Notes: Background House dust mite allergens have been shown to be a very important stimulus in the causation of asthma and triggers for the exacerbation of symptoms. Therefore, characterization of mite-derived allergens at the molecular level is an important step for the development of effective diagnostic and therapeutic approaches, as well as for epidemiological studies.Objective To clone, express and characterize at the molecular level the cysteine protease from Blomia tropicalis (Bt).Methods A full-length cDNA encoding Blo t 1 was cloned from a Bt cDNA library using a PCR and RACE-based strategy. The cDNA was PCR-amplified, sequenced and subcloned into a prokaryotic expression vector. The allergenicity of the recombinant Blo t 1 was evaluated for IgE reactivity by Western blot.Results Blo t 1 cDNA encodes a 221 amino acids polypeptide with an estimated molecular weight of 25 kDa. The recombinant protein is 35% identical to other mite cysteine proteases. Recombinant Blo t 1 (rBlo t 1) bound IgE from 62% of Bt skin test-positive serum. Dermatophagoides pteronyssinus (Dp) skin test-positive sera did not react with rBlo t 1, indicating the possible presence of unique IgE epitopes on the rBlo t 1 molecule. A three-dimensional image of Blo t 1, constructed based on predicted analysis, showed conserved secondary and tertiary structure with other cysteine proteases.Conclusion We report the cloning, expression and IgE reactivity of Blo t 1, a novel allergen from the domestic mite Blomia tropicalis (Bt), highly homologous to cysteine proteases. This putative cysteine protease, designated Blo t 1, may play a major role as an immunodominant allergen involved in dust mite-specific IgE-mediated hypersensitivity.

Notes: BIBLIOGRAFÍA

Notes: Abstract Coexisting feldspars from across 2,000 km2 of the granulite facies Oaxacan Complex, southern Mexico exhibit variable amounts of solid solution from nearly binary (Ab-An and Ab-Or) to substantially ternary (Ab-An-Or). Reintegrated analyses of 21 coarsely exsolved perthite (AF)-plagioclase (PL) pairs yield AF=Or30–63 Ab30–56An2–15 and PL=Or1–2Ab70–84An11–28. These data have been used to test existing two feldspar geothermometers for this extended composition range. For all compositions, temperature estimates show relatively little spread in value (660° to 795° C, 7 kbar) using the Haselton et al. (1983) calibration (HHHR). These temperatures are in fair agreement with estimates of 750±40° C for feldspar pairs with nearly binary compositions using the Stormer (1975) thermometer (STO). However, STO temperatures increase significantly (to 990° C) with increasing ternary solid solution in AF, suggesting that thermometers derived for binary systems are inaccurate for ternary compositions. Isotherms drawn from HHHR which take into account variable anorthite solution in alkali feldspar show that estimated temperature decreased by 50–100° C for each 5 mole percent anorthite in alkali feldspar. Experimentally determined solvus relations (Seck 1971) require feldspars with significant ternary solid solution to have crystallized or to have equilibrated at higher temperature than feldspars with more binary compositions. However, petrographic and field relations of ternary and binary feldspars in the Oaxacan Complex suggest they were all equilibrated at similar metamorphic pressures and temperatures and do not support a model where ternary feldspars have preserved higher premetamorphic temperatures. The composition of coexisting feldspars from other Precambrian granulite-facies terranes are also inconsistent with Seck's (1971) results. Hence, thermometers which fit Seck's solvus relations may not yield accurate temperatures in high grade metamorphic terranes. Parallel tie-lines for ternary and binary feldspars in the Oaxacan Complex and the consistency of inferred temperatures (HHHR) for many granulite terranes suggest that estimation of temperature using tie-line slopes rather than solvus width may yield more accurate results for these samples. Peak metamorphic conditions in the Oaxacan Complex are inferred to have been 730±50° C, 7±1 kbar. Pressure estimates from four garnet-plagioclase barometers show good agreement. Results of feldspar thermometry are consistent with diopside-forsterite equilibria in marbles which restrict T=720–765° C at P=7 kbar.

Notes: Summary The cDNAs of mammalian amino acid transporters already identified could be grouped into four families. One of these protein families is composed of the protein rBAT and the heavy chain of the cell surface antigen 4F2 (4F2hc). The cRNAs of rBAT and 4F2hc induce amino acid transport activity via systems b0,+ -like and y+L -like inXenopus oocytes respectively. Surprisingly, neither rBAT nor 4F2hc is very hydrophobic, and they seem to be unable to form a pore in the plasma membrane. This prompted the hypothesis that rBAT and 4F2hc are subunits or modulators of the corresponding amino acid transporters. The association of rBAT with a light subunit of ~40kDa has been suggested, and such an association has been demonstrated for 4F2hc. The b0,+-like system expressed in oocytes by rBAT cRNA transports L-cystine, L-dibasic and L-neutral amino acids with high-affinity. This transport system shows exchange of amino acids through the plasma membrane ofXenopus oocytes, suggesting a tertiary active transport mechanism. The rBAT gene is mainly expressed in the outer stripe of the outer medulla of the kidney and in the mucosa of the small intestine. The protein localizes to the microvilli of the proximal straight tubules (S3 segment) of the nephron and the mucosa of the small intestine. All this suggested the participation of rBAT in a high-affinity reabsorption system of cystine and dibasic amino acids in kidney and intestine, and indicated rBAT (named SLC3A1 in Gene Data Bank) as a good candidate gene for cystinuria. This is an inherited aminoaciduria due to defective renal and intestinal reabsorption of cystine and dibasic amino acids. The poor solubility of cystine causes the formation of renal cystine calculi. Mutational analysis of the rBAT gene of patients with cystinuria is revealing a growing number (~20) of cystinuria-specific mutations, including missense, nonsense, deletions and insertions. Mutations M467T (substitution of methionine 467 residue for threonine) and R270X (stop codon at arginine residue 270) represent approximately half of the cystinuric chromosomes where mutations have been found. Mutation M467T reduces transport activity of rBAT in oocytes. All this demonstrates that mutations in the rBAT gene cause cystinuria. Three types of cystinuria (types, I, II and III) have been described on the basis of the genetic, biochemical and clinical manifestations of the disease. Type I cystinuria has a complete recessive inheritance; type I heterozygotes are totally silent. In contrast, type II and III heterozygotes show, respectively, high or moderate hyperaminoaciduria of cystine and dibasic amino acids. Type III homozygotes show moderate, if any, alteration of intestinal absorption of cystine and dibasic amino acids; type II homozygotes clearly show defective intestinal absorption of these amino acids. To date, all the rBAT cystinuria-specific mutations we have found are associated with type I cystinuria (~70% of the chromosomes studied) but not to types II or III. This strongly suggests genetic heterogeneity for cystinuria. Genetic linkage analysis with markers of the genomic region of rBAT in chromosome 2 (G band 2p16.3) and intragenic markers of rBAT have demonstrated genetic heterogeneity for cystinuria; the rBAT gene is linked to type I cystinuria, but not to type III. Biochemical, genetic and clinical studies are needed to identify the additional cystinuria genes; a low-affinity cystine reabsortion system and the putative light subunit of rBAT are additional candidate genes for cystinuria.

Notes: Abstract: We explore analytically the nature of the transition to the Fulde-Ferrel-Larkin-Ovchinnikov superfluid phases in the vicinity of the tricritical point, where these phases begin to appear. We make use of an expansion of the free energy up to an overall sixth order, both in order parameter amplitude and in wavevector. We first explore the minimization of this free energy within a subspace, made of arbitrary superpositions of plane waves with wavevectors of different orientations but same modulus. We show that the standard second order FFLO phase transition is unstable and that a first order transition occurs at higher temperature. Within this subspace we prove that it is favorable to have a real order parameter and that, among these states, those with the smallest number of plane waves are preferred. This leads to an order parameter with a cos( . ) dependence, in agreement with preceding work. Finally we show that the order parameter at the transition is only very slightly modified by higher harmonics contributions when the constraint of working within the above subspace is released.

Notes: Summary Rabbits were infected successfully with two strains of human T-cell leukemia virus type I (HTLV-I), one isolated from a Colombian patient with HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP) and the other from an asymptomatic carrier. HTLV-I was repeatedly demonstrated in peripheral blood mononuclear cells (PBMNC) of infected rabbits, and the rabbits had elevated antibodies against the various structural proteins of HTLV-I. Four rabbits inoculated with HTLV-I-infected autologous lymphoid cells intravenously (i.v.) and intracerebrally (i.c.) had virus present in their PBMNC for more than 40 weeks, while those that were inoculated either with HTLV-I-infected human lymphoid cells or with autologous rabbit lymphoid cells intraperitoneally (i.p.) had episodes during which virus was not recovered from their PBMNC. The one rabbit inoculated i.p. developed antibodies to viral envelope glycoproteins earlier than did those inoculated i.v. and i.c. Rabbit lymphoid cell lines persistently infected with HTLV-I were established by cocultivating the rabbit PBMNC with HTLV-I-infected human lymphoid cells that had been irradiated or by inoculation with cell-free supernatant fluids of HTLV-I infected non-irradiated lymphoid cell cultures. HTLV-I-infected rabbit cell lines were of T-cell origin and expressed HTLV-I antigens by immunofluorescence. Electron microscopy revealed type-C retrovirus particles.

Notes: Rat calvarial cell mitogenic behavior was investigated on various biomaterials coated with Matrigel®, a basement membrane matrix containing growth factors. Low (20-40%) and high (70-90%) crystallinity hydroxyapatite (rHA and cHA), rough titanium (Ti), and tissue culture polystyrene (TP) surfaces were compared. Surface chemistry and calcium resorption of HA coatings, alkaline phosphatase activity (APA), and growth of cells were measured for Matrigel®-coated and uncoated surfaces at 2, 7, and 14 days. Gene expression for four noncollagenous bone-related proteins (osteonectin, osteopontin, alkaline phosphatase, and osteocalcin) was also investigated by reverse transcription and polymerase chain reaction up to 28 days. Ca concentration in incubating solutions increased with time for the two types of HA coatings and was always greater for rHA than cHA. Surface chemistry and coating dissolution rates were not affected by the presence of Matrigel® or cells throughout the study. APA of cells on the two HA-coated surfaces was comparably enhanced in the presence of Matrigel® and was greater than on Ti surfaces. Only HA surfaces showed an increased APA of cells with time in the presence of Matrigel®. Cell growth peaked at 7 days and was greatest for cells on the two HA surfaces and without Matrigel®. At 14 days, cell growth was comparable on the four surfaces. The presence of HA and Matrigel® enhanced cell-specific APA at 14 days. Gene expression for all four proteins investigated showed no differences between surfaces after 7 days. At 2 and 7 days, gene expression was indicative of proliferation for Ti, and of proliferation, differentiation, and mineralization for HA and TP more so without Matrigel®. The addition of this matrix significantly enhanced mitogenicity of calvarial cells on HA only after 14 days. Matrigel® eliminated differences seen between the two HA coatings. Gene expression was not enhanced or inhibited by the presence of Matrigel®. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 171-179, 1998.