The outcome of Japanese anorexia nervosa patients treated with an inpatient therapy in an internal medicine unit.

OBJECTIVE: To investigate the outcome of Japanese anorexia nervosa (AN) patients who were treated with the standard Japanese inpatient therapy. METHOD: Of the 88 female AN patients treated with our inpatient therapy between January 1997 and December 2002, 67 (76.1%) who agreed to cooperate in this study were assessed by the Global Clinical Score (GCS) at admission and follow-up, 6.3±1.8 years after discharge. Their clinical characteristics at admission and discharge were also examined. RESULTS: Four (6.0%) patients had died before follow-up. BMI was significantly increased during inpatient therapy. At follow-up, excellent, much improved, symptomatic, and poor outcomes on GCS were 57.1%, 14.3%, 14.3% and 14.3%, respectively. Younger age at admission and larger BMI at discharge were significantly associated with a better outcome. DISCUSSION: This study shows the potential for the use of this method for the treatment of AN patients in countries without specialized eating disorder units.

Evidence for clonal selection of gamma/delta T cells in response to a human pathogen.

T cells bearing gamma/delta antigen receptors comprise a resident population of intraepithelial lymphocytes in organs such as skin, gut, and lungs, where they are strategically located to contribute to the initial defense against infection. An important unsolved question about antigen-driven gamma/delta T cell responses regards the breadth of their T cell receptor (TCR) repertoire, since many specific epithelial compartments in mice display limited diversity. We have examined the diversity of TCR delta gene expression among human gamma/delta T cells from skin lesions induced by intradermal challenge with Mycobacterium leprae. We show that the vast majority of gamma/delta cells from M. leprae lesions use either V delta 1-J delta 1 or V delta 2-J delta 1 gene rearrangements and, within a given region of the lesion, display limited junctional diversity. This contrasts markedly with the extensive diversity of gamma/delta T cells from peripheral blood of these same individuals, as well as skin from normal donors. These results indicate that the gamma/delta response to M. leprae involves the selection of a limited number of clones from among a diverse repertoire, probably in response to specific mycobacterial and/or host antigens.

Interactions of human alpha/beta and gamma/delta T lymphocyte subsets in shear flow with E-selectin and P-selectin.

We have compared the ability of human alpha/beta and gamma/delta T lymphocytes to adhere to selectin-bearing substrates, an interaction thought to be essential for homing and localization at sites of inflammation. Both T cell populations form rolling adhesions on E- and P-selectin substrates under physiologic flow conditions. Although equivalent to alpha/beta T cells in binding to E-selectin, gamma/delta T cells demonstrated greater ability to adhere to P-selectin that was purified or expressed on the surface of activated, adherent platelets. Under static conditions, 80% of gamma/delta T cells and 53% of alpha/beta T cells formed shear-resistant adhesions to P-selectin, whereas only 30% of gamma/delta and alpha/beta T cells adhered to E-selectin. The enhance ability of gamma/delta T cells to adhere to P-selectin cannot be attributed to differences in expression of the P-selectin glycoprotein ligand (PSGL-1), as all alpha/beta T cells versus approximately 75% of gamma/delta T cells expressed PSGL-1. Both cell populations expressed a similar percentage of the carbohydrate antigens sialyl LewisX and cutaneous lymphocyte-associated antigen. Depletion of lymphocyte populations or T cell clones bearing these oligosaccharides with the monoclonal antibody CSLEX-1 and HECA-452, respectively, resulted in a substantial reduction in adhesion to E-selectin and slight reduction in adhesion to P-selectin under flow conditions. Treatment of cells with an endopeptidase that selectively degrades O-sialomucins such as PSGL-1, abolished P-selectin but not E-selectin adhesion. Removal of terminal sialic acids with neuraminidase or protease treatment of cells abrogated cell adhesion to both selectin substrates. These results provide direct evidence for the presence of distinct E- and P-selectin ligands on T lymphocytes and suggest that gamma/delta T cells may be preferentially recruited to inflammatory sites during the early stages of an immune response when P-selectin is upregulated.

Evidence for extrathymic changes in the T cell receptor gamma/delta repertoire.

The germline repertoire of variable genes for the TCR-gamma/delta is limited. This, together with the availability of several V delta-specific and a C delta-specific mAbs, has made it possible to assess differences in the TCR-gamma/delta repertoire in man. TCR-gamma/delta cells expressing particular V gene segments have been previously shown to be localized in different anatomical sites. In this study, analysis of TCR-gamma/delta V gene segment usage performed on subjects from the time of birth through adulthood revealed striking age-related changes in the TCR-gamma/delta repertoire in peripheral blood. V delta 1+ gamma/delta T cells predominated in thymus as well as in peripheral blood at birth and then persisted as a relatively constant proportion of CD3+ PBL. However, V delta 2+ gamma/delta T cells that constitute a small proportion of the CD3+ cells in thymus and in peripheral blood at birth, then expand and account for the major population of gamma/delta T cells in PBL in adults. No parallel postnatal expansion of V delta 2+ cells in the thymus was observed, even when paired thymus-peripheral blood specimens were obtained on subjects between the ages of 3 d and 8 yr. The subset of V delta 2+ lymphocytes that was expanded in peripheral blood expressed high levels of CD45RO suggesting prior activation of these cells, consistent with the possibility that their expansion might have resulted from exposure to foreign antigens or superantigens. In contrast, V delta 1+ T cells in PBL showed no comparable increase in relative numbers and were either negative or expressed only low levels of CD45RO. Consistent with evidence for extrathymic peripheral expansion of selective TCR-gamma/delta subsets, no link between MHC haplotype and differences in the TCR-gamma/delta V gene usage between individuals was apparent, and identical twins displayed TCR-gamma/delta variable gene segment phenotypes that were strikingly different from one another. The elements that determine the TCR-gamma/delta repertoire in individuals are not known. It is possible that both thymic selection and extrathymic factors may influence the peripheral repertoire. Recently, TCR-gamma/delta+ lymphocytes have been shown to expand markedly in peripheral lymphoid tissues and infectious lesions in response to mycobacterial antigens, and a correlation between mycobacterial responses and TCR-gamma/delta V gene usage has been shown in mice. The data presented here demonstrated peripheral age-related changes in the gamma/delta repertoire and point to the importance of extrathymic expansion of specific gamma/delta subsets in generating the human TCR-gamma/delta repertoire.

Self-recognition of CD1 by gamma/delta T cells: implications for innate immunity.

The specificity of immunoglobulins and alpha/beta T cell receptors (TCRs) provides a framework for the molecular basis of antigen recognition. Yet, evolution has preserved a separate lineage of gamma/delta antigen receptors that share characteristics of both immunoglobulins and alpha/beta TCRs but whose antigens remain poorly understood. We now show that T cells of the major tissue gamma/delta T cell subset recognize nonpolymorphic CD1c molecules. These T cells proliferated in response to CD1+ presenter cells, lysed CD1c+ targets, and released T helper type 1 (Th1) cytokines. The CD1c-reactive gamma/delta T cells were cytotoxic and used both perforin- and Fas-mediated cytotoxicity. Moreover, they produced granulysin, an important antimicrobial protein. Recognition of CD1c was TCR mediated, as recognition was transferred by transfection of the gamma/delta TCR. Importantly, all CD1c-reactive gamma/delta T cells express V delta 1 TCRs, the TCR expressed by most tissue gamma/delta T cells. Recognition by this tissue pool of gamma/delta T cells provides the human immune system with the capacity to respond rapidly to nonpolymorphic molecules on professional antigen presenting cells (APCs) in the absence of foreign antigens that may activate or eliminate the APCs. The presence of bactericidal granulysin suggests these cells may directly mediate host defense even before foreign antigen-specific T cells have differentiated.

Somatic and psychological factors related to the body mass index of patients with anorexia nervosa.

OBJECTIVE: The aim of this study was to examine somatic and psychological factors related to the body mass index (BMI) of anorexia nervosa (AN) patients. METHOD: The analysis was of 24 hospitalized AN patients from the day after admission to the 4th day. The somatic factors analyzed were duration of AN, daily food intake, eating regulatory substances in blood (acylated ghrelin, desacyl ghrelin, leptin), serum cortisol, insulin and estimated creatinine clearance (CCr). The psychological factors analyzed were depression, anxiety, Eating Disorder Inventory (EDI), and hunger/fullness feeling. Measurement of BMI and collection of blood samples were done on the morning after hospitalization. Statistical analysis was by multiple linear regression analysis. RESULTS: BMI showed a reverse correlation with desacyl ghrelin (beta=-0.486, p=0.015) and maturity fears (beta=-0.375, p=0.046), but was not associated with any other factor by multiple regression analysis. CONCLUSION: The results suggest that desacyl ghrelin and maturity fears play important roles in the prolonged malnutrition state seen in AN patients.

T-cell recognition of non-peptide antigens.

Studies of two distinct human T-cell systems have provided the exciting finding that T cells are able to recognize non-peptide antigens: gammadelta T cells have been shown to recognize isopentenyl pyrophosphate and related structures and human CD1 has been shown to present microbial lipids and lipoglycans such as mycolic acids and lipoarabinomannan to T cells. T-cell responses to these non-peptide antigens should provide a strategic target for immunologic intervention in infectious disease.

Synthesis of pyrophosphate-containing compounds that stimulate Vgamma2Vdelta2 T cells: application to cancer immunotherapy.

Human Vgamma2Vdelta2 T cells recognize nonpeptide antigens, such as isoprenoid pyrophosphomonoester intermediates, alkylamine compounds, and bisphosphonate drugs, as well as some tumor cells. Although attempts have been made to derive novel cancer immunotherapies based on the discovery of these unconventional antigens, effective therapies remain to be developed. Here, we synthesized a series of pyrophosphate-containing compounds and examined the chemical requirements for the recognition of pyrophosphomonoester antigens by gammadelta T cells. The structural analysis clearly demonstrated that a proximal methylene moiety plays a crucial role in the stimulatory activity of the antigens. For optimal gammadelta T cell proliferation, we find that the use of human serum albumin was preferred and that pyrophosphomonoesters were superior to nitrogen-containing bisphosphonate compounds. Using these techniques, we have successfully expanded gammadelta T cells from healthy donors as well as from cancer patients using one of the most active compounds, 2-methyl-3-butenyl-1-pyrophosphate (2M3B1PP). The resulting expanded gammadelta T cells exhibited potent, cytotoxic activity against a wide variety of tumor cell lines. Even gammadelta T cells from a patient with advanced liver carcinoma efficiently responded to 2M3B1PP and exhibited strong cytotoxic activity against tumor cells. The pretreatment of tumor cells with nonpeptide antigens was essential for efficient cytotoxicity via TCR-gammadelta. The present study suggests a novel strategy for cancer immunotherapy using synthetic small pyrophosphate-containing compounds and nitrogen-containing bisphosphonates.

A change in objective sleep duration is associated with a change in the serum adiponectin level of women with overweight or obesity undergoing weight loss intervention.

BACKGROUND: Although the serum adiponectin level is inversely correlated to body mass index and closely associated with obesity and related diseases, neither the impact of weight loss on the adiponectin level nor other factors that might influence the adiponectin level during weight loss intervention are well documented. OBJECTIVE: The objective of the study is to assess the change in the serum adiponectin level during weight loss intervention and to determine if sleep parameters affect the serum adiponectin level. METHODS: Ninety women with overweight or obesity aged 25 to 65 years completed a 7-month cognitive behavioural therapy based weight loss intervention that included dieting, exercise and stress management. Serum adiponectin level, body fat percent, symptoms of depression and anxiety and objective sleep parameters, assessed by actigraphy, were measured at baseline and at the end of the intervention. RESULTS: The serum adiponectin level was significantly increased after the weight loss intervention (P < 0.001). In a multiple regression analysis, the change of the adiponectin level was positively associated with the magnitude of body fat loss (ß = -0.317, P < 0.001) and an increase of sleep minutes (ß = 0.210, P = 0.043). CONCLUSION: An increase in objective sleep duration was related to a significantly increased serum adiponectin level independently of the change of body fat during the weight loss intervention.

Detection of serum antibodies to Bartonella henselae and Coxiella burnetii from Japanese children and pregnant women.

The participation of Bartonella henselae and Coxiella burnetii in the pathogenesis of fever of unknown origin (FUO) and lymphadenopathy has not been completely clarified. Prevalence of these two agents in Japanese children is also unknown. Serum IgG and IgM antibodies to B. henselae and to C. burnetii were examined by the indirect fluorescence antibody assay. Enzyme immunoassay kits were used to detect serum IgG and IgA antibodies against Chlamydia trachomatis. Out of 200 healthy normal pregnant women, two (1.0%) had serum IgG antibodies to B. henselae, four (2.0%) to C. burnetii and 49 (24.5%) to C. trachomatis. Out of 29 patients with FUO, one (3.4%) had serum IgG antibodies to B. henselae, four (13.8%) to C. burnetii and none to C. trachomatis. Out of 31 patients with cervical lymphadenopathy, three (9.6%) had serum IgG antibodies to B. henselae, two (6.5%) to C. burnetii and none to C. trachomatis. Out of 22 patients with generalized lymphadenopathy, one (4.5%) had serum IgG antibodies to B. henselae, three (13.6%) to C. burnetii and none to C. trachomatis. Prevalences of serum antibodies to C. burnetii in the patients with FUO and generalized lymphadenopathy and to B. henselae in the patients with cervical lymphadenopathy were significantly higher than those of normal pregnant women (Welch's t-test; P<0.01). These two agents may have some roles in the pathogenesis of FUO and lymphadenopathy in Japanese children.

Isolation and characterization of cell clones persistently producing teardrop-shaped particles of human immunodeficiency virus type 1.

Several cell clones producing teardrop-shaped human immunodeficiency virus type 1 (HIV-1) particles were isolated from MT-4 cells that survived HIV-1 infection after extensive cell lysis. Most of the teardrop-shaped particles contained an electron-dense core structure, and the particles could replicate in MT-4 cells. No significant structural difference in the HIV-1 proteins was observed between these cell clones and the MOLT-4 cells producing intact HIV-1 particles, although their envelope structure is morphologically abnormal.

Retinoblastoma: tissue culture lines and monoclonal antibody studies.

Retinoblastoma patients have cellular and humoral reactivity towards antigens expressed on retinoblastoma cells. We report the ultrastructural, cytogenetic, and immunologic features of four new retinoblastoma derived tissue culture cell lines. Studies with hybridoma produced monoclonal antibodies demonstrate that these lines share antigens with a previously described long-term allogeneic retinoblastoma derived tissue culture cell line, as well as with antigens on fresh retinoblastoma.

Short report: prevalence of antibodies against spotted fever, murine typhus, and Q fever rickettsiae in humans living in Zambia.

The causative agents of rickettsial diseases (Rickettsia conorii, R. typhi, and Coxiella burnetii) have been reported throughout the African continent. However, there have been no reports on epidemiologic surveys of these infections in Zambia. This study was designed to clarify the prevalence of three rickettsioses in 377 humans in Zambia. The seroprevalence of antibodies against R. conorii, R. typhi, and C. burnetii was 16.7%, 5.0%, and 8.2%, respectively. The rates of antibody positivity against R. conorii and C. burnetii were higher in the eastern (23.1% and 11.8%) and western (16.8% and 7.4%) areas of Zambia than in the northern (3.0% and 3.0%) area of this country. There was little difference among the three areas in the distribution of antibodies against R. typhi. Since cattle breeding is more extensive in the western and eastern areas than in the northern area, it is thought that cattle-breeding areas are foci of R. conorii and C. burnetii infections in Zambia.

Contrast-enhancement for the image of human immunodeficiency virus from ultrathin section by immuno electron microscopy.

A simple contrast-enhancement method is described for electron microscopic imaging of the human immunodeficiency virus (HIV) from a sample embedded in Lowicryl K4M resin, by immuneelectron microscopy. Ultrathin sections were treated with a mixture of ruthenium red dye (RR) and osmium tetroxide (OSO4). This treatment provided good contrast enhancement of the entire ultrastructural image of virus particles without the loss of immunolabelling. RR/OsO4 solution is simple to prepare and provides a better contrast than that which is achieved during conventional post-embedding immunoelectron microscopy. Treatment of ultrathin sections from low temperature-embedded samples with RR/OsO4 solution is recommended.

Disinfection potential of electrolyzed solutions containing sodium chloride at low concentrations.

Electrolyzed products of sodium chloride solution were examined for their disinfection potential against hepatitis B virus (HBV) and human immunodeficiency virus (HIV) in vitro. Electrolysis of 0.05% NaCl in tap water was carried out for 45 min at room temperature using a 3 A electric current in separate wells installed with positive and negative electrodes. The electrolyzed products were obtained from the positive well. The oxidation reduction potential (ORP), pH and free chlorine content of the product were 1053 mV, pH 2.34 and 4.20 ppm, respectively. The products modified the antigenicity of the surface protein of HBV as well as the infectivity of HIV in time- and concentration-dependent manner. Although the inactivating potential was decreased by the addition of contaminating protein, recycling of the product or continuous addition of fresh product may restore the complete disinfection against bloodborne pathogens.

Efficacy of combination antiplatelet therapy and nicardipine for chronic cerebral infarction.

To prevent recurrence of cerebral infarction (CI), the efficacy of antiplatelet therapy, when used in combination with a calcium antagonist, was examined. The study subjects were 57 chronic CI patients (40 men, 17 women; mean age, 68.5 years) who experienced either CI or its recurrence more than 3 months before the start of the study. They were randomly allocated into one of the following four groups for the 8-week study; group A--ticlopidine hydrochloride 200 mg once daily and nicardipine hydrochloride 20 mg three times daily (TID); group B--ticlopidine hydrochloride 200 mg once daily; group C--aspirin 81 mg once daily and nicardipine hydrochloride 20 mg TID; or group D--aspirin 81 mg once daily. Platelet aggregation was measured before treatment and 4 and 8 weeks after the initiation of each therapy by using adenosine diphosphate (ADP) (2 microM and 0.5 microM) and collagen (2 micrograms/mL), and evaluated in terms of percent maximum platelet aggregation. Results showed significant suppression of 2.0 microM ADP platelet aggregation in groups A, B, and C. At 0.5-microM ADP, only groups A and B showed significant platelet aggregation suppression. All groups showed significant suppression of collagen platelet aggregation. In comparing single therapy with combination therapy, groups A and B were not significantly different from one another after 4 or 8 weeks in 2-microM ADP or collagen platelet aggregation suppression. In contrast, group C had significantly greater suppression of both 2-microM ADP and collagen aggregations compared with group D. In conclusion, nicardipine hydrochloride administration with aspirin may be a useful alternative therapy for the prevention of CI recurrence.

Effective administration methods and dosages of interferon therapy for chronic hepatitis C.

The efficacy of interferon alfa (IFN-alpha) was evaluated in 127 hepatitis C virus-ribonucleic acid (HCV-RNA)-positive patients with chronic hepatitis C in relation to HCV-RNA levels and genotype. Patients were assigned to one of three groups. Patients in group A received IFN-alpha daily for 8 weeks (total dose, 336 million units [MU]); patients in group B received IFN-alpha daily for 4 weeks and then intermittently for 20 weeks (total dose, 348 MU); and patients in group C received IFN-alpha daily for 2 weeks and then intermittently for 22 weeks (total dose, 480 MU). Complete response rates in groups B and C were significantly higher than those in group A, regardless of the virus level or genotype. Complete response rates in groups B and C were similar, but in patients with a high virus level or HCV-RNA genotype II, the partial response rate in group C was significantly higher than that in group B. In conclusion, IFN-alpha was more effective after daily and then intermittent administration than after daily administration only, and a higher daily dose was necessary to be more effective in patients with high virus levels or HCV-RNA genotype II.

Propylthiouracil-induced severe hepatitis: a case report and review of the literature.

A 21-year-old woman was diagnosed as having Graves' disease in April, 1995. Thiamazole was administered; about a month later the patient had a skin rash and propylthiouracil (PTU) was given instead. Two months after commencing PTU, she rapidly developed jaundice, accompanied by severe liver damage. The drug-induced lymphocyte stimulating test was positive for PTU and she was diagnosed as having severe hepatitis induced by PTU. After pulse therapy with 500 mg of methylprednisolone was given for 3 days, liver function test results were gradually improved, and became normalized 1 1/2 months after admission. The pathology findings of the liver biopsy sample taken before administration of corticosteroid showed necrosis of hepatocytes predominantly around the central veins (i.e., zone 3 necrosis), and moderate to severe infiltration of lymphocytes and neutrophils in portal areas and lobules. Severe hepatic damage due to PTU is rare; 25 cases have been reported so far in the English-language literature. When we use PTU for patients with hyperthyroidism, we should keep in mind that severe liver damage induced by PTU can be fatal, and we should therefore diagnose it earlier by liver biopsy and lymphocyte stimulating test.

Contrast-enhanced immunoelectron microscopy for Helicobacter pylori.

Since a method of contrast enhancement for immunoelectron microscopy has not been available in bacteriology, the morphological localization of proteins of Helicobacter pylori is not well known. In this report, we established a method of contrast enhancement in immunoelectron microscopy in this organism. Immunostained ultrathin sections are stained with a mixture of alcian blue and osmium tetroxide prior to staining with uranyl acetate. This method of staining provided good contrast enhancement of the bacterial cell wall and membrane without any loss of immunolabeled gold particles on the ultrathin section.

Insulin autoimmune syndrome is the third leading cause of spontaneous hypoglycemic attacks in Japan.

From 1979 to 1981, questionnaires were sent to 2094 hospitals throughout Japan to investigate the causes of severe hypoglycemic attacks other than the administration of oral hypoglycemic agents or insulin preparations. The survey revealed three main causes for the attacks, of which the first was insulinoma, the second extrapancreatic neoplasms, and the third was insulin autoimmune syndrome (IAS), in descending order. Seven years later, a second survey was carried out, which showed the order of the three disorders as the cause of the hypoglycemic attacks to be the same as in the first survey. In both studies it was suggested that the IAS was frequently induced by thiol compounds.