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Biomedical subjects

L N Jones

Publications and source records attributed to L N Jones.

28 records · Page 2Linked to original sources

Studies of developing human hair shaft cells in vitro.

Methods for studying aspects of hair formation in vitro have been devised on the basis of isolating developing hair shaft cells. These cells were obtained using a sterile microdissection technique. Plucked anagen follicles were dissected free of surrounding tissues (inner and outer root sheaths), and presumptive hair shaft cells (including germinal epithelia) were cultured directly on mammalian fibroblasts or in media preconditioned by fibroblasts. Specimens were cultured either as dispersions or in whole tissue pieces. Trypsinized whole tissue specimens in culture were sometimes observed to form increased bulk, while dispersed cells appeared to elongate and form larger colonies. In sections of these colonies examined by transmission electron microscopy, intracellular hard keratin intermediate filaments (IFs) together with IF-matrix hard keratin complexes were observed. Radiolabelled cysteine [35S] was added to cultures (3-20 days), showing a continuing but reduced synthesis of hard keratin IF proteins (low-sulfur) over the period of study. Matrix protein (high-sulfur) production was drastically reduced after 3 days. Monoclonal antibodies directed against hair keratin IF components were used in Western transfers and immunofluorescent studies to help assess the specificity of proteins synthesized in culture. Our observations indicate that, with some refinement, the presently described methods enable preparation of hair shaft precursor cells suitable for observing certain hair-forming processes in vitro.

Cells, Cultured↗

Stroke and migraine in the Oxfordshire Community Stroke Project.

A total of 323 cases of first-ever stroke were registered in the first 2 years of the Oxfordshire Community Stroke Project. Of these patients, 244 (76%) had a stroke due to cerebral infarction. There was a past history of migraine headaches in 56 (17%) of the 323 cases of stroke and in 44 (18%) of the 244 cases of cerebral infarction. A past history of migraine headaches was no commoner in patients with stroke due to cerebral infarction than in those with stroke due to intracranial haemorrhage. One hundred and seventy-three (71%) patients with cerebral infarction had at least one risk factor for ischaemic stroke; the frequency of such risk factors was similar in patients with and without a history of migraine. In 7 (3%) of the 244 patients the cerebral infarction was presumed to be "migrainous"; however, only 3 of these 7 (1.2% of the 24) were free of risk factors for ischaemic stroke. If all 7 cases were considered migrainous, the incidence rate of first migrainous cerebral infarction was 3.36 per 100,000 per year (95% confidence limits 0.87-5.86). If only the 3 patients who were free of risk factors were included, the incidence was 1.44 per 100,000 per year (95% confidence limits 0-3.07).

Adult↗

Isolation of intermediate filament assemblies from human hair follicles.

We used developing human hair follicle cells for the isolation of hard alpha-keratin structural components. Intracellular dispersions examined by electron microscopy contained both individual alpha-keratin filaments and the tactoid-like filament assemblies observed in situ organized along subfibrillar arms of macrofibrils. The assemblies of average width 47 nm were composed of closely packed alpha-keratin filaments and originated from the initial filament arrays observed in sections of developing mammalian hair follicles. We have distinguished two types of assemblies: the para-like or hexagonally packed and the ortho-like spiral or whorl type. Axial banding extended across the width of filament assemblies, which suggested that hard alpha-keratin filaments pack in lateral register and form a lattice that contains interfilamentous bridges. We observed axial banding patterns with periods ranging from 20 to 22 nm, consistent with the 22-nm periodic structure deduced from x-ray diffraction studies and present in models proposed for hard alpha-keratin and other intermediate filaments. Preliminary biochemical studies of filaments and filament assemblies indicate that they consist of the closely related group of proteins (low-sulfur proteins) ubiquitous among extracts of hard mammalian keratins. Isolated hard alpha-keratin filament assemblies provide a new and valuable structural entity for investigating the assembly mechanisms involved in the formation of the filament-matrix framework found in hard mammalian keratin appendages.

Cytoskeleton↗

Studies on microfibrils from alpha-keratin.

alpha-Keratin microfibrils have been isolated from the presumptive cortex of developing mammalian hair follicles and purified by a centrifugation procedure. They have been obtained on a scale sufficient to enable their physical and chemical nature to be studied directly. The amino acid and protein compositions of the microfibril preparation closely resemble those of the low-sulphur protein fraction extracted from the fully keratinized fibre. High resolution electron microscopy of isolated microfibrils has indicated that they possess structural features in line with those observed in sectioned material.

Amino Acids↗

Hypertension: medical and nursing implications.

Hypertension is a major public health problem affecting 20 to 30 million people in the United States, many of whom are undetected, untreated, or inadequately treated. Because hypertension often is asymptomatic for decades, it must be systematically sought out in apparently well persons if the condition is to be treated in its earliest stages and complications prevented. With the recent advent of effective antihypertensive medications treatment is available, once the patient has been diagnosed. Physicians and nurses have a major responsibility to direct the patient into treatment and to assure the effectiveness of his follow-up care. Noncompliance is a direct challenge to everyone seeing the patient. The patient must be motivated to accept and continue therapy through education and, most important, by the establishment of a good interpersonal relationship with the nurse and/or the physician.

Aftercare↗

The isolation and characterization of alpha-keratin microfibrils.

A method of isolating alpha-keratin microfibrils which avoids the degradation previously associated with the use of chemical, physical or enzymic procedures has been developed. Electron microscope studies of the isolation procedure establish that the microfibrils originate from the presumptive cortical cells. A purification procedure, monitored by electron microscopy, has enabled microfibrils to be isolated on a scale sufficient for chemical characterization. The amino acid composition of the microfibrils is very similar to that of low-sulphur protein fractions extracted from a range of hard mammalian keratins and thus provides direct experimental evidence for the assumption that the low-sulphur proteins comprise the microfibril in alpha-keratin.

Amino Acids↗

A novel protocol to identify mutations in patients with wiskott-Aldrich syndrome.

Mutations of WASP (Wiskott-Aldrich syndrome protein) underlie the severe immunodeficiency/platelet disorder Wiskott-Aldrich syndrome (WAS) and its milder variant X-linked thrombocytopenia (XLT). The affected gene, a 12-exon structure on the X-chromosome, is expressed exclusively in blood cells. The encoded product WASP is a 502-amino-acid scaffolding protein that functions in stimulus-induced nucleation of actin filaments to form dynamic cell surface projections. To date, more than 150 mutations have been identified in 300 WAS/XLT kindred worldwide, generally through methodologies that include sophisticated exon screening steps such as single-strand conformation analysis. We report here a simpler protocol, which was designed for use in clinical settings to identify the mutations of newly diagnosed patients. The approach relies on directly sequencing amplified exons according to a staggered schedule based on statistical evaluation of previous cases. In a 2 1/2-year trial, samples from 28 consecutive patients were analyzed; these included 3 "blindly labeled" previously studied cases. The mutations that were identified include a broad spectrum (8 missense, 3 nonsense, 5 splice site mutations, 11 small insertion/deletions, 1 large deletion) and were broadly distributed (in 10 of the 12 exons). All mutations were verified and no discrepancies were encountered. Per patient, a mean of six DNA sequencing reactions and 6-7 h of staff effort sufficed for mutation identification and verification, indicating that the protocol is cost-effective. This cumulative experience demonstrates the suitability, reliability, and versatility of the new protocol.

Child, Preschool↗