Osteogenesis imperfecta: Ultrastructural and histological findings on examination of skin revealing novel insights into genotype-phenotype correlation.

Osteogenesis imperfecta (OI) is a heterogeneous group of inherited disorders of bone formation, resulting in low bone mass and an increased propensity to fracture. Over 90% of patients with OI have a mutation in COL1A1/COL1A2, which shows an autosomal dominant pattern of inheritance. In-depth phenotyping and in particular, studies involving manifestations in the skin connective tissue have not previously been undertaken in OI. The aims of the study were to perform histological and ultrastructural examination of skin biopsies in a cohort of patients with OI; to identify common and distinguishing features in order to inform genotype-phenotype correlation; and to identify common and distinguishing features between the different subtypes of OI. As part of the RUDY (Rare Diseases in Bone, Joints and/or Blood Vessels) study, in collaboration with the NIHR Rare Diseases Translational Research Collaboration, we undertook a national study of skin biopsies in patients with OI. We studied the manifestations in the skin connective tissue and undertook in-depth clinical and molecular phenotyping of 16 patients with OI. We recruited 16 patients: analyses have shown that in type 1 collagen mutation positive patients (COL1A1/ COL1A2) (n-4/16) consistent findings included: variable collagen fibril diameter (CFD) and presence of collagen flowers. Histological examination in these patients showed an increase in elastic fibers that are frequently fragmented and clumped. These observations provide evidence that collagen flowers and CFD variability are consistent features in OI due to type 1 collagen defects and reinforce the need for accurate phenotyping in conjunction with genomic analyses.

Passive absorption in a classical photonic crystal-based organic solar cell.

We study the light trapping efficiency of a bidimensional photonic crystal (PC) integrated in a classical organic multilayer solar cell. The role of the PC is to enhance light absorption in the active layer by leveraging resonant mode excitation. However the light trapping efficiency is drastically inhibited by the overall absorption of the entire multilayer, which includes absorption by the passive layers that do not contribute to the photocurrent. This study focuses on the impact of passive absorption in ITO and PEDOT, which is often neglected in the study of light trapping organic solar cell systems, despite the significant role it plays in highly absorbing devices. Indeed, we show here that the absorption enhancement in the active layer can vary between 23% and 46% depending on the optical properties of the passive layers, which are dependent on fabrication conditions. We also detail how the PC behaves with coupled parameters such as the optical indices of the passive layers, as well as the period and the air filling fraction of the PC.

Twenty-first century droughts have not increasingly exacerbated fire season severity in the Brazilian Amazon.

Biomass burning in the Brazilian Amazon is modulated by climate factors, such as droughts, and by human factors, such as deforestation, and land management activities. The increase in forest fires during drought years has led to the hypothesis that fire activity decoupled from deforestation during the twenty-first century. However, assessment of the hypothesis relied on an incorrect active fire dataset, which led to an underestimation of the decreasing trend in fire activity and to an inflated rank for year 2015 in terms of active fire counts. The recent correction of that database warrants a reassessment of the relationships between deforestation and fire. Contrasting with earlier findings, we show that the exacerbating effect of drought on fire season severity did not increase from 2003 to 2015 and that the record-breaking dry conditions of 2015 had the least impact on fire season of all twenty-first century severe droughts. Overall, our results for the same period used in the study that originated the fire-deforestation decoupling hypothesis (2003-2015) show that decoupling was clearly weaker than initially proposed. Extension of the study period up to 2019, and novel analysis of trends in fire types and fire intensity strengthened this conclusion. Therefore, the role of deforestation as a driver of fire activity in the region should not be underestimated and must be taken into account when implementing measures to protect the Amazon forest.

Carcass characteristics and meat quality of Nellore bulls submitted to different nutritional strategies during cow-calf and stocker phase.

There have been few studies realized that evaluate the effects of adopting different nutritional systems in more than one phase of cattle production on carcass and meat characteristics. This study was realized to evaluate carcass and meat characteristics from bulls submitted to different nutritional systems during two production phases. The experiment was conducted at Figueira's farm during two production phases: I (cow-calf) - 80 calves (99.6±2.72 days of age and 109.7±2.99 kg of BW) with their mothers were randomly assigned into two supplemental diets: cow-calf mineral supplement (n=40) or cow-calf creep-feeding (n=40); II (stocker) - the same 80 calves (201.2±2.11 days of age and 190.2±3.37 kg of BW) were redistributed into two production systems: stocker pasture (n=40) or stocker feedlot (SF; n=40). After, all 80 animals were kept on a pasture system (III) for 290 days, and then finished in a feedlot system (IV) for more 33 days. Then, they were slaughtered at an average 764.2±3.06 days of age and at 499.2±3.33 kg of final BW. After slaughter, the average daily gain was calculated, and the carcass and meat characteristics were measured. The statistical model design used was completely randomized in a 2×2 factorial arrangement (two treatment groups on cow-calf phase and two treatment groups on stocker phase). The single effects between the groups in each phase and the interactions between both phases (cow-calf v. stocker) were analyzed. The results were compared by Fisher's test, using the R statistical software. A cow-calf by stocker phases interaction occurred for carcass conformation and fiber diameter. For single effects, the greatest influences observed were in the stocker phase. The feedlot group was slaughtered 17 days earlier, with greater final BW (3.8%), hot carcass weight (5.7%), average daily gain (6.9%), dressing percentage (1.8%), carcass length (1.8%), carcass width (1.5%), longissimus muscle area (4.8%) and muscle depth (2.3%) than pasture group. The SF group also had influence on fat color; showing higher L* and lower b* values. These results reveal that bulls reared in feedlot at the stocker phase have higher muscle development and that the stocker phase has the greatest potential to influence carcass characteristics and meat quality.

Pregnancy outcome in ethanol-treated mice with folic acid supplementation in saccharose.

INTRODUCTION: Maternal folic acid deficiency is the most important metabolic factor in the etiology of neural tube defects (NTD) and is reduced by ethanol, which is extensively consumed by young women. OBJECTIVE: The objective of the study was to determine whether folic acid supplementation in dietary saccharose is efficient in the prevention NTD induced by ethanol in fetuses of Swiss mice. MATERIALS AND METHODS: Pregnant mice were divided into four groups of six animals each: control (C), ethanol (E), deficient-supplemented (DS), and deficient-supplemented + ethanol (DSE). Groups C and E received commercial mouse chow (containing 3 mg/kg folic acid) throughout the experiment, while groups DS and DSE received a folic acid-free diet with the addition of saccharose supplemented with folic acid (2 mg/kg folic acid) in water. Group E and DSE animals received ethanol (4 g/kg) administered intraperitoneally from the seventh to the ninth gestational day (gd) and were euthanized on the 18th gd, while groups C and DS received saline. RESULTS: Congenital anomalies were observed in groups E and DSE. The fetal weight and length of the animals in group E were lower than in groups C and DS and, in group DSE, were lower than in groups C and DS. The placental diameter of group E was smaller than that of group C, and the placental weight of group C animals was lower than that of groups E, DSE, and DS. CONCLUSION: The study demonstrated that dietary supplementation with folate in saccharose is an accessible means of consumption that could be further diffused but in an increased dose than recommended to reduce the teratogenic effects of ethanol.

P63 expression in hydropic abortion and gestational trophoblastic diseases.

Gestational trophoblastic diseases are a group of interrelated diseases of trophoblastic tissue that include partial hydatidiform mole, complete hydatidiform mole, invasive mole, choriocarcinoma, and placental site trophoblastic tumor. P63 is a p53 homologue that, in normal placentas, is expressed in the cytotrophoblast cells. The role of p63 in gestational trophoblastic diseases, however, merits further investigation. Immunohistochemistry with the p63 antibody (clone 4A4) was performed in formalin-fixed paraffin-embedded samples of hydropic abortion (n=10), partial hydatidiform mole (n=12), complete hydatidiform mole (n=12) and choriocarcinoma (n=5). P63 expression was quantitatively assessed as 0 (no stained cells), + (less than 10% positive cells), ++ (10-50% positive cells), and +++ (more than 50% positive cells). The intensity was scored as 0 (absence), + (weak), ++ (moderate), or +++ (strong). Statistical analysis was carried out by the Fisher test. In contrast to the other diagnoses, none of the choriocarcinomas analyzed exhibited p63-positive cells. There was no difference in distribution of p63 positive cells between hydropic abortion, partial hydatidiform mole, and complete hydatidiform mole. Concerning the intensity of immunostaining, there was difference only between partial hydatidiform mole and complete hydatidiform mole. According to our results, p63 might be useful to differentiate a choriocarcinoma from other gestational trophoblastic diseases. Besides, since the intensity of p63 expression was much stronger in partial hydatidiform mole and complete hydatidiform mole than in hydropic abortion, this feature may be helpful in distinguishing these two diagnoses in challenging cases.

Root growth restraint can be an acclimatory response to low pH and is associated with reduced cell mortality: a possible role of class III peroxidases and NADPH oxidases.

Low pH (<5.0) can significantly decrease root growth but whether this is a direct effect of H(+) or an active plant response is examined here. Tomato (Solanum lycopersicum cv Micro-Tom) roots were exposed directly or gradually to low pH through step-wise changes in pH over periods ranging from 4 to 24 h. Roots exposed gradually to pH 4.5 grew even less than those exposed directly, indicating a plant-coordinated response. Direct exposure to pH 4.0 suppressed root growth and caused high cell mortality, in contrast to roots exposed gradually, in which growth remained inhibited but cell viability was maintained. Total class III peroxidase activity increased significantly in all low pH treatments, but was not correlated with the observed differential responses. Use of the enzyme inhibitors salicylhydroxamic acid (SHAM) or diphenyleneiodonium chloride (DPI) suggest that peroxidase and, to a lesser extent, NADPH oxidase were required to prevent or reduce injury in all low pH treatments. However, a role for other enzymes, such as the alternative oxidase is also possible. The results with SHAM, but not DPI, were confirmed in tobacco BY-2 cells. Our results indicate that root growth inhibition from low pH can be part of an active plant response, and suggest that peroxidases may have a critical early role in reducing loss of cell viability and in the observed root growth constraint.

In vitro T-cell profile induced by BCG Moreau in healthy Brazilian volunteers.

Tuberculosis (TB) remains the world's leading cause of morbidity and mortality. Although Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only vaccine currently in use, its efficacy is highly variable. It has been suggested that early antigenic presentation is a pivotal event leading to a better immune response in TB vaccine models. To investigate this further, we compared in vitro cell-mediated immune responses in the context of early sensitization with TB (i.e. healthy adults vaccinated with BCG when they were young, HD; n = 25) to those in its absence (i.e., newborns with naïve immunity to TB, UV; n = 10) by challenging mononuclear cells with BCG Moreau. After 48 hours, CD4+ and CD8+ T cells were harvested from both groups and stained for PD-1/CD25/ FOXP3. In addition, supernatants were assayed for a broad range of cytokines using an array system. The HD group showed robust reactivity to Protein Purified Derivative and BCG while the naïve, UV group did not. Similarly, in terms of PD-1 expression and Treg cells (CD4+/CD25high(+)/FOXP3+), only the HD group showed higher levels in CD4 lymphocytes. Otherwise, only the UV group showed expression of CD25dim+ as an activation marker dependent on BCG infection. In terms of cytokines, the HD group showed higher levels of Th1 (IL-2/TNF-α/IFN-γ) and regulatory (IL-10) profiles, with monocytes, but not Tr1 cells, acting as the main source of IL-10. Taken together, our results highlight critical roles of early sensitization with TB in mounting cell-mediated immune responses.

Adenovirus and intranuclear inclusions in appendices in intussusception.

AIMS: To examine the light and electron microscopic features of appendices removed at the time of surgical reduction of intussusception in children; and to confirm that the viral inclusions seen in some of them are due to adenovirus. METHODS: A series of 39 appendices from cases of intussusception and 15 control appendices were reviewed. Light microscopic examination of haematoxylin and eosin stained sections was performed on all of them and one appendix with large numbers of inclusions was examined by electron microscopy. Non-isotopic in situ hybridisation using a biotinylated DNA probe was carried out on sections of appendix from 30 of the cases of intussusception and from the 15 controls. RESULTS: Light microscopic examination showed viral inclusions in 19 of the appendices from the cases of intussusception and in none of the controls. Electron microscopic examination showed viral particles with the typical features of adenovirus. Most of the appendices with viral inclusions in the haematoxylin and eosin stained sections also contained adenovirus DNA as shown by in situ hybridisation. CONCLUSIONS: Viral inclusions seen in appendices from cases of intussusception are caused by adenovirus. Adenovirus DNA was not demonstrable in appendices from cases of intussusception without viral inclusions and the aetiological factors involved in intussusception in these children remain unknown.

Fulminant disseminated pulmonary adiaspiromycosis in humans.

A case of fulminant disseminated pulmonary adiaspiromycosis is reported. The patient, a 35-year-old black male farm worker, presented with a four-week history of generalized weakness, unproductive cough, evening fever, and a weight loss of 8 kg. He died 12 days after hospitalization of respiratory failure due to granulomatous lung disease. The clinical and radiographic findings were indistinguishable from those of miliary tuberculosis. Microscopic examination of material obtained at autopsy revealed the large fungus characteristic of adiaspiromycosis in the center of suppurative granulomas throughout the lungs. This is believed to be the first fatal case of pulmonary adiaspiromycosis reported in humans, and it may have been occupationally acquired.

Hepatic capillariasis in children: report of 3 cases in Brazil.

Capillaria hepatica is a helminth that may cause an extremely rare condition of parasitic hepatitis. Only 29 cases have been published, 2 of them in Brazil. We report here 3 cases of children in Brazil with massive hepatic capillariasis who presented the characteristic triad of this type of infection, i.e., persistent fever, hepatomegaly, and eosinophilia. The diagnosis was made by liver biopsy. All children responded well after treatment with thiabendazole (case 1), albendazole (case 3), and albendazole in combination with a corticoid (case 2). Case 1 has been followed-up for 24 years, an event not previously reported in the literature.

Translocation (1;5)(q32;q35) in CD30+ anaplastic large cell non-Hodgkin lymphona of childhood. A case report.

The authors studied cytogenetically a case of CD30+ anaplastic large cell non-Hodgkin lymphoma previously diagnosed as malignant histiocytosis and detected a translocation involving chromosomes 1 and 5, t(1;5)(q32:q35). After comparing their findings with those from reports in the literature, they comment about the importance of breakpoint q35 on chromosome 5 and point out the importance of associating morphologic, immunoperoxidase, and cytogenetic findings to confirm the diagnosis of this tumor.

Electron-microscopic study of systemic myonecrosis due to poisoning by tropical rattlesnake (Crotalus durissus terrificus) in humans.

This article describes the ultrastructural study of skeletal muscle biopsy specimens from five patients following envenomization by tropical rattlesnake (Crotalus durissus terrificus). All the patients were bitten in the leg and the biopsy specimens were obtained from the contralateral gastrocnemius muscle in the middle of the lower leg. A wide spectrum of detailed ultrastructural changes involving muscle fibers and microvasculature was demonstrated. Essentially, such lesions included widespread necrotic myofibers intermixed with intact fibers, accompanied by changes in the endothelial lining of the intramuscular blood capillaries and small arterial vessels, reducing their lumens. Since these alterations were observed in biopsy specimens from the limb contralateral to the site of the bite, they clearly demonstrate the systemic myonecrotic action of the venom of a tropical rattlesnake. On the basis of these data, the mechanism of venom-induced myopathy is described. It is postulated that the pathogenesis of systemic myonecrosis due to poisoning by C durissus terrificus is a complex one, probably due to direct damage to cells by the myotoxins of the venom, as well as indirect effects due to ischemia.

DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells.

The objective of the present study was to develop a simplified low cost method for the collection and fixation of pediatric autopsy cells and to determine the quantitative and qualitative adequacy of extracted DNA. Touch and scrape preparations of pediatric liver cells were obtained from 15 cadavers at autopsy and fixed in 95% ethanol or 3:1 methanol:acetic acid. Material prepared by each fixation procedure was submitted to DNA extraction with the Wizard genomic DNA purification kit for DNA quantification and five of the preparations were amplified by multiplex PCR (azoospermia factor genes). The amount of DNA extracted varied from 20 to 8,640 microg, with significant differences between fixation methods. Scrape preparation fixed in 95% ethanol provided larger amount of extracted DNA. However, the mean for all groups was higher than the quantity needed for PCR (50 ng) or Southern blot (500 ng). There were no qualitative differences among the different material and fixatives. The same results were also obtained for glass slides stored at room temperature for 6, 12, 18 and 24 months. We conclude that touch and scrape preparations fixed in 95% ethanol are a good source of DNA and present fewer limitations than cell culture, tissue paraffin embedding or freezing that require sterile material, culture medium, laboratory equipment and trained technicians. In addition, they are more practical and less labor intensive and can be obtained and stored for a long time at low cost.

Rupture of the aorta due to a malpositioned tracheal cannula in a 4-month-old baby.

Complications of tracheostomy are not rare and are even more common in children. They may be acute or chronic and sometimes may have serious consequences. We report here a rare situation in which a malpositioned tracheal cannula in a false tract from the neck to the mediastinum induced necrosis and rupture of the aorta and of the left carotid artery with fatal hemorrhage.

De novo unbalanced t(11q;21q) leading to a partial monosomy 21pter-q22.2 and 11q24-qter in a patient initially diagnosed as monosomy 21.

We describe a patient in whom full monosomy 21 was initially assumed from routine GTG banded karyotyping. Re-examination with chromosome painting demonstrated an unbalanced translocation between the long arms of chromosomes 11 and 21. Fluorescence in situ hybridization (FISH) and microsatellite marker analysis revealed partial monosomy of chromosome 21 (pter-q22.2) and 11 (q24-qter). The patient was prematurely born in the 31st week of gestation and expired 3 days after delivery. She showed multiple minor anomalies, a complex cardio-vascular malformation, intestinal malrotation and cerebellar hypoplasia.

Raine dysplasia: a Brazilian case with a mild radiological involvement.

We report a preterm male infant, the first child of a young consanguineous couple, whose physical examination revealed craniofacial disproportion with microcephaly, wide fontanelles, exophthalmos, low nasal root and hypoplastic nose, long philtrum, small mouth, high arched and narrow palate, micrognathia, dysplastic, low-set and rounded ears, short neck and, arthrogryposis. Postmortem findings included hypoplastic lungs. Radiological examinations showed mild and localized increased of bone density in the cranial vault and skull base and facial bones and undermodelled in the long bones. The above findings are characteristics of Raine dysplasia but the case reported here presents a mild bone involvement with only a localized bone sclerosis and absence of prenatal fractures. We discuss the possibility that this case represents an allelic mutation of the Raine gene. The consanguinity of the parents reinforces the hypothesis of autosomal recessive inheritance for this entity.

Noonan syndrome associated with unilateral iris coloboma and congenital chylothorax in an infant.

Chylothorax and colobomas are uncommon features reported in Noonan syndrome. We describe an infant with Noonan phenotype, congenital chylothorax and a unilateral iris coloboma. The presence of these both abnormalities in the same patient has not previously been reported.

Sudden unexpected infant death syndrome in Ribeirão Preto, Brazil.

CONTEXT: Sudden Infant Death Syndrome (SIDS) is the leading cause of death in the first year of life in developed countries. Brazilian and Latin American literature lack studies on the subject, which is largely unknown among health workers. OBJECTIVE: To identify cases that could be classified as SIDS among children of less than one year of age submitted to autopsy at the Serviço de Verificação de Obitos do Interior (SVOI), in Ribeirão Preto, SP, Brazil. A retrospective analysis of all autopsies from SVOI in this age group from January 1987 to December 1990 was done. RESULTS: There were 369 autopsies of which 344 (93.2%) deaths were expected and 25 (6.8%) unexpected. From the 25 unexpected cases 16 (64%) deaths could not be explained after autopsy and from these cases only 10 were eligible for the study because they had full organ sampling. There were 7 males and 3 females and the age at death ranged from 1 to 3 months (average: 1.7 months). Two were found dead, 3 died at home, 4 died on the way to hospital and 1 died while being fed. Autopsy diagnoses were aspiration (8 cases), SIDS (1 case) and undetermined (1 case). Aspiration was not confirmed by histology and the only findings were mild pulmonary edema, subcapsular petechiae and intraparenchymatous hemorrhage in thymus. CONCLUSION: That there were 10 cases of unexpected and unexplained deaths of children less than 1 year-old during the evaluated period with characteristics similar to SIDS which should therefore be classified as such.

[Abdominal aortic thrombosis mimicking aortic coarctation in a newborn with heart failure]. / Trombose da aorta abdominal simulando coarctação da aorta em neonato com insuficiência cardíaca.

The case of a neonate in heart failure with the classical signs of coarctation of the aorta is described. Two dimension and Doppler echocardiography ruled out coarctation of the aorta and an abdominal ultrasonography detected a large thrombotic formation in the abdominal aorta, confirmed at necropsy.