A screening method for mucopolysaccharidoses with increased urinary excretion of sulfated N-acetylhexosamines

Patients with mucopolysaccharidosis have been reported to excrete elevated amounts of sulfated N-acetylhexosamines in their urine. Based on this finding, a new and simple colorimetric screening test for these disorders is presented. Chromatography of whole urine on Dowex AG 1-X8, from each of 23 normal controls, 5 patients with mucopolysaccharidosis and one patient with multiple sulfatase deficiency, was used to separate the sulfated hexosamines. The fractions eluted with 2M NaCl were analyzed according to the method of Reissig. Patients with Sanfilippo syndrome, type A, Sanfilippo syndrome, type D, Maroteaux-Lamy syndrome, Morquio syndrome, type A, and multiple sulfatase deficiency were clearly distinguished from normal controls. The procedure appeared most sensitive for Sanfilippo syndrome, type D, and multiple sulfatase deficiency, each of which involves deficient activity of N-acetylglucosamine 6-sulfate sulfatase.     

A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency

D-bifunctional protein (D-BP) plays an indispensable role in peroxisomal beta-oxidation, and its inherited deficiency in humans is associated with severe clinical abnormalities. Three different subtypes of D-BP deficiency can be distinguished: 1) a complete deficiency of D-BP (type I), 2) an isolated D-BP enoyl-CoA hydratase deficiency (type II), and 3) an isolated D-BP 3-hydroxyacyl-CoA dehydrogenase deficiency (type III). In this study, we developed a method to measure D-BP dehydrogenase activity independent of D-BP hydratase (D-BP HY) activity to distinguish between D-BP deficiency type I and type II, which until now was only possible by mutation analysis. For this assay, the hydratase domain of D-BP was expressed in the yeast Saccharomyces cerevisiae. After a coincubation of yeast homogenate expressing D-BP HY with fibroblast homogenate of patients using the enoyl-CoA ester of the bile acid intermediate trihydroxycholestanoic acid as substrate, D-BP dehydrogenase activity was measured. Fibroblasts of patients with a D-BP deficiency type II displayed D-BP dehydrogenase activity, whereas type I and type III patients did not. This newly developed assay to measure D-BP dehydrogenase activity in fibroblast homogenates provides a quick and reliable method to assign patients with deficient D-BP HY activity to the D-BP deficiency subgroups type I or type II.     

A cadmium-sensitive, glutathione-deficient mutant of Arabidopsis thaliana.

The roots of the cadmium-sensitive mutant of Arabidopsis thaliana, cad1-1, become brown in the presence of cadmium. A new cadmium-sensitive mutant affected at a second locus, cad2, has been identified using this phenotype. Genetic analysis has grown that the sensitive phenotype is recessive to the wild type and segregates as a single Mendelian locus. Assays of cadmium accumulation by intact plants indicated that the mutant is deficient in its ability to sequester cadmium. Undifferentiated callus tissue was also cadmium sensitive, suggesting that the mutant phenotype is expressed at the cellular level. The level of cadmium-binding complexes formed in vivo was decreased compared with the wild type and accumulation of phytochelatins was about 10% of that in the wild type. The level of glutathione, the substrate for phytochelatin biosynthesis, in tissues of the mutant was decreased to about 15 to 30% of that in the wild type. Thus, the deficiency in phytochelatin biosynthesis can be explained by a deficiency in glutathione.     

全自动吸乳挤奶器的研制

本文介绍一种集吸乳和挤乳为一体的新型全自动多功能吸乳挤奶器,它克服以往人工吸奶器使用不方便,效果不理想等缺点,能安全,自动,舒适地对产妇进行负压吸乳和正压挤乳,以促进乳汁的分泌和排出,提高纯母乳喂养率。     

Type VI collagen deficiency induces osteopenia with distortion of osteoblastic cell morphology

Bone consists of type I collagen as a major protein with minor various matrix proteins. Type VI collagen is one of bone matrix proteins but its function is not known. We therefore examined the effects of type VI collagen deficiency on bone. 3D-μCT analysis revealed that type VI collagen deficiency reduced cancellous bone mass. Cortical bone mass was not affected. Type VI collagen deficiency distorted the shape of osteoblasts both in the cancellous bone and in the cambium layer of periosteal region. Furthermore, type VI collagen deficiency disorganized collagen arrangement. These data indicate that type VI collagen contributes to maintain bone mass.     

2型糖尿病患者中医证型与糖脂代谢和甲状腺功能的关系

目的:探讨2型糖尿病患者中医证型与糖脂代谢和甲状腺功能的关系。方法:选择2016年1月-2017年12月期间武警宁夏总队医院收治的2型糖尿病患者104例,根据中医证型将其分为湿热困脾组23例、阴虚热盛组21例、气阴两虚组20例、阴阳两虚组22例与血瘀脉络组18例。检测并对比不同中医证型2型糖尿病患者糖脂代谢与甲状腺功能指标水平。结果:阴虚热盛组、气阴两虚组、阴阳两虚组与血瘀脉络组甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)水平低于湿热困脾组,高密度脂蛋白胆固醇(HDL-C)水平高于湿热困脾组(P0.05);阴阳两虚组TG、TC、LDL-C水平低于阴虚热盛组、气阴两虚组、血瘀脉络组(P0.05)。湿热困脾组、气阴两虚组、阴阳两虚组与血瘀脉络组空腹血糖(FPG)、餐后2h血糖(2hPPG)、糖化血红蛋白(GHb)水平低于阴虚热盛组(P0.05)。湿热困脾组、阴虚热盛组、气阴两虚组、血瘀脉络组游离甲状腺素(FT4)、游离三碘甲状腺原氨酸(FT3)水平高于阴阳两虚组,促甲状腺激素(TSH)水平低于阴阳两虚组(P0.05),各组总甲状腺素(TT4)、总三碘甲状腺原氨酸(TT3)水平总体比较差异无统计学意义(P0.05)。结论:糖脂代谢和甲状腺功能能够在一定程度上反映出2型糖尿病患者的中医证型变化,可作为2型糖尿病患者中医证型与病情变化的有效参考指标。     

Stomatal Opening as a Practical Indicator of Moisture Stress in Cotton

The response of stomata to varying soil moisture content was studied in cotton by means of a new type of porometer suitable for field work. The results showed that maximum stomatal aperture was only attained when soil moisture was near the upper limit of the so called “available range”. These results indicate the possibility of making practical use of the new type of porometer for the purpose of determining soil moisture deficiency under field conditions, thus aiding in irrigation practices.     

Radial ray deficiency and ulnar ray deficiency in two sibs.

A kindred with a diabetic mother is described with one sibling with a radial deficiency (type 1 radial dysplasia (Bayne) with a hypoplastic thumb type 3 of Blauth) and one with an ulnar ray deficiency (type 2 of Ogden or type 1 of Swanson). A metabolic cause is probable, although the diabetes was well controlled during pregnancy.     

Localization of a New Type of X-Linked Liver Glycogenosis to the Chromosomal Region Xp22 Containing the Liver α-Subunit of Phosphorylase Kinase (PHKA2)

We describe here a new type of X-linked liver glycogen storage disease. The main symptoms include liver enlargement and growth retardation. The clinical and biochemical abnormalities of this glycogenosis are similar to those of classical X-linked liver glycogenosis due to phosphorylase kinase deficiency (XLG). However, in contrast to patients with XLG, the patients described here have no reduced phosphorylase kinase activity in erythrocytes and leukocytes, and no enzyme deficiency could be found. Linkage analysis of four families with this X-linked type of liver glycogenosis assigned the disease gene to Xp22. Lod scores obtained with the markers DXS987, DXS207, and DXS999 were 3.97, 2.71, and 2.40, respectively, all at 0% recombination. Multipoint linkage analysis localized the disease gene between DXS143 and DXS989 with a maximum lod score of 4.70 at θ = 0, relative to DXS987. As both the classical XLG gene and the liver α-subunit of PHK (PHKA2) are also located in Xp22, this variant type of XLG may be allelic to classical XLG, and both diseases may be caused by mutations in PHKA2. Therefore, we propose to classify XLG as XLG type I (the classical type of XLG) and XLG type II (the variant type of XLG).     

JunD suppresses bone formation and contributes to low bone mass induced by estrogen depletion

JunD is an activator protein-1 (AP-1) component though its function in skeletal system is still not fully understood. To elucidate the role of JunD in the regulation of bone metabolism, we analyzed JunD-deficient mice. JunD deficiency significantly increased bone mass and trabecular number. This bone mass enhancement was due to JunD deficiency-induced increase in bone formation activities in vivo. Such augmentation of bone formation was associated with simultaneous increase in bone resorption while the former was dominant over the latter as accumulation of bone mass occurred in JunD-deficient mice. In a pathological condition relevant to postmenopausal osteoporosis, ovariectomy reduced bone mass in wild type (WT) mice as known before. Interestingly, JunD deficiency suppressed ovariectomy-induced increase in bone resorption and kept high bone mass. In addition, JunD deficiency also enhanced new bone formation after bone marrow ablation. Examination of molecular bases for these observations revealed that JunD deficiency enhanced expression levels of c-jun, fra-1, and fra-2 in bone in conjunction with elevated expression levels of runx2, type I collagen, and osteocalcin. Thus, JunD is involved in estrogen depletion-induced osteopenia via its action to suppress bone formation and to enhance bone resorption.     

Red cell reduced glutathione concentrations in Spanish Churra sheep

Erythrocyte reduced glutathione (GSH) levels were investigated in Spanish Churra sheep. GSH deficiency appeared in a high frequency, a clear bimodal distribution being apparent. No significant concentrations of amino acids were detected in the samples and no significant differences were found in potassium concentrations between the low-GSH and the high-GSH type animals. Such results indicate that erythrocyte GSH deficiency in Churra sheep may be similar to the ‘Merino type’ GSH deficiency. Furthermore, limited inheritance data suggested that a second type of GSH deficiency might be present also in Churra sheep.     

Partial and complete adenine phosphoribosyltransferase deficiency associated with 2,8-dihydroxyadenine urolithiasis: kinetic and immunochemical properties of APRT

We have studied adenine phosphoribosyltransferase (APRT) in the hemolysates from the families of 2,8-dihydroxyadenine urolithiasis associated with partial deficiency of APRT (the Japanese type) and complete deficiency of APRT (the null type). The APRT in the control subjects was found to be heat-stable at the physiological concentration of phosphoribosylpyrophosphate (PRPP), which was close to the value of its Km for PRPP. The APRT in the Japanese type showed 10 times higher Km values for PRPP and needed a comparably increased level of PRPP for stability in vitro. No change in red cell PRPP was found in the Japanese type of APRT deficiency. The content of APRT enzyme protein was decreased in the hemolysates of the Japanese type, probably due to its lability at the level of PRPP present in the cells. The heterozygote of the null type also had labile enzyme molecules at the physiological PRPP concentration.     

The carbohydrate-deficient glycoprotein syndromes: pre-Golgi and Golgi disorders?

The carbohydrate-deficient glycoprotein syndromes are a recentlydelineated group of genetic, multisystemic diseases with majornervous system involvement. Three distinct variants have beenrecognized and there are probably many more. They are characterizedby a deficiency of the carbohydrate moiety of secretory glycoproteins,lysosomal enzymes and probably also membranous glycoproteins.The biochemical changes are most readily observed in serum transferrinand the diagnosis is usually made by isoelectric focusing ofthis glycoprotein. The deficiency of sialic acid, in particular,results in a cathodal shift and hence the presence of abnormalisoforms of transferrin with higher isoelectric points thannormal. The basis defects are probably in the processing andsynthesis of the carbohydrate moiety of glycoproteins; thereis indirect evidence for a deficiency of asparagine-N-linkedoligosaccharide transfer in type I (endoplasmic reticulum defect)and for a deficiency of N-acetylglucosaminyltransferase II intype II (Golgi defect). From the large number of patients detectedin only a few years, it is expected that these diseases willbecome as important as, for example, the lysosomal, peroxisomalor mitochondrial disorders. Their study will undoubtedly yielda wealth of new information on the function of glycoproteinsand of their carbohydrate moiety. endoplasmic reticulum glycoproteins glycosylation Golgi sialotransferrins     

Isolation of a new serotype of simian acquired immune deficiency syndrome type D retrovirus from Celebes black macaques (Macaca nigra) with immune deficiency and retroperitoneal fibromatosis.

A new serotype of simian acquired immune deficiency syndrome (SAIDS) retrovirus (type 2) belonging to the D genus of retroviruses is associated with a SAIDS occurring spontaneously in a colony of Celebes macaques (Macaca nigra) and rhesus macaques (Macaca mulatta) at the Oregon Regional Primate Research Center. This syndrome resembles SAIDS in M. mulatta at the California Primate Research Center, which is associated with a similar type D retrovirus (type 1). However, at the Oregon Center, SAIDS is distinguished by the occurrence of retroperitoneal fibromatosis in some of the affected monkeys. Type 2 virus was isolated from seven of seven macaques with SAIDS, retroperitoneal fibromatosis, or both and from one of six healthy macaques. The new strain is closely related to SAIDS retrovirus type 1 and Mason-Pfizer monkey virus but can be distinguished by competitive radioimmunoassay for minor core (p10) antigen and by genomic restriction endonuclease cleavage patterns. Neutralization tests indicate that type 1 and type 2 SAIDS retroviruses are distinct serotypes. Therefore, separate vaccines may be necessary to control these infections in colonies of captive macaques.     

Comparative genetics of yeasts. XXIII. Unusual inheritance of toxin formation in Saccharomyces paradoxus batschinskaia

The data about cytoplasmic control of toxin formation in Saccharomyces paradoxus CBS5829 are presented. A novel determinant (KIL-k3) is probably located in the mitochondrial genome. In other mutations, adenine deficiency results in suppression of toxin formation of the K3 type. A new killer plasmid (KIL-kx) was detected in Sacch. paradoxus VKM Y-2472.     

Relationship of family income and house type to body mass index and chronic energy deficiency among urban Bengalee male slum dwellers of Kolkata, India

A cross-sectional study of 469 adult (>18 years) Bengalee male slum dwellers of Dum Dum, Kolkata, India, was undertaken to study the relationships of family income and house type with body mass index (BMI) and chronic energy deficiency. The overall frequency of chronic energy deficiency was 32.0%. Based on the World Health Organization classification, the prevalence of chronic energy deficiency among this population was high and thus the situation is serious. Overall, monthly family income was significantly positively correlated with BMI. Significant differences in mean weight, BMI and monthly family income, were observed between the two house type groups. All values were found to be significantly higher in the brick household group who also earned a comparatively higher income as evident from the mean monthly family income values. The prevalence of chronic energy deficiency was also found to be significantly higher in the bamboo-fenced household group. Subjects belonging to the lowest family income group had the lowest mean BMI and the highest rate of chronic energy deficiency while those in the highest family income group had the largest mean BMI and lowest rate of chronic energy deficiency. There was a significant family income group difference in mean BMI. There existed significant differences in chronic energy deficiency rates in family income group categories. Linear regression analyses showed that monthly family income and house type had a significant impact on BMI. Subsequent multiple regression analyses revealed that both monthly family income and house type had a significant impact on BMI, even after controlling for each other.     

A mutation in a mild form of galactosialidosis impairs dimerization of the protective protein and renders it unstable.

The lysosomal disorder galactosialidosis is caused by deficiency of the protective protein in the absence of which the activities of the enzymes beta-galactosidase and neuraminidase are reduced. Aside from its protective function towards the two glycosidases, this protein has cathepsin A-like activity. A point mutation in the protective protein gene, resulting in the substitution of Phe412 with Val in the gene product, was identified in two unrelated patients with the late infantile form of the disease. Expression in COS-1 cells of a protective protein cDNA with the base substitution resulted in the synthesis of a mutant protein that lacks cathepsin A-like activity. The newly made mutant precursor was shown to be partially retained in the endoplasmic reticulum. Only a fraction is transported to the lysosomes where it is degraded soon after proteolytic processing into the mature two-chain form. Since the mutant precursor, contrary to the wild type protein, does not form homodimers, the dimerization process might be a condition for the proper targeting and stable conformation of the protective protein. These results clarify the mechanism underlying the combined deficiency in these patients, and give new insight into the structure-function relationship of the wild type protein.     

A novel leukocyte adhesion deficiency III variant: kindlin-3 deficiency results in integrin- and nonintegrin-related defects in different steps of leukocyte adhesion

Leukocyte adhesion deficiency type III is a recently described condition involving a Glanzmann-type bleeding syndrome and leukocyte adhesion deficiency. This was ascribed to a defect of the FERMT3 gene resulting in abnormal expression of kindlin-3, a protein expressed in hematopoietic cells with a major role in the regulation of integrin activation. In this article, we describe a patient with a new mutation of FERMT3 and lack of kindlin-3 expression in platelets and leukocytes. We assayed quantitatively the first steps of kindlin-3-defective leukocyte adhesion, namely, initial bond formation, bond strengthening, and early spreading. Initial bond formation was readily stimulated with neutrophils stimulated by fMLF, and neutrophils and lymphocytes stimulated by a phorbol ester or Mn(2+). In contrast, attachment strengthening was defective in the patient's lymphocytes treated with PMA or Mn(2+), or fMLF-stimulated neutrophils. However, attachment strengthening was normal in patient's neutrophils treated with phorbol ester or Mn(2+). In addition, the patient's T lymphocytes displayed defective integrin-mediated spreading and a moderate but significant decrease of spreading on anti-CD3-coated surfaces. Patient's neutrophils displayed a drastic alteration of integrin-mediated spreading after fMLF or PMA stimulation, whereas signaling-independent Mn(2+) allowed significant spreading. In conclusion, the consequences of kindlin-3 deficiency on β(2) integrin function depend on both cell type and the stimulus used for integrin activation. Our results suggest looking for a possible kindlin-3 involvement in membrane dynamical event independent of integrin-mediated adhesion.     

Manganese is the link between frataxin and iron-sulfur deficiency in the yeast model of Friedreich ataxia

Friedreich ataxia is a human neurodegenerative and myocardial disease caused by decreased expression of the mitochondrial protein frataxin. Proteomic analysis of the mutant yeast model of Friedreich ataxia presented in this paper reveals that these cells display increased amounts of proteins involved in antioxidant defenses, including manganese-superoxide dismutase. This enzyme shows, however, lower activity than that found in wild type cells. Our results indicate that this lack of activity is a consequence of cellular manganese deficiency, because in manganese-supplemented cultures, cell manganese content, and manganese-superoxide dismutase activity were restored. One of the hallmarks of Friedreich ataxia is the decreased activity of iron/sulfur-containing enzymes. The activities of four enzymes of this group (aconitase, glutamate synthase, succinate dehydrogenase, and isopropylmalate dehydratase) have been analyzed for the effects of manganese supplementation. Enzyme activities were recovered by manganese treatment, except for aconitase, for which, a specific interaction with frataxin has been demonstrated previously. Similar results were obtained when cells were grown in iron-limited media suggesting that manganese-superoxide dismutase deficiency is a consequence of iron overload. In conclusion, these data indicate that generalized deficiency of iron-sulfur protein activity could be a consequence of manganese-superoxide dismutase deficiency, and consequently, it opens new strategies for Friedreich ataxia treatment.     

Glycogenosis type II: protein and DNA analysis in five South African families from various ethnic origins.

The molecular nature of lysosomal alpha-glucosidase deficiency was studied in five South African families with glycogenosis type II. Distinct ethnic origins were represented. Two new mutant acid alpha-glucosidase alleles were discovered. In two infantile patients from a consanguineous Indian family we found for the first time an acid alpha-glucosidase precursor of reduced size. The mutant precursor appeared normally glycosylated and phosphorylated but was not processed to mature enzyme. Abnormalities of the mRNA were not obvious, but digestion of genomic DNA with HindIII, BglII, and StuI revealed for each enzyme a fragment of increased length. Heterozygosity was demonstrated in the parents. Complete lack of acid alpha-glucosidase mRNA, as well as deficiency of precursor synthesis, was observed in two black baby girls from unrelated families. In these cases the length of all restriction-enzyme fragments was normal. Reduced enzyme synthesis but normal processing was registered in juvenile and young adult Cape colored patients. The extensive heterogeneity of glycogenosis type II is emphasized in these studies on various ethnic groups. The newly discovered mutants are valuable for the understanding of clinical diversity as a result of allelic variation.