A homozygous loss-of-function mutation in inositol monophosphatase 1 (IMPA1) causes severe intellectual disability.

The genetic basis of intellectual disability (ID) is extremely heterogeneous and relatively little is known about the role of autosomal recessive traits. In a field study performed in a highly inbred area of Northeastern Brazil, we identified and investigated a large consanguineous family with nine adult members affected by severe ID associated with disruptive behavior. The Genome-Wide Human SNP Array 6.0 microarray was used to determine regions of homozygosity by descent from three affected and one normal family member. Whole-exome sequencing (WES) was performed in one affected patient using the Nextera Rapid-Capture Exome kit and Illumina HiSeq2500 system to identify the causative mutation. Potentially deleterious variants detected in regions of homozygosity by descent and not present in either 59 723 unrelated individuals from the Exome Aggregation Consortium (Browser) or 1484 Brazilians were subject to further scrutiny and segregation analysis by Sanger sequencing. Homozygosity-by-descent analysis disclosed a 20.7-Mb candidate region at 8q12.3-q21.2 (lod score: 3.11). WES identified a homozygous deleterious variant in inositol monophosphatase 1 (IMPA1) (NM_005536), consisting of a 5-bp duplication (c.489_493dupGGGCT; chr8: 82,583,247; GRCh37/hg19) leading to a frameshift and a premature stop codon (p.Ser165Trpfs*10) that cosegregated with the disease in 26 genotyped family members. The IMPA1 gene product is responsible for the final step of biotransformation of inositol triphosphate and diacylglycerol, two second messengers. Despite its many physiological functions, no clinical phenotype has been assigned to this gene dysfunction to date. Additionally, IMPA1 is the main target of lithium, a drug that is at the forefront of treatment for bipolar disorder.

The genome of the natural genetic engineer Agrobacterium tumefaciens C58.

The 5.67-megabase genome of the plant pathogen Agrobacterium tumefaciens C58 consists of a circular chromosome, a linear chromosome, and two plasmids. Extensive orthology and nucleotide colinearity between the genomes of A. tumefaciens and the plant symbiont Sinorhizobium meliloti suggest a recent evolutionary divergence. Their similarities include metabolic, transport, and regulatory systems that promote survival in the highly competitive rhizosphere; differences are apparent in their genome structure and virulence gene complement. Availability of the A. tumefaciens sequence will facilitate investigations into the molecular basis of pathogenesis and the evolutionary divergence of pathogenic and symbiotic lifestyles.

Comparative genomic analysis of plant-associated bacteria.

This review deals with a comparative analysis of seven genome sequences from plant-associated bacteria. These are the genomes of Agrobacterium tumefaciens, Mesorhizobium loti, Sinorhizobium meliloti, Xanthomonas campestris pv campestris, Xanthomonas axonopodis pv citri, Xylella fastidiosa, and Ralstonia solanacearum. Genome structure and the metabolism pathways available highlight the compromise between the genome size and lifestyle. Despite the recognized importance of the type III secretion system in controlling host compatibility, its presence is not universal in all necrogenic pathogens. Hemolysins, hemagglutinins, and some adhesins, previously reported only for mammalian pathogens, are present in most organisms discussed. Different numbers and combinations of cell wall degrading enzymes and genes to overcome the oxidative burst generally induced by the plant host are characterized in these genomes. A total of 19 genes not involved in housekeeping functions were found common to all these bacteria.

Whole-Genome Sequence of a Bordetella pertussis Brazilian Vaccine Strain.

Despite the reduction in incidence after vaccination, pertussis disease is still considered a public health problem worldwide, mainly due to recent and potential new outbreaks. We report here the complete genome of the Bordetella pertussis Butantan strain used in the Brazilian National Immunization Program as a whole-cell pertussis antigen to compose vaccines such as DTwP (diphtheria, tetanus, and whole-cell pertussis).

Exome sequencing for mucolipidosis III: Detection of a novel GNPTAB gene mutation in a patient with a very mild phenotype.

Mucolipidosis II and III alpha/beta (ML II/III alpha/beta) are rare autosomal recessive lysosomal storage diseases that are caused by a deficiency of UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase, the enzyme responsible for the synthesis of the mannose 6-phosphate targeting signal on lysosomal hydrolases. A Brazilian patient suspected of having a very mild ML III was investigated using whole next-generation sequencing (NGS). Two mutations in the GNPTAB gene were detected and confirmed to be in trans status by parental analysis: c.1208T>C (p.Ile403Thr), previously reported as being pathogenic, and the novel mutation c.1723G>A (p.Gly575Arg). This study demonstrates the effectiveness of using whole NGS for the molecular diagnosis of very mild ML III alpha/beta patients.

Sequential production of Birbeck granules through adsorptive pinocytosis.

A case of vitiligo with inflammatory raised borders was observed electron microscopically, resulting in an interesting view of the formation of Birbeck granules in Langerhans cells. Following the formation of larger coated vesicles, which perform adsorptive pinocytosis from the cell membrane, membrane invagination of the cell occurred shown as the tubular infolding, resulting in the observation of the characteristic Birbeck granule band pattern in its interior. This phenomenon supports the theory of Hashimoto and Tarnowski (1968) that Birbeck granules are formed from the infolding of the cell membrane. In addition, our study shows the involvement of adsorptive pinocytosis in the formation of the granules. It was suggested that when the coated Birbeck granule shifts into the cell, possibly its coat is detached and the vesicle portion forms the globule of the Birbeck granule.

Chalcone and flavonol glycosides from Asarum canadense (Aristolochiaceae).

Two chalcone glycosides were isolated, together with seven known flavonol glycosides, from the leaves of Asarum canadense. The structures of the chalcone glycosides were established as chalcononaringenin 2',4'-di-O-glucoside and chalcononaringenin 2'-O-glucoside-4'-O-gentiobioside by chemical, UV, FAB MS, 1H and 13C NMR evidence.

Glucides of Cnidium monnieri fruit.

From the fruit of Cnidium monnieri Cusson (Umbelliferae), two glucides were isolated together with other known glucides. Their structures were clarified as glycerol 2-O-alpha-L-fucopyranoside and D-quinovitol (6-deoxy-D-glucitol), respectively.

Cell cycle analysis of human dermal fibroblasts cultured on or in hydrated type I collagen lattices.

The proliferation and cell cycle phase composition of human dermal fibroblasts cultured on or in type I collagen lattices (reconstituted dermis model) were examined. On collagen lattices, as compared with conventional cultures on plastic dishes, the proliferation of human dermal fibroblasts was suppressed, being arrested at about one-half the saturation density after 10 days of culture. In collagen lattices, proliferation was further suppressed, being nearly arrested within 4-7 days of culture. Cells were analyzed for cell cycle phases by two-color flow cytometry using DNA staining and S phase cell staining with FITC-conjugated antibromodeoxyuridine antibody. After 5 days of culture, the number of S phase cells on collagen lattices was 49.3% of that on plastic dishes, with an increase in G0G1 phase cells of 79.8%. In collagen lattices, the number of S phase cells was very small (4.3% of all cells), and most of the cells accumulated in G0G1 phase. These findings suggest that the cell cycle of fibroblasts is arrested at G0G1 phase by their interaction with collagen. On the basis of these results, the reconstituted dermis model using collagen lattice is considered to be analogous to the dermis in vivo with respect to cell growth and cell cycle phase composition.

Antitumor activity of an antibiotic identical with or closely related to xanthomycin A.

An antibiotic, identical with or closely related to xanthomycin A, was isolated from a soil Streptomyces. The antibiotic displayed significant therapeutic activity by i.p. administration against i.p.-implanted mouse tumors: Ehrlich carcinoma, sarcoma 180 and P388 leukemia. Less therapeutic activity was observed by i.p. injection in mice bearing s.c. solid tumors of Ehrlich carcinoma and sarcoma 180. No significant activity was found against L1210 leukemia, B16 melanoma and Lewis lung carcinoma. In vitro the antibiotic exhibited a potent cytotoxicity to human leukemia K562 and mouse lymphoblastoma L5178Y cells. DNA strand scission of PM2 phage was caused by the antibiotic in the presence of dithiothreitol.

Acquired dermal melanocytosis of the hand: a new clinical type of dermal melanocytosis.

We report a 22-year-old man with acquired dermal melanocytosis on the hand which developed without any history of previous inflammation. He had no history of contact with or oral administration of any drugs or metals which might have caused pigmentation. Histopathologically, spindle-shaped dermal melanocytes were observed running parallel to the collagen bundles and scattered in all portions of the dermis. The dopa reaction of these cells was positive. Ultrastructurally, dermal melanocytes containing numerous melanosomes (stages II to IV) which were surrounded by the extracellular sheath were observed. Differential diagnosis of other dermal melanocytosis was discussed.

Parallel arrangement, growth inhibition and cell cycle phase analysis of human dermal fibroblasts cultured in collagen lattice.

Human dermal fibroblasts were cultured in a hydrated type I collagen lattice. When collagen fibers were arranged in one direction, fibroblasts were arranged in the same direction. Cell proliferation was markedly suppressed in the collagen lattice as compared with that on plastic, with growth being arrested after day 5. No differences in proliferation were observed between aligned cells and randomly oriented cells. Flow cytometry with DNA staining was performed to analyze each phase of the cell cycle of fibroblasts. Among the 10,000 cell population, S phase cells on day 2 of culture accounted for 43% on plastic but were markedly inhibited to 25% in the lattice. On day 4, S phase cells accounted for 33% on plastic but only for 10% in the lattice. These findings suggest that cell advancement to the S phase is markedly inhibited in the collagen lattice, resulting in accumulation of most of cells in the G0G1 phase. The present study clearly showed that culture in the collagen lattice allowed alignment of fibroblasts with a definite orientation as observed in vivo and produced a status resembling that in vivo in terms of proliferation and cell cycle phase composition.

Flavonol glycosides from Asplenium foreziense and its five related taxa and A. incisum.

The flavonoids of Asplenium foreziense, A. fontanum subsp. fontanum and subsp. pseudofontanum, A. obovatum subsp. obovatum var. obovatum and var. protobillotii, A. obovatum subsp. lanceolatum, and A. incisum were isolated and identified for chemotaxonomic survey. A major constituent of all taxa was kaempferol 3-O-gentiobioside. As minor compounds, kaempferol 3,7-O-glycoside and/or kaempferol 3-O-glycoside were found in A. fontanum, A. obovatum and A. foreziense, and kaempferol 3-O-gentiobioside-4'-O-glucoside, kaempferol 3-O-glucoside and quercetin 3-O-diglucoside in A. incisum. It was suggested that A. foreziense, A. fontanum including subsp. pseudofontanum and A. obovatum including subsp. lanceolatum are not only morphologically but also chemotaxonomically related. The East Asian A. incisum was chemically and geographically different from these taxa.

[Studies on the constituents of "Trichosanthes root". IV. Constituents of roots of Trichosanthes multiloba Miq., Trichosanthes miyagii Hay. and Chinese crude drug "karo-kon"].

A mixture of alpha-spinasterol, stigmast-7-en-3-beta-ol (1) and its 3-O-beta-D-glucopyranoside (2), bryonolic acid (3), 23,24-dihydrocucurbitacin B (4), cucurbitacin B (5), cucurbitacin D (6) were obtained from the fresh roots of Trichosanthes multiloba Miq. (Cucurbitaceae). 1, 2, 4, 5 and 6 were also obtained from the fresh roots of Trichosanthes miyagii Hay. From the chinese crude drug "Karo-Kon", which was considered to be prepared from the roots of Trichosanthes kirilowii Maxim., 1, 2 and 3 were provided.

[Studies on the constituents of trichosanthes root. I. Constituents of roots of Trichosanthes kirilowii Maxim. var. japonicum Kitam].

From the fresh roots of Trichosanthes kirilowii Maxim. var. japonicum Kitam., the following substances were identified: methyl palmitate, palmitic acid, suberic acid, alpha-spinasterol, stigmast-7-en-3 beta-ol, alpha-spinasterol 3-O-beta-D-glucopyranoside, stigmast-7-en-3 beta-ol 3-O-beta-D-glucopyranoside, bryonolic acid, cucurbitacin B, cucurbitacin D, vomifoliol, ethyl alpha-L-arabinofranoside and D-glucose. The bitter taste of Trichosanthes Root was considered to be due to cucurbitacin B and D.

[Studies on the constituents of trichosanthes root. II. Constituents of roots of Trichosanthes cucumeroides Maxim].

From the fresh roots of Trichosanthes cucumeroides Maxim., three new triterpenoid glycosides with a bitter taste were obtained together with methyl palmitate, palmitic acid, alpha-spinasterol, stigmast-7-en-3 beta-ol, alpha-spinasterol 3-O-beta-D-glucopyranoside, stigmast-7-en-3 beta-ol 3-O-beta-D-glucopyranoside and vanillic acid. On the basis of the results of spectral and chemical investigations, these glycosides were characterized as 25-O-beta-D-glucopyranosyl-11-oxocucurbit-5-ene-3 beta,24(R), 25-triol 3-O-alpha-L-rhamnopyranosyl-(1----2)-beta-D-glucopyranosyl-(1----2)-beta -D- glucopyranoside, 25-O-beta-D-(6-O-acetyl)-glucopyranosyl-11-oxocucurbit-5-ene-3 beta,24(R), 25-triol 3-O-alpha-L-rhamnopyranosyl-(1----2)-beta-D-glucopyranosyl(1----2)- beta-D-glucopyranoside and 11-oxocucurbit-5-ene-3 beta,24(R), 25-triol 3-O-alpha-L-rhamnopyranosyl-(1----2)-beta-D-glucopyranosyl-(1----2)- beta-D-glucopyranoside.

[Studies on the constituents of trichosanthes root. III. Constituents of roots of Trichosanthes bracteata Voigt].

From the fresh roots of Trichosanthes bracteata Voigt., the following substances were identified: methyl palmitate, palmitic acid, suberic acid, alpha-spinasterol, stigmast-7-en-3 beta-ol, alpha-spinasterol 3-O-beta-D-glucopyranoside, stigmast-7-en-3 beta-ol 3-O-beta-D-glucopyranoside, glyceryl 1-palmitate, glyceryl 1-stearate, bryonolic acid, cucurbitacin B, isocucurbitacin B, 3-epi-isocucurbitacin B, 23,24-dihydrocucurbitacin B, 23,24-dihydroisocucurbitacin B, 23,24-dihydro-3-epi-isocucurbitacin B, cucurbitacin D, isocucurbitacin D and D-glucose. This root contains more than 6 times cucurbitacin of the root of T. kirilowii Maxim. var. japonicum Kitam.