Drosophila mutant with a transducer defect

The trp is a conditional phototransduction mutant of Drosophila. Direct electrical measurements and shot noise analysis suggest that a prolonged intense light causes in the mutant a reduction in the quantum efficiency for quantum bump production that does not arise from bleaching of the visual pigment. This effect depends on the duration of the light and only weakly on its intensity. In the normal fly, an intense blue light that shifts the visual pigment from rhodopsin to metarhodopsin, induces an excitatory process manifested by a prolonged depolarizing after potential (PDA). In the mutant, the PDA has a small amplitude and bump noise is superimposed on the response. It can thus be shown that the excitatory process underlying the PDA is also present in those trp mutants where the PDA voltage response is small or absent. It is suggested that the absence of the PDA voltage response in the mutant is probably due to a defect in an intermediate process, which links the excitatory process to the membrane conductance change.Presented at the EMBO-Workshop on Transduction Mechanism of Photoreceptors, Jülich, Germany, October 4–8, 1976     

B-type natriuretic peptide in children with atrial or ventricular septal defect: a cardiac catheterization study

In this study, we investigated the relationship between plasma B-type natriuretic peptide (BNP) levels and hemodynamics from cardiac catheterization in pediatric patients with atrial or ventricular septal defect. A total of 59 patients were studied including 80% of patients had Qp/Qs > 1.5 and 25% of patients had pulmonary hypertension. The mean BNP value and BNP z-score were 10.9?±?11.2 pg/mL and ?0.28?±?1.7 (?2.85 to 3.29), respectively. There was a statistically significant linear correlation between BNP value and the size of defects (r?=?0.303, p?=?0.002) and a trend toward to positive correlation between BNP value and Qp/Qs ratio (r?=?0.183, p?=?0.166) among all patients. To identify patients with a Qp/Qs ratio >1.5, the sensitivity and specificity were 28%, 100% in all patients at a plasma BNP cut-off point of 15 pg/mL. We concluded that a BNP > 15 pg/mL would help identify patients who need further intervention.     

B-type natriuretic peptide in children with atrial or ventricular septal defect: a cardiac catheterization study

In this study, we investigated the relationship between plasma B-type natriuretic peptide (BNP) levels and hemodynamics from cardiac catheterization in pediatric patients with atrial or ventricular septal defect. A total of 59 patients were studied including 80% of patients had Qp/Qs > 1.5 and 25% of patients had pulmonary hypertension. The mean BNP value and BNP z-score were 10.9?±?11.2 pg/mL and -0.28?±?1.7 (-2.85 to 3.29), respectively. There was a statistically significant linear correlation between BNP value and the size of defects (r?=?0.303, p?=?0.002) and a trend toward to positive correlation between BNP value and Qp/Qs ratio (r?=?0.183, p?=?0.166) among all patients. To identify patients with a Qp/Qs ratio >1.5, the sensitivity and specificity were 28%, 100% in all patients at a plasma BNP cut-off point of 15 pg/mL. We concluded that a BNP > 15 pg/mL would help identify patients who need further intervention.     

Meiotic crossing-over in nondisjoined chromosomes of children with trisomy 21 and a congenital heart defect.

We have used DNA polymorphisms to study meiotic crossovers of chromosome 21q in 27 nuclear families. Each family had a child with Down syndrome and a congenital heart defect. Twenty DNA polymorphisms on chromosome 21 were used to determine parental and meiotic origin of nondisjunction and to identify crossovers. Twenty-four cases were of maternal origin, and three were of paternal origin. Twenty-two unequivocal crossover events were identified. Sixteen crossovers were observed in 22 chromosome pairs nondisjoining at the second meiotic division. Fifty percent of crossover events in MI nondisjunction are detectable by molecular genetic means. Thus, the results suggest that, in this sample, each nondisjoined chromosome 21 pair has been involved in at least one crossover event.     

A CHO-cell mutant with a defect in cytokinesis.

In a selection procedure designed to enrich for temperature-sensitive mutant cells blocked in mitosis a CHO-cell mutant was isolated which has a defect in cytokinesis as the basis of its temperature-sensitive phenotype. Cultures of the mutant had an abnormally high percentage (ie. 34%) of polyploid cells at the permissive temperature of 34 degress C and showed further increased frequencies of polyploidy as well as many multinucleated cells at 38.5 degrees 39.5 degrees. When the mutant cells were synchronized in metaphase by Colcemid arrest and then placed into fresh medium at nonpermissive temperature, they did not divide although the completion of mitosis appeared cytologically normal. Ultrastructural examination by electron microscopy of such synchronized cells at telophase revealed no specific defects in cellular components other than failure of development of a normal midbody. The sensitivity of the mutant to cytochalasin B and to Colcemid was the same as for wild-type cells. This mutation behaved as recessive in tetraploid cell hybrids constructed by fusing the mutant with a CHO strain which was wild-type with respect to temperature sensitivty.     

Recurrent pyoderma in a family with a defect in leucocyte locomotion.

Granulocyte functions including leucocyte locomotion and chemoluminescence were studied in three generations of a family in which all male members had presented with recurrent pyoderma. While parameters of humoral immunity including serum concentrations of IgG, IgA, IgM, and IgE and of complement components C3 and C4 as well as the response of mononuclear leucocytes to mitogens proved to be within the normal range, leucocyte locomotion was found to be severely impaired in all affected subjects. Moreover, granulocyte dysfunction in male members was associated with the occurrence of a single haplotype (HLA-A2, B13, DR7). These findings suggest that the defect in leucocyte locomotion and the pyoderma might not only have been inherited in an X linked manner but might also have been linked to a gene within the inherited HLA haplotype.     

Patients with a non-dysferlin Miyoshi myopathy have a novel membrane repair defect

Two autosomal recessive muscle diseases, limb girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi myopathy (MM), are caused by mutations in the dysferlin gene. These mutations result in poor ability to repair cell membrane damage, which is suggested to be the cause for this disease. However, many patients who share clinical features with MM-type muscular dystrophy do not carry mutations in dysferlin gene. To understand the basis of MM that is not due to mutations in dysferlin gene, we analyzed cells from patients in one such family. In these patients, we found no defects in several potential candidates - annexin A2, caveolin-3, myoferlin and the MMD2 locus on chromosome 10p. Similar to dysferlinopathy, these cells also exhibit membrane repair defects and the severity of the defect correlated with severity of their disease. However, unlike dysferlinopathy, none of the conventional membrane repair pathways are defective in these patient cells. These results add to the existing evidence that cell membrane repair defect may be responsible for MM-type muscular dystrophy and indicate that a previously unsuspected genetic lesion that affects cell membrane repair pathway is responsible for the disease in the non-dysferlin MM patients.     

Immediate reconstruction of a composite mandibular defect with a regional osteomusculocutaneous flap

This is a report of an immediate reconstruction of a composite mandibular defect, using a pectoralis major osteomusculocutaneous flap incorporating a segment of the fifth rib. Viability of the transferred rib segment was established by tetracycline labeling at 2 weeks and technetium scanning at 3 months postoperatively. This clinical example supports the experimental observations of a vascular network interconnecting muscle, periosteum, and compact bone.     

Infertility in a bull with a nuclear sperm defect: A case report

A Simmental bull with a history of low fertility, both by natural service and artificial insemination, was presented for examination. Two previous semen evaluations had revealed no specific semen abnormalities that would support the breeding history. A comprehensive cytochemical analysis of the bull's ejaculate revealed a complex nuclear lesion affecting over 80% of sperm. This condition was expressed in abnormal shaping of the nuclei, with deficient distribution, condensation and stabilization of the nucleoplasm. These abnormalities were associated with various-sized intranuclear pouches or depressions. The acrosome was moderately involved and the tail was relatively free of abnormalities resulting in normal sperm motility.Two controlled breeding trials utilizing a total of 15 super-ovulated females were conducted to evaluate the bull's fertilization rate. Combined data demonstrated an 18% ($\text{23}\text{128}$) fertilization rate of recovered ova. At the same time, the fertilization rate of seven bulls classified as satisfactory potential breeders was 72% ($\text{353}\text{490}$).Data from two embryo transplant units regarding ova collected from eight donor females inseminated with semen from this bull revealed a fertilization rate of 41% ($\text{30}\text{73}$). Of the fertilized ova, 37% ($\text{11}\text{30}$) were degenerate and were not transferred. A pregnancy rate of 57% ($\text{11}\text{19}$) resulted from the transfer of 19 fertilized ova.A natural breeding pregnancy rate of 5% ($\text{2}\text{42}$) and artificial breeding pregnancy rate of 8% ($\text{15}\text{180}$) support the breeding trial results.     

Neuroacanthocytosis associated with a defect of the 4.1R membrane protein

Background

Neuroacanthocytosis (NA) denotes a heterogeneous group of diseases that are characterized by nervous system abnormalities in association with acanthocytosis in the patients' blood. The 4.1R protein of the erythrocyte membrane is critical for the membrane-associated cytoskeleton structure and in central neurons it regulates the stabilization of AMPA receptors on the neuronal surface at the postsynaptic density. We report clinical, biochemical, and genetic features in four patients from four unrelated families with NA in order to explain the cause of morphological abnormalities and the relationship with neurodegenerative processes.

Case presentation

All patients were characterised by atypical NA with a novel alteration of the erythrocyte membrane: a 4.1R protein deficiency. The 4.1R protein content was significantly lower in patients (3.40 ± 0.42) than in controls (4.41 ± 0.40, P < 0.0001), reflecting weakened interactions of the cytoskeleton with the membrane. In patients IV:1 (RM23), IV:3 (RM15), and IV:6 (RM16) the 4.1 deficiency seemed to affect the horizontal interactions of spectrin and an impairment of the dimer self-association into tetramers was detected. In patient IV:1 (RM16) the 4.1 deficiency seemed to affect the skeletal attachment to membrane and the protein band 3 was partially reduced.

Conclusion

A decreased expression pattern of the 4.1R protein was observed in the erythrocytes from patients with atypical NA, which might reflect the expression pattern in the central nervous system, especially basal ganglia, and might lead to dysfunction of AMPA-mediated glutamate transmission.     

X-linked hypogammaglobulinaemia with a late secretion defect. A family study

Three boy cousins suffering from x-linked hypogammaglobulinaemia have been described. Their disease is quite different from the classical x-linked hypogammaglobulinaemia. We present data from 15 family members of three families. The three mothers are sisters and 3 boys from 5 are suffering from immunodeficiency. HLA A, B and DR typing and in vitro diagnostics of immune functions of all family members were carried out. In the patients we could demonstrate a well functioning T cell system which seems to be regular for T cell help on PMW driven B cell maturation into cytoplasmic immunoglobulin positive (cIg+) cells of all immunoglobulin classes but we had no evidence for IgG secreting cells in a plaque forming cell (PFC) assay. The registration of spontaneous suppressor phenomena should be taken into account. If patients undergo an gamma-globulin therapy the changed suppressor effects come back to normal values but the secretion defect is unchanged. The three healthy mothers express one identical haplotype (A2 B8 DR3) which we could demonstrate in the three patients but also in one healthy boy.     

Temperature-sensitive Chinese hamster fibroblast mutant with a defect in RNA metabolism

We describe a new temperature-sensitive mutant of Chinese hamster cell fibroblasts. After a shift to the nonpermissive temperature of 40.5 degrees C, the rates of DNA, RNA, and protein synthesis declined rapidly (to < or = 50% within 12 h) and the progression of unsynchronized cells through the cell cycle was affected. We believe that DNA synthesis came to a halt after a short time, because cells no longer entered the S phase. The decrease in protein synthesis at 40.5 degrees C was shown to be a consequence of a decrease in the number of polysomes, whereas free 80S ribosomes accumulated. We concluded that the components of the protein biosynthetic machinery were intact (ribosomes and soluble factors), but synthesis was limited by a shortage of mRNA. The decline in mRNA production had a significant effect on the synthesis of proteins (e.g., heat shock proteins) translated from short-lived messages. We observed that both polyadenylated and nonpolyadenylated RNA syntheses declined at 40.5 degrees C, whereas the synthesis of small RNAs (4 to 5S) was less reduced. The argument is made that the temperature-sensitive phenotype is the result of a defect affecting mRNA synthesis.     

Local modes in a DNA polymer with hydrogen bond defect.

Vibrations of a homopolymer DNA with localized hydrogen bond defects have been examined using the recently developed decaying mode theory for long-chain polymers with local structural defects. For a poly(dA)-poly(dT) homopolymer having perturbed hydrogen bonds in one base pair, a localized mode at 63.2 cm-1 has been found. This mode has a very nearly pure H-bond stretch or "breathing" character, although the backbones do not separate. This agrees in frequency with a similar result found by other authors using a different approach. We search the full microwave frequency range for other local modes for several models of weakened H bonds. Besides the local mode with breathing characteristics, local modes with other characteristic motions were found, but only for asymmetrically perturbed bonds. We find in general that local modes are not very robust, requiring quite specific, narrow ranges in parameter space. They are also not abundant, there being only three in our most prolific model.     

Ammonia-excreting mutants ofKlebsiella pneumoniae with a pleiotropic defect in nitrogen metabolism

Enterobacterial mutants defective in the nitrogen control regulatory system (Ntr) generally display a pleiotropic phenotype with regard to expression and regulation of several enzymes and transport systems involved in the assimilation of N sources. This report describes the isolation and characterization of similar pleiotropic mutants ofKlebsiella pneumoniae that cannot be complemented byntr genes. The strains excreted ammonia, were unable to grow on a number of N sources, and contained low glutamine:2-oxoglutarate amino transferase and normal, but unmodifiable glutamine synthetase activities and a nitrogenase level largely unaffected by ammonium, but still repressible by an amino acid mixture. Genetic studies suggested that this phenotype is due to overexpression of an unknown regulatory protein.Abbreviations GS Glutamine synthetase - GOGAT Glutamate synthase - ATase Adenylyl transferase - Ntr Nitrogen regulatory system     

A high frequency and geographical distribution of MMACHC R132* mutation in children with cobalamin C defect

Cobalamin C defect is caused by pathogenic variants in the MMACHC gene leading to impaired conversion of dietary vitamin B₁₂ into methylcobalamin and adenosylcobalamin. Variants in the MMACHC gene cause accumulation of methylmalonic acid and homocysteine along with decreased methionine synthesis. The spectrum of MMACHC gene variants differs in various populations. A total of 19 North Indian children (age 0–18 years) with elevated methylmalonic acid and homocysteine were included in the study, and their DNA samples were subjected to Sanger sequencing of coding exons with flanking intronic regions of MMACHC gene. The genetic analysis resulted in the identification of a common pathogenic nonsense mutation, c.394C > T (R132*) in 85.7% of the unrelated cases with suspected cobalamin C defect. Two other known mutations c.347T > C (7%) and c.316G > A were also detected. Plasma homocysteine was significantly elevated (> 100 µmol/L) in 75% of the cases and methionine was decreased in 81% of the cases. Propionyl (C3)-carnitine, the primary marker for cobalamin C defect, was found to be elevated in only 43.75% of cases. However, the secondary markers such as C3/C2 and C3/C16 ratios were elevated in 87.5% and 100% of the cases, respectively. Neurological manifestations were the most common in our cohort. Our findings of the high frequency of a single MMACHC R132* mutation in cases with combined homocystinuria and methylmalonic aciduria may be proven helpful in designing a cost-effective and time-saving diagnostic strategy for resource-constraint settings. Since the R132* mutation is located near the last exon–exon junction, this is a potential target for the read-through therapeutics.

     

X-ray fluorescence determination of the loss of chosen electrolytes in the urine of children with a congenital cyanotic heart defect and after heart transplantation

In this paper results of the analysis of urine samples of healthy children and children with a congenital cyanotic heart defect and after heart transplantation are presented. The analysis of urine samples was carried out by X-ray fluorescence spectrometry with wavelength dispersion and using CRM urine Seronorm as a reference. It was found that for patients with a congenital cyanotic heart defect the loss of electrolytes like Na, Cl and K was increased. Moreover, urine samples of children from areas of different degree of environmental pollution were analysed. We observed (as expected) higher concentrations of heavy metals in the urine of children from ecological polluted areas.