Cobalamin F deficiency in a girl with severe skin hyperpigmentation and a homozygous LMBRD1 variant.

Cobalamin (vitamin B12) is important in gastrulation, nervous system development and haemoglobin formation. Mutations of the ABCD4 or LMBRD1 genes can lead to cobalamin-related disorders. We report a patient with disseminated skin hyperpigmentation caused by a homozygous LMBRD1 variant. Genetic disorders of cobalamin metabolism caused by variants in the ABCD4 or LMBRD1 genes should be considered in patients presenting with cutaneous hyperpigmentation. Click https://www.wileyhealthlearning.com/#/online-courses/a6ef1275-8325-4834-89d2-aa18fa31e63f for the corresponding questions to this CME article.

Unequal Impact of COL1A1 and COL1A2 Variants on Dentinogenesis Imperfecta.

Dentinogenesis imperfecta (DI) is the main orodental manifestation of osteogenesis imperfecta (OI) caused by COL1A1 or COL1A2 heterozygous pathogenic variants. Its prevalence varies according to the studied population. Here, we report the molecular analysis of 81 patients with OI followed at reference centers in Brazil and France presenting COL1A1 or COL1A2 variants. Patients were submitted to clinical and radiographic dental examinations to diagnose the presence of DI. In addition, a systematic literature search and a descriptive statistical analysis were performed to investigate OI/DI phenotype-genotype correlation in a worldwide sample. In our cohort, 50 patients had COL1A1 pathogenic variants, and 31 patients had COL1A2 variants. A total of 25 novel variants were identified. Overall, data from a total of 906 individuals with OI were assessed. Results show that DI was more frequent in severe and moderate OI cases. DI prevalence was also more often associated with COL1A2 (67.6%) than with COL1A1 variants (45.4%) because COL1A2 variants mainly lead to qualitative defects that predispose to DI more than quantitative defects. For the first time, 4 DI hotspots were identified. In addition, we showed that 1) glycine substitution by branched and charged amino acids in the α2(I) chain and 2) substitutions occurring in major ligand binding regions-MLRB2 in α1(I) and MLBR 3 in α2(I)-could significantly predict DI (P < 0.05). The accumulated variant data analysis in this study provides a further basis for increasing our comprehension to better predict the occurrence and severity of DI and appropriate OI patient management.

Evidence for an association between haptoglobin and MnSOD (Val9Ala) gene polymorphisms in essential hypertension based on a Brazilian case-control study.

Essential hypertension is a complex and multifactorial trait; genetic and environmental factors interact to produce the final phenotype. Studies have demonstrated association of hypertension with varied gene polymorphisms. However, demonstration of common genetic causes in the general population remains elusive. We investigated a possible association between hypertension and haptoglobin, angiotensin I-converting enzyme (ACE), glutathione S-transferases GSTM1 and GSTT1, MnSOD (Val9Ala), CAT (-21A/T), and GPX1 (Pro198Leu) gene polymorphisms in an urban Brazilian population group from Brasília. Although ACE has been reported to be one of the main polymorphisms associated with hypertension, we found no association with ACE's specific genotypes. However, a possible association with Hp1-1 and MnSOD Val/Ala genotypes suggests that, at least for the Brazilian population, polymorphisms related to oxidative stress should be more deeply investigated.

PIMA: A population informative multiplex for the Americas.

We describe an ancestry-informative autosomal SNP multiplex designed to be a small-scale, flexible panel that can complement uniparental markers in assessing the American variability (i.e. pre-Colombian) found in contemporary indigenous American populations. This study centered on choosing SNPs with the specific characteristics of: 1) extreme allele frequency differences between indigenous Americans and the African, European and East Asian population groups that contribute to present-day population variation in the Americas; 2) high informativeness-for-assignment In values; and 3) well-spaced genomic distribution and chromosomal separation from existing small-scale forensic ancestry marker sets. The resulting capillary electrophoresis SNaPshot single base extension test was named: PIMA (Population Informative Multiplex for the Americas), comprising 26 autosomal SNPs, a single X-chromosome SNP plus the amelogenin sex marker adapted for SNaPshot. PIMA complements the established 34plex forensic ancestry panel to provide a powerful and simple tool for the analysis of American populations, including those with admixed histories, commonly encountered in America. Comparing the results obtained with the combined marker panels of PIMA and 34plex to SNP data from a much larger ancestry panel allowed us to gauge their relative efficiency. PIMA+34plex gives equivalent power to the 314-SNP 'LACE' genomic ancestry control panel, while requiring a much smaller genotyping effort. The ancestry profiles and genetic structure of 22 populations spread across the American continent were estimated using PIMA+34plex data, and those estimates were contrasted with information provided by uniparental markers (mtDNA and Y-chromosome loci) for a small set of admixed individuals from Venezuela. Our results indicate that an American genetic component is efficiently detected in contemporary American populations using a small set of ancestry informative SNPs, and these co-ancestry estimates are consistent with the known history and demography of the Americas. The small scale and high population differentiation power of PIMA, particularly when combined with 34plex, provides a practical and powerful tool for genetic studies of American populations as well as forensic DNA analyses.

Genetic polymorphisms in oestrogen metabolic pathway and breast cancer: a positive association with combined CYP/GST genotypes.

The cytochrome P450 family (CYPs) and the glutathione S-transferase (GSTs) enzymes play an important role in the metabolism of environmental carcinogens and of oestrogen and can affect breast cancer risk. In this study we examine the role of the genes CYP1A1, CYP17, CYP2D6, GSTM1, GSTP1 and GSTT1 in breast cancer risk in Brazilian women. The study population consisted of 102 incident breast cancer cases and 102 healthy controls. Genotyping analyses were performed by PCR-based methods. A significant finding was observed between GSTP1 Ile-Val polymorphism and breast cancer risk (OR = 1.81; CI 95% = 1.04-3.16). A significant association was observed between women with 0-2 risk genotypes and those with 4 or more risk genotypes (OR = 2.42; CI 95% = 1.13-5.18) when the potential combined effects of the risk genotypes were examined. No significant differences between cases and controls were found correlating the genotypes and the clinical-histopathological parameters. In conclusion, in our population only GSTP1 was associated with breast cancer risk. However, when the genes were tested in combination, a significant association in the breast cancer risk was observed.

Cellular mechanism of the conduction abnormalities induced by serum from anti-Ro/SSA-positive patients in rabbit hearts.

In this study, IgG fractions from sera of SLE patients with anti-Ro/SSA or anti-Ro/SSA and anti-La/SSB activity were tested in Langendorff preparations of adult rabbit hearts, aiming to reproduce the cardiac manifestations observed in neonatal lupus in an experimental model. The hearts were perfused with normal Tyrode's solution for 30 min, followed by perfusion with Tyrode's containing 0.3 mg/ml of anti-Ro/SSA- (or anti-Ro/La-) positive IgG (nine sera), anti-ribonucleoprotein (RNP)-positive IgG (five sera), or IgG fractions from normal donors (five sera). In one third of the experiments done with anti-Ro/La-positive IgG, heart block was observed. With the remaining fractions, a decrease in heart rate of 17.1% was observed, but normal sinus rhythm was maintained. The IgG fractions with anti-RNP activity (five experiments) and from normal sera (six experiments) reduced heart rates by 12.9 and 3.3%, respectively, but heart block was not observed. To further characterize the cellular mechanisms involved in the conduction disturbances observed in the whole rabbit hearts, we conducted experiments with ventricular myocytes isolated from young rabbit hearts, studied by whole cell patch-clamp technique. In these experiments, the slow inward currents were analyzed during the superfusion of the cell with normal Tyrode's solution and 5 min after superfusion with Tyrode's solution containing 0.3 mg/ml of anti-Ro/SSA- (or anti-Ro/La-) positive IgG (five sera), anti-RNP-positive IgG (three sera), or IgG from normal donors (four sera). Resting and action potential amplitudes were not affected by any of the sera used. The anti-Ro/SSA IgG fraction induced a mean reduction in the peak slow inward current of 31.6%. IgG fractions with anti-RNP activity reduced slow inward current by 4.4%, whereas IgG fractions from normal donors increased this current by 3.3%. IgG-free fractions from sera of patients with anti-Ro/SSA activity did not alter the peak slow inward current. These results show, for the first time, that the presence of anti-Ro/SSA or anti-Ro/SSA and anti-La/SSB antibody activity in IgG fractions from lupus patients' sera can induce cardiac conduction disorders similar to those observed in neonatal lupus.

Genetic variability in maned wolf based on heterologous short-tandem repeat markers from domestic dog.

The maned wolf (Chrysocyon brachyurus) is the largest South American canid. Habitat loss and fragmentation, due to agricultural expansion and predatory hunting, are the main threats to this species. It is included in the official list of threatened wildlife species in Brazil, and is also protected by IUCN and CITES. Highly variable genetic markers such as microsatellites have the potential to resolve genetic relationships at all levels of the population structure (among individuals, demes or metapopulations) and also to identify the evolutionary unit for strategies for the conservation of the species. Tests were carried out to verify whether a class of highly polymorphic tetranucleotide repeats described for the domestic dog effectively amplifies DNA in the maned wolf. All five loci studied were amplified; however, one of these, was shown to be monomorphic in 69 maned wolf samples. The average allele number and estimated heterozygosity per polymorphic locus were 4.3 and 67%, respectively. The genetic variability found for this species, which is considered threatened with extinction, showed similar results when compared to studies of other canids.

Endothelium inactivation in in vitro perfused vascular beds. Comparison of methods.

In order to choose the best procedure to inactive the endothelium from vascular beds perfused in vitro, we compared four methods: perfusion with sodium deoxycholate 0.3% for 30 sec; 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate 0.3% (CHAPS) for 2.5 min; collagenase 0.2% for 15 min, and distilled water for 10 min, using the mesenteric arterial bed (MAB) of the rat. The effectiveness of the treatments used to inactivate the endothelium was assessed functionally by using acetylcholine and sodium nitroprusside and histologically using light microscopy. Phenylephrine was used to test the contractile properties of the preparations after each treatment. After collagenase, distilled water, and CHAPS treatment, a potentiated response to phenylephrine was observed, whereas sodium deoxycholate treatment did not modify phenylephrine-induced responses. Acetylcholine-induced responses were reduced by collagenase (60% reduction), CHAPS (30% reduction), and distilled water (52% reduction) treatment, and sodium deoxycholate completely abolished acetylcholine-induced responses. Except after collagenase treatment, smooth muscle relaxant responses were not altered. Medial smooth muscle cells displayed an unchanged morphology, appearing similar to those in control mesenteric arterial beds, except for collagenase and distilled water. Despite the fact that sodium deoxycholate treatment completely abolished acetylcholine-induced response, endothelial cells were still found. No treatment totally removed endothelial cells. In conclusion, we suggest that sodium deoxycholate treatment is the best procedure to inactivate endothelial cells from vascular beds perfused in vitro since it completely abolished endothelium-dependent relaxation and did not interfere with smooth muscle vasodilating and contracting properties.

Prognostic significance of silent myocardial ischemia after a first uncomplicated myocardial infarction.

Forty asymptomatic patients were studied after a first uncomplicated myocardial infarction. They were 36 men and 4 women, with a mean age of 52.6 yr; the location of myocardial infarction was in the anterior wall in 18 (45%) patients and in the inferior wall in 22 (55%). The patients were submitted to: (1) 48-h Holter monitoring, during the 2nd and 8th weeks after the acute event; (2) exercise testing during the same periods; (3) cardiac catheterization and coronary arteriography. Patients with clinical conditions associated with cardiac rhythm disturbances or repolarization abnormalities were excluded. The electrocardiographic methods identified 11 (27.5%) patients with silent myocardial ischemia. Patients with and without silent ischemia were similar in relation to sex, age, coronary risk factors, arrhythmias, left ventricular function and follow-up. Patients with silent ischemia had more inferior wall myocardial infarctions, but the difference was not statistically significant. Patients with silent ischemia had significantly more extensive coronary artery disease (45.5% multivessel disease) when compared to those without ischemia (14.8% multivessel disease) (p < 0.05). After a 2-yr follow-up, 4 (36.4%) patients with and 1 (3.4%) without silent ischemia had a coronary event (p < 0.05). Kaplan-Meier analysis demonstrated a significantly higher cumulative probability of not experiencing a new coronary event for the patients without silent ischemia (96.5%) as compared to those with silent ischemia (62.3%) (p < 0.01). Our results suggest that silent myocardial ischemia after a first uncomplicated myocardial infarction carries an adverse prognosis and should be routinely investigated.

Advanced oviductal development, transport to the preferred implantation site, and attachment of the blastocyst in captive-bred, short-tailed fruit bats, Carollia perspicillata.

The final stages of embryonic development in the oviduct, transport of the embryo to the uterus, and the initial stages of implantation have been examined in captive-bred Carollia perspicillata at the light and electron microscopic levels. Development progressed to the expanded, zona pellucida-free, blastocyst stage in the oviduct. The abundance of microvilli on the exterior of the trophoblast varied with the degree of blastocyst expansion and cell shape, and may function in part as a membrane reservoir. Cells of the blastocyst also typically contained many lipid droplets and prominent areas of cytoplasm occupied by finely granular material (probably glycogen) instead of organelles. In most females, closure of the uterine lumen occurred prior to, or around the time of, transport of the blastocyst to the usual implantation site and appeared to play a role in preventing transport of the blastocyst too far distally in the uterus. This was associated with increased endometrial edema, particularly in the fundic region of the simplex uterus, and the extravasation of many erythrocytes into the endometrial stroma. Both of these changes began while the blastocyst was still being held in the oviduct and became pronounced during implantation. Engulfment of these erythrocytes by processes of the endometrial stromal cells and their phagocytosis by macrophages was also observed. Implantation was usually initiated within narrow tubular segments, lined by endometrium, that were located between the end of each oviduct and the main cavity of the uterus, or from immediately adjacent areas of the main cavity. During the early stages of implantation, the blastocyst was clasped by the endometrium at the implantation site, and this was associated with extensive interdigitation of the microvilli of the trophoblast and adjacent uterine epithelial cells. Initial adhesion of the trophoblast, which was still cellular rather than synctial, occurred over the apical intercellular junctions of the uterine epithelial cells.

Autoradiography reveals regional metabolic differences in the endometrium of pregnant and nonpregnant mice.

The rodent endometrium undergoes remarkable modifications during pregnancy, resulting from a redifferentiation of its fibroblasts. During this modification (decidualization), the fibroblasts transform into large, polyhedral cells that establish intercellular junctions. Decidualization proceeds from the subepithelial stroma towards the deep stroma situated next to the myometrium and creates regions composed of cells in different stages of differentiation. We studied by autoradiography whether cells of these different regions have different levels of macromolecular synthesis. Radioactive amino acids or radioactive sulfate were administered to mice during estrus or on different days of pregnancy. The animals were killed 30 min after injection of the precursors and the uteri were processed for light microscope autoradiography. Silver grains were counted over cells of different regions of the endometrium and are reported as the number of silver grains per area. Higher levels of incorporation of amino acids were found in pregnant animals as compared to animals in estrus. In pregnant animals, the region of decidual cells or the region of fibroblasts transforming into decidual cells showed the highest levels of synthesis. Radioactive sulfate incorporation, on the other hand, was generally higher in nonpregnant animals. Animals without decidual cell transformation (nonpregnant and 4th day of pregnancy) showed a differential incorporation by subepithelial and deep stroma fibroblasts. This study shows that regional differences in synthetic activity exist in cells that are in different stages of transformation into decidual cells as well as in different regions of the endometrium of nonpregnant mice.

An ultrastructural observation on charquis, salted and intermediate moisture meat products.

Charqui meats are tropical intermediate moisture meat products containing 45% moisture and 15% salt with an A(w) of 0.70-0.75. Light microscopic studies of a charqui derivative popularly known as Jerked beef (JB) demonstrated considerable shrinking of muscle cells and the formation of fluid channels. The area occupied by muscle cells in JB was diminished by 30-40% in comparison with control samples. At the ultrastructural level, A-bands including the M-line disappeared indicating proteins were lost during processing. Z-lines appeared to be fragmented. In the enlarged extracellular spaces, collagen fibers retained their banding patterns although an empty space was observed surrounding these fibers. The denaturation of myofibrillar proteins during processing and the osmotic pressure caused by salting create conditions for water movement from the myofibrillar compartments to the intermyofibrillar space, then to the extracellular matrix and ultimately to the meat surface.

[Pseudoaneurysm of the left ventricle resected 20 years after acute myocardial infarction]. / Pseudoaneurisma do ventrículo esquerdo ressecado vinte anos após infarto agudo do miocárdio.

A 67 year-old-man suffered an uncomplicated myocardial infarction twenty years ago. By January 1990 an abnormal cardiac contour was noted on a chest radiography. Contrast ventriculography revealed a pseudoaneurysm of the left ventricle. The patient underwent open heart surgery and remains asymptomatic 12 months after surgery. This is the longest time interval between the myocardial infarction and successful surgery that has been reported.

[Effort angina in an adult patient with anomalous origin of the left coronary artery]. / Angina de esfórço em paciente adulto com origem anômala de artéria coronária esquerda.

Anomalous origin of the left main coronary artery from the pulmonary artery in a 50 year-old patient whose initial symptom was effort angina during the last 8 months. The clinical features and treatment are also discussed.

[Hemodynamic variable analysis in patients with systemic arterial hypertension undergoing physical exercise]. / Análise das variáveis hemodinâmicas em pacientes com hipertensão arterial sistêmica submetidos ao exercício físico.

PURPOSE: To verify the exercise-induced hemodynamic changes in moderate hypertensive patients. METHODS: Twenty nine patients were studied and they were submitted to cycloergometer supine exercise (50w and 100w) during cardiac catheterization. The hemodynamic variables were measured: cardiac index (CI), stroke volume (SV), systemic arterial resistance (SAR), pulmonary arterial resistance (PAR), wedge pressure (WP), right atrial pressure (RA), systolic arterial pressure (SP), diastolic arterial pressure (DP), mean arterial pressure (MP) and heart rate (HR). To evaluate the cardiac function, the patients were divided in two groups: GI with CI > 2.5 ml/min/m2 and GII CI < 2.5 ml/min/m2. RESULTS: During exercise, patients from GI and GII were similar-according to MAP, RA, WP, and HR. On the other hand, GI and GII exhibited different (#) behavior regarding SV, PAR and SAR. During the three exercise conditions, rest (R), 50w and 100w it was observed: a) PAR-R = 50 = 100 (GI); b) MP, PAR, WP, SV, SAR-R #50 #100 (GI); c) HR, PAR and SAR-R #50 #100 (GII). There was no correlation between the cardiac function (CI, SV) and the circulatory adjustment (PAR and SAR) or the pressure curve (SP, DP, MP) and HR. CONCLUSION: The results suggest that the cardiac function of GI patients depends, mainly, on the inotropism, while in GII patients it depends on the decrease of the afterload (PAR and SAR). Those changes may appear even in the late exercise stage (100w), with a bias to attain the GI levels. Those observations suggest functional changes in vasomotor tone of GII patients.

[Transdermal nitroglycerin in patients with stable angina]. / Nitroglicerina transdérmica em portadores de angina estável.

PURPOSE: To evaluate the nitroglycerin patches efficacy and tolerability in patients with stable angina pectoris. METHODS: One thousand and five hundred and thirty nine patients with stable angina pectoris, mean age 61.0 +/- 10.3, 891 men and 648 women were prospectively evaluated by five hundred and thirty five specialists after 5 mg or, posteriorly, if clinical necessary, 10 mg of nitroglycerin patches during 12 weeks. Clinical evaluation, electrocardiogram (ECG) and treadmill exercise were obtained on study entry and at weeks 2, 4, 8 and 12 for clinical evaluation, and at week 12 for ECG and treadmill exercise. RESULTS: A significative reduction was observed in the number of angina crisis, sublingual nitrates consumption, arterial blood pressure and on the percentage of positive treadmill exercise tests. The heart rate and nitroglycerin patches dose did not show statistical differences. The compliance of transdermal administration was excellent. CONCLUSION: The nitroglycerin patches administration was effective for stable angina pectoris with excellent patient's compliance.

[Silent ischemia after an uncomplicated myocardial infarct. A study in asymptomatic patients]. / Isquemia silenciosa após infarto do miocárdio não complicado. Estudo em pacientes assintomáticos.

PURPOSE: To verify the prognostic value of silent myocardial ischemia (SMI) after an uncomplicated myocardial infarction (MI). METHODS: Forty asymptomatic patients were studied after a first uncomplicated MI. They were submitted to 48 hour ambulatory electrocardiographic monitoring and exercise-testing, during the 2nd and 8th weeks after the acute event. Thirty-nine patients were submitted to cardiac catheterization and coronary arteriography; one patient was submitted to necropsy. The electrocardiographic study identified 11 (27.5%) individuals with SMI (group A); the other 29 patients were considered group B. RESULTS: Groups A and B were similar in relation to clinical characteristics, infarct site and ventricular function. Group A had significantly more extensive coronary artery disease when compared to group B. After a two-year follow-up, patients from group A had significantly more coronary events (36.3%) when compared to group B (3.4%). Kaplan-Meier analysis demonstrated a significantly higher cumulative probability of not experiencing a new coronary event for the group B patients. CONCLUSION: SMI may have a prognostic value after uncomplicated MI, as in other clinical manifestations of coronary artery disease.

[Variability of the heart rate and silent ischemia after myocardial infarction]. / Variabilidade da freqüência cardíaca e isquemia silenciosa após infarto do miocárdio.

PURPOSE: To show a possible relation between heart rate and silent myocardial ischemia. METHODS: Forty-nine ischemic episodes were registered in six patients during a total period of 576-hour Holter monitoring. Those patients were selected from a group of 40 asymptomatic individuals after a first uncomplicated myocardial infarction; 11 (27.5%) showed ischemia during daily activities or exercise, the six selected patients had myocardial ischemia on Holter monitoring. RESULTS: The silent episodes consisted 92% of the total ischemic burden; they lasted from 1 min 30 s to 20 min and the ST-segment depression varied from -1.1 mm to 3.3 mm. Thirty-five (72%) episodes occurred at rest or during light physical activities; nine (18.5%) occurred between 7:00 AM and 12:00 PM; eight (16.5%), between 12:00 PM and 6:00 PM; 17 (35%) between 6:00 PM and 12:00 AM and 15 (30%), between 12:00 AM and 7:00 AM. There was no significant change (more than 20%) in heart rate at the onset of ischemic episodes in relation to the heart rate 1 minute before (94.63 +/- 9.79 bpm and 99.47 +/- 10.99 bpm, respectively). Complex ventricular arrhythmias occurred in all patients and only one of them had an episode of nonsustained ventricular tachycardia related to silent ischemia. CONCLUSION: Our results suggest that there is no relation between heart rate, arrhythmias and silent ischemia.

[Severe pulmonary embolism in a single lung]. / Embolia pulmonar grave em pulmão único.

A 68 years old white man complaining of hemoptysis in the last 2 months was admitted for evaluation. He was a heavy smoker and thorax radiography as week as CT Scan showed a 5 cm mass in the right pulmonary hilum. Endoscopy with biopsy demonstrated an adenocarcinoma of middle lobe. He was operated on and a tumor was found in the middle lobe with invasion of the other two lobes and pericardium. A right pneumectomy was performed. In the 10th postoperative day the patient sustained a pulmonary thromboembolism. A pulmonary scintigraphy showed hypoperfusion of large areas of left lung. He was medically treated and had a good response. The patient was discharged in the 22nd postoperative day. Pulmonary thromboembolism in bowen more severe in patients with single lung, nevertheless the recovery of this patients was good with early diagnosis and treatment.