 + EDTA. The samples were fragmented and stained with Picrosirius. Data were analyzed by Kruskal-Wallis and Dunn (P<0.05). There was a strong correlation between the results of the canal lumen and the border of the root canal (r=0.962). Both hypochlorites at a concentration of 0.5% showed less microbial reduction compared to 2.5% and 6% (P<0.05). There was less antimicrobial activity at 15 seconds compared to 30 and 60 seconds (P<0.05). Ca(OCl)2 and NaOCl showed similar results at the same concentrations (P>0.05). In conclusion, Ca(OCl)2 caused fewer alterations to the dentin organic matrix at concentrations of 0.5% and 2.5%. Ca(OCl)2 presents antimicrobial activity similar to NaOCl, and collagen damage is concentration-dependent.

Resumo Este estudo teve como objetivo avaliar a atividade antimicrobiana do hipoclorito de cálcio (Ca(OCl)2) e do hipoclorito de sódio (NaOCl) usando a microscopia confocal a laser (MCVL) e a alteração da matriz orgânica da dentina por meio da coloração com picrosirius e microscopia óptica (MO). Amostras de dentes humanos extraídos foram infectados com Enterococcus faecalis por centrifugação da suspensão bacteriana. As amostras foram tratadas com Ca(OCl)2 ou NaOCl a 0,5%, 2,5% e 6% durante 15, 30 e 60 segundos. A MCVLe a coloração de viabilidade foram utilizadas para analisar quantitativamente as proporções de bactérias vivas/mortas no lúmen do canal e no bordo do canal radicular. Os dados foram analisados por ANOVA e teste de Fisher LSD. Para análise de MO, cem dentes bovinos foram divididos aleatoriamente em 10 grupos teste (n=10): G1- Sem tratamento; G2- 17% de EDTA; G3- 6% de NaOCl; G4- 6% de NaOCl + EDTA; G5- 0,5% de Ca(OCl)2; G6- 0,5% de Ca(OCl)2 + EDTA; G7- 2,5% de Ca(OCl)2; G8- 2,5% de Ca(OCl)2 + EDTA; G9- 6% de Ca(OCl)2; G10- 6% de Ca(OCl)2 + EDTA. As amostras foram fragmentadas e coradas com Picrosirius. Os dados foram analisados por Kruskal-Wallis e teste de Dunn (P<0,05). Houve uma forte correlação nos resultados do lúmen do canal e na borda do canal radicular (r=0,962). Ambos os hipocloritos, a uma concentração de 0,5%, mostraram menor redução microbiana em comparação com 2,5% e 6% (P<0,05). Houve menor atividade antimicrobiana aos 15 segundos em comparação com 30 e 60 segundos (P<0,05). Em conclusão, o Ca(OCl)2 e o NaOCl apresentaram resultados semelhantes nas mesmas concentrações (P>0,05). O Ca(OCl)2 causou menos alterações na matriz orgânica da dentina em concentrações de 0,5% e 2,5%. O Ca(OCl)2 apresenta atividade antimicrobiana semelhante ao NaOCl, e os danos ao colágeno dependem da concentraçã

A new species of Orientatractis (Nematoda: Cosmocercoidea: Atractidae) parasite of Yellow-Spotted Amazon River Turtle, Podocnemis unifilis Troschel, 1848 (Testudines: Podocnemididae) in Brazilian Amazon.

The current study describes a new species of nematode, Orientatractis matosi n. sp. (Atractidae), from the stomach and large intestine of Podocnemis unifilis Troschel, 1848 (Testudinidae), collected in the Tocantins and Xingu rivers in the Brazilian state of Pará. The new species was described using light and scanning electron microscopy and differs from its congeners mainly in the number and distribution of the male caudal papillae and by having a shorter right spicule. Orientatracits matosi n. sp. is the eleventh species of the genus Orientatractis Petter, 1966, the third reported from the Amazon, and the second described from P. unifilis.

A NEW SPECIES OF PNEUMOATRACTIS (COSMOCERCOIDEA: ATRACTIDAE) FROM TWO SPECIES OF FRESHWATER TURTLES FROM THE BRAZILIAN AMAZON.

The monotypic genus PneumoatractisBursey, Reavill, and Greiner, 2009 was erected to allocate nematodes collected from the lungs of the freshwater turtle Podocnemis unifilis Troschel. During a helminthological survey of parasites from freshwater turtles of Tocantins and Xingu rivers, Pará State, eastern Amazon, Brazil, we found nematodes parasitizing the stomach and large intestine of Po. unifilis and Podocnemis expansa Schweigger. We assigned them to a new species of Pneumoatractis, which is described herein. Pneumoatractis gibbonsae n. sp. resembles Pneumoatractis podocnemis by the morphology of the oral opening, the position of the excretory pore, and the lanceolate shape of the spicules, but differs from it in males by having 10 pairs of caudal papillae plus 1 unpaired anterior to cloacal lip, different length of the right spicule, and short gubernaculum; it differs in females by the distances of vulva and anus from the posterior end respectively. We found the new species in a different infection site from that of the type species. Thus, this is the second species of Pneumoatractis described in Po. unifilis and the first in Po. expansa.

Calcium oxalate crystal macropattern and its usefulness in the taxonomy of Baccharis (Asteraceae).

This study provides a comprehensive account of the various types of calcium oxalate crystals found in the genus Baccharis and assesses the exceptional value of crystal macropatterns for the taxonomy of the genus. The morphotype, occurrence, and chemical composition of the crystals found in the stems and leaves are studied. The 44 species included in this study were selected based on a broad phylogeny-based sampling covering seven subgenera and 31 sections. These species were chosen to represent all the main phylogenetic lineages of Baccharis; thus, the sampling also represents a comprehensive coverage concerning evolutionary significance for such a large and environmentally and economically important plant group. The samples were analyzed by light microscopy, scanning electron microscopy (SEM), and energy-dispersive x-ray spectroscopy (EDS). Several morphotypes of crystals, including druses, crystal sand, styloids and prisms, were present. Based on their chemical composition, the crystals were classified as pure calcium oxalate, mixtures of oxalates and sulfates, and mixtures of oxalates, sulfates, and silica. The crystal macropatterns observed in this study aid in species identification and provide novel data for the taxonomy of Baccharis. RESEARCH HIGHLIGHTS: Most species of Baccharis have a specific crystalline pattern. Each species produces a crystal morphotype or a set of morphotypes specific to it. The crystals observed are formed by calcium oxalate.

Optimization of Testicular Fixation-Embedding Techniques for Improved Evaluation of Mammalian Spermatogonial Morphology and Function.

Numerous methods have been successfully used to evaluate mammalian spermatogonial biology However, the conventional light microscopy assays present a challenge in precisely identifying spermatogonial phenotypes, which can result in discrepancies between molecular and morphological findings. Such precise association could lead to a more robust interpretation of spermatogonial activity in steady-state spermatogenesis, which may facilitate the translation from basic research to clinical applications. In this chapter, we present two histological processing methods that enable a comprehensive analysis of spermatogonial morphology and function, involving fixation of mammalian testicular tissue in glutaraldehyde and embedding in plastic resin. These techniques have proven to be effective in light microscopy studies.

Instant polarized light microscopy pi (IPOLπ) for quantitative imaging of collagen architecture and dynamics in ocular tissues.

Collagen architecture determines the biomechanical environment in the eye, and thus characterizing collagen fiber organization and biomechanics is essential to fully understand eye physiology and pathology. We recently introduced instant polarized light microscopy (IPOL) that encodes optically information about fiber orientation and retardance through a color snapshot. Although IPOL allows imaging collagen at the full acquisition speed of the camera, with excellent spatial and angular resolutions, a limitation is that the orientation-encoding color is cyclic every 90 degrees (π/2 radians). In consequence, two orthogonal fibers have the same color and therefore the same orientation when quantified by color-angle mapping. In this study, we demonstrate IPOLπ, a new variation of IPOL, in which the orientation-encoding color is cyclic every 180 degrees (π radians). Herein we present the fundamentals of IPOLπ, including a framework based on a Mueller-matrix formalism to characterize how fiber orientation and retardance determine the color. The improved quantitative capability of IPOLπ enables further study of essential biomechanical properties of collagen in ocular tissues, such as fiber anisotropy and crimp. We present a series of experimental calibrations and quantitative procedures to visualize and quantify ocular collagen orientation and microstructure in the optic nerve head, a region in the back of the eye. There are four important strengths of IPOLπ compared to IPOL. First, IPOLπ can distinguish the orientations of orthogonal collagen fibers via colors, whereas IPOL cannot. Second, IPOLπ requires a lower exposure time than IPOL, thus allowing faster imaging speed. Third, IPOLπ allows visualizing non-birefringent tissues and backgrounds from tissue absorption, whereas both appear dark in IPOL images. Fourth, IPOLπ is cheaper and less sensitive to imperfectly collimated light than IPOL. Altogether, the high spatial, angular, and temporal resolutions of IPOLπ enable a deeper insight into ocular biomechanics and eye physiology and pathology.

Morphology and morphometry of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) through light and scanning electron microscopy / Morfologia e morfometria de Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) através da microscopia de luz e microscopiaeletrônica de varredura

Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).

Paratanaisia bragai é um trematódeo digenético que atinge a maturidade sexual nos ductos coletores de aves domésticas e silvestres, enquanto os moluscos Subulina octona e Leptinaria unilamellata atuam como seus hospedeiros intermediários no Brasil. O presente estudo analisou a morfologia e morfometria de P. bragai. Amostras adultas do parasito foram coletadas de rins de Columba livia naturalmente infectada, fixadas e preparadas para observação na microscopia de campo claro e microscopia de luz de contraste de interferência diferencial e microscopia eletrônica de varredura. O parasite possui corpo alongado e achatado, com uma ventosa oral subterminal localizada na extremidade anterior do corpo, conforme observado por todas as técnicas utilizadas. A coloração do parasito com hematoxilina-eosina permitiu observar a faringe, localizada posteriormente à ventosa oral, as glândulas vitelogênicas, que são extracecais e estendem-se anteriormente à região pré-ovariana e posteriormente à região mediana do corpo, e os cecos intestinais paralelos às glândulas vitelinas. A presença e funcionalidade do acetábulo são pontos controversos na literatura, mas foi observado em todos os espécimes analisados por microscopia eletrônica de varredura, com diâmetro maior de 38.36 ± 6.96 (28.77 - 45.39) e diâmetro menor de 31.59 ± 7,04 (21.75 - 38.16). Próximo ao acetábulo foram observadas escamas no tegumento do parasito. Escamas com (1 - 5) divisões de lâmina foram identificadas. No poro genital, foi possível visualizar o cirro evertido com formato de roseta. O poro excretor (primeiro registro morfométrico) é dorsal e subterminal, com diâmetro maior de 12.27 ± 9.16 (5.79 - 18.75) e diâmetro menor de 3.95 ± 1.49 (2.89 - 5.00).

Morphology and morphometry of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) through light and scanning electron microscopy / Morfologia e morfometria de Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) através da microscopia de luz e microscopiaeletrônica de varredura

Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).

Paratanaisia bragai é um trematódeo digenético que atinge a maturidade sexual nos ductos coletores de aves domésticas e silvestres, enquanto os moluscos Subulina octona e Leptinaria unilamellata atuam como seus hospedeiros intermediários no Brasil. O presente estudo analisou a morfologia e morfometria de P. bragai. Amostras adultas do parasito foram coletadas de rins de Columba livia naturalmente infectada, fixadas e preparadas para observação na microscopia de campo claro e microscopia de luz de contraste de interferência diferencial e microscopia eletrônica de varredura. O parasite possui corpo alongado e achatado, com uma ventosa oral subterminal localizada na extremidade anterior do corpo, conforme observado por todas as técnicas utilizadas. A coloração do parasito com hematoxilina-eosina permitiu observar a faringe, localizada posteriormente à ventosa oral, as glândulas vitelogênicas, que são extracecais e estendem-se anteriormente à região pré-ovariana e posteriormente à região mediana do corpo, e os cecos intestinais paralelos às glândulas vitelinas. A presença e funcionalidade do acetábulo são pontos controversos na literatura, mas foi observado em todos os espécimes analisados por microscopia eletrônica de varredura, com diâmetro maior de 38.36 ± 6.96 (28.77 - 45.39) e diâmetro menor de 31.59 ± 7,04 (21.75 - 38.16). Próximo ao acetábulo foram observadas escamas no tegumento do parasito. Escamas com (1 - 5) divisões de lâmina foram identificadas. No poro genital, foi possível visualizar o cirro evertido com formato de roseta. O poro excretor (primeiro registro morfométrico) é dorsal e subterminal, com diâmetro maior de 12.27 ± 9.16 (5.79 - 18.75) e diâmetro menor de 3.95 ± 1.49 (2.89 - 5.00).

Usefulness of Trichoscopy over Hair Light Microscopy in Menkes Disease.

Menkes disease (MD) is a rare X-linked recessive neurodegenerative disorder caused by mutations in the ATP7A gene, with a high mortality rate within the first 3 years of life. It typically affects males and is characterized by impaired copper distribution and malfunction of several copper-dependent enzymes. Patients develop progressive muscle hypotonia associated with neurological damage and hair shaft dysplasia - particularly pili torti. Pili torti is usually very subtle in the first 3 months of life and gradually increases during the first year. Light microscopy examination in search for pili torti requires the observation of more than 50 hair shafts. In contrast, trichoscopy with a hand-held dermatoscope allows to easily identify the hair shaft defect. We report a case of a Hispanic male infant with MD in whom we show that trichoscopy is superior to hair light microscopy in revealing pili torti.

Why human prepuce is a valuable resource in science? Histological, ultrastructural, immunohistochemical and statistical analysis / ¿Por qué el prepucio humano es un recurso valioso en la ciencia? Análisis histológicos, ultraestructurales, inmunohistoquímicos y estadísticos

SUMMARY: This research was to examine the histological and ultrastructural characteristics of prepuce samples, as well as vimentin and S100 protein localization and statistical analysis. Urologists have long struggled with the prepuce, which is used to treat a variety of urethral problems. Skin biopsies were collected from the prepuce at the moment of circumcision and processed for light microscopy, electron microscope examination, immunohistochemical techniques, and statistical analysis in a total of six boys. Histologically, the prepuce epidermis displayed focal spiky ridges, which are saw-toothed interspersed with sulci, slight hyperpigmentation, looser connective tissue and plentiful vascular components. Immunohistochemically, the existence of melanocytes and Langerhans cells in the epidermis, as well as smooth muscles in the dermis, was stained positively for vimentin. Also, there was a positive reactivity of the Langerhans cells in the epidermis and around Meissner's corpuscles in the dermis for S100 protein staining. Ultrastructurally, the prepuce's intercellular gaps were widened, melanocytes rested on a folded basement membrane, and desmosomal content was reduced, with a prominent active euchromatic nucleus. Cytoplasmic projections were distended and elongated, and the interstitial blood vessels were surrounded by endothelial cells and rested on a basement membrane. There were also minimal collagen fibers in the interstitium. The prepuce's histological and ultrastructural features, as well as immunohistological studies using vimentin and S100 protein as intermediate filaments and statistical analysis, all demonstrated that it is a useful scientific resource.

RESUMEN: El presente trabajo de investigación se realizó para examinar las características histológicas y ultraestructurales de las muestras de prepucio, así como la localización y el análisis estadístico de la vimentina y la proteína S100. Los urólogos han intentado trabajar durante mucho tiempo con el prepucio, que se usa para tratar una variedad de problemas uretrales. Se recolectaron biopsias de piel del prepucio de seis niños en el momento de la circuncisión y se procesaron para microscopía óptica, examen con microscopio electrónico, técnicas inmunohistoquímicas y análisis estadístico. Histológicamente, la epidermis del prepucio mostraba crestas puntiagudas focales, intercaladas con surcos, hiperpigmentación leve, tejido conectivo más laxo y abundantes componentes vasculares. Inmunohistoquímicamente, la existencia de melanocitos y células dendríticas epidérmicas (células de Langerhans), así como músculo liso en la dermis, se tiñeron positivamente para vimentina. Además, hubo una reactividad positiva de las células dendríticas epidérmicas en la epidermis y alrededor de los corpúsculos del tacto (de Meissner) en la dermis para la tinción de la proteína S100. Ultraestructuralmente, los espacios intercelulares del prepucio se ensancharon, los melanocitos descansaban sobre una membrana basal plegada y el contenido desmosómico se redujo, con un núcleo eucromático activo prominente. Las proyecciones citoplasmáticas estaban distendidas y alargadas, y los vasos sanguíneos intersticiales estaban rodeados por células endoteliales y descansaban sobre una membrana basal. También había fibras de colágeno mínimas en el intersticio. Las características histológicas y ultraestructurales del prepucio, así como los estudios inmunohistológicos utilizando vimentina y proteína S100 como filamentos intermedios y el análisis estadístico, demostraron que es un recurso científico útil.

Sequential SEM-EDS, PLM, and MRS Microanalysis of Individual Atmospheric Particles: A Useful Tool for Assigning Emission Sources.

In this work, the particulate matter (PM) from three different monitoring stations in the Monterrey Metropolitan Area in Mexico were investigated for their compositional, morphological, and optical properties. The main aim of the research was to decipher the different sources of the particles. The methodology involved the ex situ sequential analysis of individual particles by three analytical techniques: scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS), polarized light microscopy (PLM), and micro-Raman spectroscopy (MRS). The microanalysis was performed on samples of total suspended particles. Different morphologies were observed for particles rich in the same element, including prismatic, spherical, spheroidal, and irregular morphologies. The sequential microanalysis by SEM-EDS/PLM/MRS revealed that Fe-rich particles with spherical and irregular morphologies were derived from anthopogenic sources, such as emissions from the metallurgical industry and the wear of automobile parts, respectively. In contrast, Fe-rich particles with prismatic morphologies were associated with natural sources. In relation to carbon (C), the methodology was able to distinguish between the C-rich particles that came from different anthopogenic sources-such as the burning of fossil fuels, biomass, or charcoal-and the metallurgical industry. The optical properties of the Si-rich particles depended, to a greater extent, on their chemical composition than on their morphology, which made it possible to quickly and accurately differentiate aluminosilicates from quartz. The methodology demonstrated in this study was useful for performing the speciation of the particles rich in different elements. This differentiation helped to assign their possible emission sources.

Virtual Microscopy as a Learning Tool in Brazilian Medical Education.

Virtual microscopy (VM) is a widely used teaching method in Medical Education in many developed countries. In Brazil, however, this is not the case for most medical schools, considering Brazilian social inequality and uneven access to technology. Recently, the Covid-19 pandemic has also challenged Universities to seek and make a transition toward more effective methods of full-time online education. Thus, the main goal of this work was to verify student's perception and academic performance, assessed upon VM implementation in a Brazilian Medical School. Ribeirao Preto Medical School students answered a 26-question survey with regards to optical microscopy (OM) and VM. Academic performance was compared between participants that were (year of 2019) or were not (year of 2015) exposed to VM. Taken the results together, subjective impressions such as handling, suitability, learning effectiveness, and pleasure using the tools, have shown a higher score for virtual microscopy (median = 29), when compared to optical microscopy (median = 24) with a P-value < 0.001 by Wilcoxon rank test, upon measurement using an ordinal scale. Regarding academic performance, no statistically significant differences were found between groups (P-value = 0.38, Cohen's d = 0.19). Therefore, VM proved to be adequate to the Brazilian medical education in light of Brazilian social contexts and Covid-19 pandemic.

The use of photobiomodulation therapy or LED and mineral trioxide aggregate improves the repair of complete tibial fractures treated with wire osteosynthesis in rodents.

The repair of large bone defects is lengthy and complex. Both biomaterials and phototherapy have been used to improve bone repair. We aimed to describe histologically the repair of tibial fractures treated by wiring (W), irradiated or not, with laser (λ780 nm, 70 mW, CW, spot area of 0.5 cm2, 20.4 J/cm2 (4 × 5.1 J/cm2, Twin Flex Evolution®, MM Optics, Sao Carlos, SP, Brazil) per session, 300 s, 142.8 J/cm2 per treatment) or LED (λ850 ± 10 nm, 150 mW, spot area of 0.5 cm2, 20.4 J/cm2 per session, 64 s, 142.8 J/cm2 per treatment, Fisioled®, MM Optics, Sao Carlos, Sao Paulo, Brazil) and associated or not to the use of mineral trioxide aggregate (MTA, Angelus®, Londrina, PR, Brazil). Inflammation was discrete on groups W and W + LEDPT and absent on the others. Phototherapy protocols started immediately before suturing and repeated at every other day for 15 days. Collagen deposition intense on groups W + LEDPT, W + BIO-MTA + LaserPT and W + BIO-MTA + LEDPT and discrete or moderate on the other groups. Reabsorption was discrete on groups W and W + LEDPT and absent on the other groups. Neoformation varied greatly between groups. Most groups were partial and moderately filed with new-formed bone (W, W + LaserPT, W + LEDPT, W + BIO-MTA + LEDPT). On groups W + BIO-MTA and W + BIO-MTA + LaserPT bone, neoformation was intense and complete. Our results are indicative that the association of MTA and PBMT (λ = 780 nm) improves the repair of complete tibial fracture treated with wire osteosynthesis in a rodent model more efficiently than LED (λ = 850 ± 10 nm).

Clinical, trichoscopy, and light microscopic findings in hypohidrotic ectodermal dysplasia: Report of 21 patients and a review of the literature.

INTRODUCTION: Hypohidrotic ectodermal dysplasia (HED) is a genetic condition typified by alterations in skin structures including sweat glands, hair, nails, and teeth. Hair findings in HED have been poorly characterized in larger series. OBJECTIVE: To characterize scalp and hair findings of patients with HED clinically and with trichoscopy and light microscopy. METHODS: A cross-sectional study in 21 pediatric HED patients was performed using available clinical and scalp dermatoscopic images, as well as pulled-hair samples for clinical evaluation, trichoscopic, and light microscopic analyses. RESULTS: Seventeen out of 21 patients (81%) were men. Twenty patients had straight hair. Sixteen patients had decreased hair density, 6 of whom had hair loss mainly in the temporal and occipital regions. Fourteen patients had hair whorls. On trichoscopy, we observed: single-hair follicular units (n = 19, 90%), scalp hyperpigmentation (n = 13, 62%), variable diameter of the hair shafts (n = 12, 57%), perifollicular scales (n = 8, 38%), scalp erythema (n = 8, 38%), and short curly pigtail hairs (n = 6, 29%). On light microscopy, findings included: hair shafts with irregular diameter (n = 7, 33%), heterogeneous hair color (n = 6, 29%), trichoptilosis (n = 2, 10%), and pili torti (n = 1, 5%). CONCLUSIONS: In this series, hair findings in HED were similar to those described in previous studies. However, we describe two new clinical and two trichoscopic findings: decreased hair density mainly in the temporal and occipital regions, oblique upwards occipital hair follicles orientation, angled hairs, and short curly pigtail hairs. These heterogeneous findings may reflect the multiple factors and signaling pathways that can be affected in these syndromes.

Morphological and anatomical insights into de novo shoot organogenesis of in vitro 'Delite' rabbiteye blueberries.

Blueberries are valued for their taste and their high nutritional benefits, including their antioxidant and anti-inflammatory properties. In vitro culturing is an alternative method for clonal propagation, and has been used in many biotechnological studies. Most blueberry research is concentrated on highbush and lowbush taxa (Vaccinium corymbosum and Vaccinium angustifolium respectively), with only limited investigations of rabbiteye cultivars (Vaccinium virgatum) that are more suitable for subtropical climates and regions with warmer winters as a result of climate change. There is therefore a need to determine in vitro protocols for that species and group of cultivars. We examined here adventitious shoot regeneration in the 'Delite' rabbiteye blueberry cultivar. Leaf explants were cultured in vitro in Woody Plant Medium (WPM), and the effects of different thidiazuron (TDZ) concentrations, the orientation of the leaf (adaxial or abaxial surface in contact with the medium), and two portions of the leaf segment (basal or apical) were examined. De novo shoot development was studied using light and scanning electron microscopy. All concentrations of TDZ used showed similar survival and regeneration rates; 0.5 µM TDZ showed high efficiency in regenerating adventitious shoots (100%, with 57 adventitious shoots/explant), as did the adaxial surface in contact with the medium using either the apical or the basal portion of the leaf (97% shoot regeneration, 47.5 adventitious shoots/explant). Anatomical analyses showed direct and indirect organogenesis. The shoots developed leaf primordia with stomata, trichomes, and well-developed vascular tissues, with further elongation and rooting of the plants. We therefore describe here a high-efficiency regeneration method through de novo shoot organogenesis using TDZ in foliar explants of rabbiteye blueberry, with direct and indirect organogenesis.

Intrapuparial Development of Hemilucilia semidiaphana (Diptera: Calliphoridae) and Its Use in Forensic Entomology.

In forensic entomology, the minimum postmortem interval (PMImin) estimative is usually based on the oldest immature recovered from a local of death. The time spent by fly immatures in the intrapuparial period comprises more than 50% of their complete life cycles. An accurate estimate of the duration of this period will improve PMImin estimates. The blow fly Hemilucilia semidiaphana (Rondani 1850) was found in six criminal cases in the city of Curitiba. Even though there is data on the morphology of the larval instars and developmental rate of H. semidiaphana, the intrapuparial period has not been investigated. Here, we provide a detailed description of the intrapuparial morphological changes of H. semidiaphana, which might be useful to estimate minimum PMI. Samples of H. semidiaphana in the intrapuparial period were obtained from immatures reared on an artificial diet in incubators adjusted to 25°C or 20°C temperature regimes. Blow fly puparia of H. semidiaphana were fixed at intervals of 3 and 6 h until emergence of the adult. The external morphological traits of sampled immatures were analyzed using light microscopy. Our analysis of the intrapuparial period of H. semidiaphana provided 21 traits from which nine were age informative. These nine characteristics divide the developmental time (144 h at 25°C and 192 h at 20°C) into smaller sections. The developmental data provided, together with the time line allows a practical way to make interspecific comparisons as well as to estimate the age of H. semidiaphana based on the intrapuparial development.

Optimal Preparation of Tissue Sections for Light-Microscopic Analysis of Phloem Anatomy.

In order to successfully analyze and describe any plant tissue, the first challenge is preparation of good anatomical slides. The challenge is even greater when the target tissue has heterogeneous characteristics, such as the phloem where soft and stiff tissues occur side by side. The goal of this chapter is to present a detailed protocol containing various techniques for optimal preparation of phloem tissue samples for light microscopic analysis. The process typically involves the steps of fixation, softening, embedding, sectioning, staining, and mounting. The protocol can be applied to make samples of phloem and surrounding tissues of stems and roots, from woody to herbaceous plants.