Rubella infection: a cause of foetal wastage.

A study was conducted on 180 women with 2 or more obstetric losses totalling 410 foetal wastages. Control cases included 100 women without obstetric losses. Significant titre of immunoglobulin M against rubella was found in 17.77% patients with 2 or more obstetric losses in contrast to 9% of control patients (p less than 0.005). Significant titre of immunoglobulin M was observed in cases of abortions (5.75%), preterm deliveries (12.67%), macerated stillbirths (8.19%), fresh stillbirths (8.69%) and neonatal deaths (8.34%).


Clinically malignant scalp and underlying bone tumours diagnosed by aspiration cytology: a study of 121 cases.

Fine needle aspiration biopsy cytology was employed in 121 cases of clinically malignant scalp and underlying bone tumours from July 1984 to March 1991. The age of the patients ranged from 2 to 76 years. There were 48 benign cases and 73 malignant lesions. The cytological findings correlated with histopathological analysis except in 5 cases in benign tumours (diagnostic accuracy being 89.6%) and in 5 cases in malignant tumours (diagnostic accuracy being 93.2%). The clinical history, the radiological parameters and the cytological analysis helped to avoid false positive report. No complication was encountered in this procedure.


Morphological changes in diseased gall bladder: a study of 415 cholecystectomies at Aligarh.

Morphological changes of gall bladder were studied in 415 cholecystectomy specimens. There was a preponderance of females (male to female ratio--1:6.5). The mean age of the cases was 43.6 years. Most of the cases (63.4%) were in the 4th and 5th decades of life. The average duration of illness was 2.8 years. Associated cholelithiasis was present in 85.3% cases. Gall-stones were of mixed variety in 78.2% cases, cholesterol type in 15.3% cases and both types were present in 6.5% cases. Chronic cholecystitis was the main histological diagnosis (50.8%). Other lesions observed were adenomyomatosis (8.2%), adenomatous hyperplasia (10.1%), granulomatous cholecystitis (4.1%), cholesterosis (2.7%), acute cholecystitis (4.1%), acute on chronic infection (10.8%), sub-acute cholecystitis (2.4%) and carcinoma gall bladder (6.8%). The frequency of Rokitansky-Aschoff's sinuses was closely related with the degree of inflammatory response. In 13 (6.2%) cases the diagnosis of chronic follicular cholecystitis was made. All the cases of cholesterosis were multiparous females and of younger age. Of the malignant lesions, adenocarcinoma was the commonest (96.4%).


Cervical spine injury: how easy to miss!

Acute injuries of the cervical spine are not uncommonly missed on the initial examination. In a study of 48 patients of acute injury of the cervical spine, the diagnosis was missed initially in 8 patients. The common reasons for missed diagnosis had been, head injury in 3 patients, polytrauma in 2, inadequate or improper radiologic examination in 2 and erroneous diagnosis in one case. Neurologic status was worsened in one patient because of delayed diagnosis.


Processing of structural polypeptides of infectious flacherie virus of the silkworm, Bombyx mori: VP1 and VP4 are derived from VP0.

Infectious flacherie virus of the silkworm, Bombyx mori, contains five major polypeptides termed VP0, VP1, VP2, VP3, and VP4 in the order of descending molecular weight. Immunoblot analysis with specific antisera against each of these structural polypeptides showed that antisera against VP1 and VP4 unequivocally reacted with VP0 as well as their homologous structural polypeptides. Limited proteolysis of VP0 and VP1 by Staphylococcus aureus V8 protease gave several common polypeptide fragments. Amino acid sequence analysis showed that VP0 and VP4 shared a common NH2-terminal amino acid sequence. These results indicate that VP1 and VP4 are generated from VP0 and that VP4 occupies the NH2-terminal portion of VP0.


Relationship of hemolymph phenol oxidase and mosquito age in Aedes aegypti.

Monophenol oxidase (MPO) and diphenol oxidase (DPO) activity in hemocytes and cell-free plasma perfused from 7-, 14-, 21-, and 28-day-old Aedes aegypti mosquitoes were compared. A progressive decrease of enzyme activity was detected as mosquito age increased, and this decrease was significant in both hemocytes and cell-free plasma when mosquitoes were 28 days old as compared with that found in 7-day-old mosquitoes. There was no significant difference in total hemolymph protein as mosquito age increased. Although this decreased MPO and DPO activity might be partially responsible for the reduced immune response against filarial worms previously reported for older mosquitoes, other factors undoubtedly play a significant role.


Effect of azadirachtin on the development of Trypanosoma cruzi in different species of triatomine insect vectors: long-term and comparative studies.

Studies were carried out on the effects of azadirachtin on the course of Trypanosoma cruzi infection in the gut of different triatomine vector species. In Rhodinus prolixus the development of T. cruzi clone Dm28c decreased in a dose-dependent manner, and the ED50 of this inhibitory effect was 0.25 microgram azadirachtin/ml bloodmeal. Using this insect, we conducted a long-term experiment which showed that azadirachtin (1.0 microgram/ml bloodmeal) completely blocks the development of T. cruzi even 120 days after treatment with the drug, and after four infectant meals. Similarly, the elimination of T. cruzi in feces and urine was also completely blocked over a period of 50 days after infection in insects treated with azadirachtin. Fifth-instar larvae of R. prolixus, Triatoma infestans, and Dipetalogaster maximus, infected with different clone/strains of T. cruzi, displayed drastic inhibition of trypanosome development when treated with azadirachtin. The discussion of these results focuses on the possibility that azadirachtin may act directly on gut physiology and/or indirectly through the neurosecretory system.


Internalization of Ia molecules into Birbeck granule-like structures in murine dendritic cells.

Dendritic cells isolated from the draining lymph nodes of mice sensitized epicutaneously with hapten are potent antigen-presenting cells and contain Birbeck granules and cored tubules characteristic of antigen-activated epidermal Langerhans cells. We used immunogold labeling and transmission electron microscopy to follow the internalization of Ia molecules in these antigen-presenting cells. We found that Ia molecules were internalized into Birbeck granule-like structures in the antigen-activated dendritic cells. Computer reconstruction of serial sections of the dendritic cells demonstrated that these structures span the cytoplasm from the cell membrane to the nuclear membrane and are associated with lysosomes. The internalization of Ia molecules into these structures supports the hypothesis that the Birbeck granule-like structures are derived from the cell membrane and are involved in the antigen-processing/presenting function of the dendritic cells.


Identification and characterization of a cell surface proteoglycan on keratinocytes.

Proteoglycans fill the intercellular space between keratinocytes but their structure and function are not well understood. We have identified and partially characterized one intercellular proteoglycan on human keratinocytes, for which we propose the name epican (epidermal intercellular proteoglycan). Monoclonal antibodies (MoAb) were generated from a mixture of keratinocyte proteoglycans. One, designated MoAb17, identified the core protein of an intercellular proteoglycan that had an apparent mobility of greater than 250 kDa on Western blots. The core protein itself had an apparent mobility of 180 kDa following deglycosylation with trifluoromethanesulfonic acid. Enzymatic deglycosylation revealed that most core protein molecules were substituted with heparan sulfate but that some carried chondroitin sulfate instead. Smaller forms of the core protein were more abundant in tissue-culture medium than in cell extracts. This proteoglycan was localized by immunofluorescence to the intercellular space of the epidermis and the surface of keratinocytes in vitro, particularly at cell-cell contacts. MoAb17 did not react with protoglycans extracted from other skin cells, nor did it bind to basement membranes or connective tissue. Comparison of Western immunoblots using MoAb17 and antibodies to core proteins of other proteoglycans suggested that epican is not related to syndecan but is a member of the CD44 family.


Topography of proteoglycan and glycosaminoglycan free chain expression in 3T3 fibroblasts and human keratinocytes.

Synthesis of heparan sulfate-free chains by human keratinocytes is upregulated during terminal differentiation. The cellular location of this product class and the significance of the differentiation effect are unknown. Differential plasma membrane shearing with cationized colloidal silica was used to evaluate the compartmentalization of the heparan and chondroitin sulfate free chains and their respective proteoglycans in 3T3 fibroblasts and human keratinocytes. The method exploits the topologic segregation of plasma membranes of adherent cells into ventral, dorsal, and intracellular domains and the selective binding of the silica to the dorsal membranes, which by shearing can be separated from ventral membranes adherent to the substratum. Analysis of membrane preparations from sheared cells that had been prelabeled with [35S]-sulfate revealed the proteoglycans to be predominantly ventral, at which location a matrix binding function could be accommodated. Proteoglycans were also recovered from dorsal and intracellular membranes, suggesting active trafficking between intra- and extra-cellular sites. In contrast, the major fraction of heparan and chondroitin sulfate free chains was either cytosolic or associated with intracellular membranes, with the remaining approximately 20% segregated to dorsal and ventral membranes. These results suggest different cellular functions for the proteoglycans and glycosaminoglycan free chains. The partial localization of the free chains to peripheral membranes is compatible with our prior hypothesis that they arise by processing of precursor proteoglycans on cell surfaces. Following this origin, the free glycosaminoglycan polymers could be available to bind ligands such as cytokines prior to transport to intracellular sites of action.


Syndecan-1, a cell-surface proteoglycan, changes in size and abundance when keratinocytes stratify.

In epidermis, keratinocytes in the basal cell layer differentiate, lose their attachment to the underlying extracellular matrix, and form extensive intercellular adhesions as they stratify. The alterations in cell-matrix and cell-cell adhesion required for keratinocyte stratification result from changes in the expression of numerous adhesion molecules. Syndecan-1, a member of a family of cell-surface proteoglycans, is known to bind cells to interstitial matrix. Syndecan-1 localizes to specific layers of mouse epidermal keratinocytes; its expression is modest in the basal layer, heavy in the suprabasal layers, but absent from the most superficial, terminally differentiated layers. This layer-specific difference suggests that syndecan-1 expression changes with keratinocyte differentiation. To assess this hypothesis, syndecan-1 expression was evaluated before and after calcium-induced stratification and differentiation. Cells growing as an unstratified monolayer express a higher molecular mass form of syndecan-1 than do stratified cells (modal relative mass of 160 kD versus 110 kD). This structural difference is due to larger and more heparan sulfate chains on syndecan-1 from monolayer cells. In addition, the amount of cell-surface syndecan-1 changes with stratification; stratified cultures show approximately 2.5 times more syndecan-1 per cell than do unstratified cultures, but do not significantly change the level of syndecan-1-specific mRNA. Thus, the structure and amount of syndecan-1 may be regulated to meet the changing adhesive requirements of stratifying keratinocytes.


Clinical, laboratory, and histopathologic indicators of the development of progressive acute graft-versus-host disease.

Graft-versus-host disease (GvHD) is the major cause of morbidity and mortality following bone marrow transplantation (BMT). The goal of this study of 69 cyclosporin-treated, allogeneic BMT patients was to identify early clinical, laboratory, or histopathologic indicators of the development of progressive, fatal GvHD. Peak values within 100 d of allogeneic BMT for total bilirubin, stool volume in a day, clinical stage of cutaneous GvHD (based on extent of rash), and overall clinical stage of GvHD (based on a combination of graft-versus-host reactions in the skin, liver, and gastrointestinal tract) were most useful (p less than 0.05, by logistic regression) in identifying those patients with clinically progressive and fatal GvHD. Peak values for each of these parameters were reached an average of 40 d or less after BMT. Each unit increase in peak clinical stage of rash (e.g., stage 2 versus stage 3) was associated with an odds ratio incremental risk of 5.8 for clinical progression of GvHD, and each tenfold increase in peak total bilirubin (e.g., 2 mg/dl versus 20 mg/dl) or stool output in a day (e.g., 100 cm3/d versus 1000 cm3/d) was associated with an incremental risk of 8.4 and 10.6, respectively, for a fatal outcome from GvHD. Number of exocytosed lymphocytes and dyskeratotic epidermal keratinocytes (DEK) per linear millimeter of epidermis, the presence of follicular involvement, and the degree of dermal perivascular lymphocytic infiltration in 121 skin biopsy specimens were not associated with the development of progressive or fatal GvHD. Pretransplant total body irradiation was associated (p = 0.03, by Mann-Whitney U testing) with an increased number of DEK in skin biopsy specimens taken less than 20 d after BMT. This study demonstrates that monitoring of total bilirubin, stool output, extent of rash, and overall clinical stage of GvHD is most useful during the first 40 d after BMT in formulating the prognosis of early acute GvHD in allogeneic BMT patients receiving cyclosporin.


Enhanced IL-4 production and IL-4 receptor expression in atopic dermatitis and their modulation by interferon-gamma.

The in vivo and in vitro immunomodulatory effects of interferon gamma (IFN-gamma) treatment on peripheral blood mononuclear cells (PBMC) from patients with atopic dermatitis (AD) and elevated IgE levels were studied. As part of a double-blind placebo-controlled clinical trial, 14 AD patients were treated with IFN-gamma (n = 7) or saline (n = 7) for 12 weeks. To assess the in vivo effects of IFN-gamma treatment on interleukin (IL)-4-dependent lymphocyte function, we assessed the proliferation of AD PBMC in response to IL-4. Prior to IFN-gamma treatment, AD PBMC had proportionately decreased proliferative responses to IL-4 when compared to IL-2. After 12 weeks of in vivo treatment with IFN-gamma, there was an increase of IL-4- but not IL-2-induced lymphocyte proliferation in seven of eight AD patients. To further study the immunologic basis of these observations, we studied the expression of IL-4 receptor (IL-4R) mRNA and the production of IL-4 by PBMC from AD patients. PBMC from AD patients expressed higher levels of IL-4R mRNA and produced significantly higher amounts of IL-4 than normal controls (p less than 0.05). More importantly, the in vitro addition of IFN-gamma caused significant reduction in both IL-4R and mRNA expression and IL-4 production of PBMC from AD and non-atopic controls. These data indicate that AD is characterized by an in vivo overstimulation of the IL-4-IL-4R pathway. The poor proliferative responses of untreated AD PBMC to exogenous IL-4 may be due to increased levels of endogenous IL-4 production with constant occupancy on the IL-4R. Furthermore, in vivo and in vitro treatment with IFN-gamma down-regulates this pathway.


Transforming growth factor-beta up-regulates the expression of the genes for beta 4 integrin and bullous pemphigoid antigens (BPAG1 and BPAG2) in normal and transformed human keratinocytes.

Three distinct proteins, namely, beta 4 integrins, and the 230-kDa (BPAG1) and 180-kDa (BPAG2) bullous and pemphigoid antigens, have been shown to co-localize with hemidesmosomes at the dermal-epidermal basement membrane zone. In this study, we examined the expression of the corresponding genes in cultures of normal and transformed human epidermal keratinocytes. The expression of these genes was detected by Northern and in situ hybridizations, and the expression of beta 4 integrins was also demonstrated by indirect immunofluorescence. The results indicated clearly detectable expression of all three genes in normal keratinocytes, whereas extremely low or undetectable levels of expression were noted in two transformed cell lines. Addition of TGF-beta 1 or TGF-beta 2 (10 ng/ml) up-regulated mRNA levels for all three proteins (up to 4.6 times). The increase by TGF-beta 1 was particularly striking in keratinocyte cultures incubated in the presence of low (0.15 mM) Ca++, and somewhat less pronounced in the presence of high (1.2 mM) Ca++. The increase in beta 4 integrin synthesis was also documented by enhanced immunosignal of the corresponding epitopes. These results indicate that the three hemidesmosomal genes studied here are all responsive to TGF-beta. These observations, together with previous data on the effects of TGF-beta on other components of the skin, suggest that this cytokine may play a role in the development and repair of the cutaneous basement membrane zone.


Characterization, expression, and immunohistochemical localization of 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase in human skin.

Three beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase (3 beta-HSD) catalyses an obligatory step in the biosynthesis of all classes of hormonal steroids, namely, the oxidation/isomerization of 3 beta-hydroxy-5-ene steroids into the corresponding 3-keto-4-ene steroids in gonadal as well as in peripheral tissues. Because humans are unique with some primates in having adrenals that secrete large amounts of the steroid precursors dehydropiandrosterone (DHEA) and its sulfate (DHEA-S) and its exceptionally large volume makes the skin an important site of steroid biosynthesis, we have isolated and characterized cDNA clones encoding 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase from a human skin lambda gt11 library. The longest clone obtained contains the entire coding sequence for type I 3 beta-HSD (372 amino acids) as well as an additional 131 nucleotides in the 5'-untranslated region. The insert of 1647 bp containing the entire coding region has been inserted in a pCMV expression vector and transfected into human cervical carcinoma cells (HeLa). The expressed enzyme efficiently catalyzes the transformation of pregnenolone, DHEA, and dihydrotestosterone into progesterone, 4-androstenedione, and 5 alpha-androstane-3 beta, 17 beta-diol, respectively. Using the enzyme expressed in HeLa cells, we have shown cyproterone acetate, a progestin used in the treatment of acne and hirsutism, as well as norgestrel and norethindrone, two steroids widely used as oral contraceptives, to be relatively potent inhibitors, with Ki values of 0.38 microM, 1.3 microM, and 1.2 microM, respectively. Immunohistochemical localization of 3 beta-HSD, illustrated by using an antibody raised against human placental 3 beta-HSD, shows that the enzyme is localized in sebaceous glands.


Epidermal keratinocyte self-renewal is dependent upon dermal integrity.

The epidermis is a major site of self-renewal in which there is constant replacement by cell division in the basal layers of cells lost by desquamation in the superficial layers. Such a tissue is therefore likely to contain stem cells and in this study we have examined the role of the dermis in the maintenance of epidermal self-renewal. We have developed a mouse model to address the question of whether the maintenance of epidermal self-renewal is dependent, as in the hemopoietic system, upon a heterologous cell type. Intact epidermis separated from dermis at the dermo-epidermal junction or epidermis derived from disaggregated epidermal cells, can reconstitute a stratified squamous epithelium when grafted onto the lumbo-dermal fascia of the mouse or onto an experimentally induced granulation tissue bed. However, we have shown that, after grafting, the clonogenic capacity of the keratinocytes declines sharply and the colonies that are produced are incapable of self-renewal in vitro. Although initially hyperplastic, these epidermal grafts assume an atrophic appearance after 40-70 d and this may be related to the loss of self-renewal observed in vitro. With both experimental murine grafts and clinical grafts the failure of keratinocytes to self-renew can be alleviated, partially, by the presence of the dermis in full-thickness or split-thickness grafts, which implies that the dermis has a functional role in epidermal stem cell maintenance. The relevance of these observations to the clinical experience with cultured autologous keratinocyte sheets as wound dressings to patients is discussed.


Percutaneous absorption of flurbiprofen in the hairless rat measured in vivo using 19F magnetic resonance spectroscopy.

The objective of this investigation was to develop a new methodology using 19F-magnetic resonance spectroscopy (MRS) to measure the in vivo percutaneous absorption of flurbiprofen through hairless rat skin. A 2% W/V flurbiprofen gel (Klucel HF, hydroxypropyl cellulose 1.5% to 2% W/V) containing isopropyl alcohol, water, and propylene glycol (55:35:10 v/v/v) was prepared. A 2-mg dose (100 mg of gel) was applied to the skin of the lower back of an anesthetized hairless rat, contained with a rubber o-ring, and occluded with a lexan plastic cover slip. The animal was placed on an MR surface coil (3.5-cm diameter tuned to 19F) and measurements taken continuously over approximately 3 h in 10-min intervals with a 2-tesla GE CSI nuclear magnetic resonance (NMR) spectrometer. One measures the disappearance of MR signal intensity per interval, which directly relates to the percent of drug disappearance over time, which in turn was converted to a flux value. The flux of flurbiprofen in vivo was found to be 95 +/- 22 micrograms/cm2/h. This is approximately four times greater than the flux of flurbiprofen through excised human skin reported by Akhter and Barry (22 +/- 14 micrograms/cm2/h). This new in vivo method measures drug disappearance and can be readily transferred to man. This method may be adapted to study other fluorine compounds or other nuclei with magnetic properties. It avoids exposure of a patient or animal to the radiation used in x-ray fluorescence methods or to 14C- or 3H-radiolabeled drugs.


Ultraviolet A irradiation stimulates collagenase production in cultured human fibroblasts.

This study was designed to investigate the biochemical mechanisms responsible for the connective tissue changes seen in actinically damaged skin, which is characterized histologically by diminution and ultrastructural alterations of collagen fibrils and deposition of elastotic material in the papillary dermis. We hypothesized that ultraviolet light could stimulate synthesis of interstitial collagenase in the skin, resulting in collagen degradation. Monolayer cultures of human fibroblasts or keratinocytes were irradiated with ultraviolet A (UVA) or ultraviolet B (UVB) radiation and interstitial collagenase or its inhibitor, TIMP (tissue inhibitor of metalloproteinases) assessed in the conditioned medium with Western immunoblots 24 h after irradiation. Northern blot analysis of the irradiated fibroblasts with a cDNA probe representing collagenase was also performed. Cell viability was greater than 90% with all doses of UV radiation studied. A dose-related increase in immunoreactive collagenase was detected in the medium of fibroblasts irradiated with 0-10 J/cm2 of UVA radiation as well as a parallel increase in the collagenase mRNA in the irradiated cells. UVA radiation stimulated collagenase synthesis in both neonatal and adult fibroblasts. TIMP production in UVA-irradiated fibroblasts increased to a lesser degree than did collagenase and its increase did not parallel the increase in collagenase. UVB (0-100 mJ/cm2) did not stimulate collagenase production by fibroblasts. In contrast to the stimulation of collagenase production by fibroblasts, a slight decrease in immunoreactive collagenase was seen in UVA-irradiated keratinocytes. These data suggest that direct stimulation of collagenase synthesis by human skin fibroblasts by UVA radiation may contribute to the connective tissue damage induced by ultraviolet radiation leading to photoaging.


Differential effects of structurally unrelated chemical irritants on the density of proliferating keratinocytes in 48 h patch test reactions.

Alterations in the proliferative capacity of human epidermis following topical exposure to structurally unrelated chemical irritants were investigated, with the aim of improving our understanding of the cellular changes that take place during the development of irritant contact dermatitis (ICD). Healthy volunteers were patch tested for 48 h with the following six irritants and their appropriate vehicle and occlusion controls: 5% sodium lauryl sulphate (SLS), 0.5% benzalkonium chloride, 80% nonanoic acid (NAA), 0.02% dithranol, 0.8% croton oil, and 100% propylene glycol (PG). After the degree of inflammation induced was visually graded, biopsy samples were removed and the dividing keratinocytes were identified immunocytochemically by using the monoclonal antibody Ki-67, with quantification being performed on the basis of the number of positive cells/100 basal keratinocytes. Statistically significant increases in the density of proliferating cells occurred in the reactions to SLS, NAA, and PG, whereas, in contrast, dithranol caused a marked decrease in the number of dividing keratinocytes. Overall, the density of proliferating keratinocytes did not show a linear relationship with the visually assessed intensity of inflammation, indicating that the changes observed were related to the chemical nature of the individual irritants and their specific biochemical interactions with the keratinocytes, rather than being the consequence of a generalized inflammatory response. Differential release of epidermal cytokines and mediators by the six irritants may account for these varying states of keratinocyte proliferation. Application of the Spearman rank coefficient of correlation revealed that the changes in mitotic activity of keratinocytes were unrelated either to the total density of leukocytes infiltrating the epidermis and dermis, or to the individual densities of the major phenotypic classes of inflammatory cells present. This makes it unlikely that the localized release of cytokines by infiltrating leukocytes is, by itself, the primary factor in the alteration in epidermal cell kinetics seen in ICD. Our results provide a further demonstration of the diverse actions of different chemical irritants on human skin and emphasize the need to regard ICD as a heterogeneous disorder.


Cultured human melanocytes from black and white donors have different sunlight and ultraviolet A radiation sensitivities.

Short-term and long-term survival of cultured neonatal foreskin melanocytes from black and white individuals were assessed following a single exposure to simulated sunlight or ultraviolet A (UVA) radiation. Melanocytes from black individuals contained significantly more melanin than melanocytes from white individuals (p less than 0.05). Black and white melanocytes had similar survival profiles following simulated sunlight exposure, whereas black melanocytes were significantly more resistant to UVA cytotoxicity than melanocytes from white subjects (p less than 0.05) at UVA doses above 15 J/cm2. There was no difference in unscheduled DNA synthesis in the black or white melanocytes following simulated sunlight exposure and no unscheduled DNA synthesis was measurable following melanocyte exposure to UVA radiation. Low-dose UVA (1 or 5 J/cm2) was mitogenic to both black and white melanocytes. By analysis of co-variance, the melanin content of melanocytes of black and white subjects was significantly (p less than 0.05) associated with susceptibility to UVA killing; melanocytes with high melanin content had high resistance to UVA cytotoxicity and those with low melanin content had low resistance to UVA cytotoxicity. From these data we suggest that the higher melanin content of melanocytes of black subjects confers increased resistance to UVA damage. This is likely to be of importance in epidermal photodamage.


Skin color measurements in terms of CIELAB color space values.

The principles of color measurement established by the Commission International d'Eclairage have been applied to skin and the results expressed in terms of color space L*, hue angle, and chroma values. The distribution of these values for the ventral forearm skin of a sample of healthy volunteers is presented. The skin-color characteristics of a European subgroup is summarized and briefly compared with others. Color differences between individuals were identified in terms of one, two, or all three color-space parameters. Because the method is quantitative and the principles internationally recognized, these color-space parameters are proposed for the unambiguous communication of skin-color information that relates directly to visual observations of clinical importance or scientific interest.


Allergic contact dermatitis releases soluble factors that stimulate melanogenesis through activation of protein kinase C-related signal-transduction pathway.

Phenylazo-naphthol (PAN) allergy induces visibly well-defined and late-appearing hyperpigmentation of brownish yellow guinea pig skin in clear contrast to dinitrochlorobenzene (DNCB) allergy, which has very low incidence of hyperpigmentation. Skin extract from PAN allergy at 20-29 d post-challenge exhibited marked melanogenic stimulatory effects (3H2O release and 14C-thiouracil incorporation) when added to cultured guinea pig melanocytes. The time course in the appearance of melanogenic factor was definitely consistent with the induction pattern of visible pigmentation. By contrast, the addition of DNCB-challenged skin extract demonstrated no significant stimulating effect on melanogenesis in either assay system on any of the post-challenge days tested. Assay of intracellular inositol 1,4,5-trisphosphate formed through incubation with the melanocytes demonstrated that the PAN-allergy skin extract at day 28, which contains definite melanogenic factors, stimulated the formation of inositol 1,4,5-trisphosphate that occurs around 50 seconds in contrast to no or little increase with extracts obtained at days 0 and 1 post-challenge. Gel chromatographic analysis revealed that the PAN-allergy skin extract at day 28 contained a newly generated melanogenic fraction with a molecular weight of approximately 9000 Da which was also capable of stimulating DNA synthesis and activating the signal-transduction process (inositol trisphosphate formation) when added to guinea pig melanocytes. Both stimulations of melanogenesis and DNA synthesis by the 9000 Da fraction were completely abolished by the prior and simultaneous addition of protein kinase C (PKC) inhibitor (H-7) or its down-regulatory agent, phorbol 12,13-dibutyrate (PdBu). Taken together, these results suggest that PAN allergy provides a new mechanism of hypermelanization in which endogenous factors synthesized within skin induce the activation of signal-transduction pathways such as phosphoinositide turnover through ligands-receptor binding, resulting in the stimulation of melanocytes possibly through the activation of PKC.


Expression of integrin alpha 6 beta 4 in junctional epidermolysis bullosa.

The integrin alpha 6 beta 4 is a member of the integrin family of adhesion receptors. The integrin alpha 6 beta 4 is preferentially expressed in stratified squamous epithelia, where it is localized in hemidesmosomes. A reduced number of rudimentary hemidesmosomes is often found in skin from patients with junctional epidermolysis bullosa (JEB). In this study we have investigated the expression of alpha 6 beta 4 in skin specimens of patients with junctional (one non-lethal, two lethal) and dystrophic (two) epidermolysis bullosa, using immunofluorescent (IF) staining with five different monoclonal antibodies against the alpha 6 and beta 4 subunits. The intensity of IF staining of the integrin alpha 6 beta 4 and bullous pemphigoid antigen (BPA) was unreduced along the epidermal basement membrane zone (EBMZ) of all EB patients, compared to that in skin of healthy human controls. However, in the skin of two patients with lethal (Herlitz) JEB, who did not express GB3, IF staining of integrin alpha 6 beta 4 and BPA showed a "stitchy" discontinuous linear pattern along the EBMZ with interruptions at the borders of adjoining basal keratinocytes. The same results were obtained by immunoelectron microscopy. They corresponded with freeze-induced partial cell detachment from the basement membrane at the ultimate baso-lateral edge of the basal keratinocytes in lethal JEB skin. The basal lamellipodia at that location almost completely lacked tonofilaments and hemidesmosomes. Furthermore, in JEB there was a split between the intra- and extracellular epitopes of the integrin alpha 6 beta 4 receptor, whereas the integrin remains intact in salt-split skin. This suggests that the defect is in alpha 6 beta 4 itself or perhaps its ligand.


Expression of 92-kDa type IV collagenase mRNA by eosinophils associated with basal cell carcinoma.

Metalloproteinases are thought to be important for tumor invasion and metastasis. We used in situ hybridization with 35S-labeled cRNA probes to localize sites of expression for 92-kDa type IV collagenase mRNA in sections of nodular basal cell carcinoma. Positive signal for 92-kDa type IV collagenase mRNA was detected in eosinophilic granulocytes within inflammatory infiltrates surrounding the tumor nodules. Eosinophils, however, were not adjacent to tumor cells, suggesting that metalloenzyme production by these granulocytes in this disease may be targeted more to stromal components than to remodeling or destruction of the basement lamina. The identity of the eosinophils was confirmed by cell morphology and specific histochemical staining. No resident or other migratory cells were positive for enzyme mRNA in these samples. Signal specificity for in situ hybridization was shown by a duplication of the results with complementary oligomeric probes and by a lack of signal in sections hybridized with a sense RNA probe or nonspecific oligomer. No signal for 92-kDa type IV collagenase mRNA was detected in circulating eosinophils or in eosinophils associated with Hodgkin's lymphoma. These data suggest that eosinophils migrate into the dermis and express type IV collagenase in response to basal cell carcinoma and that this process may have a role in tumor growth.


Control of human sebocyte proliferation in vitro by testosterone and 5-alpha-dihydrotestosterone is dependent on the localization of the sebaceous glands.

Androgens stimulate the activity of sebaceous glands in vivo. In this study the in vitro effect of androgens on the proliferation of cultured human sebocytes derived from facial and non-facial skin were assessed. Human sebocytes from sebaceous glands isolated from the face and the upper and lower legs of five individuals were cultured in vitro with or without testosterone or 5-alpha-dihydrotestosterone (5 alpha-DHT) at different concentrations (10(-11)-10(-5) M). Cell proliferation was assessed in 96-well culture plates using a fluorometric assay. Testosterone and 5 alpha-DHT stimulated the proliferation of human facial sebocytes in a significant dose-dependent manner. In our system 5 alpha-DHT exhibited the strongest effect; on the contrary, the proliferation of non-facial sebocytes was inhibited by testosterone, whereas 5 alpha-DHT enhanced their growth. The stimulatory effect of 5 alpha-DHT was more prominent on facial than on non-facial sebocytes. These results provide first evidence that the effect of testosterone and 5 alpha-DHT on the proliferation of cultured human sebocytes may depend on the localization of the sebaceous glands at different skin regions.


Prevalence and characteristics of pharyngeal group A beta-hemolytic streptococci in US Navy recruits receiving benzathine penicillin prophylaxis.

US military recruits receive benzathine penicillin prophylaxis because of endemicity of group A beta-hemolytic streptococcal (GABHS) infections. GABHS prevalence in Navy recruits receiving single-dose benzathine penicillin prophylaxis was assessed during spring and fall 1989 by culturing throat specimens from randomly selected groups of approximately 230 men before and 2, 4, and 7 weeks after prophylaxis and from men with pharyngitis diagnosed at sick call. Of 60 GABHS isolates, 75% were serotype M-3. The pharyngitis rate increased from 0.18% in the spring to 1.55% in the fall with a concurrent increase in serotype M-3 prevalence from 35% to 91%. The GABHS prevalence rate was three- to fourfold lower after prophylaxis. There were no cases of acute rheumatic fever (ARF) despite predominance of M-3, a rheumatogenic serotype. It was concluded that penicillin prophylaxis continues to be effective for control of GABHS infections and prevention of ARF in Navy recruits.


Genetic diversity in T1M1 group A streptococci in relation to clinical outcome of infection.

Genetic diversity was found at high frequency downstream of the emm1 gene among T1M1 group A streptococci (GAS) isolated in Scandinavia during a recent epidemic. Clonal variation was also seen in the speA and speB genes but at much lower frequency; no variation was detected in the speC gene. Erythrogenic toxin A was found to be expressed at low levels in all strains; erythrogenic toxins B and C were produced in high amounts. All strains were found to harbor the speA, speB, and speC genes, regardless of the amount of toxin produced. No correlation was found between one specific T1M1 clone and the more serious infections when isolates from bacteremic patients (fatalities or survivors), those with uncomplicated infections, and healthy carriers were compared. Similar results were obtained in a family study in which 3 family members were found to be asymptomatic carriers of the same GAS T1M1 clone as in the bacteremic patient, defined by genotypic and phenotypic experiments.


Immunogenicity of Haemophilus influenzae type b conjugate vaccine in allogeneic bone marrow recipients.

A randomized study was conducted in 40 allogeneic marrow recipients to compare the immunogenicity of two Haemophilus influenzae type b (Hib) vaccines (either the Hib capsular polysaccharide [Hib-CPS] or tetanus toxoid-conjugated Hib-CPS [Hib-CPS-T]). A second injection consisted of Hib-CPS-T. Before immunization, 3 patients had serum antibody levels > 1 microgram/mL. After the first injection, the response was better after Hib-CPS-T than after Hib-CPS but lower than in normal subjects; a number of patients lacked any IgG antibody response, especially after Hib-CPS. Of patients who received two injections of Hib-CPS-T, 85% achieved an antibody concentration > or = 1 microgram/mL. Hib-CPS-T induced a response in IgG2-deficient patients whereas Hib-CPS alone did not. IgG antibodies predominantly belonged to the IgG1 subclass. The antibody response was better in patients immunized late after graft. This study shows that Hib-CPS-T is more immunogenic than Hib-CPS in marrow recipients.


Evidence that inactivated oral cholera vaccines both prevent and mitigate Vibrio cholerae O1 infections in a cholera-endemic area.

In a randomized, placebo-controlled field trial of B subunit-killed whole cell (BS-WC) and killed whole cell only (WC) inactivated oral cholera vaccines in rural Bangladesh, active surveillance of selected neighborhoods during the first year after vaccination identified 127 Vibrio cholerae O1 infections among 3285 three-dose recipients. For each vaccine, protective efficacy was greater against symptomatic (57%, P < .05 for BS-WC; 58%, P < .05 for WC) than against asymptomatic infections (46%, P < .05 for BS-WC; 32%, P = .09 for WC), and protection against each grade of infection was demonstrable for both the classical and El Tor biotypes. Although vaccine protection against symptomatic infections was evident in both young children and older persons, only persons vaccinated at age > 5 years were protected against asymptomatic infections. These results suggest that the inactivated oral vaccines acted both to protect against intestinal colonization by V. cholerae O1 and to interrupt the pathogenic sequence of established infections.


Toxicity in neuronal cells caused by cerebrospinal fluid from pneumococcal and gram-negative meningitis.

To identify neurotoxic factors in meningitis, a neuronal cell line (HN33.1) was exposed to cerebrospinal fluid (CSF) obtained from rabbits with pneumococcal meningitis or Escherichia coli meningitis or 2 h and 6 h after meningitis was induced by proinflammatory bacterial products (pneumococcal cell walls, endotoxin). CSF from all types of meningitis induced similar degrees of cytotoxicity. When a soluble tumor necrosis factor (TNF) receptor that completely blocked TNF-mediated toxicity at 10(-7) M was used, all toxicity in meningitis caused by E. coli, endotoxin, or pneumococcal cell wall administration (2 h afterwards) was mediated by TNF. In contrast, CSF from animals with meningitis caused by live pneumococci or pneumococcal cell wall injection (6 h afterwards) retained cytotoxicity in the presence of the TNF receptor. Thus, in established pneumococcal meningitis, but not in the other forms of meningitis, TNF is not the only component toxic in this neuronal cell line.


Frequencies of lipopolysaccharide core types among clinical isolates of Escherichia coli defined with monoclonal antibodies.

Mouse monoclonal antibodies (MAbs) specific for the lipopolysaccharide (LPS) core types R1, R2, and R3 of Escherichia coli and a cross-reactive MAb that binds to the LPS core of almost all E. coli were used in ELISA to determine the frequency of cores resembling R1, R2, and R3 in strains of E. coli isolated from clinical samples (blood and urine specimens) and from the feces of asymptomatic individuals. Of the 180 wild-type isolates, 123 were assigned to R1 core type, 14 to R2, and 18 to R3. Twenty-five wild-type E. coli isolates could not be assigned to a particular core type and may have either an R4 or K12 core or a previously unrecognized core type. R1 core type was associated with O types 1, 4, 6, 8, and 18 and with K1 or K5 capsules. R3 was associated with O15. O75 isolates could be of either R1 or R2 core type.


Characterization of Staphylococcus aureus isolates with decreased susceptibility to vancomycin and teicoplanin: isolation and purification of a constitutively produced protein associated with decreased susceptibility.

"Derivative isolates" with 4- to 8-fold and 8- to 16-fold increases in MICs of vancomycin and teicoplanin, respectively, were selected from 2 susceptible clinical isolates of Staphylococcus aureus by serial incubation in low-level vancomycin. A protein of approximately 39 kDa was demonstrable in the cytoplasmic fraction and occasionally in the membrane fraction by SDS-PAGE of both derivatives. This protein was purified by DEAE chromatography, preparative SDS-PAGE, and electroelution. Derivative bacteria were larger on transmission electron microscopy, had thicker cell walls, and had changes in colony morphology on solid media. Further evidence for cell wall reorganization included loss of phage and capsular typing, decreased susceptibility to lysostaphin/lysozyme killing, and changes in condition for detection of optimal coagulase activity. The mechanism of decreased susceptibility to glycopeptide antibiotics among S. aureus derivative isolates is uncertain. The production of the approximately 39-kDa cytoplasmic protein and cell wall reorganization may mediate changed affinity of glycopeptide-peptidoglycan binding or impairment of glycopeptide access to its cell wall target.


Potentiation of Helicobacter pylori vacuolating toxin activity by nicotine and other weak bases.

About 50% of Helicobacter pylori isolates produce a vacuolating toxin in vitro, which may be an important determinant of virulence. Because ammonium salts potentiate H. pylori toxin activity, the effect of other weak bases upon toxin activity was determined. Vacuolation of HeLa cells was quantitated using a neutral red uptake assay. As expected, ammonium chloride, trimethylamine, triethanolamine, and nicotine each induced vacuolation of HeLa cells when tested independently. In addition, each of these weak bases potentiated H. pylori vacuolating toxin activity, whereas sodium chloride or sodium hydroxide did not. Sequential incubation of cells with toxin followed by nicotine resulted in potentiation of vacuolation, whereas sequential incubation in the reverse order did not lead to potentiation. Monensin inhibited the formation of vacuoles by either H. pylori vacuolating toxin or nicotine. The potentiation of H. pylori toxin activity by ammonia and nicotine may contribute to gastroduodenal mucosal injury associated with this infection.


Sialylation and human neutrophil killing of group C Neisseria meningitidis.

This study describes the association of lipooligosaccharide (LOS) and capsule sialylation with the survival of 25 serogroup C meningococcal strains in phagocytosis assays. Eleven strains isolated from children were of diverse protein serotypes or were nontypeable; 14 were serotype 2b:P1.2 and were isolated from children during or immediately after a focal epidemic in Texas. Degree of endogenous LOS sialylation and amount of sialic acid capsule were associated with each other and with susceptibility to killing by neutrophils for the non-2b:P1.2 strains. The 2b:P1.2 strains as a group had significantly greater survival in the presence of neutrophils than did the non-2b:P1.2 strains. The susceptibility of these strains to killing by neutrophils was not associated with endogenous LOS sialylation or amount of capsule. These data suggest that many virulent strains evade neutrophil killing, either by sialylation or another mechanism. Evasion of neutrophil killing might enhance a strain's epidemic potential.


Recombinant interleukin-8 induces changes in cytosolic Ca2+ in human neutrophils.

Activation of polymorphonuclear leukocytes (PMNL) by most soluble stimulants is associated with a marked increase in cytosolic free Ca2+ ([Ca2+]i). Interleukin-8 (IL-8), a monocyte-derived neutrophil chemotactic factor and potent neutrophil-activating cytokine, effectively enhanced the resting free [Ca2+]i within human PMNL in a dose-dependent manner (maximal effect with 100 ng/mL). The increase in [Ca2+]i was substantially (55%) inhibited in the absence of extracellular Ca2+. Thus, the increase was due to extra- and intracellular cooperative mobilization of Ca2+, as supported by the reduced effect of IL-8 on [Ca2+]i after quenching with Mn2+. Granulocyte-macrophage colony-stimulating factor and interferon-gamma failed to induce a change in [Ca2+]i, suggesting that they may operate through different signal pathways. Pretreatment with Bordetella pertussis toxin largely inhibited the IL-8-induced change in [Ca2+]i. Thus, IL-8-induced cooperative mobilization of intra- and extracellular Ca2+ leads to a net Ca2+ influx into the cytoplasm through a process mediated by a guanosine triphosphate-binding protein.


Phagolysosomal alkalinization and the bactericidal effect of antibiotics: the Coxiella burnetii paradigm.

Most infections due to intracellular bacteria respond poorly to antibiotic treatment. The chemical conditions within the subcellular site of bacteria may change antibiotic activity. Coxiella burnetii multiplies within phagolysosomes. The antimicrobial activity of antibiotics combined with the lysosomotropic agents amantadine (1 microgram/mL), chloroquine (1 microgram/mL), and ammonium chloride (1 mg/mL), which alkalinized Coxiella burnetii-containing phagolysosomes from pH 4.8 to 5.3, 5.7, and 6.8, respectively, was evaluated. Percentages of residual viable bacteria (RVB) in cell cultures were significantly reduced after exposure to combinations of doxycycline (4 micrograms/mL) with amantadine (RVB = 18.2% +/- 8.7%, P < .05), chloroquine (RVB = 0.64% +/- 0.38%, P < .01), or ammonium chloride (RVB = 0.29% +/- 0.17%, P < .01); the same was seen with pefloxacin (1 microgram/mL) with chloroquine (RVB = 27.6% +/- 10.8%, P < .05) or ammonium chloride (RVB = 3.72% +/- 1.1%, P < .05). Such bactericidal activity correlated with increased phagolysosomal pH, as determined by Pearson's correlation coefficient, suggesting that phagolysosomal alkalinization is critical for the bactericidal effect of antibiotics.


Identification of a mannoprotein fraction from Candida albicans that enhances human polymorphonuclear leukocyte (PMNL) functions and stimulates lactoferrin in PMNL inhibition of candidal growth.

Mannoprotein fractions of Candida albicans were assayed for their effects on the anticandidal activity of human polymorphonuclear leukocytes (PMNL). One fraction, MP-F2, enhanced PMNL inhibition of candidal growth in vitro as potently as bacterial lipopolysaccharide (LPS), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-8. MP-F2-mediated PMNL activation was manifested on yeast and mycelial forms of the fungus, required the integrity of the mannan, and was due to an increase in the actual number of phagocytic PMNL rather than to a greater ingestion of fungal cells by each individual neutrophil. While not inducing augmented O2 production or degranulation of azurophilic granules, MP-F2 strongly stimulated the release of lactoferrin. Lactoferrin inhibited candidal growth in the absence of PMNL, and anti-lactoferrin antibodies reversed both this inhibition and the PMNL activation by MP-F2, GM-CSF, and LPS. Thus, PMNL may be activated by relevant candidal mannoproteins, and release of lactoferrin may add to other antimicrobial mechanisms of PMNL for the control of candidal infections.


Expression and characterization of a cDNA clone encoding an immunodominant surface glycoprotein of Pneumocystis carinii.

Most studies of antigens of Pneumocystis carinii have focused on an abundant, immunogenic 95- to 140-kDa surface glycoprotein referred to as gpA. Expression cloning of gpA from P. carinii obtained from ferrets resulted in isolation of colinear fragments of gpA cDNA encoding approximately 87 kDa of the core protein. Northern hybridization detected an abundant, single species of gpA-specific mRNA of 3600 nucleotides. Southern hybridization revealed gpA-specific bands only in P. carinii-infected lung genomic DNA, suggesting that gpA cDNA did not result from induction of a host lung gene. Antiserum raised against a fragment of recombinant gpA detected P. carinii cysts and isolated native P. carinii gpA, indicating retention of epitopes between the nonglycosylated recombinant gpA and glycosylated native gpA. The deduced amino acid sequence is hydrophilic and contains 12 potential N-linked glycosylation sites and 47 cysteine residues, consistent with the surface orientation of gpA on the organism and other known characteristics of the native molecule.


Epidemiology of visceral leishmaniasis in northeast Brazil.

Epidemiologic aspects of the relationship between infection with Leishmania chagasi and development of clinical visceral leishmaniasis (VL) were studied in all children < 11 years old in a defined, endemic, rural area of the state of Ceará in northeast Brazil. Antileishmanial antibodies were measured in the same subjects by ELISA on six occasions between May 1987 and August 1989. Seroconversion was documented during this period in 108 children, with a cumulative annual incidence of 4.6%. Twelve (11.1%) of these children developed VL. Age < 4 years, hematocrit < 33%, and living in the mountains predicted the development of clinically apparent VL after seroconversion. Despite a high percentage of dogs serologically positive in the region (38%), there was no increased risk of infection for children living in the same household with dogs. Since children in households with a prior case of VL had a threefold increased risk of infection, human-sandfly-human transmission might have been important.


Migration of 75Se-methionine-labeled Schistosoma japonicum in normal and immunized mice.

The patterns of migration and attrition of Schistosoma japonicum larvae were studied in a mouse model. Control and immunized mice were challenged with 100 S. japonicum cercariae tagged with 75Se-labeled methionine. Skin, lungs, liver, and other organs were analyzed by compressed organ autoradiography for the presence of larvae that appeared as reduced silver foci. The pattern of migration of S. japonicum was similar in mice with primary infection and in mice immunized with irradiated cercariae. Skin was not a site of attrition after primary infection nor after immunization. Attrition occurred after migration to the lungs and continued until after migration to the liver in mice with primary infection, while in immunized mice attrition occurred before lung migration and continued at a faster rate than in normal mice. In both control and immunized mice, the lungs and liver were the major sites of attrition.


Incidence of zidovudine-resistant human immunodeficiency virus isolated from patients before, during, and after therapy.

The zidovudine sensitivity of 372 isolates of human immunodeficiency virus (HIV) obtained from 237 patients before, during, and after treatment with zidovudine was examined. Virus resistant to > 0.5 micrograms/mL (1.87 microM) zidovudine was isolated from most patients (93%) after 36 months of therapy. Zidovudine-sensitive virus was isolated from 5 of 15 patients who had ended antiretroviral therapy but had previously shed resistant virus. The emergence of sensitive virus after end of therapy appeared to be influenced by both the duration of treatment and the time off drug. Patients with resistant virus tended to have low CD4 cell counts and HIV antigenemia at the commencement of therapy, suggesting that these two factors are important in the development of drug resistance.


Diagnosis of vertical human immunodeficiency virus type 1 infection by whole blood culture.

The qualitative and quantitative performance of a human immunodeficiency virus type 1 (HIV-1) whole blood culture method was assessed for use in the diagnosis of vertical HIV-1 infection. This method requires < 1 mL of whole blood and allows for the quantification of blood virus load. HIV-1 was isolated from 36 (95%) of 38 whole blood specimens from 27 HIV-1-infected children compared with 0 of 16 whole blood specimens from 16 uninfected children. HIV-1 titers were significantly higher in severely symptomatic children (2450 TCID/mL) than in less symptomatic children (179 TCID/mL). This simple quantitative culture assay may be useful for the early diagnosis of vertical HIV-1 infection, the assessment of blood virus load, and the evaluation of antiretroviral therapies.


Initial low CD4 lymphocyte counts in recent human immunodeficiency virus infection and lack of association with identified coinfections.

Initial CD4 lymphocyte counts were studied in 244 patients with human immunodeficiency virus (HIV) seroconversion. The CD4 cell counts at initial presentation after seroconversion were normally distributed (mean, 579/mm3; SD, 252). The mean percentage of CD4 cells was 26.1% (SD, 5.6). CD4 cell counts were < 500/mm3 in 41% and < 200/mm3 in 4%. The mean calculated duration of HIV infection was 7.7 months, which was not significantly different between the highest and lowest CD4 count quartiles (8.1 vs. 7.9). Age, sex, race, and serologic evidence of toxoplasmosis, cytomegalovirus, hepatitis B, syphilis, and varicella-zoster virus were not associated with initial low CD4 cell counts; however, never-married men were significantly overrepresented in the lowest quartile. These findings suggest that extensive CD4 lymphocyte depletion is common in early HIV infection and that frequent screening is necessary to identify newly infected patients who would benefit from antiretroviral therapy.


Immune activation during measles: beta 2-microglobulin in plasma and cerebrospinal fluid in complicated and uncomplicated disease.

Beta 2-microglobulin (beta 2m) is a small protein that forms the light chain of the class I major histocompatibility molecule and is also present in soluble form in serum and cerebrospinal fluid (CSF). Measles is associated with immune activation and evidence of immunologic abnormalities that persist for several weeks. To assess further the immunologic changes occurring during measles, beta 2m was measured in plasma and CSF. beta 2m became elevated during measles before the onset of the rash and was highest during the rash. Elevations persisted for several weeks and correlated well with levels of soluble interleukin-2 receptor and neopterin and less well with soluble CD8. CSF beta 2m was elevated in postmeasles encephalomyelitis. Plasma levels of beta 2m did not correlate with spontaneous proliferation of peripheral blood mononuclear cells (PBMC) or with in vitro production of beta 2m by cultured PBMC. The data suggest that increases in beta 2m in measles correlate better with cytokine production than with cell proliferation.


Genotyping of Chlamydia trachomatis from a trachoma-endemic village in the Gambia by a nested polymerase chain reaction: identification of strain variants.

Direct amplification of the major outer membrane protein (MOMP) gene by polymerase chain reaction (PCR) was used to identify Chlamydia trachomatis in eye swabs from clinically active cases of endemic trachoma in a Gambian village. Chlamydial DNA was detected in 51% of 96 subjects with clinically active disease and in 5% of 37 clinically negative individuals. The PCR detection was combined with typing, using nested primers to variable sequences (VS) 1, 2, and 4 of the MOMP genes to distinguish between trachoma genotypes A, B, and C, respectively. Genotypes A and B were detected in the village, with some individuals harboring both genotypes within the same eye. DNA sequencing revealed strain variants of both genotypes. Typing of genotype and strain variants is now in progress to study trachoma transmission within the village.


Evaluation of polymerase chain reaction, tuberculostearic acid analysis, and direct microscopy for the detection of Mycobacterium tuberculosis in sputum.

Tuberculosis remains a major global cause of morbidity and mortality. There is an urgent need for improved bacteriologic diagnosis of Mycobacterium tuberculosis infection. Three methods for rapid identification of M. tuberculosis in sputum samples (direct microscopy, gas chromatography-mass spectrometry [GC-MS], and polymerase chain reaction [PCR]), were compared with culture on Lowenstein-Jensen medium. Growth of M. tuberculosis was observed in 38 of 145 sputum samples. Detection of acid-fast bacilli by direct microscopy gave a sensitivity of 66% and a specificity of 100%. Detection of tuberculostearic acid by GC-MS gave a sensitivity of 55% and a specificity of 87%. Amplification by PCR of a fragment of the insertion sequence IS6110 gave a sensitivity of 95% and a specificity of 93% compared with culture and a corrected specificity of 99% compared with both culture and clinical data. This study indicates that PCR can be adapted for clinical use and is the method of choice for rapid diagnosis of pulmonary tuberculosis.


An analysis of the quantitative relationship between oral temperature and severity of illness in experimental shigellosis.

The relationship between oral temperature and other parameters of illness was examined in 139 adult volunteers infected experimentally with Shigella sonnei. In subjects developing clinical disease, peak temperature correlated positively with total number of signs and symptoms other than fever (rxy = .71, P < .001), stool volume (rxy = .41, P < .001) and number of stools produced during the illness (rxy = .46, P < .001). Peak temperature correlated negatively with incubation period (rxy = -.34, P = .007) but exhibited no apparent correlation with duration of illness. The average oral temperature during illness correlated positively with number of other signs and symptoms of infection but not with stool volume or stool number. These results suggest that in clinical investigations involving S. sonnei, and perhaps with other pathogenic microorganisms, oral temperature is a useful quantitative marker for estimating disease severity.


Serum concentrations of penicillin, doxycycline, and ciprofloxacin during prolonged therapy in rhesus monkeys.

Concentrations of penicillin, doxycycline, and ciprofloxacin were measured by bioassay in sera of rhesus monkeys treated with these drugs for inhalation anthrax. Antibiotic doses were determined on the basis of published serum concentration data from humans and comparative body surface area calculations for humans and rhesus monkeys. The antibiotics were well tolerated. Serum peak and trough concentrations of penicillin, doxycycline, and ciprofloxacin, respectively, averaged 2.7 and 0.8, 1.31 and 0.26, and 1.22 and 0.14 microgram/mL. These were within the range usually observed with standard oral doses in humans, and peak concentrations in all monkeys exceeded the MICs for 90% of Bacillus anthracis strains.


Transfer of pheromone-inducible plasmids between Enterococcus faecalis in the Syrian hamster gastrointestinal tract.

Pheromone-responsive plasmids are common to Enterococcus faecalis, transfer at high frequency in vitro, and carry cytolysin and other gene products implicated in the pathogenesis of enterococcal infection. A Syrian hamster model of enterococcal intestinal overgrowth was used to test for transfer of three isogenic plasmids differing in conjugative and cytolytic phenotypes. Transconjugants were found in 8 (44%) of 18 and 6 (35%) of 17 hamsters given donor strains containing cytolytic (pAM714) and noncytolytic (pAM771) pheromone-responsive plasmids. Of the 14 hamsters from which transconjugants were isolated from stool, 9 (64%) had transconjugants 1 day after donor strain inoculation. The frequency of transfer (mean +/- SD) for pAM714 and pAM771 was 1.4 +/- 2.2 x 10(-1) and 2.9 +/- 4.2 x 10(-2) transconjugants/donor, respectively (P > .20). Transconjugants were not recovered from hamsters receiving a cytolytic, nonconjugative plasmid (pAM930; transfer frequency < 2 x 10(-5) transconjugants/donor). Pheromone-responsive plasmid transfer between E. faecalis strains occurs at high frequency in the gastrointestinal tract of hamsters and may be one means by which enterococcal resistance and virulence factors disseminate.


Isolation and characterization of 115 street rabies virus isolates from Ethiopia by using monoclonal antibodies: identification of 2 isolates as Mokola and Lagos bat viruses.

There were 115 isolates of rabies viruses recovered by tissue culture technique from 119 animal brains collected in Ethiopia. By using 17 selected antinucleocapsid monoclonal antibodies (MAbs), 113 isolates were classic street rabies viruses (serotype 1). An isolate of feline origin (Eth-16) was a Mokola virus (serotype 3) and another isolate (Eth-58, obtained from a rabid dog) was serotype 2 (Lagos bat virus). None of the 16 antiglycoprotein MAbs used neutralized the Eth-16 isolate, whereas Eth-58 was neutralized by 1 (TERA543). Antirabies vaccines prepared from Pitman-Moore and Pasteur virus strains protected mice against homologous challenge, but neither was protective against the 2 rabies-related virus isolates. The isolation of Mokola and Lagos bat viruses from domestic animals in eastern Africa is of public and veterinary concern mainly due to lack of effective vaccines against these agents and the difficulty of proper diagnosis.


Long-term results on the quadrangular osteotomy.

The results of a follow-up study of 17 patients who underwent a quadrangular osteotomy, are presented. The indication for a quadrangular osteotomy includes a hypoplastic maxilla with retruded infra-orbital rims and infra-orbital area, but with normal nose projection. Almost all patients were satisfied with the result. Relapse in a horizontal direction appeared to be approximately 12%; however, considerable relapse was seen in the vertical direction (61.7%-158%). A major problem during operation appeared to be the high rate of fractures of the infra-orbital wing. Sensory loss in the area of the infra-orbital nerve occurred in 79% of the operated sides. Four patients needed surgical correction of conditions that should be considered complications resulting from the osteotomy, including a case of partial ischaemic necrosis of the premaxilla in a BCLP patient.


Computerized cephalometric orthognathic surgical simulation, prediction and postoperative evaluation of precision.

A new computerized, cephalometric, orthognathic surgical program (TIOPS) has been evaluated in surgical simulation, prediction and postoperative assessment of precision. Records of 10 consecutive patients admitted for orthognathic surgical treatment were analysed and prediction plans produced by computerized surgical simulation. Predicted and postoperative positions of maxilla and mandible were compared with linear and angular measurements. No statistically significant differences between predicted and postoperative positions could be demonstrated (p greater than 0.05).


Occlusion of the incisal canal with bone chips. A procedure to facilitate insertion of implants in the anterior maxilla.

In 4 patients, who had lost one or both central maxillary incisors due to trauma, the incisal canals were filled with autogenous cancellous bone harvested from the chin. After a healing period of 4-5 months implants were inserted. At the time of implant surgery in all cases the canal appeared to be replaced by cancellous bone and the implants were placed partially into the grafted area. After another 6 months abutments were connected and crowns made. After follow-up of between 12 and 15 months no fixture has been lost.


Posterior dislocation of mandibular condyle into external auditory canal. A case report.

A case is reported of a 49-year-old edentulous patient who suffered a posterior dislocation of the right fractured mandibular condyle causing a fracture of the tympanic plate. This was associated with a right transverse fracture of the petrous bone and an intact tympanic membrane. Some of the problems related to this condition are discussed.


Applications of the lateral vastus muscle flap.

The lateral vastus muscle flap has the potential to become one of the most appropriate free flaps for reconstructive surgery in the oral maxillofacial area in selected cases. The flap is relatively easy to harvest and gives rise to little donor site morbidity. The quality of the large diameter vessels allows for relatively easy microsurgical anastomosis. Nerve anastomosis is possible, which makes it extremely suitable for tongue reconstruction.


Neoplastic disease in the head and neck of patients with AIDS.

Immunosuppression increases the risk of developing malignancies. In immunosuppression due to human immunodeficiency virus (HIV) disease the common head and neck tumors are Kaposi's sarcoma and non-Hodgkin's lymphoma. Squamous cell carcinoma has also been reported. Kaposi's sarcoma is the commonest neoplastic disease in AIDS. The incidence of lymphoma is rapidly increasing. This article reviews the incidence, clinical presentation and management of these diseases in the head and neck in AIDS patients.


Investigating a lingual thyroid.

During a 15-year-period, in a thyroid investigation centre in Sri Lanka comprising 16,593 cases, 8 lingual thyroids were seen. All patients were female. Radioisotope studies and scintiscanning performed during the investigations of these cases are presented. All subjects were euthyroid at presentation and the clinical courses of 5 cases during follow-up are discussed.


Giant ossifying fibroma. Case report on a bimaxillary presentation.

Ossifying fibroma is a slow-growing, benign neoplasm, but some lesions behave aggressively, reaching massive proportions, thus demanding special treatment. The following case report holds particular interest, because of the simultaneous occurrence of an active ossifying-cementifying fibroma in the maxilla and mandible with the maxillary lesion attaining enormous size.


Necrotizing fasciitis of the neck and chest. Report of a case.

A case is presented of necrotizing fasciitis of the neck and chest characterized by rapid progressive necrosis of subcutaneous tissue, fascia and skin. The diagnosis and management is discussed.


Effect of marrow perforation on the sheep temporomandibular joint.

The effect of surgically perforating the mandibular condyle to allow synovial fluid to contact the marrow was examined in 5 sheep temporomandibular joints. The surgical defect showed replacement of the marrow with fibro-osseous tissue and subcortical cysts. A vertical, central osteophyte emerged from the perforation, causing attenuation or perforation of the disc and temporal surface proliferation. These changes were radiographically and histologically similar to advanced osteoarthritis. This supports the concept that intraarticular micro or macrofracture may result in osteoarthritis.


[Activation of aerobic bacteria used as BOD measurement of seawater].

Waste of seawater used for cooling water for machines in some kinds of industries in a littoral district in Sakai City, comes under the application of BOD control of the pollution control ordinance in Osaka Prefecture. But unfortunately the BOD measurement method of seawater sample has not been established. In order to establish the measurement method, we studied the strength of activation of aerobic bacteria in the incubation water used and the dilution water. From our results it was proved that we can use the incubation seawater consisting of more than 10(3) cells/ml general bacteria count and more than 0.50 microgram/l ATP (Adenosin triphosphate) as the incubation water and the artificial seawater as the dilution water.


[Recurrent case of subacute necrotizing lymphadenitis in young man].

A 10-year-old male was admitted to our hospital because of lymphadenopathy and fever. Biopsy of a cervical lymph node demonstrated nodular necrotic foci containing many macrophages phagocyting the karyorrhexic nuclei. He was diagnosed as subacute necrotizing lymphadenitis (SNL) and all the symptoms disappeared spontaneously within a month. Five year later, lymphadenopathy recurred, and re-biopsy showed the same histological findings and he also recovered spontaneously. Although the prognosis of SNL is generally excellent and the majority of the patients recover without any treatment, recurrences are very rare, especially in young male patient.


[Correlation between hepatitis B virus infection and chronic liver disease in Okinawa].

In Okinawa prefecture, prevalence of hepatitis B surface antigen (HBsAg) among blood donors is 3.5% and is twice as high as the average for the whole of Japan (1.5%), and is the highest in Japan (p less than 0.005). In contrast, mortality rates of both liver cirrhosis (LC) and primary liver cancer (PLC) in Okinawa are the lowest in Japan. Many epidemiological studies have shown that the positive rate of HBsAg correlates with mortality rate of PLC. To elucidate the cause of this epidemiological discrepancy, cross-sectional seroepidemiological studies and a prospective clinical study were conducted. In the cross-sectional studies, the following results were obtained; (1) Positive rate of HBsAg among patients with LC in Okinawa was 15.2% and lower than the average for the whole of Japan (23.4%). A similar comparison among patients with hepatocellular carcinoma showed 24.4% in Okinawa Vs. 31.4% in the whole of Japan. (2) The age-specific hepatitis B e antigen positive rate among 829 HBsAg positive health examinees tend to decrease with increase in age; 50% in less than 20 years old age group, 15.7% in third decade and 2-3% or less in 30 or more age group. Of the 829, 431 HBsAg positive subjects were referred our liver out-patient clinic. Then, of the 431, 27 (6.3%) were diagnosed or suspected as having chronic hepatitis and one (0.2%) was diagnosed as having cirrhosis. Of the 431, 381 (88.4%) were diagnosed as healthy HBsAg carrier, the great majority (94.0%) of whom had positive reaction of anti-HBe antibody and normal values of both GOT and GPT.(ABSTRACT TRUNCATED AT 250 WORDS)


[Clinical and epidemiological aspects of enteritis due to Salmonella hadar. I. Isolation of S. hadar from sporadic diarrhea--clinical and bacteriological study].

Before 1983, S. hadar was seldom isolated from man, animals, food or the environment in Japan: only one strain having been isolated from man and one from the environment. In subsequent years there has been a progressive increase in the number of isolations. S. hadar is now one of the commonest serotypes isolated from cases of sporadic diarrhea in Shizuoka Prefecture (Table 1). However, the epidemiology of S. hadar is not clearly understood. Reports on the clinical features of S. hadar gastroenteritis are also scarce. We examined the clinical symptoms of 15 cases of S. hadar gastroenteritis. S. hadar was encountered in patients of all ages. Infants and young children below 10 years of age constituted 47% of all cases. Seventy per cent of young children below 10 years of age experienced fever of more than 39 degrees C. This incidence was significantly higher than that of general Salmonella gastroenteritis observed in our previous study, in which fever of more than 38 degrees C was noted in 61% of the children. We found one case of presumptive person-to-person spread. No other household contacts of index patients suffered from diarrhea during the same period. Systematic examination for Salmonella contamination was performed for poultry farms, broiler chickens, broiler processing plants and meat on the market. 259 Salmonella strains were isolated from 1197 samples. S. hadar accounted for 37.1% (96) of all isolations (259). A drug resistance test was performed for 51 strains of the diarrhea cases and 67 strains of the environment. The pattern of the distribution of MICs of 9 drugs was similar in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)


[Clinical and epidemiological aspects of enteritis due to Salmonella hadar. II. Environmental contamination by Salmonella hadar in Shizuoka Prefecture--studies on the feasibility of reducing S. hadar infection].

A systematic examination was performed for environmental food contamination by Salmonella in poultry farms, broiler chickens, broiler processing plants and meat on the market. Salmonella was isolated from 219 of 1197 samples and the serotypes showed a wide distribution. (Table 1-1). S. hadar accounted for 37.1% (96) of all isolations (259). The contamination rate of broiler chickens on arrival at broiler processing plants is relatively low. However, in the broiler processing plants, containers, processing machinery, cooling water and slaughters were highly contaminated by Salmonella, S. hadar being the most prominent serotype in the plants. 64% of chicken meat on the market was contaminated by Salmonella, S. hadar being the second most prominent serotype. 11% of the pork and none of the beef or horseflesh was contaminated by Salmonella. These results indicate that poultry is the main source of S. hadar infection in humans. However, no S. hadar was isolated from cultures of 119 samples of feed for chickens from each delivery (Table 1-1). Thus, as the main source of infection by S. hadar of broiler chickens, an association with the feed seems to be ruled out. S. hadar was isolated at three of 18 poultry farms within Shizuoka Prefecture. Follow-up studies were performed at the three poultry farms which revealed that in two of them, Salmonella was completely eradicated on completion of disinfection. In the other one farm, which is still being disinfected, various strains of Salmonella including S. hadar still survived. We conclude that the main cause of the problem is the magnification of contamination of Salmonella-free material during the process at the broiler processing plants.(ABSTRACT TRUNCATED AT 250 WORDS)


[Genetic identification of 25 Legionella species by modified microdilution plate hybridization, and its evaluation with isolated strains].

Modified microdilution plate hybridization was used for genetic identification of 25 Legionella species. The cell wall of the isolate was broken down by glass beads. And the DNA was extracted, labeled photo-reactive biotin, and hybridized with reference DNAs immobilized in microdilution wells. Hybridized DNAs were detected by colormetric method. Each type strain of 25 Legionella species was clearly differentiated by this method. Among 103 clinical and environmental Legionella strains, 97 strains were genetically identified by this method. 60 strains were identified as the same species as by the conventional method. Concerned with ten strains that had been serologically identified as L. bozemanii, two strains isolated from the human lung were genetically identified as L. bozemanii, but the remaining environmental 8 strains were identified as L. anisa. Among the 35 strains of legionellae that had been unidentified at species level, by physiological and serological tests, 27 strains were genetically identified as L. pneumophila, 1 strains as L. feeleii, and another, as L. anisa, but 6 strains were not identified. We found that three strains among the 6 strains belong to a single species of genus Legionella other than the 25 reference species used in this study.


[Study of the anti Borrelia burgdorferi antibody of hunters in Hokkaido].

We examined the sera of 587 hunters in Hokkaido (Japan's northernmost island) for the antibody to Borrelia burgdorferi (B. burgdorferi) by enzyme immunoassay, clarified the conditions related to antibody positivity in these subjects according to region, and studied the effects of factors such as age and lifestyle on the antibody titer. In contrast with an anti-B. burgdorferi antibody positive rate of 7.1% in control sera, that in the hunters' sera was 16.0%. Among those positive for the anti-B. burgdorferi antibody, the antibody positive rate in sera excluding those testing positive in the serological test for syphilis was 5.5% in the controls, and 15.4% in the hunters, the latter rate being significantly higher (p less than 0.05). In both hunters and control groups, the antibody-positive rate tended to be higher in older subjects, but the antibody titer showed no correlation with their age, or the duration of their hunting experience. Examination of the hunters' occupations revealed a tendency toward high titers in those engaged in dairy farming. The antibody positivity of those who went gathering edible wild plants was significantly higher than those did not (p less than 0.05). These observations suggested that the high antibody-positive rate in hunters may have been due largely to the effect of activities other than hunting as sources of infection by Borrelia.


[In vitro antimicrobial activity of DR-3355, a new quinolone antibacterial agent, against clinical isolates of enteritis-causing bacteria].

We determined the minimum inhibitory concentration (MIC) of DR-3355, a newly developed quinolone-derivative antibacterial agent, against clinical isolates of various bacterial species from enteritis patients, and compared them with those of ofloxacin (OFLX), ciprofloxacin (CPFX), nalidixic acid (NA), ampicillin (ABPC), kanamycin (KM). MIC90 of DR-3355 against 94 strains of Shigella spp. and 5 strains of Escherichia coli, 36 strains of Salmonella spp., 22 strains of Vibrio cholerae, 5 strains of Vibrio parahaemolyticus, and 19 strains of Campylobacter jejuni were 0.05, 0.10, 0.0025, 0.39, and 0.78 micrograms/ml, respectively. These values were 1/2 of that of OFLX, and two times of that of CPFX. MIC90 of DR-3355, OFLX and CPFX against C. jejuni were 0.78 micrograms/ml. MIC90 of DR-3355 against isolates from enteritis patients except for Vibrio spp., were 1/30 to 1/60 of those of NA, ABPC, and KM.


[Significance of the combined treatment with isepamicin and piperacillin in an in vitro model for complicated cystitis operated by automatic simulator apparatus for urinary concentration].

Isepamicin (ISP) and piperacillin (PIPC) were shifted to the urinary concentration by employing an in vitro complicated cystitis model operated by a computer-controlled automatic simulator for urinary concentration, and administrated to bacteria in the urinary bladder model (Pseudomonas aeruginosa: P. aeruginosa, initial cell concentration: 10(7) cfu/ml). In this case, effects by single treatment with ISP or PIPC on cell number curves were examined. Further, significance of the combined treatment with ISP and PIPC were investigated by changing the order of each treatment. And following the results were obtained. 1. In a single treatment with PIPC the cell concentration was minimum (10(4) cfu/ml) at 9th hour after its treatment and thereafter, regrowth to the same level as the initial concentration was observed at 16th hour. 2. In the case of single treatment with ISP, the cell concentration became minimum (10(2) cfu/ml) at 13th hour after the treatment and raised to the same concentration as the initial one at 25th hour. 3. In the combined treatment, the cell concentration was minimum (less than 10(1) cfu/ml) at 26th hour in the case of prior treatment with ISP. Thereafter, regrowth was observed and the cell concentration at 42nd hour reached to the initial cell concentration. 4. In simultaneous treatment with ISP and PIPC, the cell concentration at 24th hour was minimum (10(1) cfu/ml) and reached to the same level as the initial one after regrowth. From these results, it was found that the combined treatment with ISP and PIPC caused more reduction of the cell concentration than either single treatment. Further, regrowth of the cells was suppressed for longer duration.(ABSTRACT TRUNCATED AT 250 WORDS)


[Evaluation of chlamydial IgM antibodies for clinical diagnosis].

Chlamydia trachomatis (C. trachomatis), C. psittaci, and C. pneumoniae are now well established as pathogens of respiratory infections including pneumonia. Serum samples from 223 infants and children with pneumonia, 31 patients with adult inclusion conjunctivitis, 16 parents of babies with neonatal inclusion conjunctivitis and others were tested for IgM antibodies to Chlamydiae. Diagnostic kits for chlamydial IgM antibodies (SeroELISA and IPAzyme) have been also evaluated for their diagnostic value. It was found that detection of specific IgM antibodies with SeroELISA has a diagnostic value in chlamydial pneumonias.


[Identification of mycobacteria by the acridinium-ester labeled DNA probes for M. tuberculosis and M. avium-intracellulare complex in culture and its clinical application].

The detectability of mycobacterium in culture by non-isotopic, chemiluminescent DNA probes for Mycobacterium tuberculosis (M. tuberculosis) and M. avium-intracellulare complex (MAC) was evaluated and compared with that by 125I-labeled DNA probe for the same mycobacteria. The sensitivity and specificity of the AE-DNA probes for MAC were 97.2% and 100%, respectively, for the conventional method and were both 100% for the 125I-labeled DNA probes. The detection limits of the AE-DNA probes tests for both M. tuberculosis and MAC were 10(5)-10(6) CFU/tube which were almost the same as those of the 125I-labeled DNA probes tests. Because the procedure is simple, rapid (it can be completed within 90 min), and safe (it does not use radioisotopes), it can be easily performed in any clinical laboratory.


[Four cases of respiratory tract infections caused by Corynebacterium pseudodiphtheriticum].

Four cases of respiratory tract infections caused by Corynebacterium pseudodiphtheriticum were reported. The first two patients developed pneumonia with Corynebacterium pseudodiphtheriticum during steroid therapy used against their underlying diseases. The other two patients had acute exacerbation of chronic pulmonary diseases caused by C. pseudodiphtheriticum. These four patients improved by antibiotic therapy. Though nondiphtheria corynebacteria are regarded as "normal flora" when they are isolated from sputum, they should be recognized as potential pathogens.


[A case of miliary tuberculosis (miliary TB) accompanied with adult respiratory distress syndrome (ARDS) in a patient with Cushing's syndrome].

A 74-year-old housewife was admitted to the hospital with complaints of high fever and general fatigue. The physical examinations on admission showed no particular findings except for mild hepatomegaly, but laboratory findings showed severe liver dysfunction, active inflammation and negative tuberculine test. On the 4th day, she suddenly complained of severe respiratory distress. A chest X-ray film demonstrated surprising changes in comparison with that taken on admission. On suspicion of adult respiratory distress syndrome (ARDS) associated with military tuberculosis (Miliary TB), administration of Methylpredonisolone (1000 mg a day for 3 days) in addition to antituberculous drugs was immediately started. With this therapy she was recovered from such ill condition, but the general exhaustion and slight fever continued. We suspected that her condition might be due to adrenocortical involvement of Miliary TB and hormonal examinations were performed. Unexpectedly, Cushing's syndrome was suspected on the basis of the following; high level of plasma cortisol without normal daily variation, normal ACTH level, an absent response to the Dexamethasone suppression test. Computed tomography revealed left side adrenal mass. During these examinations, renal dysfunction probably due to Miliary TB grew gradually worse and she died of renal failure on the 56th day. Necropsy revealed disseminated tuberculosis involving the lungs and the liver, but the adrenal glands were not examined.


[A case of HBsAg positive liver cirrhosis who died after withdrawal of steroid].

A 42-year-old male was admitted with subarachnoidal hemorrhage. Dexamethasone 224 mg was used to reduce brain edema. His operation was successful without blood transfusion. No remarkable signs and symptoms were found except HBsAg positive and mild GPT elevation during his admission. He was discharged on the 33rd day. But 2 weeks later, he felt general fatigue and became worse day by day. He was re-admitted on the 75th day. Several therapies were given but he died of hepatic failure on the 85th day. The autopsy showed liver cirrhosis with massive necrosis. We believed that the steroid-withdrawal-phenomenon caused excessive immunological response and this process caused his hepatic failure leading to death.


[Enteropathogenicity and enteropathogenic toxin production of Vibrio mimicus].

Enteropathogenicity and enteroreactive-toxins were examined in 66 strains of Vibrio mimicus and the following results were obtained. Frequencies of enteropathogenic strains judged by the result of suckling mouse tests were 11/13 (85%) for clinical isolates and 37/53 (70%) for fish or environmental isolates. Frequencies of preservation of cholera toxin gene and NAG-ST gene were 2 and 15%, respectively, for 48 enteropathogenic strains, and 0 and 6%, respectively, for 18 non-enteropathogenic strains. Frequencies of production of NAG-rTDH, FAF and hemolysin were 4, 63 and 100%, respectively, for 48 enteropathogenic strains, and 6, 50, and 100%, respectively, for 18 non-enteropathogenic strains. No correlation between serovar and enteropathogenicity was observed in the suckling mouse test. Six out of 12 enteropathogenic strains produced hemolysin in ligated rabbit ileal loop, while 1 out of 12 non-enteropathogenic strains did so under the same condition. A significant inhibition of fluid accumulation in the ligated rabbit ileal loop test with viable cells was noted in rabbits immunized with hemolysin of non-O1 V. cholerae. These results suggest that approximately two-thirds of environmental isolates are enteropathogenic and that hemolysin is the most important toxin in the enteropathogenic mechanism of V. mimicus strains.


[Aztreonam or gentamicin combined with piperacillin as empiric antibiotic therapy during neutropenia of patients with hematologic diseases].

Fourty-two febrile episodes of 32 patients with hematologic disease during neutropenia were treated with two randomly assigned antibiotic combinations of either piperacillin plus gentamicin or piperacillin plus aztreonam. Eleven of the 22 febrile episodes treated with piperacillin plus gentamicin and 12 of the 20 febrile episodes treated with piperacillin plus aztreonam responded. Addition of cefamandole to non-responders improved the outcome in 2 of the 16 febrile episodes. Mean nadir leucocyte count, age, sex, and underlying disease were not significantly different in both groups. Side effects were tolerable in both groups, although 1 patient treated with piperacillin plus gentamicin showed severe renal impairment. Piperacillin plus aztreonam is as effective as piperacillin plus gentamicin as an empiric antibiotic combination in the treatment of febrile episodes with hematologic disease during neutropenia.


[O serogroup and virulence factors of motile Aeromonas].

A total of 182 isolates of motile Aeromonas from patients with diarrhea and environmental sources was investigated for hemolytic activity to rabbit erythrocyte and cytotoxicity to HeLa 229 cell. Furthermore, the relation between O serogroup and virulence factors, which were lethal to mouse and autoagglutination, were investigated. There were many strains possessing both the hemolytic and cytotoxic activities in A. hydrophila from overseas traveller's diarrhea, suggesting that these activities were associated with intestinal pathogenecity. Although there was a clear correlation between hemolytic and cytotoxic activities in A. hydrophila from overseas traveller's diarrhea, the correlation was not found in A. hydrophila from domestic cases of diarrhea and A. sobria from overseas traveller's diarrhea. Especially, some A. hydrophila isolates from domestic case of diarrhea produced only hemolysin. These results indicated that there was a difference in specificity between the toxins accounted for hemolytic and cytotoxic activities, and more than two different toxins were developed. O serogroups 11, 34, 14, 16, and 35 in that order were the most frequent serogroups. About half of O11 and O34 strains possessed lethal activity to mouse. Autoagglutination phenomenon did not seem to be associated with the lethal activity. In O11 strains, high cytotoxic titer was more frequently found in lethal activity positive-strains than in the activity negative-strains, suggesting that cytotoxicity contributed preferentially to lethal activity to mouse. But such a correlation was not found in O34 strains, so other virulence factors than hemolysin and cytotoxin may be associated with the lethal activity.


[A clinical study on postantibiotic effect (PAE) and its application to chemotherapy for complicated cystitis with an automatic simulator of urinary drug concentration].

In an in vitro complicated cystitis model, the concentrations of the urinary antimicrobial agents were determined using a computer-controlled automatic urine concentration simulator. The effects on the bacterial count curves showing the presence or absence of PAE in antimicrobial agents were studied by comparing the times required for regrowth to the concentration at the initial inoculation, i.e., effective regrowth time (ERT). The following results were obtained. 1. When beta-lactam antimicrobial agents (such as AMPC and CFIX) with no PAE against the gram-negative rods were tested, the ERT of the gram-negative rods were about two hours shorter than that of the gram-positive coccus. 2. When new quinolone antimicrobial agents (such as OFLX) and aminoglycosides (such as ISP) that possess PAE against both the gram-positive and negative organisms were used there was no difference between ERT of the gram-negative rods and gram-positive coccus. Therefore, it was demonstrated that the presence or absence of PAE is also reflected in the cell number curve in the case of this in vitro model, more closely related to clinical cases, when the antibiotics is simulated in urinary concentration shifting.


Proteus penneri isolated from the pus of a patient with epidural abscess.

P. mirabilis and P. vulgaris are the two wellknown species in the genus Proteus. P. myxofaciens and P. penneri are recent additions to the genus. We isolated P. penneri from the pus of a patient with suppurative otitis media and an epidural abscess. The characteristics of the organism, including morphology, staining, physiology and biochemistry, were studied. Clinical microbiological laboratories should suspect P. penneri in the case of as Proteus strain that is negative for indole, salicin and esculin, but otherwise resembles P. vulgaris. Proteus penneri, formerly known as Proteus vulgaris indole-negative or as Proteus vulgaris biogroup 1, was named by Hickman et al in 1932. Little information about human infection by this organism is available. In 1982, Hickman and co-workers studied 20 strain of P. penneri which were isolated from clinical specimens (urine, stool, etc.) in the USA. However, its clinical significance, until recently, was unknown. We isolated a strain of P. penneri from the pus of a patient with suppurative otitis media and an epidural abscess on June 10 and 15, 1989. This paper concerns the problems encountered in identifying this organism and its clinical significance.


Misleading serological identification of Legionella anisa as Legionella bozemanii.

Identification of six Legionella species, which we previously identified by serological test as Legionella bozemanii (L. bozemanii), was performed by DNA-DNA hybridization using a commercial DNA-DNA hybridization kit (Kobayashi Pharm. Co., Japan) introduced by Ezaki et al. All strains were identified as Legionella anisa (L. anisa), this being the first identification of L. anisa in Japan. Conventional laboratory tests were performed following the DNA-DNA hybridization. In this study the results of biochemical examination obtained, corresponded closely with those described in previous reports, but the oxidase reaction was very weak and varied according to the age of the culture, indicating the unreliability of this test in our case. All strains examined under long wave ultraviolet (UV) light (366 nm) revealed a blue-white fluorescence, the intensity of which ranged from strong to weak. Serological identifications were performed by both the slide agglutination test (SAT) and indirect immunofluorescent assay (IFA). SAT using commercially available antiserum (Denka Seiken., Japan) supposedly specific for L. bozemanii showed cross-reaction between L. bozemanii and L. anisa. Hyperimmune rabbit antisera prepared in this study for both L. bozemanii and L. anisa, from which cross-reactive antibodies were removed by the absorption of each antigen, reacted only with homologous antigens. IFA using a commercially available antiserum and hyperimmune rabbit antiserum previously described, gave positive reactions with each strain.


[Studies of serological diagnosis for invasive candidiasis].

Serological diagnosis was examined for deep mycotic infections, especially candidiasis. Candida antigen and antibody researches were made by means of CAND-TEC (RAMCO Co., Ltd) and passive hemagglutination test (PHA) (Roche Co., Ltd), respectively, and simultaneous determinations were made of D-arabinitol, a fungal metabolite and (1----3)-beta-D-glucan, fungal parietal component. 1) For normal subjects, 1 of 173 cases (0.6%), 5 of 200 cases (2.5%) and 7 of 157 cases (4.5%) showed greater than or equal to 1:4, greater than or equal to 320 x and greater than or equal to 11 mumol/l for CAND-TEC, Candida antibody and D-arabinitol, respectively, and values above these were judged positive. 2) Of the 171 patients with pyrexia refractory to general antibiotic agents, who were treated at our Department and related institutions from November 1988 to March 1990, 41 with obvious Candida infections were CAND-TEC, Candida antibody, D-arabinitol and (1----3)-beta-D-glucan positive in 33 (80.5%), 21 (51.2%), 29 (70.7%) and 13 cases (56.6%), respectively. At least one item-positive patient was as high as 97.4%. On the other hand, in the group having no Candida infection at all, 1 of 57 cases on CAND-TEC (1.7%) and 10 of 55 cases on Candida antibody (18.2%) were positive, indicating significant differences from the surely Candida-infectious group. 3) Comparative examination of CAND-TEC and other testing methods revealed a correlation of CAND-TEC with D-arabinitol in cases showing 1:4 and 1:8 less than or equal to, but not other significant difference.(ABSTRACT TRUNCATED AT 250 WORDS)


[Comparison of polymerase chain reaction and enzyme immunoassay (Chlamydiazyme) in detection of Chlamydia trachomatis from male urethritis].

A method using polymerase chain reaction (PCR) was compared to an enzyme immunoassay (Chlamydiazyme) for detection of Chlamydia trachomatis by testing a reference strain and clinical specimens. Two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2 were used as primers for the PCR. A DNA fragment of 242 bp specific for C. trachomatis was amplified by the PCR, when DNA of greater than or equal to 10(2) C. trachomatis was used as template for the PCR. A chlamydial antigen was detected by Chlamydiazyme, when greater than or equal to 2.6 x 10(3) C. trachomatis were applied for the enzyme immunoassay. The PCR method was 26 times more sensitive than Chlamydiazyme in detection of C. trachomatis. The PCR method and Chlamydiazyme were carried out to examine 74 urethral swabs obtained from male patients with urethritis for detection of C. trachomatis. In 45 of 74 specimens, the DNA fragment of C. trachomatis was amplified by the PCR, and in 41 of 74, the chlamydial antigen was detected by Chlamydiazyme. The detection rate of the PCR method (60.8%) was higher than that of Chlamydiazyme (55.4%). The positive coincidence rate of the PCR method to Chlamydiazyme was 100% (41/41) and negative coincidence rate was 87.9% (29/33). The overall coincidence rate between the two methods was high (94.6%). Thus, the PCR method was more sensitive than Chlamydiazyme for detection of C. trachomatis and specific for diagnosis of chlamydial urethritis.