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Biomedical subjects

L Johnson

Publications and source records attributed to L Johnson.

At least 325 records · Page 18Linked to original sources

Chronic intracellular leucocytoclastic bacterial vitritis. A transmission electron microscopic study of the monocytes.

Vitritis, the presence of leucocytes in the acellular ocular vitreous, often accompanies ocular vasculitis (V). Non-cultivatable intracellular mollicutes (M), i.e. cell wall deficient bacteria, readily identified by transmission electron microscopy (TEM), are well known plant vascular pathogens. Recently similar intracellular polymorphonuclear leucocytes (PMNL) and lymphocytes (L) parasitising/destroying and mouse ocular and lethal systemic V producing M-like bacteria were reported to be a common cause of human ocular V. In this TEM restudy of 8 human and 3 mouse VM induced chronic ocular V materials severe nuclear and cytoplasmic alterations associated with generalized cytoskeletal nuclear-anchoring 0.005-0.010 micron particles, 0.01-0.05 micron diameter branching filaments and tubules, and trilaminar membrane bound complex internal structure containing 0.1-1.9 micron spherules, all indistinguishable from VM, were observed within 1-3% of the monocytes in all 11 specimens. The results indicate that VM also parasitise and destroy both human and mouse monocytes. VM induced alterations in monocytes, PMNL, and L are compared, and the monocyte nuclear damage/Rifampin beneficial effect relationship in VM induced ocular V is discussed.

Bacterial Infections↗

Efficacy of systemic phenytoin in the treatment of junctional epidermolysis bullosa.

Oral phenytoin has been shown to be effective in the treatment of recessive dystrophic epidermolysis bullosa (EB). To determine the possible efficacy of this drug in junctional forms of this disease, we treated four children (two with generalized atrophic benign EB [GABEB] and two with Herlitz disease) with two 16- to 20-week treatment periods with phenytoin separated by an eight- to 12-week drug-free period. Phenytoin was administered to achieve serum levels similar to those previously proved effective in treating recessive dystrophic EB. Serial counts were made of lesions (blisters, crusts, and erosions); surface areas of granulation tissue, when present, were also serially measured. In addition, changes in the percentage of surface area involved with scarring as well as changes in extracutaneous disease activity were similarly assessed. Both patients with GABEB showed excellent response during each course of phenytoin, with average reductions in lesion counts of 70% and 38%. In addition, subjectively, the healing times also appeared to be reduced. During the "wash-out" period, total lesional counts returned toward pretreatment levels. Both patients with GABEB subsequently have received phenytoin for an additional two years off of protocol, with continued excellent response. In contrast, the conditions of both patients with the Herlitz variant worsened while taking phenytoin, with increases in overall lesional counts of 31% and 72%. Also, the surface areas of exuberant granulation tissue either remained unchanged or increased during the course of phenytoin treatment. We conclude that phenytoin is effective in at least some patients with the GABEB subset of junctional EB, whereas no efficacy has been noted in patients with the more severe Herlitz variant.

Administration, Oral↗

Primary structure of an analog of crustacean pigment-dispersing hormone from the lubber grasshopper Romalea microptera.

An octadecapeptide capable of inducing pigment dispersion in the chromatophores of the fiddler crab Uca pugilator has been isolated from lyophilized heads of the lubber grasshopper Romalea microptera. This pigment-dispersing factor (PDF) was purified by gel filtration, ion-exchange chromatography, partition chromatography, and reversed-phase high performance liquid chromatography. Automated gas-phase sequencing, followed by the identification of the carboxyl-terminal amide, established the primary structure of this PDF as Asn-Ser-Glu-Ile-Ile-Asn-Ser-Leu-Leu-Gly-Leu-Pro-Lys-Leu-Leu-Asn-Asp-Ala- NH2. This structure was confirmed by chemical synthesis and by demonstrating that the synthetic and native PDF displayed identical chromatographic behavior and biological activity. The Romalea PDF is structurally related to the crustacean pigment-dispersing hormones (PDHs), which are also octadecapeptides. The sequence of grasshopper PDF shows 78% homology with beta-PDH (from the crabs U. pugilator and Cancer magister) and 50% homology with alpha-PDH (from the prawn Pandalus borealis). This study provides the first direct chemical evidence for the structural relatedness of insect PDF to the crustacean PDHs, thus identifying them as an authentic family of arthropod peptides.

Amino Acid Sequence↗

Acute, whole-body microwave exposure and testicular function of rats.

Male Sprague-Dawley rats were exposed for 8 h to continuous-wave microwave radiation (MWR, 1.3 Ghz) at a mean specific absorbed dose rate of 9 mW/g. MWR exposure and sham-irradiation took place in unidirectionally energized cylindrical waveguide sections, within which the animals were essentially unrestrained. The MWR treatment in this setting was determined to yield an elevation of deep rectal temperature to 4.5 degrees C. The animals were taken for analysis at 6.5, 13, 26, and 52 days following treatment, which corresponded to .5, 1, 2, and 4 cycles of the seminiferous epithelium. Net mass of testes, epididymides, and seminal vesicles; daily sperm production (DSP) per testis and per gram of testis; and the number of epididymal sperm were determined. The levels of circulating follicle-stimulating hormone (FSH) and leutinizing hormone (LH) were derived via radioimmunoassay of plasma samples taken at the time of sacrifice. Despite the evident acute thermogenesis of the MWR at 9 mW/g, no substantial decrement in testicular function was found. We conclude that, in the unrestrained rat, whole body irradiation at 9 mW/g, while sufficient to induce evident hyperthermia, is not a sufficient condition for disruption of any of these key measures of testicular function.

Animals↗

DNA deletions in mild and severe Becker muscular dystrophy.

The DNA of 33 patients diagnosed as suffering from Becker muscular dystrophy (BMD) has been probed with cloned DNA sequences from Xp21, known to reveal DNA deletions in patients suffering from the more severe Duchenne muscular dystrophy (DMD). Two BMD cases showed clear deletions. A third case gave aberrant band sizes, which further analysis showed to be caused by a small deletion. This suggests that deletions in DXS164 occur approximately as frequently in BMD as they do in DMD. Of the two cases showing large deletions, one is at the severe end of the Becker clinical spectrum, whilst the other is a classical Becker-type dystrophy. The fact that loci defined by probes commonly deleted in classical DMD patients are also deleted in BMD patients of varying severity is strong additional evidence that these disorders are allelic, and further justifies the use of probes with defined linkage relationships to DMD also being used for counselling in BMD families.

Adolescent↗

Effects of age, season and active immunization against estrogen on serum prolactin concentrations in stallions.

Prolactin concentrations in stallion serum were measured by radioimmunoassay based on antiserum generated against equine prolactin and radioiodinated canine prolactin. Prolactin concentrations in serum collected from 152 stallions at a slaughterhouse were higher (P less than .01) in summer than in winter (5.7 +/- .15 vs 2.0 +/- .17 ng/ml). Moreover, there was an effect of age (P less than .02) in the analysis of variance; there was no interaction between age and season. In general, prolactin concentrations increased with age up to 3 to 5 years. Samples of serum collected from five control and five estrogen-immunized stallions from 6 to 26 months of age also were assessed for prolactin concentration. Active immunization against estrogen had no effect (P greater than .10) on prolactin concentrations. In contrast, whether analyzed by age or by month of the year, prolactin concentrations varied with time (P less than .01) in these stallions. Averaged over both groups, concentrations were high during summer and low during winter and were also gradually increasing with time as indicated by a linear trend (P less than .01) in the data. It is concluded that serum prolactin concentrations in stallions 1) are seasonal, as in many other seasonally breeding species, 2) in general are highest in older stallions and lowest in young stallions and 3) are not affected by active immunization against estrogens.

Aging↗

Quality assurance in home health agencies: the role of the clinical pharmacist.

Drug therapy for home care patients, most of whom are elderly, is becoming increasingly complex. In addition, these patients are usually left to their own resources to manage drug therapy. Clinical pharmacy services are therefore a vital component to assuring and maintaining positive therapeutic outcomes in home care. The Visiting Nurse Association (VNA) of Baltimore includes clinical pharmacy in its array of existing home health care services. The VNA system is described and two case studies are presented that illustrate how clinical pharmacy services can directly affect quality of care.

Aged↗

A receptor-binding region in human choriogonadotropin/lutropin beta subunit.

Synthetic fragments have not been widely used thus far to evaluate structure-activity relations in the glycoprotein hormones. We prepared a series of peptides representing the intercysteine "loop" sequence (residues 38-57) in human choriogonadotropin (hCG) and lutropin (hLH) beta subunits, anticipating that it might be oriented toward the surface and accessible to receptors. The peptides were characterized chemically and tested for bioactivity by binding to rat ovarian membrane receptor and stimulation of Leydig cell testosterone production. The hCG beta-(38-57) and hLH beta-(38-57) peptides inhibited binding of 125I-labeled hCG half-maximally at 1.51 X 10(-4) and 2.03 X 10(-5) M, respectively, while other peptide hormones and fragments from elsewhere in the beta subunit were inactive. Both peptides stimulated testosterone production, with half-maximal responses at 3.55 X 10(-5) M (hCG) and 2.18 X 10(-5) M (hLH). By radioimmunoassay with an antibody to thyroglobulin-conjugated hCG beta-(38-57) peptide, native hCG and beta subunit were highly reactive, as were the reduced and carboxymethylated subunit and peptide. Helical-wheel projection predicted an amphipathic region in the N-terminal portion of the 38-57 sequence, and circular dichroic measurements showed an increase in ordered structure, especially alpha-helix, when the 38-57 peptides were transferred from an aqueous to a more lipophilic (90% trifluoroethanol) environment. These results indicate that the 38-57 region of beta subunit is exposed on the surface and constitutes a component in the receptor-binding domain for hCG and hLH. A region of amphipathic-helical structure in the 38-57 sequence may promote hormone-receptor interactions in a manner proposed for several other peptide hormones.

Amino Acid Sequence↗

Seminiferous tubules and daily sperm production in older adult men with varied numbers of Leydig cells.

Previous studies of adult men have failed to reveal a relationship between numbers of Leydig cells in the testes and rates of sperm production, perhaps because of a functional excess of these cells in younger men. Hence, a possible relationship between Leydig cell numbers and sperm production was sought in 50 older men, aged 50-90 years, in whom the Leydig cell population had been depleted by age-related attrition. When these men were sorted by increasing numbers of Leydig cells per man into two, three, or five groups, no difference could be found between or within these groups when daily sperm production per man (DSP); seminiferous tubular volume, diameter, or length; or seminiferous epithelial volume was examined. Furthermore, no significant correlation could be detected between Leydig cell numbers and DSP in these 50 men. The only relationship between numbers of Leydig cells and spermatogenesis appeared to be a threshold effect, in that men with fewer than 60 million Leydig cells (4 in this study) had drastically reduced DSP. Men with few Leydig cells tended to have larger Leydig cells, and the increased size was due to more cytoplasm instead of nucleoplasm. There were weak but significant positive correlations between total Leydig cell cytoplasm per man and DSP and between average size of a Leydig cell and DSP. These findings suggest that a relationship may exist between sperm production and the amount of cytoplasm containing testosterone-producing organelles in surviving Leydig cells of older men.

Aged↗

Partial purification and characterization of dynein adenosine triphosphatase from bovine sperm.

Outer dynein arm polypeptides that possess Mg+2-adenosine triphosphatase (ATPase) activity have been extracted from the flagellar axonemes of demembranated bovine sperm. Electron microscopy of intact and salt-extracted sperm demonstrates a relatively selective removal of the outer dynein arms. The salt extract contains a specific ATPase activity of 55 nmoles inorganic phosphate (Pi)/min/mg protein. Sucrose density gradient centrifugation of this extract results in a 6-fold increase in specific activity of ATPase (333 nmole/Pi/min/mg protein), which sediments as a single 13S peak. Concomitant with the increase in specific activity, there is enrichment of three high molecular weight polypeptides (Mr greater than 300,000) characteristic of dynein heavy chains. ATPase activities in the initial extract and in the 13S peak are inhibited by concentrations of vanadate and erythro-9-[3-2-(hydroxynonyl)]adenine similar to those that inhibit ATPase activity in sea urchin sperm dynein. These findings indicate that outer arm dynein ATPase can be extracted and partially purified from bovine sperm.

Adenosine Triphosphatases↗

Effect of photoperiod on the size of the Leydig cell population and the rate of recruitment of Leydig cells in adult Syrian hamsters.

The number of Leydig cells was determined by stereologic procedures in adult Syrian hamsters housed in long days (14L:10D) to maintain testicular activity (active), in short days (5L:19D) for 12-13 wk to induce testicular regression (photoperiod-induced regressed), or in short days for a period of 21 wk or more to allow spontaneous gonadal recrudescence (spontaneously recrudesced). Testes were removed, sliced, fixed, embedded in Epon 812, and observed by bright-field microscopy. Testicular and seminal vesicle weights, plasma testosterone concentration, total Leydig cell volume per testis, and volume of single Leydig cell were greater (p less than 0.01) in active and recrudesced animals than in regressed animals. The density of Leydig cells was greater in the regressed testes, but the total number per testis was not influenced by photoperiod. In Experiment 2, the rate of recruitment of Leydig cells was determined in 5 adult hamsters exposed to long days (active) or 5 hamsters whose testes were regressed by exposure of animals to short days for 13 wk followed by long-day exposure to initiate testicular growth (photoperiod-induced recrudescing). Hamsters were injected for 3 days/wk for 3 wk with tritiated thymidine, 0.5 or 1 microCi/g body weight. Testes were fixed and tissues prepared, as above, and processed for autoradiography. Again, the photoperiod did not influence the number of Leydig cells per testis. Labeling of Leydig cell nuclei revealed that recruitment of new Leydig cells occurred at approximately 1.3% per day in recrudescing testes but also occurred at approximately 0.6% per day in active testes. Without change in the total number of Leydig cells, new Leydig cells were added continually to the existing population in adult hamsters with either recrudescing or active testes.

Animals↗

Quantification of human spermatogenesis: germ cell degeneration during spermatocytogenesis and meiosis in testes from younger and older adult men.

Germ cell degeneration during spermatocytogenesis and meiosis was investigated to explain the age-related decline in daily sperm production (DSP). Numbers of Types A-dark, A-pale, and B-spermatogonia, potential daily sperm production per g parenchyma (PDSP) based on type B-spermatogonia, early primary spermatocytes, and late primary spermatocytes, and DSP per g based on early spermatids were determined in 15 men aged 20 to 48 yr (mean +/- SEM, 33 +/- 2 yr) and 15 men aged 52 to 90 yr (65 +/- 3 yr). Testes obtained within 15 h of death (largely due to trauma or heart failure) were perfused vascularly with glutaraldehyde. The number of each cell type per g parenchyma was calculated as the product of the percentage of nuclei in the parenchyma times a correction factor for section thickness and nuclear diameter divided by the volume of a single nucleus of that cell type. Paired testicular weight was lower (p less than 0.01) in older men (33 +/- 3 g) than in the younger men (49 +/- 3 g). Younger and older men had similar numbers of A-dark, A-pale, and B-spermatogonia per g parenchyma. PDSP based on late primary spermatocytes and DSP based on early spermatids were lower (p less than 0.01) in older men than in younger men. In younger men, PDSP was similar (p greater than 0.05) between B-spermatogonia and late primary spermatocytes, whereas DSP measured at the spermatid level was abruptly lower than that estimated from younger cell types. Older men showed reduction in PDSP between early and late primary spermatocytes, with further reduction occurring in DSP at the spermatid level.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

The application of linkage analysis to genetic counselling in families with Duchenne or Becker muscular dystrophy.

A total of 278 families of probands with Duchenne or Becker muscular dystrophy has been ascertained and offered genetic counselling. Linkage studies have been performed in these families using polymorphic DNA markers identifying loci linked to Duchenne and Becker muscular dystrophy. The clinical features of the probands are discussed: there was marked intrafamilial resemblance in the severity of the disease. We estimate that a complete study of potential carriers in these families would require analysis of samples from approximately 1400 subjects. The results of linkage studies tended to move women's carrier risk estimates (based on CK and pedigree data) towards the extremes of the risk categories, providing a more definitive risk estimate for 81% of the women who were previously in the middle range of carrier risk probabilities. About 70% of the families had only one affected member. Linkage analysis altered carrier risk estimates in 95% of sisters and aunts of index cases, but only affected estimates of the mother's carrier risks in about 11% of isolated cases. Even where linkage studies were not helpful in elucidating carrier risks, information could usually be obtained for use in prenatal diagnosis if required. We have assessed the attitudes to pregnancy and prenatal diagnosis of women at risk of being carriers of Duchenne or Becker muscular dystrophy and report 17 pregnancies in these women.

Amniocentesis↗

Effects of pulse-modulated microwave radiation and conventional heating on sperm production.

The effects on testicular function of pulse-modulated microwave radiation (PM MWR, 1.3 GHz) and of conventional heating were studied in the rat. Anesthetized adult males (Sprague-Dawley, 400-500 g) were treated then killed at specific intervals with respect to the 13-day cycle of the seminiferous epithelium. PM MWR at 7.7 mW/g (90 min) yielded a modest decline in daily sperm production (DSP) that derived primarily from effects on primary spermatocytes. PM MWR at 4.2 mW/g was ineffective. The mean intratesticular temperature during the former reached 40 degrees C and did not exceed 38 degrees C during the latter. MWR considerably in excess of 7.7 mW/g yielded decrements in virtually all germ cell types, with primary spermatocytes again being most markedly affected. Using conventional heating, intratesticular temperatures in excess of 39 degrees C for 60 min were required for significant decrements in DSP. Levels of circulating follicle-stimulating hormone and of leutinizing hormone were resistant to either treatment. We conclude that the damage threshold and the differential sensitivity of immature germ cells to PM MWR can be adequately explained by the consequent macroscopic heating.

Animals↗

Effect of seasonal changes in Leydig cell number on the volume of smooth endoplasmic reticulum in Leydig cells and intratesticular testosterone content in stallions.

Testes from 47 adult (4-20 years) stallions obtained in November-January (non-breeding season) and 41 adult stallions obtained in May-July (breeding season) were perfused with glutaraldehyde, placed in osmium and embedded in Epon 812. Percentage Leydig cell cytoplasm or nuclei in the testis was determined by point counting of 0.5 micron sections under bright-field microscopy. Testes from 6 randomly selected horses per season were processed for electron microscopy. The volume (ml) of SER/testis was calculated from the % SER in the cytoplasm % Leydig cell cytoplasm, and parenchymal volume. Number of Leydig cells was calculated from the % nuclei, parenchymal volume, histological correction factor, and volume of single nucleus. Intratesticular testosterone content was determined from the contralateral testis by radioimmunoassay. The volume of SER/g and testosterone/g tended to be higher in the breeding than non-breeding season. Leydig cell number/g, volume of SER/testis, testosterone/testis, and Leydig cell number/testis were significantly greater in the breeding than in the non-breeding season. Volume of SER/testis and testosterone/testis were related significantly to the cell number/testis, and SER/testis was related (P less than 0.05) to testosterone/testis. These results emphasize the importance of seasonal changes in the number of Leydig cells on the amount of SER available to produce testosterone and on testosterone content/testis in the stallion.

Animals↗

Chronic leucocytoclastic bacterial vitritis. A lymphocyte transmission electron microscopic study.

Ultrastructurally unique and distinctive non-cultivable human and mouse uveitis and vitritis producing bacteria (B) that parasitise, differentiate into both cell walled and cell wall deficient variants within, and destroy polymorphonuclear leucocytes (PMNL) were recently reported to be commonly present in the vitreous of chronic idiopathic vitritis patients. In this transmission electron microscopic restudy of the lymphocytes in 8 human and 3 mouse chronically inflamed ocular specimens, all of which had demonstrated those B within PMNL, in each specimen about 3-15% were larger atypical variants and 0.5-1.0% also harbored those B. The earliest detectable B, the 0.005-0.010 micron spherical elemental particles, had a paranuclear cytoskeletal location, where they produced cytoskeletal lysis and nuclear envelope and chromatin lesions. From that site the elemental particles proliferated, elongated, branched, and enlarged into tightly coiled 0.03-0.05 micron in diameter cell wall deficient tubules and elaborated ultrastructurally distinctive 0.5-0.7 micron cell walled B, eventually replacing the cytoskeleton and destroying the lymphocyte.

Animals↗