Decreased survival in vivo of diamide-incubated dog erythrocytes. A model of oxidant-induced hemolysis.

Erythrocytes from patients with chronic hemolytic variants of glucose-6-phosphate dehydrogenase (G-6-PD) deficiency have structural membrane protein abnormalities accompanied by decreased cell membrane deformability which we postulate represent the consequences of oxidant-induced membrane injury. To evaluate the pathophysiologic significance of oxidant-induced membrane injury, we studied the in vitro and in vivo effects of the thiol-oxidizing agent, diamide, on dog erythrocytes. In vitro incubation of dog erythrocytes with 0.4 mM diamide in Tris-buffered saline for 90 min at 37 degrees C resulted in depletion of GSH, formation of membrane polypeptide aggregates (440,000 and > 50,000,000 daltons) and decreased cell micropipette deformability, abnormalities similar to those observed in the erythrocytes of patients with chronic hemolytic variants of G-6-PD deficiency. In addition, diamide-incubated cells had increased viscosity and increased membrane specific gravity, but no change in ATP. Reinjection of 51Cr-labeled, diamide-incubated cells was followed by markedly shortened in vivo survival and splenic sequestration. Further incubation of diamide-incubated cells in 4 mM dithiothreitol reversed the membrane polypeptide aggregates, normalized micropipette deformability, decreased cell viscosity, prolonged in vivi survival, and decreased splenic sequestration. These studied demonstrate that diamide induces a partially reversible erythrocyte lesion which is a useful model of oxidant-induced membrane injury. They suggest that oxidant-induced erythrocyte membrane injury plays an important role in the pathophysiology of chronic hemolysis which accompanies some G-6-PD variants.

CA-15.3, TPA and MCA as markers for breast cancer.

Serum concentrations of CA-15.3, tissue polypeptide antigen (TPA) and mucinous-like carcinoma-associated antigen (MCA) were measured in 327 women: 81 controls, 93 patients with benign breast disease, 46 patients recently diagnosed with breast cancer and 107 patients during breast cancer follow-up. CA-15.3 was elevated in 16% of the controls, in 29% of the patients with benign breast disease, in 65% of the breast cancer patients and in 74% of the follow-up patients. TPA was elevated in 4%, 11%, 36% and 75%, respectively. The corresponding figures for MCA were 10%, 8%, 30% and 64%. The highest sensitivity for cancer detection (74%) was obtained with a combination of CA-15.3 and TPA, while the specificity of this panel was 75%. The negative predictive value of these combined tests was 93%. MCA scored lower values, being only 30% sensitive. The CA-15.3/TPA panel may increase sensitivity compared with single marker tests and provide additional information for clinical evaluation.

Association between the serotonin 2C receptor gene and tardive dyskinesia in chronic schizophrenia: additive contribution of 5-HT2Cser and DRD3gly alleles to susceptibility.

RATIONALE: Tardive dyskinesia (TD) is a longterm adverse effect of dopamine receptor blockers. The dopamine D3 receptor gene (DRD3) ser9gly polymorphism has been previously associated with susceptibility to TD. Serotonin receptor antagonism has been proposed as a common mechanism contributing to the low extra-pyramidal effects profile of atypical antipsychotic drugs. OBJECTIVES: To examine the association of a functional polymorphism in the 5-HT2C receptor gene (HT2CR) with TD and the joint contribution of HT2CR and DRD3 to susceptibility. METHODS: Case control association analysis of allele and genotype frequencies among schizophrenia patients with (n=55) and without TD (n=60), matched for antipsychotic drug exposure and other relevant variables, and normal control subjects (n=97). Parametric analyses of the contribution of 5-HT2Cser and DRD3gly alleles to dyskinesia scores. RESULTS: We found a significant excess of 5-HT2Cser alleles in schizophrenia patients with TD (27.2%) compared to patients without TD (14.6%) and normal controls (14.2%; chi2=6.4, df 2, P=0.03) which was due to the female patients (chi2=8.6, df 2, P=0.01). Among the female TD patients there was an excess of cys-ser and ser-ser genotypes (chi2= 11.9, df 4, P=0.02). Analysis of covariance (ANCOVA), controlling for age at first antipsychotic treatment, revealed a significant effect of 5-HT2C genotype on orofacial dyskinesia (OFD) scores (F=3.47, df 2, P=.03). In a stepwise multiple regression analysis, 5-HT2C and DRD3 genotype (5-HT2Cser and DRD3gly allele carriage) respectively contributed 4.2% and 4.7% to the variance in OFD scores. CONCLUSIONS: These findings support a small but significant contribution of the HT2CR and DRD3 to susceptibility to TD, which is additive in nature.

A rare short allele of the serotonin transporter promoter region (5-HTTLPR) found in an aggressive schizophrenic patient of Jewish Libyan origin.

The serotonin transporter gene (SLC6A4), which plays a key role in the serotonergic pathway in the brain, is a candidate for mediating genetic susceptibility to various psychiatric disorders. There are two predominant alleles in the polymorphic promotor region [5-HT transporter gene-linked polymorphic region (5-HTTLPR)] of this gene: a long and a short allele with 16 and 14 repeat units, respectively. The short allele has lower activity and is associated with several psychiatric disorders and personality traits. We identified and sequenced a novel allele with 13 repeat units, 23 base pairs shorter than the common short allele. This unique allele was detected in a schizophrenic patient of Jewish Libyan origin. The patient exhibited extreme aggressive behavior and committed suicide after several attempts. The novel short allele was not detected in 172 healthy control subjects and 361 patients with various mental disorders. The presence of a very short unique allele in a severely aggressive schizophrenic patient may reflect a specific effect on the particular phenotype, although it is unlikely that this allele has a major contribution to susceptibility to schizophrenia. The role of the allele in serotonin transport and possible association with disease phenotype should be further investigated.

Intermittent, luteal phase nefazodone treatment of premenstrual dysphoric disorder.

Three outpatients who fulfilled full DSM-IV diagnostic criteria for premenstrual dysphoric disorder (PDD) were successfully treated with intermittent (luteal phase) nefazodone. They received the medication at low doses of up to 100 mg/day (50 mg b.i.d.), for 2 weeks through the luteal phase of the menstrual cycle only. All the patients reported a marked symptomatic improvement, including full remission of their emotional symptoms, and two achieved in addition full remission of their somatic symptoms. Side-effects reported during the treatment were mild. The use of luteal phase nefazodone seems to be a promising treatment strategy for the management of PDD. It offers advantages over daily dosing throughout the menstrual cycle, such as reduced incidence and severity of side-effects, and avoids the stigma that may accompany the continuous use of psychopharmacological treatment, with the advantage that compliance may be improved.

Dose-dependent olanzapine-associated leukopenia: three case reports.

Leukopenia and agranulocytosis are well reported and dangerous haematological side-effects associated with the use of typical and atypical antipsychotics. These potentially life-threatening phenomena have led to treatment discontinuation and the consequent reemergence of psychiatric symptoms. We report three cases of patients who developed leukopenia during olanzapine treatment. In each case, the leukopenia was dose-dependent. Reduction in the dose of olanzapine was followed by normalization of the white blood count which allowed continuation of the medication. These cases suggest the possibility that, in some patients, leukopenia or agranulocytosis during olanzapine treatment might be dose-related. Thus, olanzapine dose reduction may permit treatment continuation where this is clinically indicated. In our cases, haematological side-effects were satisfactorily controlled by dose reduction without allowing the reemergence of psychiatric symptoms. This clinical management may offer an alternative to treatment suspension. A careful monitoring of the white blood count is obviously recommended. Olanzapine may be considered a potential and safer treatment for a this specific group of patients.

LSD-induced hallucinogen persisting perception disorder treatment with clonidine: an open pilot study.

A pilot open study was conducted in order to evaluate the efficacy of clonidine in the treatment of LSD-induced hallucinogen persisting perception disorder (HPPD). Eight patients fulfilled entrance criteria. All complained of HPPD for at least 3 months and were drug free at least 3 months. They received fixed low doses of clonidine, 0.025 mg, three times a day for 2 months. They were evaluated by the Clinical Global Impression Scale (CGI) and a self-report scale on the severity of symptoms (graded 0-5). Patients scored an average of 5.25 (SD = 0.46) on the CGI and 4 on the self-report scale at baseline, indicating marked psychopathology. One patient dropped out at week 3 and a second patient dropped out at week 5. Of the six patients remaining at the end of 2 months, the average CGI score was 2.5 (SD = 0.55) and the self-report scale score was 2, indicating mild symptomatology. LSD-related flashbacks associated with excessive sympathetic nervous activity may be alleviated with clonidine in some patients.

Olanzapine treatment in patients with typical and atypical neuroleptic-associated agranulocytosis.

Two cases are reported of patients who developed agranulocytosis after treatment with typical and atypical antipsychotics. The patients were successfully treated with olanzapine. We suggest the consideration of olanzapine as a safer treatment in patients who require immediate continuation of antipsychotic medication and have a prior history of classic and atypical neuroleptic-induced agranulocytosis.

Effects of acute, low-dose UVB radiation on the induction of contact hypersensitivity to diphenylcyclopropenone in man.

Healthy volunteers (n = 14, age range 20-31 years, mean 23) were irradiated on the inside of the left forearm on four consecutive days with their individual minimal erythemal dose of ultraviolet B (UVB) prior to sensitization in the same skin area with a 2% solution of diphenylcyclopropenone (DPCP). The reaction patterns were compared with 14 alopecia areata patients (age range 16-69 years, mean 40) starting topical immunotherapy with DPCP, sensitized without prior UVB treatment. Primary allergic reactions occurred in ten volunteers and in four alopecia areata patients. Patch testing on the upper back with serial dilutions of DPCP (1% to 10(-8)%) showed minimal dermatitis-eliciting concentrations ranging from 1 to 10(-4)% (mean 0.19%) in the volunteers as compared with 10(-1) to 10(-8)% (mean 0.025%) in the alopecia areata patients. Two patterns were discernible within the volunteers with respect to the intensity of the primary allergic and elicitation reactions. Ten volunteers reacted in a similar way to the alopecia areata patients, whereas four probands demonstrated very high minimal dermatitis-eliciting concentrations and overall less severe reactions. The DPCP-specific T-cell response using blood macrophages and B lymphocytes as antigen-presenting cells was measured in an in vitro assay in two alopecia areata patients and two volunteers having similar skin reactions as well as in two volunteers with overall less severe skin reactions. B lymphocytes from the alopecia areata patients and the volunteers with similar skin reactions induced a significant DPCP-specific T-cell proliferation exceeding the responses obtained using macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)

Pulsed alexandrite laser technology for noninvasive hair removal.

OBJECTIVE: Determination of the efficacy of pulsed Alexandrite Laser technology for rapid noninvasive hair removal. SUMMARY BACKGROUND DATA: Although previous studies have already shown that Ruby lasers are capable of noninvasive hair removal, a technology for the substantial increase of treatment speed is of great interest. METHODS: We have used a 2 msec free running pulsed Alexandrite (lambda = 755 nm) laser operated at a repetition rate of up to 5 pps at energy fluences of 25-40 J/cm2 to treat a wide range of body sites on 126 patients in conjunction with a fiber delivery system and a transparent target ruler. A transparent gel was used as epidermal heat sink. The study lasted 15 months. Pretreatment as well as follow-up hair count per cm2 was performed to determine the level of success. Treatments were repeated when 1-2 mm growth was observed. RESULTS: The average hair count before the second treatment was found to be close to 65% of the pretreatment count. The average hair count 3 months after the last treatment, was found to be lower than 12%. The interval between treatments ranged from 4 weeks to 3 1/2 months. CONCLUSIONS: The 2 msec pulsed Alexandrite laser technology is effective for the removal of unwanted hairs, ranging from fair to dark, except when hairs are absent in the shaft depending on the stage of their growth cycle. This results in the necessity of a few treatments or touchups. Adverse effects are minimal and transient.

[Immunogenicity and stability of an aluminum-free liposomal hepatitis A vaccine (Epaxal Berna)]. / Immunogenität und Stabilität eines aluminiumfreien liposomalen Hepatitis-A-Impfstoffs (Epaxal Berna).

The high immunogenicity of the liposomal hepatitis A vaccine (Epaxal Berna) after a single dose and after a booster dose one year later has been confirmed in several studies with healthy adult volunteers: 95-100% and 96-100% seroconversion (> or = 20 mIE/ml) after 1 and 12 months respectively, as well as a booster effect in 100% of the cases after revaccination. The tolerability of this new, alum-free vaccine has been excellent with 6-25% local and 0-13% mild systemic reactions after a dose of 0.5 ml. Stability testing with and without detergent indicated partial internalization of the hepatitis A virions in the phospholipid bilayer of the liposome vesicles with storage. Immunization of 10 healthy adult volunteers with vaccine stored for 32 months at 4 degrees C showed, however, that the duration of storage has no influence on immunogenicity and tolerability of the vaccine.

Immunogenicity of new virosome influenza vaccine in elderly people.

The safety and immunogenicity of a new virosome influenza vaccine was compared to commercial whole-virus vaccine and subunit vaccine in elderly people. The virosome vaccine was made by extracting the haemagglutinin from influenza virus and incorporating it into the membrane of liposomes composed of phosphatidylcholine (PC) and phosphatidylethanolamine (PE). 126 residents of a nursing home, aged 63-102, were randomised to receive one of the vaccines. All three were well tolerated and caused a significant rise in the geometric mean anti-haemagglutinin inhibiting (HAI) antibody titre to the 3 vaccine components (H1N1 Singapore, H3N2 Beijing, and B/Yamagata). The virosome formulation caused the highest geometric mean titres in addition to significantly (p = 0.039-0.0016) higher rates of more than four-fold or more titre rises to all 3 vaccine components. The percentage of those immunised who achieved protective levels of antibody (HAI > or = 40) was significantly (p = 0.035-0.0017) higher for the H1N1 and B/Yamagata strains following immunisation with virosome formulation. Participants with non-protective baseline titres to the H1N1 or B/Yamagata strains were more likely (p = 0.0049-0.006) to achieve protective levels of antibodies after immunisation with the virosome vaccine. Immunisation with the virosome formulation did not result in a significant rise in anti-PC or anti-PE antibodies.

Immunogenicity and protectivity of a new liposomal hepatitis A vaccine.

In this study we investigated a new liposomal hepatitis A vaccine (Epaxal) developed by the Swiss Serum and Vaccine Institute clinically and immunologically using a one dose priming schedule and a booster injection after 1 year. This vaccine contains formalin inactivated hepatitis A virus particles attached to phospholipid vesicles together with influenza virus haemagglutinin. Two doses of the vaccine were administered at months 0 and 12 in 117 volunteers. Blood samples were drawn at days-7, 14 and 28 and after 6, 12 and 13 months, local and systemic reactions were monitored by means of questionnaires. Immunogenicity was evaluated as usual by the determination of anti-HAV from the collected sera using the ELISA technique. In order to evaluate the protective efficacy of the vaccine induced antibodies a sample of 25 sera mainly from vaccinees showing low ELISA titres was additionally analysed by means of a virus NT. The vaccine was excellently tolerable and highly immunogenic. Seroconversion evaluated by ELISA was 97 and 99%, respectively, 14 and 28 days after the first dose and 100% after the second dose. NT titres were well correlated with ELISA titres and showed similar seroconversion rates even in the early phase of immunization. The results of this study show that with two doses of the liposomal hepatitis A vaccine administered at months 0 and 12 early protection within 14 days and long lasting immunity can be achieved.

Rapid antibody response after vaccination with a virosomal hepatitis a vaccine.

BACKGROUND: This study was designed to assess the early antibody kinetics after a priming dose, and the extent of the antibody increase after a booster dose of an inactivated virosomal hepatitis A virus (HAV) vaccine (Epaxal). PATIENTS AND METHODS: This was an open, uncontrolled study in 30 healthy subjects. The vaccine was injected intramuscularly on day 1 and month 12. Serum antibody titers were measured by ELISA on day 1 (pre dose) and at various time points thereafter until month 12 (pre-booster dose). After the booster dose, antibody titers were measured at various intervals until month 24. Neutralizing antibody titers were measured in 12 subjects a number of times during the 1st month by an antibody neutralization assay. Titers > or = 10 mIU/ml were considered seroprotective. RESULTS: ELISA antibody titers showed a rapid increase post vaccination. By day 15, 96% of subjects were seroprotected, which increased to 100% by day 22 (n = 27 evaluable subjects, aged 18-43 years; 13 male, 14 female). All subjects achieved seroprotective HAV-neutralizing antibody titers by day 11 (n = 12). The booster vaccination at month 12 resulted in a strong response in all subjects, with a sustained anti-HAV antibody titer (1,155 mIU/ml) at month 24. Both the priming and booster doses were well tolerated. CONCLUSION: Primary vaccination with this virosomal HAV vaccine is well tolerated and induces a rapid HAV-neutralizing antibody response resulting in seroprotection in all subjects within 10 days. In addition, the booster vaccination results in prolonged seroprotective antibody levels.

Single-dose intramuscular ketorolac versus diclofenac for pain management in renal colic.

A double-blind controlled study was designed to compare the effective- ness of a single intramuscular dose of 60 mg ketorolac with that of 75 mg diclofenac in the treatment of renal colic and to monitor side effects. Fifty-seven patients completed the study, 27 in the ketorolac group and 30 in the diclofenac group. Effectiveness of treatment was monitored by pain relief reported on a 4-point verbal scale at different time points. At 60 minutes 77.8% and 86.6% (P = 0.4) of patients, and at 120 minutes 81.5% and 96.6% (P = .1 5) of patients, reported significant pain relief following ketorolac and diclofenac doses, respectively. Both groups had an equal 92% significant pain relief at discharge from the emergency department. Both drugs were well tolerated by the patients. Ketorolac therefore, seems as effective as diclofenac in the treatment of renal colic.

Increased membrane binding of erythrocyte catalase in hereditary spherocytosis and in metabolically stressed normal cells.

Normal red blood cell (RBC) membranes were compared with (1) RBC membranes from six patients with hereditary spherocytosis (HS), (2) normal membranes after hemolysis of the RBC in the presence of calcium, or (3) normal membranes after incubation of RBC for 24 hr in phosphate-buffered saline containing calcium without added glucose. When compared with normal controls, polyacrylamide gel electrophoresis with sodium dodecyl sulfate (PAGE SDS) of all three preparations showed an increase in membrane binding of globin and protein band 4.5 (60,000 molecular weight). In an attempt to identify band 4.5, 14 enzymes were assayed in the RBC membranes. Of these, catalase and lactate dehydrogenase were increased in membranes from HS RBC and from normal cells exposed to calcium. Only catalase, however, was present in sufficient quantity and had the correct subunit molecular weight on PAGE SDS and calcium-dependent membrane binding to account for an appreciable portion of 4.5. Caralase was further identified with a component of band 4.5 by double immunodiffusion using a specific anti-catalase antibody.

Study of a kindred with hereditary spherocytosis and glyceraldehyde-3-phosphate dehydrogenase deficiency.

A patient with hereditary spherocytosis (HS) was found to have glyceraldehyde-3-phosphate dehydrogenase (G3PD) deficiency by electrophoresis of the isolated red cell membranes on polyacrylamide gels with sodium dodecyl sulfate (PAGE SDS) as demonstrated by a diminished band 6 (G3PD) and confirmed by specific enzyme assay. Thirteen members of his family were studied: four were normal, two had HS alone, three had G3PD deficiency alone, and four had both HS and G3PD deficiency. G3PD deficient kindred members were probably heterozygous, since their red cell enzyme, while qualitatively normal, was present in half normal amounts. The G3PD deficiency alone was asymptomatic, and there was no evidence that the combination of HS with G3PD deficiency increased the clinical severity of the disease. However, G3PD deficiency, when combined with HS, was associated with an increase in protein band 4.5 on PAGE SDS. This band was also increased by incubation of normal red cells without glucose, and appeared to be a protein absorbed to the membrane as a consequence of metabolic stress. Hence, red cells with the combined abnormalities of both HS and G3PD deficiency showed signs of the exceptional metabolic stress to which they were exposed.

Induction of antigen-specific T suppressor cells by soluble Paracoccidioides brasiliensis antigen.

In naturally acquired paracoccidioidomycosis, patients have depressed in vivo and in vitro cell-mediated immune (CMI) responses to Paracoccidioides brasiliensis antigen. In addition, it has been reported that these patients have significant levels of circulating paracoccidioidal antigen in their sera. The primary purpose of this investigation was to assess the effects of P. brasiliensis antigen on the CMI responses in a mouse model. On the basis of findings with other fungal agents, we predicted that circulating paracoccidioidal antigen may be inducing suppressor cells which modulate the CMI response. In this study, we show (i) that a soluble P. brasiliensis culture filtrate antigen (Pb.Ag) emulsified in complete Freund adjuvant and injected subcutaneously into mice induces reasonably high levels of delayed-type hypersensitivity (DTH) in CBA/J mice; (ii) that Pb.Ag elicits DTH reactions specific for P. brasiliensis when injected into footpads of immunized mice; and (iii) that an intravenous injection of Pb.Ag induces a population of lymph node and spleen cells which, upon adoptive transfer, suppress the afferent limb of the DTH response to paracoccidioidal antigen. The afferent suppressor cells can be detected in spleens as early as 5 days after Pb.Ag treatment, are present in significant numbers by 7 days in both spleens and lymph nodes, and are virtually absent by 14 days. In contrast, at 14 days after antigen injection, efferent suppressor cells were detected in spleens and lymph nodes. The Pb.Ag-induced afferent suppressor cells specifically inhibit the antiparacoccidioidal DTH response. They are nylon wool-nonadherent cells, and their activity is abrogated by anti-Thy-1 and complement treatment, indicating that they are T lymphocytes. The phenotype of these afferent suppressor T cells is L3T4+ Lyt-1+2- I-J+. The Pb.Ag-specific suppressor cells described in this paper are similar to the Ts1 cells in the azobenzenearsonate, 4-hydroxy-3-nitrophenyl acetyl, and cryptococcal models of suppression of the DTH response and to the afferent suppressor cells in the dinitrofluorobenzene contact sensitivity system.