Search PubMed⌕ Search

Biomedical subjects

S Stein

Publications and source records attributed to S Stein.

At least 55 records · Page 3Linked to original sources
PubMed1998

Inhibition of HIV-1 replication by a Tat RNA-binding domain peptide analog.

The peptidic compound, N-acetyl-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Cys(biotin)-NH2 (Tat10-biotin), contains the 9-amino acid sequence from the basic domain of the Tat protein responsible for specific interaction with TAR RNA. The cysteine residue provides an attachment site for biotin, which acts as a cellular uptake enhancer. Tat10-biotin binds a fragment of TAR RNA (deltaTAR) avidly and specifically, as measured in an electrophoretic gel shift assay. Tat10-biotin inhibited tat gene-induced expression of a stably transfected chloramphenicol acetyl transferase (CAT) reporter gene linked to the HIV-1 long terminal repeat (LTR) in a model cell assay, but did not inhibit phorbol ester-induced expression of CAT, thereby demonstrating a Tat-dependent mechanism of inhibition. Inhibition of HIV-1 replication after acute infection of MT2 cells was demonstrated by absence of HIV-induced syncytium formation and cytotoxicity, as well as by suppression of reverse transcriptase production. These results suggest that a peptide or peptide mimetic capable of competing with the TAR RNA-binding domain of Tat protein might be useful as a therapeutic agent for AIDS.

Anti-HIV Agents↗
PubMed1998

Space geodetic observations of nazca-south america convergence across the central andes

Space geodetic data recorded rates and directions of motion across the convergent boundary zone between the oceanic Nazca and continental South American plates in Peru and Bolivia. Roughly half of the overall convergence, about 30 to 40 millimeters per year, accumulated on the locked plate interface and can be released in future earthquakes. About 10 to 15 millimeters per year of crustal shortening occurred inland at the sub-Andean foreland fold and thrust belt, indicating that the Andes are continuing to build. Little (5 to 10 millimeters per year) along-trench motion of coastal forearc slivers was observed, despite the oblique convergence.

Journal Article↗
PubMed1998

Very early-onset bulimia nervosa: report of two cases.

Bulimia nervosa is very rare in children below the age of 14 years, and no reliable reports of prepubertal bulimia nervosa have been published. We describe two cases of early-onset bulimia nervosa who presented before the age of 14 years, and with premenarchal onset in one patient. Both girls demonstrated high levels of the risk factors known to play a part in the etiology of bulimia nervosa. Implications of these cases regarding the etiology and occurrence of bulimia nervosa in younger adolescents are discussed.

Adolescent↗
PubMed1998

Presence of an N-terminal polyhistidine tag facilitates stable expression of an otherwise unstable N-terminal domain of mouse tissue inhibitor of metalloproteinase-1 in Escherichia coli.

The active N-terminal domain of the mouse tissue inhibitor of metalloproteinases-1 is a 14.1-kDa polypeptide with three disulfide bonds. When expressed using a T7 system in Escherichia coli, this truncated protein, in contrast to the WT protein, was found only in trace amounts in the cell. However, when the coding sequence was placed downstream of a 60-bp sequence that encoded an in-frame histidine-rich "tag," the fusion product (NF.TIMP*His) was expressed in considerably increased abundance. WT.TIMP-1 was expressed in abundance with or without the tag. The mRNAs encoding the various forms of TIMP were present in similar amounts in all four cases. NF.TIMP*His, renatured and purified on a nickel affinity column, was found to be about 10-fold less effective than native human TIMP-2 at inhibiting cleavage of collagen type I by human fibroblast collagenase. A thrombin cleavage site in the tag was susceptible to cleavage by low levels of a contaminating proteinase.

Animals↗
PubMed1998

CMIX, a paired-type homeobox gene expressed before and during formation of the avian primitive streak.

We cloned a chicken homeobox gene closely related to the Xenopus Mix. 1 gene. CMIX is expressed early in embryogenis in a sickle-shaped area in the posterior zone of the blastoderm. With the beginning of gastrulation, CMIX transcripts are found in the primitive streak primordium, then in the young and medium-sized streak, however not in the mesoderm after its emergence. In the fully-extended streak, CMIX is restricted to its middle, i.e. the prospective ventral mesoderm. CMIX RNA is undetectable by whole-mount in-situ analysis in later stages. We compare CMIX expression to the early pattern of the brachyury gene.

Amino Acid Sequence↗
PubMed1998

Immunoglobulin M capture assay for serologic confirmation of early Lyme disease: analysis of immune complexes with biotinylated Borrelia burgdorferi sonicate enhanced with flagellin peptide epitope.

We previously reported on the efficacy of the enzyme-linked immunoglobulin M capture immune complex (IC) biotinylated antigen assay (EMIBA) for the seroconfirmation of early Lyme disease and active infection with Borrelia burgdorferi. In earlier work we identified non-cross-reacting epitopes of a number of B. burgdorferi proteins, including flagellin. We now report on an improvement in the performance of EMIBA with the addition of a biotinylated form of a synthetic non-cross-reacting immunodominant flagellin peptide to the biotinylated B. burgdorferi B31 sonicate antigen source with the avidin-biotinylated peroxidase complex detection system used in our recently developed indirect IgM-capture immune complex-based assay (EMIBA). As in our previous studies, the enzyme-linked immunosorbent assay (ELISA) reactivities of antibodies liberated from circulating ICs (by EMIBA) were compared with those of antibodies in unprocessed serum (antibodies found free in the serum, thus as an IgM-capture ELISA, but not EMIBA, because the antibodies were not liberated from ICs), the sample usually used in standard ELISAs and Western blot assays. The addition of the flagellin epitope enhanced the ELISA signal obtained with untreated sera from many Lyme disease patients but not from healthy controls. In tests with both free antibodies and ICs, with or without the addition of the flagellin epitope to the sonicate, we found the most advantageous combination was IC as the source of antibodies and sonicate plus the flagellin epitope as the antigen. In a blinded study of sera obtained from patients with early and later-phase Lyme disease, EMIBA with the enhanced antigenic preparation compared favorably with other serologic assays, especially for the confirmation of early disease.

Antibodies, Bacterial↗
PubMed1998

Haploinsufficiency of MSX1: a mechanism for selective tooth agenesis.

Previously, we found that the cause of autosomal dominant selective tooth agenesis in one family is a missense mutation resulting in an arginine-to-proline substitution in the homeodomain of MSX1. To determine whether the tooth agenesis phenotype may result from haploinsufficiency or a dominant-negative mechanism, we have performed biochemical and functional analyses of the mutant protein Msx1(R31P). We show that Msx1(R31P) has perturbed structure and reduced thermostability compared with wild-type Msx1. As a consequence, the biochemical activities of Msx1(R31P) are severely impaired, since it exhibits little or no ability to interact with DNA or other protein factors or to function in transcriptional repression. We also show that Msx1(R31P) is inactive in vivo, since it does not display the activities of wild-type Msx1 in assays of ectopic expression in the limb. Furthermore, Msx1(R31P) does not antagonize the activity of wild-type Msx1 in any of these assays. Because Msx1(R31P) appears to be inactive and does not affect the action of wild-type Msx1, we propose that the phenotype of affected individuals with selective tooth agenesis is due to haploinsufficiency.

Amino Acid Substitution↗
PubMed1997

Females in proestrus state maintain splenic immune functions and tolerate sepsis better than males.

OBJECTIVES: To determine: a) whether the cell-mediated immune response during sepsis differs in females vs. males; and b) whether the survival rate in females is different than in males after a septic insult. DESIGN: A prospective, randomized animal study. SETTING: University research laboratory. SUBJECTS: Male and female proestrus C3H/HeN mice. INTERVENTIONS: After anesthesia, male and proestrus female mice underwent cecal ligation puncture to induce sepsis. The mice were killed at 24 hrs after the onset of sepsis. MEASUREMENTS AND MAIN RESULTS: Splenocyte proliferation, as well as splenocyte interleukin (IL)-2 and IL-3 release, was determined by bioassay. In additional studies, survival rate after septic challenge was measured over 10 days. Splenocyte proliferative capacity and splenocyte IL-2 and IL-3 release were markedly decreased in male, but not in female, septic mice. Furthermore, the survival rate of septic female proestrus mice was significantly higher than in comparable male mice. CONCLUSIONS: These results support the concept that the immune response of females differs from males, and that females are immunologically better positioned to meet the challenge of sepsis.

Adolescent↗
PubMed1997

Clinical, biochemical, and molecular investigations of a genetic isolate of growth hormone insensitivity (Laron's syndrome).

We have characterized the GH receptor mutation that is responsible for extreme short stature and GH insensitivity in a Bahamian genetic isolate. Heights of affected individuals ranged from -4.0 to -6.3 SD. Like others with Laron's syndrome, they had normal to high serum GH concentrations and low serum insulin-like growth factor I concentrations. Circulating levels of GH-binding protein activity were below limits of detection. Amplification of exons 2-7 and screening with single strand conformational polymorphism analysis located an abnormality in exon 7. Sequencing identified homozygosity for a C to T transition in the third position of codon 236. Reverse transcription and PCR amplification of complementary DNA from lymphocytes showed that this same sense mutation generated a new splice donor site 63 bp 5' to the normal exon 7 splice site. This novel site was used to the exclusion of the normal site in homozygotes. Both normal and variant messenger ribonucleic acid species were detected in heterozygotes. The predicted protein lacks 21 amino acids, including those defining the WS-like motif of the GH receptor extracellular domain. The high frequency of Laron's syndrome in this isolated island population probably reflects the introduction of the G236 splice mutation by a settler early in the 300-yr history of English settlement.

Adolescent↗
PubMed1997

Prone postioning and low-volume pressure-limited ventilation improve survival in patients with severe ARDS.

STUDY OBJECTIVES: Investigating the effect of low-volume pressure-limited ventilation and repeated prone positioning on the short-term course and outcome in patients with severe ARDS. SETTING: Level 1 trauma center of a university hospital. PATIENTS: Twenty-five patients suffering from ARDS with a lung injury score (LIS) > or = 2.5 admitted consecutively to our ICU from January 1992 to December 1994. METHODS: Mechanical ventilation with peak inspiratory pressure limitation to 35 mbar, irrespective of hypercapnia and prone positioning to achieve adequate oxygenation. SCORING AND MEASUREMENTS: Patient assessment with LIS, APACHE (acute physiology and chronic health evaluation) II score, injury severity score, and multiple organ failure score. Blood gas analyses and estimation of static compliance were repeated at least every 4 h during the treatment period. PaO2/FIO2 (fraction of inspired oxygen) ratio, alveolo-arterial oxygen difference, and intrapulmonary shunt were calculated according to standard equations. The best values taken from each 4-h period during the investigation were used to evaluate the best possible performance of the lung within this interval and to investigate the entire course. RESULTS: Mean predicted mortality based on the APACHE II score was 35.4+/-15.2%. Three of the 25 patients (12%) died. However, none was related to respiratory failure. No pneumothorax occurred. Sixteen patients, lacking any contraindication for prone positioning, responded positively to this change in position, each to a different individual degree. CONCLUSION: We assume that our low mortality in patients with severe ARDS might be due mainly to low-volume pressure-limited ventilation and prone positioning. This simple strategy seems to allow successful treatment for patients with severe ARDS.

APACHE↗
PubMed1997

Special problems of the asthmatic patient.

Environmental factors and physical activity present special problems for the asthmatic patient. Bronchoconstriction due to workplace exposure is one of the most common forms of occupational lung disease and has similar pathogenic mechanisms to nonoccupational asthma. Diagnosing occupational asthma may be difficult because the association between symptoms during or after work and a causative agent may be obscure. Nevertheless, it is important to recognize occupational asthma because even trace amounts of the causative agent may trigger bronchoconstriction, and the affected employee must almost always avoid exposure. Bronchoconstriction induced by exercise has affected the lifestyles of many asthmatic patients and has created special problems for numerous accomplished athletes. Although exercise-induced asthma (EIA) characteristically occurs immediately after cessation of exertion, a recent study suggests that it may sometimes begin during sustained exertion. Asthmatic patients who participate in winter sports are particularly susceptible to EIA. Vocal cord dysfunction in athletes who participate in intense competition can mimic EIA. In addition to beta-agonists used immediately before exercise, newer drugs that show promise in the treatment of EIA include the leukotriene inhibitors and furosemide. Asthmatic patients are susceptible to extremes in atmospheric pressure and have increased risk of barotrauma and arterial gas embolism during scuba diving. During longer stays at high altitude, however, asthmatic patients appear to have attenuated bronchial responsiveness.

Adrenergic Agonists↗
PubMed1997

Humane and practical implications of using carbon dioxide mixed with oxygen for anesthesia or euthanasia of rats.

A series of studies was undertaken to determine whether CO2 can be used as a humane as well as practical agent for euthanasia or anesthesia of rats. Human volunteers rated the degree of discomfort associated with breathing 50 to 100% CO2 mixed with oxygen. Increasing concentrations of CO2 were judged as progressively more noxious, from "highly unpleasant" for 50% CO2 to "painful" for 100% CO2. The practical aspects of anesthesia and euthanasia with 50 to 100% CO2 were studied, using male Sprague Dawley rats. Time to anesthesia and death were inversely related to CO2 concentration, as were the frequency and severity of adverse reactions, including seizures and hemorrhaging from the nose. The severity of edema and hemorrhage, which were observed on histologic examination of the lungs of all rats euthanized with CO2, were greatest in the animals exposed to the lowest concentrations. There were no significant effects of CO2 concentration on time to recumbency or recovery, and there were no significant effects of precharging versus not precharging the chamber on any of the parameters studied. It was concluded that, although CO2 can be used in a humane manner, the concentrations that are least likely to cause pain and distress are associated with the longest times to anesthesia and death, highest incidence of unwanted side effects, and most severe histologic changes in the lungs. Acceptably humane and reasonably practical euthanasia or anesthesia can be achieved using a nonprecharged chamber and a low gas flow rate so that conscious animals are never exposed to CO2 concentrations > 70%.

Adult↗
PubMed1996

Differential activation of the clustered homeobox genes CNOT2 and CNOT1 during notogenesis in the chick.

CNOT2, a newly identified homeobox gene, is physically linked to the CNOT1 gene in the chicken genome. The two chicken genes represent two different subgroups of the Not gene family, the first including CNOT1 and the Xenopus genes XNot1 and XNot2, and the second CNOT2 and the zebrafish floating head gene. The overall expression pattern of CNOT2 in Hensen's node, notochord, neural plate, tailbud, and epiphysis resembled the CNOT1 pattern. However, several significant differences occurred: CNOT2 expression was much stronger and more widespread in the pregastrulation embryo, it showed an additional, transient domain on the anterior intestinal portal, and lacked expression on the early anterior neural folds and the anterodistal limb bud. We studied CNOT expression by transplanting parts of the primitive streak into growing embryos or by explanting them into tissue culture. CNOT gene expression from young nodes was maintained in vivo, but required in vitro the addition of retinoic acid. The generation of differentiated notochord structures could only be obtained, if either older node grafts were used in vitro or young node grafts were transplanted close to the primary axis in vivo. We conclude that CNOT expression in the anterior streak is not enough for notochord differentiation, but further influences are necessary. A Not-related gene has previously been isolated from Drosophila melanogaster and its expression was detected in the posterior brain and the neuroblasts (Dessain and McGinnis, 1993. Adv. Dev. Biochem. 2, 1-55). The correspondence between Not gene-expressing cells in the nervous system of Drosophila and the early neuroectoderm in the chick and its implication for a phylogenetic relationship between neuroectoderm and the notochord is discussed.

Amino Acid Sequence↗
PubMed1996

Multi-well ELISA based on independent peptide antigens for antibody capture. Application to Lyme disease serodiagnosis.

Novel procedures for the use of peptides as antibody-capture reagents in the ELISA format have been investigated. Epitope sequences from known immunodominant antigens of Borrelia burgdorferi were selected by screening peptide libraries with sera from patients with Lyme disease. Several epitope peptides were synthesized and immobilized, separately, on the ELISA plate as haptens on bovine serum albumin. Based on a comparative analysis of serum samples, it appears that peptide antigens can be used as effectively as a whole cell lysate to discriminate between Lyme disease and non-Lyme disease sera, thus avoiding dependence on bacterial sonicates which vary from passage to passage. Further improvements in epitope design for enhancement of accuracy in serodiagnosis are discussed.

Amino Acid Sequence↗
PubMed1996

Hybridization properties of oligodeoxynucleotide pairs bridged by polyarginine peptides.

The hybridization properties of a series of probes, based on two 9mer oligodeoxynucleotides (designated as I and II) having an appended oligoarginine chain (Rn) to produce peptide-oligonucleotide conjugates or peptide-bridged oligonucleotide pairs (e.g. Rn-I or II-Rn-I), were investigated. For the double-linked probes, we found that the peptide bridge induces the two 9mers to bind complementary single-stranded DNA or RNA targets with substantially enhanced thermal stability. The resulting hybrid with complementary DNA was found to assume a 1:1 complex in the B conformation as judged by UV mixing curves and CD spectroscopy. Complexes of single or double-linked probes with complementary RNA exhibited sensitivity to RNase H digestion. The influence of the identity and chirality of the repeating unit in the bridge, the length of the bridge, the gap size and the salt concentration on the hybridization properties of this new class of oligonucleotide probes was also studied. Our data reveal that these compounds exhibit properties that should prove useful in the development of antisense strategies.

Amino Acid Sequence↗
PubMed1996

[Added work of breathing, respiratory pattern and determination of ventilator weaning readiness in inspiratory pressure support and and automatic tube compensation].

We measured the ventilatory pattern and additional work of breathing (WOBadd) at three different levels of inspiratory pressure support [IPS 5, 10, 15 mbar above positive end-expiratory pressure (PEEP)] and in a new ventilatory mode, automatic tube compensation (ATC), in nine operative patients without lung injury nine patients ventilated for several following acute respiratory insufficiency (ARI). In ATC, endotracheal tube resistance is compensated automatically by means of closed-loop control of the calculated tracheal pressure. Pressure support in this mode, i.e. airway pressure above PEEP, is equal to the actual flow-dependent pressure drop across the endotracheal tube (ETT). Airway pressure rises at the beginning of inspiration and falls towards the end. As the tube resistance of ETT seriously hinders expiration and can cause desynchronization between ventilator and patient, airway pressure is reduced below PEEP during expiration in the same way as it is increased during inspiration. The result is a near-constant tracheal pressure at PEEP both during inspiration and during expiration. This mode could be best termed as "electronic extubation". The most striking difference between the postoperative patients and the ARI patients was their minute ventilation (17.8 +/- 1.85 l/min in ARI patients vs 7.3 +/- 3.1 l/min in the postoperative patients). In the postoperative patients augmentation of IPS from 5 to 15 mbar induced a steady increase in tidal volume (VT) and a consecutive decrease in respiratory rate (rr) compared with ATC (VTATC,postop = 463 +/- 78 ml; rrATC,postop = 16 +/- 4 min-1; VTIPS5.postop = 505 +/- 79 ml; rrIPS5,postop = 15 +/- 4 min-1; VTIPS10,postop = 562 +/- 86 ml; rrIPS15,postop = 14 +/- 4 min-1; VTIPS15.postop = 660 +/- 151 ml; rrTPS15,postop = 12 +/- 4 min-1), whereas the augmentation of IPS of 5 and 10 mbar in the ARI patients could not compensate for the increase in rr and the decrease in VT, after switching from ATC to IPS (VTATC,ARI 724 +/- 308 ml, rrATC,ARI = 24 +/- 6 min-1; VTIPS5,ARI = 649 +/- 315 ml; rrIPS5,ARI = 27 +/- 8 min-1; VTIPS10,ARI = 653 +/- 353 ml; rrIPS10,ARI = 25 +/- 8 min-1: Even IPS 15 was not able to reestablish VT at the values observed during ATC (VTIPS15,ARI = 680 +/- 312 ml). During ATC WOBadd was small in both postoperative and ARI patients (WOBadd,ATC,postop = 93 +/- 36 mJ/l, WOBadd,ATC,ARI = 116 +/- 72 mJ/l). In the postoperative patients, an inspiratory pressure support of 5 mbar was not sufficient to compensate WOBadd compared with ATC. However, IPS 10 and 15 mbar were able to compensate for WOBadd (WOBadd,ATC5.postop WOBadd,IPS5,postop = 189 +/- 77 mJ/l; WOBadd,IPS10,postop = 55 +/- 30 mJ/l; WOBadd,IPS15,postop = 21 +/- 11 mJ/l). In the ARI patients an IPS 5, 10 or 15 mbar was not sufficient to compensate for WOBadd (WOBadd,IPS 5,ARI = 1126 +/- 262 mJ/l; WOBadd,IPS 10,ARI 863 +/- 253 mJ/l; WOBadd,IPS15,ARI 763 +/- 298 mJ/l). Under ATC, WOBadd was only 15% of WOBadd under IPS of 15 mbar. All but two patients were successfully extubated after the investigation. These two patients were not extubated because they were dependent on an FIO2 > 0.5. Our results strongly indicate that ventilatory dependence in ARI patients may be caused by the ETT rather than by mechanical dysfunction of the lung. ATC is a very helpful mode to use in distinguishing between ventilatory failure caused by ETT and real ventilatory dependence.

Adult↗