Novel DNMT3A germline mutations are associated with inherited Tatton-Brown-Rahman syndrome.

Tatton-Brown-Rahman syndrome (TBRS) was recently described in 13 isolated cases with de novo mutations in the DNMT3A gene. This autosomal dominant condition is characterized by tall stature, intellectual disability and a distinctive facial appearance. Here, we report six cases of inherited TBRS caused by novel DNMT3A germline mutations. The affected individuals belong to two sib-ships: four from an Old Order Amish family in America and two from a French Canadian family in Canada. All of them presented with characteristic features of TBRS, including dysmorphic facial features, increased height, intellectual disability, and variable additional features. We performed clinical exome sequencing and identified two mutations in the DNMT3A gene, a c.2312G>A (p.Arg771Gln) missense mutation in the Amish family and a c.2296_2297delAA (p.Lys766Glufs*15) small deletion in the French Canadian family. Parental DNA analysis by Sanger sequencing revealed that the Amish mutation was inherited from the healthy mosaic father. This study reflects the first cases with inherited TBRS and expands the phenotypic spectrum of TBRS.

Alternative method for determination of contaminated heparin using chiral recognition.

Since 2008 a significant amount of work has focused on the development of methods to analyze contaminated heparin. This work focuses on utilizing heparin's ability to serve as a chiral selector as a means for determining contamination. Specifically, the effect of contamination on the separation of pheniramine and chloroquine enantiomers was explored. Separations were conducted using heparin contaminated with chondroitin sulfate at varying levels. For each pair of enantiomers, electrophoretic mobility and resolution were calculated. For pheniramine enantiomers, an increase in contamination leads to a decrease in the electrophoretic mobility and resolution. A linear relationship between contamination level and electrophoretic mobility of the pheniramine enantiomers was observed for the entire contamination range. A linear relationship was also found between contamination level and resolution of the enantiomers between 0 and 70 percent contamination. For the separation of chloroquine enantiomers, it was found that at low levels of contamination, the resolution of enantiomers was increased due to the secondary interaction between the chloroquine enantiomers and the chondroitin sulfate. Results of this study illustrate the potential of using chiral recognition as a means to determine heparin contamination as well as the improvement of the chiral resolution of chloroquine with the additional of low levels of chondroitin sulfate A.

Association of the GABRD gene and childhood-onset mood disorders.

The chromosome 1p36 region was previously indicated as a locus for susceptibility to recurrent major depressive disorder based on a linkage study in a sample of 497 sib pairs. We investigated the gamma-aminobutyric acid A (GABA(A)) delta receptor subunit gene, GABRD, as a susceptibility gene to childhood-onset mood disorders (COMD) because of substantial evidence implicating GABAergic dysfunction in mood disorders and the position of this gene near the 1p36 linkage region. Using a sample consisting of 645 Hungarian families with a child/adolescent proband diagnosed with a mood disorder with the onset of the first episode before age 15, we found some evidence for the association of two polymorphisms located within the gene, rs2376805 and rs2376803, as well as significant evidence for biased transmission of the haplotypes of these two markers (global chi(2) test for haplotypes = 12.746, 3 df, P = 0.0052). Furthermore, significant evidence of association was only observed in male subjects (n = 438) when the results were analyzed by sex (chi(2) = 9.000 1 df, P = 0.003 for rs2376805). This was in contrast with the previous linkage findings, as LOD scores exceeding 3 were only in female-female pairs in that study. These findings point to the GABRD gene as a susceptibility gene for COMD; however, this gene may not explain the previous linkage finding.

Recent findings on the pathogenesis of bronchial asthma.

In the first part of this series of papers (Székely and Pataki, 102) the pathogenesis of asthma was approached as a pathological antigen-antibody complex induced vago-vagal axon reflex. In the next part (103) the contribution of individual hormonal predisposition, the environmental and the most frequent allergizing factors have been reviewed. In the first section of this last (third) part of the review the genetic factors contributing to the asthma are surveyed. In this field a great progress has been made during the last decade, a lot of genes have been pinpointed which contribute to the heredity of the disease. In the second section of this last paper on the etiology of asthma an attempt is made to summarize the previously reviewed data and some new ones. Actually a new hypothesis is proposed that beyond the multitude of genetic, environmental and hormonal factors the underlying biochemical mechanism is simple: the disequilibrium of two functionally opposing second messenger systems in the airways: the Ca i ++ liberating PLC-PKC cascade and the Ca i ++ level reducing cAMP mediated one with preponderance of the former.

Recent findings on the pathogenesis of bronchial asthma. Part I. Asthma as a neurohumoral disorder, a pathological vago-vagal axon reflex.

The novel data on the pathogenesis of asthma are summarized in this three-part review. Its immunological background is well established but it is more than an immunological disorder. Multiple lines indicate that both peripheral and central neural mechanisms are also involved in the pathogenesis of asthma. In the present first part of the review asthma is described as vago-vagal axon reflex brought about by multiple positive feed-back mechanisms, receptor upregulation, wind-up, phenotypic switch and formation of a pathological conditioned reflex. In the coming second part the main dispositional (mostly hormonal) and external contributing factors are reviewed, while the third part deals with the role of inheritance, i.e., with gene alleles leading to enhanced production of mediators of asthma.

A PET study on the characterization of partially reversible radiogenic lower motor neurone disease.

OBJECTIVE: To investigate the pathomechanism of the rare radiogenic lower motor neurone disease (LMND) on the basis of a case history involving a partial functional recovery. PATIENT: A 31-year-old seminoma patient received postoperative para-aortic and para-iliac telecobalt irradiation with a biologically effective dose of 88 Gy(2) (44 Gy in 2 Gy fractions/day, with an estimated alpha/beta of 2 Gy) delivered to the spinal cord following a single cycle of chemotherapy. LMND developed 4 months after the completion of radiotherapy. The patient exhibited flaccid paraparesis of the lower extremities (without sensory or vegetative signs), followed by a worsening after further chemotherapy, due to pulmonary metastatization. A gradual spontaneous functional improvement commenced and led several years later to a stabilized state involving moderately severe symptoms. METHODS: In the 15th year of the clinical course, magnetic resonance imaging (MRI) and positron emission tomography (PET) with [(18)F]fluorodeoxyglucose (FDG) and [(11)C] methionine were conducted. Four lines of experiments (clonogenic assay using fibroblasts isolated from a skin biopsy sample of the patient, comet assay, micronucleus assay, and the testing of chromosome aberrations after in vitro irradiation of peripheral blood samples) were performed in a search for an increased individual radiosensitivity. RESULTS: MRI investigations failed to reveal any pathological change. PET demonstrated an increased FDG accumulation, but a negligible [(11)C] methionine uptake in the irradiated spinal cord segments. The radiobiological investigations did not indicate any sign of an increased individual radiosensitivity. CONCLUSIONS: We suggest that the observed partial functional recovery and stabilization of the symptoms of radiogenic LMND may be explained by the higher than normal density of sodium channels expressed along the demyelinated axons of the restored conduction. The increased energy demands of this type of conduction are proved by a higher metabolic rate (increased FDG uptake) of the irradiated spinal cord segments without a substantial regenerative process (lack of detectable protein synthesis).

The "endorphin story": a subjective account.

The contribution of the researchers of IDR to some early discoveries on endorphins is recounted. Exact determination of the amino acid sequence of beta-lipotropin, discovery of the analgesic action of beta-endorphin in rats, demonstration of the partial cross-tolerance between morphine and beta-endorphin, description of the elevation of the pain threshold upon inhibition of the enzymatic breakdown of endogenous opioids by bacitracin, demonstration of inhibition of opiate tolerance and dependence by alpha-MSH and Met-enkephalin, development of the second strongest enkephalin analogue, D-Met2,Pro5-enkephalinamide, recognition of some factors conferring analgesic activity to synthetic enkephalin analogues, etc. prove that IDR has played an active role in the early history of endorphin research.

Short-term changes in renal function after extracorporeal shock wave lithotripsy in children.

PURPOSE: To our knowledge despite numerous studies the biological effect of extracorporeal shock wave lithotripsy (ESWL, Dornier Medical Systems, Inc., Marietta, Georgia) on the function of the immature kidney has not yet been evaluated. We determined the short-term effect of ESWL on renal function in children. MATERIALS AND METHODS: In a 5-year period 65 children were treated for 77 kidney stones by ESWL and followed regularly at our department. In 16 children the short-term effect of shock wave energy was studied by measuring blood parameters (sodium, potassium, urea, creatinine and C-reactive protein), urinary electrolytes (sodium, potassium and creatinine), urinary enzyme activity (aspartate transaminase, alanine transferase, alkaline phosphatase and lactate dehydrogenase) and the excretion of beta 2-microglobulin. Samples were obtained immediately before and 2 hours after ESWL, and on days 1, 2, 8, 15, 30 and 90 after treatment. RESULTS: Morphological changes in the kidneys were not detected by ultrasound during followup. No significant changes were noted in overall renal function, serum parameters or urine electrolytes. A significant elevation in the excretion of aspartate transaminase, alkaline phosphatase, lactate dehydrogenase and beta 2-microglobulin was observed, indicating proximal tubular dysfunction and cell destruction. Enzyme levels returned to baseline within 15 days. CONCLUSIONS: Our results demonstrate that shock wave energy induces transient functional damage of tubular function in children. Minimizing the kV. and number of shocks may decrease the deleterious effect. When considering functional regeneration time, the minimal interval between 2 shock wave treatments should be at least 15 days. The long-term effect needs further investigation.

[Discovery of the analgesic action of beta-endorphin: a subjective account]. / Az endorphinok fájdalomcsillapító hatásának felfedezése: ahogy azt az elfogult farmakológus látja.

The contribution of the research workers of the Institute for Drug Research to some early discoveries in the field of endorphins is reviewed. The author takes pride in the fact the analgesic action of beta-endorphin was described in his laboratory concurrently with an American and a British group, respectively. The other main findings of his laboratory include demonstration of the cross-tolerance between beta-endorphin and morphine, description of the elevation of the pain threshold upon inhibition of the breakdown endogenous opioids by intracerebroventricular administration of bacitracin, attenuation of morphine tolerance by alpha-MSH and Met-enkephalin, pharmacological development and clinical examination of a highly potent enkephalin analogue, (D-Met2,Pro5)-enkephalinamide, coded as GYKI 14,238.

AMPA receptor antagonists, GYKI 52466 and NBQX, do not block the induction of long-term potentiation at therapeutically relevant concentrations.

The involvement of alpha-amino-3-hydroxy-5-methylizoxazole-4-propionic acid (AMPA) receptors in induction of long-term potentiation (LTP) was examined in rat hippocampal slice preparation. Using conventional extracellular recording, excitatory postsynaptic potentials (EPSPs) and population action potentials (PSs), evoked by low-frequency stimulation of the Schaffer collateral-commissural fibres, were recorded in the CA1 region. The effects of a competitive AMPA receptor antagonist, 6-nitro-7-sulfamoylbenzo(f)quinoxaline-2, 3-dione (NBQX), and that of a non-competitive blocker, 1-(4-aminophenyl)-4-methyl-7,8-methylendioxy-5H-2,3-benzodiazepine (GYKI 52466) have been examined. 0.25-0.5 microM of NBQX and 20-40 microM of GYKI 52466 did not suppress the induction of LTP. LTP was attenuated only at the highest concentrations tested (1 microM NBQX or 80 microM GYKI 52466). These in vitro concentrations, however, exceed the brain levels needed for in vivo anticonvulsant action. Furthermore, even at the highest concentrations both compounds suppressed only the expression but not the induction of LTP. Namely after their washout LTP reappeared. Thus, at pharmacologically relevant concentrations these AMPA receptor antagonists apparently do not suppress LTP, a cellular mechanism underlying memory formation. These experiments suggest that in clinical practice AMPA receptor blockade may have some advantage over N-methyl-D-aspartate receptor antagonism, which is accompanied by severe memory impairment.

High frequency of metastatic Leydig cell testicular tumours.

690 patients were treated for testicular tumour in the course of 18 years. The histology of 7 cases showed Leydig cell tumour. In 4 of the 7 cases, a metastatic process leading to death was observed. At the time of diagnosis, 5 patients were found to have low stage, whereas 2 of the patients had advanced lymphatic involvement. The hematogenous and lymphatic metastases proved to be resistant to chemotherapy. Contrary to the major part of the literature, retroperitoneal lymphadenectomy should be performed with this histological type for the exact pathological staging immediately following orchiectomy.

Differential modulation of the GYKI 53784-induced inhibition of AMPA currents by various AMPA-positive modulators in cerebellar Purkinje cells.

The effects of various (S)-alpha-amino-3-hydroxy-5-methyl-4-izoxazole-propionate (AMPA) receptor modulators on AMPA-induced whole-cell currents were compared in isolated rat cerebellar Purkinje cells. The positive modulators, aniracetam, cyclothiazide, 1-(1, 3-benzodioxol-5-ylcarbonyl)-piperidine (1-BCP), and 1-(quinoxaline-6-ylcarbonyl)-piperidine (BDP-12), dose-dependently potentiated the steady-state component of AMPA currents. The negative modulator, (-)1-(4-aminophenyl)-4-methyl-7, 8-methylenedioxy-4,5-dihydro-3-methylcarbamoyl-2,3-benzodiazepine (GYKI 53784), dose-dependently suppressed AMPA responses. Its concentration-response curve was shifted to the right in a parallel fashion by all positive modulators, indicating a competitive type of interaction. However, the relative potencies of the positive modulators were different with regard to the enhancement of AMPA responses and the reversal of GYKI 53784-induced inhibition, respectively. It is supposed that positive modulators act at multiple allosteric sites and that they interact with GYKI 53784 at only one of these sites.

[Percutaneous embolization of hemorrhaging renal arteriovenous fistula]. / Vérzö renalis arteriovenosus fistula perkután embolizációja.

A 67 year old male patient with macroscopic hematuria was treated by percutaneous intraarterial embolization with Gianturco-coil (Cook). The patient had a percutaneous stone removal from the left kidney two weeks before the bleeding. The emergency angiography showed an arteriovenous shunt in the left kidney. A superselective embolization was performed. There was no bleeding on the control angiography. The embolization was performed without complication.

Diprotin A, an inhibitor of dipeptidyl aminopeptidase IV(EC 3.4.14.5) produces naloxone-reversible analgesia in rats.

The dipeptidyl aminopeptidase IV (DP IV) inhibitor Diprotin A produces a full, dose-dependent, short-lasting and naloxone-reversible analgesia in the rat tail-flick test when given intracerebroventricularly, with an ED50 of 295 nmol/rat but it has no direct opioid agonist activity in the longitudinal muscle strip of guinea-pig ileum bioassay. Two of the potential DP IV substrates, morphiceptin and endomorphin 1, identified recently in bovine brain were also analgesic given by similar route. The action of endomorphin 1 was more potent (ED50 = 7.9 nmol/rat) and slightly but significantly more sustained than that of Diprotin A. Diprotin A neither potentiated nor prolonged the effect of a marginally analgesic dose of endomorphin 1. The distinct time course and the lack of potentiation indicate that in the analgesic effect of Diprotin A in rats the protection of a brain Tyr-Pro-peptide other than endomorphin 1 is involved.

Total body irradiation before bone marrow transplantation. Technique and acute toxicity.

PURPOSE: To evaluate the total body irradiation methods in the National Institute of Oncology between January 1984 and February 1998. PATIENTS AND METHODS: One hundred and twenty-four patients underwent total body irradiation prior to bone marrow transplantation in the last 15 years. A special cobalt unit has been used, the dose rate was 6 to 8 cGy/min in the midline of the abdomen. The source-midline distance (SMD) was 340 cm and the field size was 80 x 200 cm. The dose calculation was done on the basis of a tissue-phantom ratio curve measured in total body irradiation conditions and effective tissue thickness (ETT). Between 1984 and 1992 the beam direction was horizontal, the patients laid in lateral position. In 11 cases the total dose to the abdominal midline was 10 Gy in 1 fraction. From 1986 the fractionation changed to 4 x 3 Gy in 4 days. Within individual lung shielding the average lung dose was 8.5 Gy. In 44/124 cases the order of conditioning treatment was chemo-radiotherapy. Since 1992 vertical beams were used, and the patients (80/124) laid in prone/supine position. The fractionation remained the same but radio-chemotherapy regime has been used. RESULTS: The irradiation in prone position proved to be safer than lateral because of smaller patient motion and it resulted in a more accurate positioning of lung shielding too. In all cases, the acute side effects (headache, nausea, vomiting) were moderate. Using radio-chemotherapy the acute side effects during the total body irradiation were uncommon and well tolerable. CONCLUSION: Our technique with the large source-midline distance, vertical beam direction and the supine/prone position is stable, convenient and safe to produce homogeneous dose distribution and ensures accurate and reproducible lung shielding.

Report from the front lines: rural Medicaid primary care providers as gatekeepers of rural health service networks.

This study of primary care practitioners in Louisiana's Medicaid managed care program, Community Care, focused on the program's implementation and perceived impact. PCPs were interviewed to assess their perceived responsibilities as gatekeepers and how well they believed the system facilitated or impeded their fulfillment of those responsibilities, to examine issues of access and quality, and to determine their knowledge and use of local referral networks. Findings indicate that although they appreciated the financial stability derived from having a larger patient base and understood their roles well, they were concerned with several facets of the program, including perceived limitations imposed on their medical decision making, an added burden of paperwork, and a lack of both downward and upward communication. They also believed the referral network of available providers for medical and social services to be more limited for their Medicaid patients than for their private patients. Policy implications are discussed.