Biallelic SCN10A mutations in neuromuscular disease and epileptic encephalopathy.

OBJECTIVES: Two consanguineous families, one of Sudanese ethnicity presenting progressive neuromuscular disease, severe cognitive impairment, muscle weakness, upper motor neuron lesion, anhydrosis, facial dysmorphism, and recurrent seizures and the other of Egyptian ethnicity presenting with neonatal hypotonia, bradycardia, and recurrent seizures, were evaluated for the causative gene mutation. METHODS AND RESULTS: Homozygosity mapping and whole exome sequencing (WES) identified damaging homozygous variants in SCN10A, namely c.4514C>T; p.Thr1505Met in the first family and c.4735C>T; p.Arg1579* in the second family. A third family, of Western European descent, included a child with febrile infection-related epilepsy syndrome (FIRES) who also had compound heterozygous missense mutations in SCN10A, namely, c.3482T>C; p.Met1161Thr and c.4709C>A; p.Thr1570Lys. A search for SCN10A variants in three consortia datasets (EuroEPINOMICS, Epi4K/EPGP, Autism/dbGaP) identified an additional five individuals with compound heterozygous variants. A Hispanic male with infantile spasms [c.2842G>C; p.Val948Leu and c.1453C>T; p.Arg485Cys], and a Caucasian female with Lennox-Gastaut syndrome [c.1529C>T; p.Pro510Leu and c.4984G>A; p.Gly1662Ser] in the epilepsy databases and three in the autism databases with [c.4009T>A; p.Ser1337Thr and c.1141A>G; p.Ile381Val], [c.2972C>T; p.Pro991Leu and c.2470C>T; p.His824Tyr], and [c.4009T>A; p.Ser1337Thr and c.2052G>A; p.Met684Ile]. INTERPRETATION: SCN10A is a member of the voltage-gated sodium channel (VGSC) gene family. Sodium channels are responsible for the instigation and proliferation of action potentials in central and peripheral nervous systems. Heterozygous mutations in VGSC genes cause a wide range of epileptic and peripheral nervous system disorders. This report presents autosomal recessive mutations in SCN10A that may be linked to epilepsy-related phenotypes, Lennox-Gastaut syndrome, infantile spasms, and Autism Spectrum Disorder.

A novel mutation of IL1RN in the deficiency of interleukin-1 receptor antagonist syndrome: description of two unrelated cases from Brazil.

OBJECTIVE: Monogenic autoinflammatory diseases are disorders of Mendelian inheritance that are characterized by mutations in genes that regulate innate immunity and whose typical features are systemic inflammation without high-titer autoantibodies or antigen-specific T cells. Skin and bone inflammation in the newborn period have been described in 3 of these autoinflammatory disorders: neonatal-onset multisystem inflammatory disease, Majeed syndrome, and deficiency of interleukin-1 (IL-1) receptor antagonist (DIRA) syndrome. This study was undertaken to present the characteristics of the DIRA syndrome in 2 cases from Brazil, and describe a novel mutation in IL1RN. METHODS: Two unrelated Brazilian patients were evaluated for the clinical signs and symptoms of these 3 disorders, and peripheral blood samples were assessed for mutations in NLRP3, LPIN2, and IL1RN by DNA resequencing analysis. A mutation in IL1RN that encodes a mutant protein was identified, and the expression and function of this mutant protein were compared to those of the wild-type protein. RESULTS: Both patients presented with pustular dermatitis resembling generalized pustular psoriasis, recurrent multifocal aseptic osteomyelitis, and elevation in the levels of acute-phase reactants, all of which are features most consistent with the DIRA syndrome. Chronic lung disease was observed in 1 of the patients, and jugular venous thrombosis was observed in the other patient. Both patients showed a partial response to corticosteroid therapy, and 1 patient experienced an initial improvement of dermatitis with the use of acitretin. Both patients were homozygous for a novel 15-bp (in-frame) deletion on the IL1RN gene. The mutated protein expressed in vitro had no affinity with the IL-1 receptor, and stimulation of the patients' cells with recombinant human IL-1α or IL-1ß led to oversecretion of proinflammatory cytokines, similar to the findings obtained in previously reported patients. CONCLUSION: The presence of the same homozygous novel mutation in IL1RN in 2 unrelated Brazilian patients suggests that this genetic variant may be a founder mutation that has been introduced in the Brazilian population.

Golgi-type I and Golgi-type II neurons in the ventral anterior thalamic nucleus of the adult human: morphological features and quantitative analysis.

The morphological and quantitative features of neurons in the adult human ventral anterior thalamic nucleus were studied in Golgi preparations. Two neuronal types were found and their quantitative features were studied. Golgi-type I neurons were medium to large cells with dense dendritic trees and dendritic protrusions and short hair-like appendages. They have somatic mean diameter of 30.8 microm (+/-9.4, n = 85). They have an average 100.3 dendritic branches, 48.97 dendritic branching points, and 58.85 dendritic tips. The mean diameters of their primary, secondary, and tertiary dendrites were 3.1 microm (+/-1, n = 80), 1.85 microm (+/-0.8, n = 145), and 1.5 microm (+/-0.4, n = 160), respectively. Golgi-type II neurons were small to medium cells with few sparsely branching dendrites and dendritic stalked appendages with or without terminal swellings. They have somatic mean diameters of 22.2 microm (+/-5.8, n = 120). They have an average 33.76 dendritic branches, 16.49 dendritic branching points, and 21.97 dendritic tips. The mean diameters of their primary, secondary, and tertiary dendrites were 1.6 microm (+/-0.86, n = 70), 1.15 microm (+/-0.55, n = 118), and 1 microm (+/-0.70, n = 95), respectively. These quantitative data may form the basis for further quantitative studies involving aging or some degenerative diseases that may affect cell bodies and/or dendritic trees of the Golgi-type I and/or Golgi-type II thalamic neurons.

Serum sex hormones in premenopausal women with coronary heart disease.

OBJECTIVES: The objectives of this study were to investigate the relationship between the incidence of coronary heart diseases in premenopausal women and plasma level of total and free testosterone, estrogen, androstenedione and sex hormone binding globulin with coronary risk factors: fasting plasma concentration of glucose, triglyceride, total cholesterol, high and low density lipoproteins. METHODS: The study was conducted in Faculty of Medicine, Jordanian University of Science/Irbid Jordan and Technology and department of cardiology in Queen Alia Heart Institute/Amman Jordan during the period from April 2003 to March 2004. Serum sex hormones levels were measured in fifty-three premenopausal women; 25 women with coronary heart disease who had hypertension and/or diabetes mellitus and 28 women without coronary heart disease. Ages ranged from 34 to 48 years. Blood samples were collected just before performing coronary angiography and serum was obtained and frozen at -70 degrees C until use. RESULTS: We found that the mean triglyceride, fasting blood sugar total cholesterol, and low density lipoprotein concentration were significantly higher in coronary heart disease patients than in patients with normal coronaries. We also found that the low levels of sex hormone binding globulin and high levels of free testosterone are associated with development of coronary heart disease. No significant correlation could be established between other plasma sex hormones level and coronary heart disease. CONCLUSION: In young women, in the presence of coronary risk factors and normal level of serum estrogen, the high levels of serum free testosterone and low levels of serum sex hormone binding globulin are associated with development of atherosclerosis and increased incidence of coronary heart disease.

Homozygous AMN mutation in hereditary selective intestinal malabsorption of vitamin B12 in Jordan.

OBJECTIVE: Juvenile megaloblastic anemia is a rare and often hereditary disorder of cobalamin absorption, transport or intracellular metabolism. Several syndromes present with megaloblastic anemia such as congenital megaloblastic anemia due to intrinsic factor defect and juvenile megaloblastic anemia with proteinuria due to defects in the cubilin or the amnionless protein. METHODS: We identified a large kindred with juvenile megaloblastic anemia. Four genes, GIF, CUBN, TCN1, and TCN2, was previously excluded from being responsible for the syndrome of this family who was discovered in Irbid, Jordan, during the year 1999. At that time, the amnionless (AMN) gene was not yet known to implicate in megaloblastic anemia. In this study, we screened the AMN for mutations in the Ohio State University, Iowa, United States of America. In addition, follow-up testing was carried out in the University of Iowa in 2004. RESULTS: We identified a homozygous splice site mutation in the patients. This mutation was previously detected in families from Turkey and Tunisia. It is suspected to be a founder mutation of Middle Eastern origin. CONCLUSION: Molecular testing for this specific mutation in cases of Middle Eastern origin is a valuable tool for presymptomatic diagnosis, carrier identification and perhaps prenatal diagnosis.

Bcl-2 gene rearrangement in Jordanian follicular and diffuse large B-cell lymphomas.

OBJECTIVE: Follicular lymphoma (FL), a common subtype of non-Hodgkin's lymphoma (NHL) in the West, represents a rare subtype in Jordan. Bcl-2 gene rearrangement plays a crucial role in the biology of the vast majority of FL and a substantial number of diffuse large B-cell lymphoma (DLBCL) in the West; but its presence has not been studied in Jordan. Our aims are to document if bcl-2 gene rearrangement exists in Jordanian FL and DLBCL, and if present to determine whether its frequency among these lymphomas is different from the West and therefore may be responsible for some of the epidemiological differences seen between Jordan and the West. METHODS: The study was conducted in the year 2001 using polymerase chain reaction (PCR), to detect bcl-2 gene rearrangement in paraffin sections in 5 FL and 23 DLBCL cases diagnosed at the Department of Pathology at Jordan University of Science and Technology, Irbid, Jordan. Two sets of primers including the major breakpoint region (MBR) and the minor cluster region (MCR) were used. RESULTS: Amplifiable DNA was extracted from all cases. Bcl-2 gene rearrangement was seen among 4 (80%) of 5 FL cases, and 8 (35%) of 23 DLBCL cases. The majority of the rearrangements involved the MBR; however, one fourth of cases (one of 4 FL; 2 of 8 DLBCL) with bcl-2 rearrangement involved the MCR. CONCLUSION: Bcl-2 gene rearrangement was seen in the vast majority of Jordanian FL cases and approximately one third of all DLBCL cases. These figures are similar to those reported in the West, and therefore bcl-2 gene rearrangement does not help in explaining the epidemiological differences of NHL between Jordan and the West. The presence of bcl-2 gene rearrangement in DLBCL may define a subset of lymphomas that may be biologically and clinically unique and different from the rest of DLBCL.

Familial Mediterranean fever mutation frequencies and carrier rates among a mixed Arabic population.

OBJECTIVE: Familial Mediterranean Fever (FMF) is an autoinflammatory periodic disorder characterized by febrile and painful attacks due to inflammation involving the serosal membranes. The gene implicated in this disorder, MEFV, has been cloned and mutations in its coding regions have been identified. We aimed at identifying the frequency of MEFV mutations and carrier frequency in a mixed Arabic population. METHODS: We identified 29 probands from 29 unrelated sibships segregating the disorder and representing the affected individual cohort. We screened 200 anonymous deoxyribonucleic acid (DNA) samples, representing a healthy adult cohort, for the mutations found to be common in the affected individual cohort. We also, screened anonymous DNA samples from 4 Arabic countries, namely, Egypt (231), Syria (225), Iraq (176) and the Kingdom of Saudi Arabia (107) thus enlarging our healthy adult cohort. The study was carried out between 1999 and 2002 at Jordan University of Science and Technology, Irbid and the University of Jordan, Amman, Jordan. RESULTS: Out of the 58 alleles of the 29 probands, only 31 mutations were identified and M694V and V726A are the most common. The mutation E148Q was the most common among the healthy adult cohort, but was not present in affected individuals. The collective mutant allele frequency "q" was 0.101. The expected carrier rate was 18.1% (one in 5.5) while the observed carrier rate was 18.4% (one in 5.4). CONCLUSION: E148Q has reduced penetrance and thus, a proportion of the individuals genetically affected with FMF remain asymptomatic. M694I and M680I are more prevalent in the affected individuals cohort, which points to their higher penetrance. The overall carrier rate is one in 5, but the selective heterozygote advantage could not be demonstrated in this study due to the relatively small sample size.

Linkage analysis of a large inbred family with congenital megaloblastic anemia.

OBJECTIVE: Megaloblastic anemia during infancy and early childhood often reflects a hereditary disorder of cobalamin's absorption, transport, or intracellular metabolism. There are 3 well defined autosomal recessive syndromes manifesting with megaloblastic anemia due to defects in cobalamin absorption or transport, namely congenital pernicious anemia, Imerslund-Grasbeck syndrome and Transcobalamin II deficiency. The genes responsible for the 3 disorders are gene intrinsic factor (GIF), MGA1 and TCN2, as well as the gene for Transcobalamin I, TCN1 are mapped or cloned, or both. METHODS: We describe the clinical picture of 7 patients from 3 sibships, belong to one large inbred family who presented with megaloblastic anemia during infancy. The mode of inheritance follows an autosomal recessive pattern and the syndrome was completely reversed by parentral vitamin B12 therapy. The ascertainment of the family was carried out in 1998 in the Princess Rhama Children's Hospital, which is affiliated with Jordan University of Science and Technology, Jordan. We performed linkage analysis in this family for genes or regions involved in the above mentioned disorders. RESULTS: The genes implicated in the etiology of the previously mentioned disorders were excluded from being responsible for the disorder in this family. CONCLUSION: The exclusion of the involvement of GIF, MGA1, TCN1 and TCN2 in this family suggests that another gene and its product, involved in cobalamin absorption or transport, remains to be identified. A genome-wide search of the gene implicated in this family may give some insight on that gene, and its function.

Zinc in normal and pathological human prostate gland.

OBJECTIVE: This study is conducted to detect quantitatively zinc in the nuclei and cytoplasm of epithelial cells of the prostate from normal, acute prostatitis, benign prostatic hyperplasia and adenocarcinoma. METHODS: Prostatic tissues from normal, acute prostatitis, benign prostatic hyperplasia and adenocarcinoma were obtained from patients and processed for zinc detection using x-ray microanalysis technique. The samples were collected over a period of 2-3 years and were processed at Jordan University of Science and Technology, Irbid, Jordan. RESULTS: Zinc was increased and decreased both in the nuclei and the cytoplasm of the glandular epithelium of benign prostatic hyperplasia and adenocarcinoma of the prostate. Although zinc was increased in the nuclei and cytoplasm of epithelial cells of acute prostatitis, it was not significant. CONCLUSION: These findings might be caused by factors affecting the zinc metabolic pathway directly or through the zinc bound protein metallothionein. In addition, these findings could be used in diagnosing different prostatic pathological conditions and advancing prostatic tumors and those with similar histopathological profiles.

Nuclear zinc in liver of cadmium and zinc treated rats.

X-ray microanalysis was performed to detect quantitatively, the variation of the nuclear zinc in the liver cells of rats. The nuclear zinc concentration showed statistical decrease and increase in response to cadmium and zinc treatments, respectively. The results suggest that the liver responds differently to cadmium and zinc treatments. The difference in response to either treatment may reflect different mechanisms of zinc transport and metabolism in the liver. The difference in binding affinity of metallothionein (MT) may suggest the involvement of Mt in the metabolism and transport of zinc, an effect, which may be modified by treatment.