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H Wood

Publications and source records attributed to H Wood.

18 recordsLinked to original sources

Evolutionary conserved nucleotides within the E.coli 4.5S RNA are required for association with P48 in vitro and for optimal function in vivo.

E.coli 4.5S RNA is homologous to domain IV of eukaryotic SPR7S RNA, the RNA component of the signal recognition particle. The 4.5S RNA is associated in vivo with a 48kD protein (P48), which is homologous to a protein component of the signal recognition particle, SRP54. In addition to secondary structural features, a number of nucleotides are conserved between the 4.5S RNA and domain IV of all other characterised SRP-like RNAs from eubacteria, arachaebacteria and eukaryotes. This domain consists of an extended stem-loop structure; conserved nucleotides lie within the terminal loop and within single-stranded regions bulged from the stem immediately preceding the loop. This conserved region is a candidate for the SRP54/P48 binding site. To determine the functional importance of this region within the 4.5S RNA, mutations were introduced into the 4.5S RNA coding sequence. Mutated alleles were tested for their function in vivo and for the ability of the corresponding RNAs to bind P48 in vitro. Single point mutations in conserved nucleotides within the terminal tetranucleotide loop do not affect P48 binding in vitro and produce only slight growth defects. This suggests that the sequence of the loop may be important for the structure of the molecule rather than for specific interactions with P48. On the other hand, nucleotides within the single-stranded regions bulged from the stem were found to be important both for the binding of P48 to the RNA and for optimal function of the RNA in vivo.

Alleles

Signal-sequence recognition by an Escherichia coli ribonucleoprotein complex.

Hydrophobic signal-sequences direct the transfer of secretory proteins across the inner membrane of prokaryotes and the endoplasmic reticulum membranes of eukaryotes. In mammalian cells, signal-sequences are recognized by the 54K protein (M(r) 54,000) of the signal recognition particle (SRP) which is believed to hold the nascent chain in a translocation-competent conformation until it contacts the endoplasmic reticulum membrane. The SRP consists of a 7S RNA and six different polypeptides. The 7S RNA and the 54K signal-sequence-binding protein (SRP54) of mammalian SRP exhibit strong sequence similarity to the 4.5S RNA and P48 protein (Ffh) of Escherichia coli which form a ribonucleoprotein particle. Depletion of 4.5S RNA or overproduction of P48 causes the accumulation of the beta-lactamase precursor, although not of other secretory proteins. Whether 4.5S RNA and P48 are part of an SRP-like complex with a role in protein export is controversial. Here we show that the P48/4.5S RNA ribonucleoprotein complex interacts specifically with the signal sequence of a nascent secretory protein and therefore is a signal recognition particle.

Bacterial Proteins

Detection of occult lymph node metastases in malignant melanoma.

An accurate prediction of the prognosis of melanoma is dependent on the ability to determine the presence or absence of malignant cells in lymph nodes. Our laboratory has developed a method for screening lymph node sections for the presence of micrometastatic disease. The nodes obtained from regional lymphadenectomies are dissected free from surrounding tissue and divided into halves. One-half is subjected to routine histopathological evaluation and the other one-half is disrupted into a single cell suspension and placed into cell culture. Lymph nodes from 19 patients with malignant melanoma were evaluated by tissue culture. Tumor cells were isolated in 125 of the 173 nodes examined. In contrast, only 14 of these nodes were initially found positive for melanoma by routine pathological evaluation. All lymph nodes positive for melanoma by pathological evaluation were also culture positive. Examination by cell culture has led to an upgrade in the stage of disease in 58% of the histologically node-negative patients. In addition, 43% of the patients who were histologically node negative but culture positive have recurred, suggesting some clinical correlation. These results indicate that the tissue culture method for detecting micrometastatic lymph node disease may prove more reliable and sensitive than current techniques for determining the presence of occult melanoma.

Antigens, Neoplasm

Induction of c-fos mRNA in cerebral cortex by excitotoxin stimulation of cortical inputs: involvement of N-methyl-D-aspartate receptors.

In this study we have characterized the induction of c-fos mRNA in cerebral cortex in response to unilateral kainate injection into the nucleus basalis. This treatment is associated with an intense stimulation of the ascending pathway and the subsequent induction of ornithine decarboxylase (ODC) enzyme activity and ODC mRNA in ipsilateral cerebral cortex which is sensitive to treatment with MK-801 and dihydropyridine antagonists. Unilateral injection of kainate into nucleus basalis caused a marked induction of c-fos mRNA in ipsilateral cortex which was detectable at 1 h, reached a maximal value at 8 h where c-fos mRNA levels were 16 times those in unoperated animals and then returned to control values by 24 h. However, the early induction of c-fos mRNA at 1 h was not related to a specific effect of kainate since at this time point, sham-operated animals also showed a significant increase in the level of c-fos mRNA in ipsilateral cerebral cortex. No significant induction of c-fos mRNA was detected in ipsilateral cortex in sham-operated animals at 4 and 8 h after injection of vehicle. Treatment with the N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801 (3 mg/kg) significantly attenuated the response obtained at 4 h and 8 h after kainate injection by 73% and 55% respectively, but did not influence the level of c-fos mRNA induced at 1 h. Delaying administration of MK-801 by 30 min reduced the effectiveness of this treatment on the response obtained at 4 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Excitotoxin lesion of nucleus basalis causes a specific decrease in Go mRNA in cerebral cortex. Sensitivity to MK-801.

Lesions of the ascending cholinergic pathway from nucleus basalis are known to have profound effects on cortical function. In particular, a substantial potentiation of carbachol-stimulated polyphosphoinositide turnover is detected from 1 day after lesion and is maintained for several days before returning to normal by 1 month. In this study the effect of this lesion was investigated on levels of three G-protein alpha-subunit mRNAs. Excitotoxin lesion of the nucleus basalis caused a selective reduction in the levels of Go alpha mRNA in cerebral cortex ipsilateral to the lesion, Gs alpha and Gi alpha mRNA being unaffected. The maximal effect was obtained at 3 days after lesion where levels of Go alpha mRNA were decreased by 40% compared to sham-operated animals. Levels of Go alpha mRNA returned to normal values by 28 days. Treatment with MK-801 caused a significant attenuation of the decrease in Go alpha mRNA, indicating the involvement of NMDA receptors in this response.

Animals

Induction of ornithine decarboxylase mRNA in cerebral cortex in response to kainate lesion of nucleus basalis: involvement of NMDA receptors.

Excitotoxin lesion of nucleus basalis with kainate or ibotenate results in degeneration of cholinergic neurones and a subsequent cholinergic deficit in cerebral cortex ipsilateral to the lesion. This lesion is accompanied by a massive increase in ornithine decarboxylase (ODC) activity in ipsilateral cortex, which was further investigated in this study by assay of levels of ODC mRNA in cerebral cortex after lesion. Injection of kainate into the nucleus basalis induced a significant increase in ODC mRNA in ipsilateral cerebral cortex which was maximal at 8 h after lesion and declined to control levels by 72 h. This induction showed clear regional specificity and was completely prevented by injection of an N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801, at a dose of 1 mg/kg, 2 h after excitotoxin lesion. This study indicates that excitotoxin lesion causes an early and transient increase in ODC mRNA that is mediated by NMDA receptors and may represent a physiological response to injury.

Animals

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England

Lysosomal elastase and cathepsin G in beige mice. Neutrophils of beige (Chediak-Higashi) mice selectively lack lysosomal elastase and cathepsin G.

A profound decrease in activities of the two lysosomal serine proteinases, elastase, and cathepsin G, was found in neutrophils of four independent beige mutants. Elastase and cathepsin G activities were assayed with the specific synthetic substrates MeO-Suc-Ala-Ala-Pro-Val-MCA and Suc-Ala-Ala-Pro-Phe-pNA, respectively. The defect is intrinsic to cells of beige mice, since transplantation of bone marrow from normal to mutant mice restored normal proteinase activity, and normal mice transplanted with beige marrow produced neutrophils with a deficiency of proteinase activity. The loss of elastase and cathepsin G activity was confirmed by separation of [3H]diisopropylfluorophosphate-labeled proteins on denaturing gels, which also revealed that other serine proteinases are at normal levels in beige neutrophil extracts. The deficiency of lysosomal proteinase activity appears specific, in that four other common neutrophil lysosomal enzymes, plus the spectrum of major neutrophil proteins are not affected by the beige mutation. The deficiency of proteinase activity is likely not the primary genetic alteration of the beige mutation, since more than one proteinase is affected, and heterozygous F1 mice have normal rather than intermediate levels of both proteinases. The lowered proteinase activity may contribute to the high susceptibility of beige mice and Chediak-Higashi patients to infection.

Animals

Norfloxacin in the therapy of uncomplicated gonorrhea.

In an open study, 70 patients with uncomplicated anogenital Neisseria gonorrhoeae infection were evaluated to determine the efficacy and safety of a single oral dose of norfloxacin (800 mg). Norfloxacin cured all 31 male urethral and 25 endocervical infections. All 63 isolates of N. gonorrhoeae tested were inhibited by 0.05 microgram of norfloxacin per ml.

Child

The Shiers arthroplasty of the knee.

Thirty-one patients with rheumatoid arthritis were reviewed after Shiers arthroplasty of the knee joint for which the main indication was intractable pain. Seven patients had the operation done to both knees. This review was done to assess the long-term results two to seven years later. All patients were clinically and radiologically assessed, and our results showed that pain and instability recurred within eighteen months. Thus it is likely that a prosthesis which allows rotation, and in which the bearing surfaces are metal on plastic, will replace the Shiers prosthesis.

Arthritis, Rheumatoid

The use of footprints in assessing the results of operations for hallux valgus. A comparison of Keller's operation and arthrodesis.

One hundred and seventy feet have been reviewed after operations for hallux valgus; eighty-five had had arthrodesis of the first metatarso-phalangeal joint and eighty-five had had Keller's operation. Footprints were made in order to assess the patterns of weight-bearing on the big toe and on the lesser metatarsal heads. After arthodesis the big toe bore weight in 80 per cent compared with 40 per cent after Keller's operation. The ability to bear weight on the big toe is related to the presence of metatarsalgia and excessive weight-bearing on the lesser metatarsal heads. These complicaitons were seen more commonly after Keller's operation (particularly when more than one-third of the phalanx had been excised) than after arthrodesis.

Arthrodesis

Improved diagnosis of gonorrhea in women.

Single endocervical cultures are estimated to be 85-95% reliable in the diagnosis of gonorrhea; a properly obtained rectal culture increases this yield by 5%. In this prospective trial, the increased diagnostic yields from two first-day endocervical cultures and second-day endocervical and rectal cultures were examined. The value of cervical gram smears was also reviewed. During a six-month period, 748 women were evaluated on day 1, with 195 returning for second-day cultures. The mean increased diagnostic yields were 2.2% for the second first-day endocervical culture, 2.9% for second-day cultures, and 4.0% for rectal cultures.

Adolescent