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Biomedical subjects

J Herold

Publications and source records attributed to J Herold.

At least 37 records · Page 2Linked to original sources
PubMed1997

Effective amplification of 20-kb DNA by reverse transcription PCR.

Polymerase chain reaction has been applied to the amplification of long DNA fragments from a variety of sources, including genomic, mitochondrial, and viral DNAs. However, polymerase chain reaction amplification from cDNA templates produced by reverse transcription has generally been restricted to products of less than 10 kilobases. In this paper, we report a system to effectively amplify fragments up to 20 kilobases from human coronavirus 229E genomic RNA. We demonstrate that the integrity of the RNA template and the prevention of false priming events during reverse transcription are the critical parameters to achieve the synthesis of long cDNAs. The optimization of the polymerase chain reaction conditions enabled us to improve the specificity and yield of product but they were not definitive. Finally, we have shown that the same reverse transcription polymerase chain reaction technology can be used for the amplification of extended regions of the dystrophin mRNA, a cellular RNA of relatively low abundance.

Base Sequence↗
PubMed1997

Identification and subcellular localization of a 41 kDa, polyprotein 1ab processing product in human coronavirus 229E-infected cells.

The translation products of the human coronavirus (HCV) 229E open reading frames 1a and 1b, the polyproteins 1a and 1ab, are processed by virus-encoded proteinases. One of the key enzymes in this process is a chymotrypsin-like enzyme, the 3C-like proteinase. In this study we have identified an ORF 1b-encoded, 41 kDa processing product in HCV 229E-infected cells by using a monoclonal antibody with defined specificity. We show that this polypeptide is released from polyprotein 1ab by 3C-like proteinase-mediated cleavage of the peptide bonds Gln-6110/Gly-6111 and Gln-6458/Ser-6459. Also, we have investigated the subcellular localization of the 41 kDa processing product. Immunofluorescence microscopy revealed a punctate, perinuclear distribution of the 41 kDa polypeptide in infected cells and an identical subcellular localization was observed for three additional pp1ab-derived polypeptides. In contrast, the virus nucleocapsid protein showed a homogeneous cytoplasmic localization.

Antibodies, Monoclonal↗
PubMed1996

Sub total unilateral oculomotor nerve palsy in a Le Fort I osteotomy.

A 35-year-old man developed a pupil sparing palsy of the oculomotor nerve after a standard Le Fort I osteotomy during which there had been more bleeding than usual. No definite cause was found but we suggest that it was caused by ischaemia of the nerve secondary to local injury by haematoma or instrumentation. The anatomical basis for this is discussed.

Adult↗
PubMed1996

Aspergillosis of the paranasal sinuses. A case report and radiographic review.

Aspergillosis of the paranasal sinuses is uncommon; however, its incidence in recent years has shown a marked increase, and it is believed that it may account for a number of cases of nonspecific sinusitis. It may present in one of four forms, one of which is the noninvasive Aspergillus mycetoma. One such case affecting the maxillary sinus is reported, and reference is made to its possible sequelae and management. The significance of its presentation as a radiodense focus within the antrum on plain radiographs and computed tomography is discussed, as is the use of magnetic resonance imaging for excluding aspergillosis from the differential diagnosis.

Adult↗
PubMed1995

Extraction of high quality DNA from bloodstains using diatoms.

A simple method is described for the extraction of high quality DNA for PCR amplification. The DNA was extracted by using Chelex-100 ion exchange resin or a special cell lysis buffer containing proteinase K. For further purification the DNA was bound to silica in the presence of a chaotrophic agent. Hence it is possible to unlimitedly wash the bound DNA and inhibitory substances are removed. By using diatoms as a source of silicates, this method is very economical and can therefore be used as a routine method.

Blood Stains↗
PubMed1995

Characterization of a human coronavirus (strain 229E) 3C-like proteinase activity.

The RNA polymerase gene of human coronavirus (HCV) 229E encodes a large polyprotein that contains domains with motifs characteristic of both papain-like cysteine proteinases and proteinases with homology to the 3C proteinase of picornaviruses. In this study, we have, first, expressed the putative HCV 229E 3C-like proteinase domain as part of a beta-galactosidase fusion protein in Escherichia coli and have shown that the expressed protein has proteolytic activity. The substitution of one amino acid within the predicted proteinase domain (His-3006-->Asp-3006) abolishes, or at least significantly reduces, this activity. Amino-terminal sequence analysis of a purified, 34-kDa cleavage product shows that the bacterial fusion protein is cleaved at the dipeptide Gln-2965-Ala-2966, which is the predicted amino-terminal end of the putative 3C-like proteinase domain. Second, we have confirmed the proteolytic activity of a bacterially expressed polypeptide with the amino acid sequence of the predicted HCV 229E 3C-like proteinase by trans cleavage of an in vitro translated polypeptide encoded within open reading frame 1b of the RNA polymerase gene. Finally, using fusion protein-specific antiserum, we have identified a 34-kDa, 3C-like proteinase polypeptide in HCV 229E-infected MRC-5 cells. This polypeptide can be detected as early as 3 to 5 h postinfection but is present in the infected cell in very low amounts. These data contribute to the characterization of the 3C-like proteinase activity of HCV 229E.

Amino Acid Sequence↗
PubMed1995

[Comparison of transdermal with oral hormone substitution: a multicenter study with a new matrix patch].

166 postmenopausal patients, aged 45-65 years at baseline, with moderate to severe climacteric symptoms were randomly allocated to an open-label, multi-center study which compared the efficacy, safety and acceptance of a transdermal 17 beta-estradiol matrix patch with an oral form of estrogen replacement therapy. In a cyclic sequential regimen, the transdermal system delivered 0.05 mg of estradiol/day. Oral dosages of conjugated equine estrogens were 0.625 mg/day. An oral progestin was also given for 11 days in each cycle for each group. A statistically significant reduction compared to baseline in the primary efficacy parameter, the mean number of hot flashes, occurred with a decrease in each group from 6 per day at baseline to 1 per day at 12 weeks; there was no statistically significant difference between the two groups. The incidence and severity of other postmenopausal symptoms, particularly sweating, difficulty in concentration and palpitations were reduced to a greater extent in the patch group without revealing a significant inter-group difference in the total symptom score. There were also no statistically significant differences in the mean serum estradiol and FSH concentrations between the two treatment groups after 12 weeks of therapy. A similar number of adverse events was observed in both groups. The most frequent adverse events were breast pain and under oral estrogen therapy, gastrointestinal complaints and weight increase. Skin irritation or dermatitis occurred infrequently in the patch group. In summary, the matrix patch represents at least as effective a therapy for postmenopausal symptoms as a standard oral estrogen.

Administration, Cutaneous↗
PubMed1995

[Nosocomial myiasis in Central Europe].

The authors describe a case of nosocomial myiasis of cavities caused by larvae of the fly Lucilia sericata (Meigen 1926), in a patient hospitalized at the resuscitation unit after a motor car accident where he suffered fractures of the skull and other injuries. On the fourth day in hospital in the nasal and oral cavity of the patient larvae of the 1st and 2nd instar of the mentioned species were detected. The ova were laid in hospital. L. sericata is the most frequent causal agent of tissue, cavity, ocular and urogenital human myiasis in Central Europe. In Czechoslovakia so far seven cases were recorded, incl. three nosocomial ones. Myiases usually attack immobile, injured or severely ill people in summer. It is important to draw attention to the hazard of myiases under exceptional conditions.

Adult↗
PubMed1994

Disseminated intravascular coagulopathy presenting as a bleeding tooth socket.

Disseminated intravascular coagulopathy (DIC) was diagnosed during haematological investigation of a patient with a persistent bleeding tooth socket. On further clinical examination an abdominal aortic aneurysm was found. No other cause for the DIC was detected. The DIC was treated and the socket healed. The patient refused to have definitive surgery for the aneurysm.

Aged↗
PubMed1993

An 'elaborated' pseudoknot is required for high frequency frameshifting during translation of HCV 229E polymerase mRNA.

The RNA polymerase gene (gene 1) of the human coronavirus 229E is approximately 20 kb in length and is located at the 5' end of the positive-strand genomic RNA. The coding sequence of gene 1 is divided into two large open reading frames, ORF1a and ORF1b, that overlap by 43 nucleotides. In the region of the ORF1a/ORF1b overlap, the genomic RNA displays two elements that are known to mediate (-1) ribosomal frameshifting. These are the slippery sequence, UUUAAAC, and a 3' pseudoknot structure. By introducing site-specific mutations into synthetic mRNAs, we have analysed the predicted structure of the HCV 229E pseudoknot and shown that besides the well-known stem structures, S1 and S2, a third stem structure, S3, is required for a high frequency of frameshifting. The requirement for an S3 stem is independent of the length of loop 2.

Amino Acid Sequence↗
PubMed1993

Nucleotide sequence of the human coronavirus 229E RNA polymerase locus.

The nucleotide sequence of the human coronavirus 229E (HCV 229E) RNA polymerase gene and the 5' region of the genome has been determined. The polymerase gene is comprised of two large open reading frames, ORF1a and ORF1b, that contain 4086 and 2687 codons, respectively. ORF1b overlaps ORF1a by 43 bases in the (-1) reading frame. The in vitro translation of SP6 transcripts which include HCV 229E sequences encompassing the ORF1a/ORF1b junction show that expression of ORF1b can be mediated by ribosomal frame-shifting. The predicted translation products of ORF1a (454,200 molecular weight) and ORF1a/1b (754,200 molecular weight) have been compared to the predicted RNA polymerase gene products of infectious bronchitis virus (IBV) and murine hepatitis virus (MHV) and conserved structural features and putative functional domains have been identified. This analysis completes the nucleotide sequence of the HCV 229E genome.

Amino Acid Sequence↗
PubMed1993

Characterization of the human coronavirus 229E (HCV 229E) gene 1.

The sequence of the HCV 229E gene 1 has been determined and compared with the homologous sequences of the murine hepatitis virus and the avian infectious bronchitis virus. The coding sequence of gene 1 is 20,273 nucleotides in length. Within this coding region are two large open reading frames, ORF 1a (4,086 codons) and ORF 1b (2,687 codons) which overlap by 40 nucleotides. In the overlapping region, the genomic RNA can be folded into a pseudoknot structure, an element which is known to mediate -1 ribosomal frame-shifting in other coronaviruses. Assuming that -1 frame-shifting occurs at the HCV sequence UUUAAAC (nucleotides 12,514-12,520), the ORF 1a - ORF 1b product is predicted to be 6,758 amino acids in length. Our sequence analysis of the HCV 229E gene 1 has revealed a high degree of similarity within the ORF 1b of HCV, MHV and IBV, whereas ORF 1a is much less conserved. Elements which are believed to be necessary for specific (e.g. frame-shifting) and general (e.g. NTP-binding/helicase) transcriptional functions have been identified. This study completes the genomic sequence of HCV 229E which is 27.27 kb long and one of the largest known RNA genomes.

Base Sequence↗
PubMed1993

Molecular analysis of the human coronavirus (strain 229E) genome.

The nucleotide sequence of the human coronavirus strain 229E (HCV 229E) has been determined. This article describes the organization of the virus genome, the predicted viral gene products and the mechanisms which regulate viral gene expression. This information provides a basis to investigate the biology and pathogenesis of HCV.

Amino Acid Sequence↗
PubMed1986

Employment, attitudes toward employment, and women's health.

The relationships between self-reported general health, employment, and attitudes toward the employment of married women have been analyzed for a representative sample of married, middle-aged women in the United States. The cross-sectional data indicate that women who were in the labor force had better health than women who were out of the labor force. In addition, women whose labor force status was compatible with their attitudes toward employment tended to have better health than women for whom there was a discrepancy between labor force status and attitudes. Analysis of the longitudinal data indicate that several causal mechanisms contributed to the relationships observed in the cross-sectional data. For the women with favorable attitudes toward employment, it appears that being a housewife had more detrimental effects on health than being employed. In contrast, for the women with unfavorable or neutral attitudes toward employment, it appears that employment status did not affect health. For healthy women, being employed may have contributed to more favorable attitudes toward employment. Healthy women were more likely than unhealthy women to stay in the labor force. Thus, it appears that there are multiple causal relationships linking employment status, attitudes toward employment and women's health.

Adaptation, Psychological↗
PubMed1985

Part-time employment and women's health.

The relationships between part-time employment and self-reported health are analyzed for a national probability sample of middle-aged women. Overall, there was a tendency for full-time workers to have the best health, part-time workers to have an intermediate level of health, and women who were not in the labor force to have the poorest health. However, the pattern varied by race and marital status. For married black women, part-time workers reported poorer health than full-time workers. This appeared to be due in part to the lower socioeconomic status of the black part-time workers. For married white women, there was little or no difference in health or socioeconomic status between part-time and full-time workers. Additional hypotheses and relevant evidence are presented.

Black or African American↗
PubMed1982

How valid are self-report measures for evaluating relationships between women's health and labor force participation?

For a sample of white women aged 45-64, women who were out of the labor force had poorer self-reported health and higher mortality than women who were in the labor force. It has been hypothesized that women who are out of the labor force may tend to exaggerate their poor health in self-report data. However, no evidence of bias of this type was found in an analysis of the relationships between self-reported health and subsequent mortality. The validity of self-reports of illness as a reason for not seeking work has been assessed using data for a sample of 30-44 year old women who were out of the labor force. Over 90% of the women who gave illness or disability as their main reason for not seeking work had previous or contemporaneous independent, self-report evidence of poor health. The findings of this study and previous evidence indicate that poor health reduces the likelihood that a woman will join the labor force, and this is a major reason why women who are not in the labor force have poorer health than those who are in the labor force.

Employment↗