Miracle-dz/UniFAR-FTUs huggingface dataset:
Fibrillins Can Co-assemble in Fibrils, but Fibrillin Fibril Composition Displays Cell-specific Differences*

Fibrillins are microfibril-forming extracellular matrix macromolecules that modulate skeletal development. In humans, mutations in fibrillins result in long bone overgrowth as well as other distinct phenotypes. Whether fibrillins form independent microfibrillar networks or can co-polymerize, forming a single microfibril, is not known. However, this knowledge is required to determine whether phenotypes arise because of loss of singular or composite functions of fibrillins. Immunolocalization experiments using tissues and de novo matrices elaborated by cultured cells demonstrated that both fibrillins can be present in the same individual microfibril in certain tissues and that both fibrillins can co-polymerize in fibroblast cultures. These studies suggest that the molecular information directing fibrillin fibril formation may be similar in both fibrillins. Furthermore, these studies provide a molecular basis for compensation of one fibrillin by the other during fetal life. In postnatal tissues, fibrillin-2 antibodies demonstrated exuberant staining in only one location: peripheral nerves. This surprising finding implicates distinct functions for fibrillin-2 in peripheral nerves, because a unique feature in humans and in mice mutant for fibrillin-2 is joint contractures that resolve over time.
Detection of Epidemic USA300 Community-Associated Methicillin-Resistant Staphylococcus aureus Strains by Use of a Single Allele-Specific PCR Assay Targeting a Novel Polymorphism of Staphylococcus aureus pbp3

ABSTRACT In recent years, the dramatic increase in community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections has become a significant health care challenge. Early detection of CA-MRSA is important because of its increased virulence associated with the arginine catabolic mobile element (ACME), Panton-Valentine leukocidin (PVL), and other toxins that may contribute to disease severity. In particular, the USA300 epidemic clone has emerged and now represents the cause of as much as 98% of CA-MRSA skin and soft tissue infections in the United States. Current diagnostic assays used to identify CA-MRSA strains are based on complex multiplex PCRs targeting the staphylococcal cassette chromosome mec (SCCmec) DNA junction, a multitude of genes, and noncoding DNA fragments or on a number of lengthy sequence-typing methods. Here, two nucleotide polymorphisms, G88A and G2047A, that were found to be in strict linkage disequilibrium in the S. aureus penicillin-binding protein 3 (pbp3) gene were also found to be highly associated with the USA300 clone of CA-MRSA. Clinical isolates that contained this pbp3 allele were also positive for the presence of SCCmec type IV, the ACME, and the PVL toxin gene and matched the t008 or t121 molecular spa types, which are associated specifically with the USA300 CA-MRSA clone. A single allele-specific PCR targeting the G88A polymorphism was developed and was found to be 100% sensitive and specific for the detection of USA300 CA-MRSA and 91.5% sensitive and 100% specific for the detection of all CA-MRSA isolates in this study.
Correlates of Self-Reported Coercive Parenting of Preschool-Aged Children at High Risk for the Development of Conduct Problems

Objective: This study examines the correlates of coercive parenting in a high-risk sample of 305 three-year-old children likely to develop later conduct problems. As parental coercion has been identified as a significant risk factor for future psychopathology, the study sought to identify modifiable inter and intra-personal factors most closely associated with coercion. Method: Key variables known to place young children at future risk, such as maternal mood states, current child behaviour problems, demographic characteristics such as low income, past mental health problems and parents' sense of competence, were analyzed based on parent-report measures and clinical interviews. Correlational and heirachical regression analysis identified key predictors of coercion. Results: Three variables emerged as the strongest predictors of maternal coercion: selfefficacy, child behaviour and maternal depression. Demographic factors contributed little to the model. Conclusions: Enhancing parental self-efficacy, especially specific parenting tasks with disruptive young children has the potential to make a significant contribution toward prevention of future conduct problems.
Human Rhinovirus Type 16: Mutant V1210A Requires Capsid-Binding Drug for Assembly of Pentamers To Form Virions during Morphogenesis

ABSTRACT Our laboratory has previously reported isolation of human rhinovirus type 16 (HRV16) mutants which depend on WIN 52035 to grow. A rapid rise of progeny virus infectivity occurred when drug was added late in growth cycles, suggesting that the drug-dependence lesion was at the step of virus assembly (W. Wang et al., J. Virol. 72:1210-1218, 1998). Here, we report that capsid subunits, 5S protomers and 14S pentamers, of a drug-dependent mutant were produced normally in the absence of drug, but mutant 80S empty capsids and 150S provirions were not formed, maturation cleavage of provirions (VP0 → VP2 + VP4) did not occur, and the unassembled mutant capsid subunits were degraded with a half-life of 15 min. Drug was not required by mutant virus for attachment, uncoating, RNA synthesis and protein synthesis, and polyprotein processing except maturation cleavage. The requirement of drug for assembly of mutant pentamers to form provirions and the rapid assembly of preformed subunits (synthesized in the absence of drug) after drug addition suggested that after native pentamers (P5) have been formed they must be converted to an assembly active state (P5∗), possibly by a conformational change induced by the binding of drug. We propose that pocket factor plays the same role in wild-type virus. In addition, we also report the construction and the properties of a full-length cDNA clone of HRV16, pR16.11, which produces in vitro transcripts with infectivity similar to that of virion RNA. This cDNA clone is available at the American Type Culture Collection.
Small signal modeling of illuminated MOSFET for RF application

Recent development in the device technology has led to the potential research in the area of optical characterization of FET devices. It is mainly because of the high gain, speed, power handling capacity and high packing density of IC viz. OEIC, MMIC, VLSI, etc. It needs rigorous mathematical modeling to characterize the device and obtain the various parameters to get the operational behavior of the device working in various operating conditions. MOSFET is one of the device of interest. A small signal analytical model of short channel MOSFET under dark and illumination is presented. Model is mainly based on CV characteristics of MOSFET. In the present paper to carry out the AC analysis under illumination, the optical radiations are made to incident on semi insulating gate of the MOSFET and its effect on capacitance is observed. EKV capacitance model is developed in MATLAB. Results shows that the significant change in the various intrinsic capacitances of the device under illumination. The device is expected to behave as a photodetector, optical transceiver and microwave devices etc
Characterization of a periplasmic 3':5'-cyclic nucleotide phosphodiesterase gene, cpdP, from the marine symbiotic bacterium Vibrio fischeri

Vibrio fischeri, a marine bacterium that forms a bioluminescent symbiosis with certain fish and squids, exhibits the unusual attribute of growth on 3':5'-cyclic AMP (cAMP), apparently through the activity of a 3':5'-cyclic nucleotide phosphodiesterase (3':5'-CNP) with exceptionally high activity. The V. fischeri 3':5'-CNP is located in the periplasm, a novel cellular location for this enzyme in bacteria. To gain insight into the physiological function of this enzyme, we cloned the gene (designated cpdP) encoding it from V. fischeri MJ-1. This is the first bacterial 3':5'-CNP gene to be cloned. Sequencing and analysis of the 1.26-kb cpdP locus revealed a single open reading frame specifying a protein of 330 amino acid residues, including a 22-amino-acid leader peptide. The putative cpdP promoter contained a reasonable -10 promoter region (TATTAT) but contained no obvious -35 region; instead, a 12-bp inverted repeat (TTAAATATTTAA) occurred just upstream of this location. A possible rho-independent transcriptional terminator with a calculated free energy of -21.2 kcal.mol-1 (ca. -88.7 kJ.mol-1) followed the CpdP protein coding sequence. The predicted subunit molecular weight of 33,636 for the mature CpdP protein (36,087 less 2,451 for the leader peptide) was consistent with the molecular weight of 34,000 estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The deduced amino acid sequence of the CpdP protein exhibited 30.3% identity with that of the low-affinity 3':5'-CNP (PDE1) of Saccharomyces cerevisiae and 33.6% identity with that of the extracellular 3':5'-CNP of Dictyostelium discoideum. The residue identities clustered in two regions, residues 100 to 146 and 238 to 269, which contained 30 of the 33 amino acids conserved in all three proteins, 4 of which were histidines. A gene replacement mutant of V. fischeri MJ-1 containing a 0.45-kb BglII deletion within the cpdP gene lacked periplasmic 3':5'-CNP activity and did not grow on cAMP, confirming for V. fischeri the relationship among cpdP, synthesis of the periplasmic 3':5'-CNP, and growth on cAMP. The mutant exhibited no obvious sensitivity to high extracellular concentrations of cAMP (5 and 10 mM), suggesting that the enzyme does not play a role in defense against extracellular cAMP.
Mechanisms of Osteoclast Dysfunction in Human Osteopetrosis: Abnormal Osteoclastogenesis and Lack of Osteoclast‐Specific Adhesion Structures

Osteoclasts from a patient affected by osteopetrosis were examined in vivo and in vitro. Iliac crest biopsy revealed an osteosclerotic pattern, with prominent numbers of osteoclasts noted for hypernuclearity and incomplete adherence to the bone surface. A population comprising tartrate‐resistant acid phosphatase (TRAP)‐positive, multinucleated and mononuclear cells, and alkaline phosphatase‐positive stromal fibroblasts was obtained in vitro from bone marrow. Mononuclear TRAP‐positive precursors spontaneously fused in culture to form giant osteoclast‐like cells. These cells expressed the osteoclast marker MMP‐9 and calcitonin receptor, and lacked the macrophage marker, Fc receptor. Expression and distribution of c‐src, c‐fms, and CD68, and response to steroid hormones relevant to osteoclast differentiation and function were apparently normal, whereas cell retraction in response to calcitonin was impaired. TRAP‐positive multinucleated cells did not form osteoclast‐specific adhesion structures (clear zone, podosomes, or actin rings). Bone resorption rate was severely reduced in vitro. Focal adhesions and stress fibers were observed en lieu of podosomes and actin rings. Adhesion structures contained low levels of immunoreactive vitronectin receptor, most of this integrin being retained in cytoplasmic vesicles. These data provide the first characterization of abnormal differentiation and function of human osteopetrotic osteoclast‐like cells.
Erythromycin attenuates metalloprotease/anti-metalloprotease imbalance in cigarette smoke-induced emphysema in rats via the mitogen-activated protein kinase/nuclear factor-κB activation pathway

The present study investigated whether erythromycin (ERY) reduces cigarette smoke (CS)-induced emphysema in rats and aimed to determine the anti-inflammatory effect of ERY, which may identify potential treatments for chronic obstructive pulmonary disease. Furthermore, the current study focused on the potential effects on the imbalance between matrix metalloprotease (MMP) and anti-MMP activity, the phosphorylation of mitogen-activated protein kinases (MAPKs) and the nuclear factor-κB (NF-κB) signaling pathway. Wistar rats were divided into the following three groups (n=12 each): control (ERY vehicle only, without any CS exposure), CS (animals were exposed to CS for 12 weeks) and CS + ERY (animals were exposed to CS for 12 weeks and received 100 mg/kg/day ERY). The recruitment of inflammatory cells into the bronchoalveolar lavage fluid (BALF) and the histopathology of lung tissue from all groups was evaluated to grade the severity of the emphysema. The expression of MMP-2, MMP-9 and tissue inhibitor of metalloproteinase-1 was evaluated by immunohistochemistry and western blotting. The activation of MAPKs, NF-κB and inhibitor of NF-κB (IκBα), in lung tissues was examined by western blotting. Treatment with ERY resulted in fewer inflammatory cells and cytokines in the BALF, and fewer emphysema-associated changes in the lungs compared with control. The stimulus of CS promoted the phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and p38, but not c-Jun NH2-terminal kinase, thereby inducing the activation of the ERK/MAPK signaling pathway in rats. Furthermore, CS exposure increased the expression of NF-κB and decreased the expression of IκBα. The levels of phosphorylated ERK1/2 and p38 were significantly reduced in rats with CS-induced emphysema when treated with ERY compared with the CS group. The results of the present study therefore indicate that oral administration of ERY may suppress CS-induced emphysema by regulating inflammatory cytokines and the MMP/anti-MMP imbalance via the MAPK/NF-κB pathway.
A triple mutation in the second transmembrane domain of mouse dopamine transporter markedly decreases sensitivity to cocaine and methylphenidate

Previously, we reported that Phe105 in transmembrane domain 2 of the mouse dopamine transporter (DAT) is crucial for high‐affinity cocaine binding. In the current study, we investigated whether other residues surrounding Phe105 also affect the potency of cocaine inhibition. After three rounds of sequential random mutagenesis at these residues, we found a triple mutant (L104V, F105C and A109V) of mouse DAT that retained over 50% uptake activity and was 69‐fold less sensitive to cocaine inhibition when compared with the wild‐type mouse DAT. The triple mutation also resulted in a 47‐fold decrease in sensitivity to methylphenidate inhibition, suggesting that the binding sites for cocaine and methylphenidate may overlap. In contrast, the inhibition of dopamine uptake by amphetamine or methamphetamine was not significantly changed by the mutations, suggesting that the binding sites for the amphetamines differ from those for cocaine and methylphenidate. Such functional but cocaine‐insensitive DAT mutants can be used to generate a knock‐in mouse line to study the role of DAT in cocaine addiction.
Immune mechanisms and protective antigens of Vibrio cholerae serogroup O139 as a basis for vaccine development

We have characterized 11 isolates of Vibrio cholerae O139 Bengal with regard to properties deemed to be relevant for development of a vaccine against O139 cholera. For most strains two colony variants, A and B, which are nonhemolytic and hemolytic, respectively, were detected on blood agar. The A and B variants were associated with high- and low-level production of soluble hemagglutinin-protease, respectively. However, on Luria-Bertani agar both types formed opaque colonies, which has been shown to be associated with capsule formation. Interestingly, under the stationary tube-shaken flask culture conditions in yeast extract-peptone water medium which were used to stimulate the production of cholera toxin (CT) and toxin-coregulated pili, B variants constitutively produced CT and TcpA, two ToxR-regulated proteins, at 28 and 37 degrees C, whereas the production of these proteins by A variants was downregulated at the higher temperature. One of the strains, 4260B, having a well-exposed O antigen and capsule and the capacity to produce large amounts of TcpA, CT, and mannose-sensitive hemagglutinin pili but minimal amounts of the proteolytic soluble hemagglutinin, was selected to produce antibacterial antisera and as a challenge strain in protection studies using the rabbit ileal loop model. Rabbit antisera to live, heat-killed, or formalin-killed O139 vibrios or to purified O139 lipopoly-saccharide (LPS) as well as monoclonal antibodies (MAbs) to O139 LPS agglutinated all O139 isolates. However, when A and B variants of strain 4260 were tested for sensitivity to vibriocidal activity of these antibody preparations, only the B variant was killed. All of the antisera against live or killed O139 vibrios conferred passive protection against fluid accumulation induced by the challenge strain. The protective effects of the antisera were correlated to anti-LPS antibody titers rather than to titers against whole bacteria that had been grown for toxin-coregulated pilus expression. This protection was considerably higher than that conferred by antisera to classical, EI Tor, or recombinantly produced (classical) CT or CTB. Furthermore, MAbs to O139 LPS and CTB-CT exhibited a strong synergistic protection against O139 challenge irrespective of the level of sensitivity of challenge strains to O139 LPS MAbs in vibriocidal assays in vitro.
Disposition of warfarin enantiomers and metabolites in patients during multiple dosing with rac-warfarin.

1. The disposition of warfarin enantiomers and metabolites has been studied in 36 patients receiving chronic rac-warfarin therapy, titrated to approximately the same anticoagulant response. 2. A stereoselective h.p.l.c. assay was employed to determine the concentrations of (R)- and (S)-warfarin, (R,S)-warfarin alcohol and (S)-7-hydroxywarfarin in plasma and 24 h urine samples. The concentrations of (R)-7-hydroxywarfarin, (S,S)-warfarin alcohol and (R)-6- and (S)-6-hydroxywarfarin were also determined in urine samples. The fractions unbound of warfarin enantiomers were determined using equilibrium dialysis. 3. Wide variability was observed in daily dose requirements (mean 6.1 mg; range: 2.5-12 mg), in plasma concentrations of (S)-warfarin (0.48 mg l(-1); 0.11-1.02 mg l(-1)), (R)-warfarin (0.87 mg l(-1); 0.29-1.82 mg l(-1)), (R,S)-warfarin alcohol (0.31 mg l(-1); 0.02-0.72 mg l(-1)) and (S)-7-hydroxywarfarin (0.25 mg l(-1); 0.07-0.37 mg l(-1)) and the percentage unbound of (S)-warfarin (0.53%; 0.29%-0.82%) and (R)-warfarin (0.54%; 0.26%-0.96%). 4. The mean plasma clearances of warfarin enantiomers were 7.5 1 day-1 per 70 kg (2.5-22.1) for (S)-warfarin and 3.6 1 day-1 per 70 kg (1.6-8.8) for (R)-warfarin. There was a significant correlation between the estimated formation clearance of (S)-7-hydroxywarfarin and the clearance of (S)-warfarin, which accounted for much of the variability in the latter. (ABSTRACT TRUNCATED AT 250 WORDS)
An analytical and structural database provides a strategy for sequencing O-glycans from microgram quantities of glycoproteins.

A sensitive, rapid, quantitative strategy has been developed for O-glycan analysis. A structural database has been constructed that currently contains analytical parameters for more than 50 glycans, enabling identification of O-glycans at the subpicomole level. The database contains the structure, molecular weight, and both normal and reversed-phase HPLC elution positions for each glycan. These observed parameters reflect the mass, three-dimensional shape, and hydrophobicity of the glycans and, therefore, provide information relating to linkage and arm specificity as well as monosaccharide composition. Initially the database was constructed by analyzing glycans released by mild hydrazinolysis from bovine serum fetuin, synthetic glycopeptides, human glycophorin A, and serum IgA1. The structures of the fluorescently labeled sugars were determined from a combination of HPLC data, mass spectrometric composition and mass fragmentation data, and exoglycosidase digestions. This approach was then applied to human neutrophil gelatinase B and secretory IgA, where 18 and 25 O-glycans were identified, respectively, and the parameters of these glycans were added to the database. This approach provides a basis for the analysis of subpicomole quantities of O-glycans from normal levels of natural glycoproteins.
Morphology of retinal ganglion cells in a New World monkey, the marmoset Callithrix jacchus

We studied the morphology of retinal ganglion cells in a diurnal New World primate, the marmoset Callithrix jacchus. This species is of interest as a model for primate vision because it has good behavioural visual acuity, and the retina and subcortical visual pathways are very similar to those of Old World monkeys and humans. Ganglion cells were labelled by placing small crystals of the carbocyanin dye Dil into the optic fibre layer, or by intracellular injection of neurobiotin. Two main classes of ganglion cell were labelled. We call these Group A cells and Group B cells: they are respectively homologous to parasol and midget cell classes. Group A and Group B cells show similar patterns of dye coupling, dendritic stratification and dendritic field size as their counterparts in Old World monkeys and humans. A third group of cells, which we call Group C, is morphologically heterogeneous. Examples corresponding to wide‐field ganglion cell types described in Old World primates were encountered. One subgroup of C cells has a morphology very similar to that of the small bistratified (blue‐on) cell described in macaque retina, suggesting that this functional pathway is common to all primates. As for other New World monkeys, the marmoset shows a sex‐linked polymorphism of cone pigment expression, such that all males are dichromats and the majority of females are trichromats. No systematic differences in Group B cells were seen between male and female retinas, suggesting that trichromacy is not accompanied by specific changes in ganglion cell morphology. © 1996 Wiley‐Liss, Inc.
Differential effects of petit mal anticonvulsants and convulsants on thalamic neurones: calcium current reduction

1 Succinimide derivatives can be either convulsant (tetramethylsuccinimide (TMS)), or anticonvulsant (ethosuximide (ES); α‐methyl‐α‐phenylsuccinimide (MPS)). ES, an anticonvulsant succinimide, has previously been shown to block calcium currents of thalamic neurones, while the convulsant succinimide TMS blocks γ‐aminobutyric acid (GABA) responses in a similar fashion to the convulsant pentylenetetrazol (PTZ). 2 Using voltage‐clamp techniques, we analysed the effects of the anticonvulsant succinimides ES and MPS and the convulsants TMS and PTZ on calcium currents of acutely isolated thalamic relay neurones of the rat. 3 MPS and ES reduced low‐threshold calcium current (LTCC) in a voltage‐dependent manner, without affecting steady‐state inactivation. MPS was less potent than ES (IC50 of 1100 vs 200 μm) but greater in efficacy (100% maximal reduction vs 40% for ES). 4 PTZ had no effect on calcium currents, and TMS only reduced LTCC at very high concentrations, and did not occlude MPS effects when applied concurrently. 5 These results, which demonstrate that anticonvulsant, but not convulsant, succinimides block LTCC, provide additional support for the hypothesis that LTCC reduction is a mechanism of action of the anticonvulsant succinimides related to their effects in petit mal epilepsy.