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J Cohen

Publications and source records attributed to J Cohen.

At least 469 records · Page 26Linked to original sources
PubMed1997

Immunization against the murine malaria parasite Plasmodium yoelii using a recombinant protein with adjuvants developed for clinical use.

Mice vaccinated with a recombinant protein containing the two EGF-like modules of Plasmodium yoelii merozoite surface protein-1 in liposomes or combined with the formulations SBAS2.1 and SBAS2, were protected against a lethal malaria infection. The protection achieved with these adjuvants developed for clinical use was as good as or better than that achieved with Freund's adjuvant. A parasite-specific response was needed for protection. Analysis of the immunoglobulin sub-class response showed that MSP-1-specific IgG1, and to a lesser extent IgG2a and IgG2b, were induced, suggesting that these antibodies were important for protection. Mice passively immunized with serum or purified IgG from vaccinated mice had delayed onset of parasitemia and were able to control the infection.

Adjuvants, Immunologic↗
PubMed1997

A permutation test for the robust sib-pair linkage method.

The robust sib-pair method introduced by Haseman & Elston (1972) is one of the most widely circulated allele-sharing methods for linkage analysis. The procedure evaluates linkage by significance testing of a regression coefficient and, hence, a standard t-test has traditionally been applied despite known violations of the statistical assumptions underlying the test. We present a permutation based reference distribution for the estimate of the regression coefficient that is motivated by genetic principles rather than by standard regression testing procedures. The permutation test approximates Mendelian co-segregation under the null hypothesis of no linkage, making it a very natural approach. Theory and simulations show that the conventional t-test approximates the permutation test quite well, even when dependent sib pairs are used for analysis. These results thus indirectly address concerns over the t-test. To illustrate the permutation test using real data we applied the procedure to two lipoprotein systems that have been well characterized.

Animals↗
PubMed1997

The Apo(a) gene is the major determinant of variation in plasma Lp(a) levels in African Americans.

The distributions of plasma lipoprotein(a), or Lp(a), levels differ significantly among ethnic groups. Individuals of African descent have a two- to threefold higher mean plasma level of Lp(a) than either Caucasians or Orientals. In Caucasians, variation in the plasma Lp(a) levels has been shown to be largely determined by sequence differences at the apo(a) locus, but little is known about either the genetic architecture of plasma Lp(a) levels in Africans or why they have higher levels of plasma Lp(a). In this paper we analyze the plasma Lp(a) levels of 257 sibling pairs from 49 independent African American families. The plasma Lp(a) levels were much more similar in the sibling pairs who inherited both apo(a) alleles identical by descent (IBD) (r = .85) than in those that shared one (r = .48) or no (r = .22) parental apo(a) alleles in common. On the basis of these findings, it was estimated that 78% of the variation in plasma Lp(a) levels in African Americans is attributable to polymorphism at either the apo(a) locus or sequences closely linked to it. Thus, the apo(a) locus is the major determinant of variation in plasma Lp(a) levels in African Americans, as well as in Caucasians. No molecular evidence was found for a common "high-expressing" apo(a) allele in the African Americans. We propose that the higher plasma levels of Lp(a) in Africans are likely due to a yet-to-be-identified trans-acting factor(s) that causes an increase in the rate of secretion of apo(a) or a decrease in its catabolism.

Adolescent↗
PubMed1997

Numerical taxonomy on data: experimental results.

We consider the problem of fitting an n x n distance matrix D by a tree metric T. This problem is NP-hard for most reasonable distance functions between D and T. Recently, an approximation algorithm was presented (Agarwala et al., 1996) which achieves a factor of 3 approximation to the L infinity best fitting tree. We call this method the Single Pivot (SP) heuristic. Within the biology community, the so-called Neighbor-Joining (NJ) heuristic (Saitou and Nei, 1987) has wide acceptance. In this paper, we introduced a new Double Pivot (DP) heuristic, which is an extension of the SP heuristic, and show that DP outperforms NJ on biological and random data.

Classification↗
PubMed1997

Genetic approach to regulated exocytosis using functional complementation in Paramecium: identification of the ND7 gene required for membrane fusion.

Paramecium is a unicellular organism that possesses a specialized pathway for regulated secretion that is amenable to genetic studies. Numerous mutations affecting the process have been isolated over the years, among which is a subclass blocking the terminal step of fusion of the secretory granule with the plasma membrane. We report herein the cloning by functional complementation of one such gene, ND7. The 506-amino acid polypeptide encoded by ND7 is predicted to be a type I integral membrane protein with a highly charged cytosolic domain featuring amphipathic and coiled-coil regions. This structure is compatible with the physiological data on the mutant nd7-1 suggesting that the protein is anchored in the membrane of the secretory granule and that it may interact with other proteins. This work presents the first identification by a genetic approach of a novel gene involved in regulated secretion and establishes Paramecium as a powerful model system for the genetic dissection of this process.

Amino Acid Sequence↗
PubMed1997

Treatment-related chromosome abnormalities in human embryos.

Mosaicism was studied in good quality embryos from four different centres in order to assess the effects of follicular induction and exposure to laboratory conditions on chromosomal status. The donated embryos were fully biopsied and analysed by fluorescence in-situ hybridization using probes for chromosomes X, Y, 13, 18 and 21, simultaneously. The number of abnormal cells present indicated the division at which mosaicism first occurred (4/4 cells at first division, 2/4 cells at second, 2/8 at third). The rate of mosaicism in embryos from different centres varied greatly (P < 0.001). Most of the mosaic embryos were obtained before 1991. In one clinic increased mosaicism was found in embryos obtained before 1991 when compared to embryos obtained thereafter. The results suggest that certain culture conditions and/or hormonal stimulation protocols may induce chromosomal abnormalities and partly explain differences in pregnancy rates between in-vitro fertilization centres.

Biopsy↗
PubMed1997

Cryopreservation of single human spermatozoa.

A procedure is described that allows cryopreservation and efficient post-thaw recovery of either a single or a small group of human spermatozoa. This is achieved by injecting them into cell-free human, mouse or hamster zonae pellucidae before the addition of cryoprotectant. The method involves a combination of physical micromanipulation procedures and glycerol-mediated cryoprotection. Zonae were tracked by positioning them in straws between two small air bubbles prior to freezing. Spermatozoa from poor specimens were cryopreserved and their fertilizing ability after thawing was compared with that of fresh spermatozoa from fertile men. Human eggs used for fertilization testing were either 1 day old or in-vitro matured. Only 2% of the frozen zonae were lost and >75% of spermatozoa cryopreserved in this manner were recovered and prepared for intracytoplasmic sperm injection. The feasibility of cryopreserving a single spermatozoon was assessed. Fifteen motile spermatozoa were frozen in 15 zonae, of which 14 were recovered after thawing. Ten were injected into spare eggs, of which eight became fertilized. Spermatozoa recovered mechanically from human zonae fertilized the same proportion of oocytes as fresh fertile control spermatozoa. The recovery and fertilization rates with spermatozoa frozen in animal zonae were 87 and 78% respectively. The fertilization rate was marginally higher (P < 0.05) than that for spermatozoa frozen in human zonae, perhaps because the latter may have acrosome reacted more frequently. The zona pellucida appears to be an ideally suited sterile vehicle for storage of single spermatozoa.

Animals↗
PubMed1997

Ambient air and its potential effects on conception in vitro.

Incidences of chemical air contamination (CAC) are common in assisted reproductive technology, but not reported in peer review format. Justified fear of car and industrial emissions clearly exists among reproductive specialists, but standards for air contents and gaseous emission limits have not been reported. Here, we describe air sampling methods and assay systems which can be applied to any laboratory or laboratory item. It was found that unfiltered outside air may be cleaner than high efficiency particulate air filtration (HEPA) filtered laboratory air or air obtained from incubators, due to accumulation of volatile organic compounds derived from adjacent spaces or specific laboratory products such as compressed CO2, sterile Petri dishes and other materials or devices known to release gaseous emissions. Specific groups of products such as anaesthetic gases, refrigerants, cleaning agents, hydrocarbons and aromatic compounds such as benzene and toluene are described. The latter were shown to accumulate specifically in incubators. Isopropyl alcohol was the most dominant product found, though it was not used by the laboratory staff. Concentrations of this agent were low in incubator air, indicating that it was probably absorbed by the water in the pan or by culture medium. Measures to counter CAC are proposed, including the use of activated carbon filters and oxidizing material placed in the central air handling systems, in separate free-standing units or even inside the incubators.

Air Pollution, Indoor↗
PubMed1997

Comparative effects of clindamycin and ampicillin on superantigenic activity of Streptococcus pyogenes.

We have tested the ability of Streptococcus pyogenes to produce streptococcal pyrogenic exotoxin A (SPEA) and superantigenic activity in the presence of sub-inhibitory concentrations of ampicillin and clindamycin. After 6 h of culture, SPEA production by S. pyogenes was higher in broths containing ampicillin (715.7 +/- 296.4 pg/10(6) cfu) than in broths containing clindamycin (167.1 +/- 31.9 pg/10(6) cfu), a difference that was not significant (P = 0.25). Promitogenic activity of bacterial supernatants was also greater in ampicillin-treated cultures (467.5 +/- 17.2 ccpm/10(6) cfu) than in clindamycin-treated cultures (169.2 +/- 8.9 ccpm/10(6) cfu), a difference that was highly significant (P = 0.0001). The data support the use of clindamycin in the treatment of streptococcal toxic shock-like syndrome, in order to inhibit superantigen synthesis.

Ampicillin↗
PubMed1997

Preoperative evaluation of patients with primary hyperparathyroidism: role of high-resolution ultrasound.

Unilateral parathyroid exploration with adenoma removal and identification of a normal parathyroid gland is an accepted surgical approach for the treatment of patients with primary hyperparathyroidism. This study prospectively evaluated the ability of high-resolution sonography to localize adenomas preoperatively and the effect of this localization on operative time and cost in 37 previously untreated patients undergoing surgical treatment for primary hyperparathyroidism. The sensitivity and positive predictive value of the sonographic examination were 84% and 90%, respectively. A true positive sonographic examination decreased surgical time by an average of 40 minutes and reduced operative charges by $564, including the cost of the test ($156). In experienced hands high-resolution sonography can be a cost-effective means of localizing parathyroid adenomas where unilateral exploration is considered an accepted surgical approach.

Adenoma↗
PubMed1997

Molecular epidemiology of human rotaviruses in Santiago, Chile.

BACKGROUND: Protective immunity against rotavirus infection is directed against antigenic epitopes on the outer capsid proteins VP7 and VP4. Our aim was to characterize the epidemiology of rotavirus antigenic types over time in Santiago, Chile. METHODS: We prospectively obtained 2097 stool samples for rotavirus testing, VP7 (G1 to G4) and VP4 (P4, P6, P8, P9) typing from children with diarrhea evaluated in emergency rooms of 5 base hospitals of Santiago. In addition 256 rotavirus-positive samples collected between 1985 and 1987 in the north health care area of Santiago were studied. RESULTS: Of 995 rotavirus-positive samples obtained 825 (82%) were typable for 1 or more VP7 types. G1 represented 81% of the G-typed samples during 1993 through 1995 and 77% during 1985 through 1987, predominating in all health care areas. G2 was next most common in all 5 areas, representing 6 to 23% of typed samples, with 1 area, the Southeast concentrating a significantly higher number of G2 infections. G2 declined from 35% of rotavirus-positive samples in 1993 to 0% in 1995 (P < 0.001), and from 25% to 2% in the north health care area from 1985 to 1987 (P < 0.001). G4 was uncommon and significantly more prevalent in 1985 through 1987 than in 1993 through 1995 (7% vs. 3%, P = 0.015). G3 was not detected. G1P8 (53%) and G2P4 (16%) combinations were by far the most commonly detected G-P associations. CONCLUSIONS: In Santiago, Chile, rotavirus antigenic type G1P8 has been highly prevalent and G2P4 has circulated in cycles. Differences in epidemiology of rotavirus antigenic types worldwide may prove to be relevant in efficacy of rotavirus vaccines.

Child↗
PubMed1997

Synergy and specificity in induction of gene activity by proinflammatory cytokines: potential therapeutic targets.

This review focuses on two important routes of gene activation mediated by proinflammatory cytokines, which serve as a paradigm for mechanisms of specificity and synergy in cytokine regulated gene activity: STAT (signal transducers and activators of transcription)-related gene activation and the pathways initiated by proinflammatory cytokines leading to the activation of the NF-kappa B family of transcriptional activators. The proinflammatory cytokine cascade is involved in both the normal immune response and in the pathogenesis of several disease states. An understanding of specificity and synergism in these pathways offers an opportunity to develop novel therapeutic drugs for the selective manipulation of these disease processes. Such potential targets are examined in this review.

Animals↗
PubMed1997

Cell lines susceptible to infection are permeabilized by cleaved and solubilized outer layer proteins of rotavirus.

It has previously been shown that trypsinized triple-layered particles of rotavirus induce destabilization of liposomes and membrane vesicles in the absence of Ca2+, a condition which leads to solubilization of the outer capsid proteins of the virus. In this work, we have studied the relationship between outer capsid solubilization and permeabilization of membrane vesicles, monitoring particle and vesicle size simultaneously by changes in light scattering. Permeabilization of intact cells induced by solubilized outer capsid proteins was monitored by following the rate of entry of ethidium bromide into the cells. Solubilized outer capsid proteins separated from double-layered particles induced vesicle permeabilization. Solubilization of the outer capsid preceded and was required for vesicle or cell permeabilization. Membrane damage induced by rotaviral outer proteins was not repaired upon addition of 1 mM Ca2+ to the medium. Rotavirus infection and cell permeabilization were correlated in six different cell lines tested. This phenomenon might be related to the mechanism of virus entry into the cell. We propose a new model for rotavirus internalization based on the permeabilizing ability of outer capsid proteins and the cycling of trapped calcium in the endosomal compartment.

Capsid↗
PubMed1997

Solubilized and cleaved VP7, the outer glycoprotein of rotavirus, induces permeabilization of cell membrane vesicles.

It has been previously shown that rotavirus triple-layered particles induce permeabilization of liposomes and membrane vesicles. These effects were mediated by one or both of the solubilized outer-capsid proteins, VP4 and VP7. Permeabilization was dependent on trypsin treatment of the viral particles, suggesting that VP4 was involved. To analyse the respective roles of the outer-capsid proteins in this permeabilization process, we have used membrane vesicles loaded with carboxyfluorescein and virus-like particles derived from insect cells co-expressing various sets of capsid proteins. Virus-like particles containing VP2, VP6 and VP7 (VLP2/6/7) are as efficient in permeabilizing vesicles as triple-layered particles. As with double-layered particles, virus-like particles made of VP2 and VP6 had no effect on vesicle permeabilization. Permeabilization of membrane vesicles required trypsinization of the VP7 solubilized from VLP2/6/7. These results show that solubilized and trypsinized VP7 is able to induce membrane permeabilization, independently of the presence of VP4.

Antigens, Viral↗
PubMed1997

The role of nitric oxide in experimental murine sepsis due to pyrogenic exotoxin A-producing Streptococcus pyogenes.

Nitric oxide (NO) produced by inducible NO synthase (iNOS) mediates hypotension in endotoxemia. In this study, NO induction by a toxin-producing Streptococcus pyogenes isolate, H250, and by recombinant streptococcal pyrogenic exotoxin A (rSPEA) has been examined, both in vitro and in vivo. Streptococcal supernatants, but not rSPEA, induce production of nitrite by murine macrophages when both are coincubated with gamma interferon. Intraperitoneal injection of rSPEA did not cause significant production of NO. However, an elevated level of nitrate in serum was detected in a model of streptococcal fasciitis due to live H250. iNOS was localized to Kupffer cells, hepatocytes, and renal tubular cells by immunostaining. Administration of a NOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA), reduced peak concentrations of nitrate in serum but did not affect survival. NO is induced by H250, both in vitro and in vivo, mainly via SPEA-independent mechanisms. In this model, iNOS is expressed predominantly in the liver. Furthermore, in this model L-NMMA is not protective.

Animals↗
PubMed1997

In vivo and in vitro phosphorylation of rotavirus NSP5 correlates with its localization in viroplasms.

NSP5 (NS26), the product of rotavirus gene 11, is a phosphoprotein whose role in the virus replication cycle is unknown. To gain further insight into its function, we obtained monoclonal antibodies against the baculovirus-expressed protein. By immunoprecipitation and immunoblotting experiments, we showed that (i) NSP5 appears in many different phosphorylated forms in rotavirus-infected cells; (ii) immunoprecipitated NSP5 from rotavirus-infected cells can be phosphorylated in vitro by incubation with ATP; (iii) NSP5, produced either by transient transfection of rotavirus gene 11 or by infection by gene 11 recombinant vaccinia virus or baculovirus, can be phosphorylated in vivo and in vitro; (iv) NSP5 expressed in Escherichia coli is phosphorylated in vitro, and thus NSP5 is a potential protein kinase; and (v) NSP5 forms dimers and interacts with NSP2. The intracellular localization of NSP5 in the course of rotavirus infection and after transient expression in COS7 cells has also been investigated. In rotavirus-infected cells, NSP5 is localized in viroplasms, but it is widespread throughout the cytoplasm of transfected COS7 cells. NSP5 produced by transfected COS7 cells did not acquire the multiphosphorylated forms observed in rotavirus-infected COS7 cells. Thus, there is a tight correlation between the localization of NSP5 in the viroplasms and its protein kinase activity in vivo or in vitro. Our results suggest that cellular or viral cofactors are indispensable to fully phosphorylate NSP5 and to reach its intracellular localization.

Animals↗