Psychometric evaluation of the German Version of the Supportive Care Needs Survey for Partners and Caregivers (SCNS-P&C-G) of cancer patients.

This study aimed for psychometric validation of the German version of the Supportive Care Needs Survey for Partners and Caregivers (SCNS-P&C-G). In- and outpatients with lung, urological and gastrointestinal cancer at Heidelberg University Hospital in Germany and in each case one relevant caregiver were asked to complete a set of questionnaires assessing their unmet needs together with distress, depression, anxiety and caregiver strain. In addition, medical data of the patients were collected. Fully completed questionnaires were received from 188 pairs of patients and their caregivers. Using exploratory factor analysis, four domains of unmet needs were identified with an appropriate variance explanation (58.7%) and acceptable (>0.70) internal consistencies (α = 0.95 to 0.76) for each domain. Convergent validity was found with respect to significant positive correlations (>0.40) of the SCNS-P&C-G domains with caregivers' anxiety, depression and strain. Although poorer health status of the patient indicated more unmet caregiver needs, this finding was not consistent for all need domains. Overall, associations were only moderate to weak pointing out the necessity of a separate screening for caregivers' needs. The findings of this study support that the SCNS-P&C-G is an appropriate research instrument to assess caregivers' needs on different domains throughout the disease trajectory.

Geographically predominant genotypes of Aspergillus terreus species complex in Austria: s microsatellite typing study.

Aspergillus terreus species complex is recognized as a frequent agent of invasive aspergillosis in Tyrol. The reason for this specific epidemiological situation is unclear. Aspergillus terreus strains isolated from environmental and clinical sources were genotyped using a novel panel of short tandem repeats and were evaluated for virulence. Three major endemic genotypes collected from the Inn region and its side valleys were found to cause the majority of invasive A. terreus infections. All of these genotypes were of the same mating type, which suggests that a mating barrier is present between these geographically well-adapted strains which is found to persist for at least 11 years. The three major genotypes were prevalent in both human infections and the environment. No major differences in virulence were observed using Galleria mellonella as model. Our data suggest a specific environmental exposure being responsible for the high incidence of A. terreus infections in Innsbruck, the Inn valley and side valleys (Tyrol, Austria).

Mixed colony formation in vitro by the heterogeneous compartment of multipotential progenitors in human bone marrow.

The physiology of the human haemopoietic primitive progenitor populations can be studied in normal and disease states by clonal in vitro cultures in which the primitive progenitor cells proliferate and differentiate to form mixed colonies. For many applications it is essential that such assays detect a high proportion of primitive progenitor cells. We describe an in vitro assay which detects a high incidence of human CD34+ multipotential progenitor cells. Bone marrow mononuclear cells (MNC) or selected CD34+ cells were plated at low cell concentrations in semisolid agar cultures with synergizing growth factor combinations. The optimum growth factor combination of conditioned medium from Mia PaCa-2 cells (Mia-CM), recombinant granulocyte-macrophage colony-stimulating factor and recombinant stem cell factor (SCF) supported the formation of macroscopic (> or = mm) colonies (97% of which were multilineage), at an average incidence of 250/10(5) MNC. The colony-forming cells (human colony-forming unit, type A) detected, showed a low cycling status (7.3%) and the macroscopic colonies had a high replating efficiency (46%), reflecting their probable primitive nature. This assay should prove invaluable, for studies on the regulation of proliferation of the multipotential compartment and in studies involving the assessment of these cells in transplantation and neoplastic disease.

Effect of deoxycytidine on 2-chloro-deoxyadenosine-mediated growth inhibition of normal human erythroid and myeloid progenitor cells.

The cytotoxic effect of chlorodeoxyadenosine (CdA) on lymphocytes and monocytes requires phosphorylation by the enzyme deoxycytidine kinase and can be antagonized by coadministration of deoxycytidine (dCyt), a competitive substrate of deoxycytidine kinase. It has also been shown for lymphocytes that coadministration of 3-aminobenzamide (3-ABA), an inhibitor of the enzyme poly-(ADP ribose) synthetase, is activated by CdA-mediated DNA strand breaks, consumes intracellular nicotinamide-dinucleotide (NAD) and can antagonize the lethal effect of CdA. Recent in vitro studies have shown that not only growth of lymphocytes and monocytes, but also colony formation by erythroid and myeloid progenitors derived from normal human bone marrow, is inhibited by CdA in a dose-dependent manner. In this study we examined the effect of various doses of dCyt (10(-6) to 10(-3) M) on CdA-mediated growth inhibition of erythroid and myeloid progenitor cells in vitro. Our results show that colony formation by human bone marrow-derived progenitor cells--CFU-E (colony-forming unit erythroid), BFU-E (burst-forming unit erythroid) and CFU-GM (colony-forming unit granulocyte/macrophage)-in semisolid medium is protected by a high, but clinically achievable and non-toxic, concentration of dCyt (> 10(-4) M) against the inhibitory effects of coadministered high concentrations of CdA. The protective effect of dCyt was markedly different on the various subclasses of progenitor cells, however. Thus, with coadministration of 10(-4) M dCyt, the CFU-E colony formation could be restored to almost 100% despite the presence of high concentrations of CdA (160 nM) compared to control cultures, whereas the colony formation of BFU-E and CFU-GM was restored to only 50%. At a concentration of 10(-3) M dCyt, colony formation of BFU-E and CFU-GM was raised to 80% of control cultures even in the presence of high concentrations of CdA (160 nM). Further experiments in which 3-ABA was coadministered to CdA-treated cultures showed that in all concentrations tested (0.3 to 5 mM) 3-ABA was not able to prevent CdA-mediated cytotoxicity on bone marrow progenitors. Based on these studies, we suggest that the CdA toxicity on CFU-E is mainly mediated by phosphorylation by deoxycytidine kinase, whereas additional mechanisms may be operative in BFU-E and CFU-GM. Considerable biochemical differences seem to exist between hematopoietic stem cells on the one hand and lymphocytes and monocytes from peripheral blood on the other.

Growth-inhibitory effect of 2-CdA on myeloid CFU-GM progenitors in normal human long-term bone marrow cultures and its compensation by G-CSF or IL-3.

As neutropenia is a common side effect of treatment with 2-chlorodeoxyadenosine (2-CdA), we investigated the myelosuppressive action of 2-CdA in Dexter-type human long-term bone marrow cultures (LTBMCs). LTBMCs were incubated with varying doses of 2-CdA (5 to 20 nM/L) during the first week. At 20 and 10 nM/L 2-CdA, we found a marked reduction in colony-forming unit-granulocyte/macrophage (CFU-GM) production throughout the culture period of 7 weeks (maximum reduction to 3.5% of untreated control cultures with 20 nM/L and to 27.2% with 10 nM/L, respectively). Even the lowest 2-CdA dose tested (5 nM/L) strongly reduced the number of CFU-GM progenitors during the first 3 weeks (maximum reduction to 52.4% of untreated controls), but this effect was transient, and values had recovered to normal within in 5 weeks. 2-CdA was also shown to cause a dose-dependent decrease in long-term culture-initiating cell (LTCIC) detections after 5 weeks in culture (49.6% of control cultures with 10 nM/L 2-CdA and 14% with 20 nM/L 2-CdA, respectively). When 2-CdA was added to LTBMCs initiated on preformed irradiated stromal feeder layers, similar results on CFU-GM production were obtained, indicating that the effects observed were not secondary to effects on the formation of a supportive layer. In addition, IL-6-concentrations in the supernatant of LTBMCs measured at various intervals after the addition of fresh medium with or without 2-CdA showed no significant decrease in cultures treated with 2-CdA. As neutropenia has been shown to be associated with a small but significant risk of fatal infection, we subsequently investigated the reversal potential of the 2-CdA effect by addition of recombinant human granulocyte colony-stimulating factor (rhG-CSF) or rh interleukin-3 (rhIL-3). The weekly addition of 100 ng/mL rhG-CSF counteracted the 2-CdA-mediated decrease in CFU-GM numbers during the entire period of 7 weeks, reaching statistical significance from weeks 3 to 7 (p < 0.05). Addition of rhIL-3 (100 ng/mL) showed an enhancement of CFU-GM output in 2-CdA-treated cultures that resulted in their numbers exceeding those in control cultures (without 2-CdA) from weeks 1 to 5 (p < 0.05) with a maximum increase of 5.1-fold over the parallel control value at week 3.(ABSTRACT TRUNCATED AT 400 WORDS)

The effects of creatine supplementation on thermoregulation and isokinetic muscular performance following acute (3-day) supplementation.

AIM: The purpose of this investigation was to determine the effects of 3 d of creatine supplementation on thermoregulation and isokinetic muscular performance. METHODS: Fourteen males performed two exercise bouts following 3 d of creatine supplementation and placebo. Subjects exercised for 60 min at 60-65% of VO2max in the heat followed by isokinetic muscular performance at 60, 180, and 300°·s(-1). Dependent variables for pre- and postexercise included nude body weight, urine specific gravity, and serum creatinine levels. Total body water, extracellular water and intracellular water were measured pre-exercise. Core temperature was assessed every 5 min during exercise. Peak torque and Fatigue Index were used to assess isokinetic muscular performance. RESULTS: Core temperature increased during the run for both conditions. Total body water and extracellular water were significantly greater (P<0.05) following creatine supplementation. No significant difference (P>0.05) was found between conditions for intracellular water, nude body weight, urine specific gravity, and serum creatinine. Pre-exercise scores for urine specific gravity and serum creatinine were significantly less (P<0.05) versus post-exercise. No significant differences (P>0.05) were found in peak torque values or Fatigue Index between conditions for each velocity. A significant (P<0.05) overall velocity effect was found for both flexion and extension. As velocity increased, mean peak torque values decreased. CONCLUSION: Three d of creatine supplementation does not affect thermoregulation during submaximal exercise in the heat and is not enough to elicit an ergogenic effect for isokinetic muscle performance following endurance activity.

An enzyme immunoassay for the determination of low levels of IgA in human serum.

An enzyme immunoassay (EIA) has been developed which facilitates the detection of low levels of immunoglobulin A in human serum. IgA is captured by an anti-IgA antibody linked to micron-sized polyacrylamide beads and subsequently detected by an anti-IgA horseradish peroxidase conjugate. The standard curve is linear in the region between 25-1000 ng/ml IgA. The assay is particularly suited to measure IgA antibodies in sera from IgA-deficient individuals and IgA contaminants in blood products.

An immunoassay for the determination of total IgA subclass antibodies in human serum.

An enzyme immunoassay capable of determining total IgA1 and IgA2 concentrations in human serum has been developed. Subclass-specific monoclonal antibodies are bound to polyacrylamide bead-conjugated anti-mouse immunoglobulin antibodies. Bound IgA is detected with an anti-IgA peroxidase conjugate and the standard curve is linear in the region 0.25 - 2.0 micrograms/ml. Coefficient of variation values range from 0.24 - 5.77% for the IgA1 standard curve and from 0.86 - 5.92% for the IgA2 standard curve. Inter-assay variation for the IgA1 and IgA2 control sample values were 8.2% and 13.4%, respectively.

A rapid procedure for the purification of IgA1 and IgA2 subclasses from normal human serum using protein G and jackfruit lectin (jacalin) affinity chromatography.

Immunoglobulin A (IgA) from pooled normal human sera was purified using antibody and protein G affinity chromatography and gel filtration high-pressure liquid chromatography (HPLC). This high purity product was separated into IgA1 and IgA2 subclasses utilizing the agarose-bound lectin 'jacalin'. Evaluation of product homogeneity by immunological testing confirmed greater than 95% purity. The total IgA1 and IgA2 recovered from sera was approximately 26% of the initial antibody present.

Differentiation of primary and secondary cutaneous B-cell lymphoma by Southern blot analysis.

Malignant B-cell lymphomas represent a heterogenous group of lymphoreticular disorders that involve the skin in about 20% of reported cases. Skin involvement may be primary or secondary (ie, the result of hematogenous spread). Primary cutaneous B-cell lymphomas (PCBCLs) are thought to take a comparatively favorable course, respond readily to nonaggressive treatment, and lack evidence of extracutaneous spread. Nine primary B-cell lymphomas (7 centrocytic or centroblastic follicular, 1 immunoblastic, 1 centroblastic), three secondary (follicular) cutaneous B-cell lymphomas (SCBCLs) and two pseudolymphomas were studied. Staging revealed that bone marrow was involved only in SCBCLs. Centrocytes were detected in blood smear preparations of all SCBCLs. All lymphomas were treated with local irradiation. Patients with primary centroblastic and immunoblastic cutaneous lymphomas and those with secondary lymphomas received additional chemotherapy. Pseudolymphomas were treated by simple excision. Patients were monitored on average for 55 months. During this period, no patients with PCBCLs exhibited cutaneous relapses or hematogenous spread. In contrast, all patients with SCBCLs experienced cutaneous relapses. Peripheral blood, bone marrow, and skin samples from all patients were subjected to Southern blot analysis using a JH probe. Clonal rearrangement was found in all skin samples investigated except specimens from pseudolymphomas. Peripheral blood and bone marrow samples were positive in SCBCLs (the rearrangement pattern was different from that of the skin samples for two of the three patients), whereas it was negative in all PCBCLs and pseudolymphomas. In conclusion, Southern blot analysis of peripheral blood may be useful in differential diagnosis of PCBCLs and SCBCLs and a prognostic marker. Furthermore, these data confirm the comparatively favorable clinical course of PCBCLs and suggest that in these cases, local irradiation can be considered adequate treatment, whereas SCBCLs require additional systemic therapy.

Interhospital transport of the patient on extracorporeal cardiopulmonary support.

Portable extracorporeal cardiopulmonary support systems have enhanced the resuscitation and support of moribund patients outside of the operating room environment. The literature documents the successful application of emergency cardiopulmonary support within the hospital setting. Clinicians have reported the use of helicopter and ground ambulance to transport patients requiring intraaortic balloon counterpulsation and fixed-wing transport of neonates requiring extracorporeal membrane oxygenation. As medical transport capabilities extend the sphere of tertiary care to outlying medical facilities, there is a role for extracorporeal cardiopulmonary support in the initial stabilization and safe transport of critically ill patients, via air or ground ambulance. Potentially, the early application of life-sustaining technology can lower mortality and morbidity in patients with a survivable pathology. This is a report on the experience with the resuscitation and interhospital transport of patients on extracorporeal cardiopulmonary support.

Cytotoxicity of derivatives from dehydrocrotonin on V79 cells and Escherichia coli.

New derivatives from dehydrocrotonin (DHC, compound I), with the same anti-ulcerogenic properties but less toxicity were synthesised by reducing the cyclohexenone moiety of DHC with NaBH(4) (compound II), by reducing the cyclohexenone and lactone moieties with LiAlH(4) (compound III) and by transforming the lactone moiety into an amide (compound IV) using dimethylamine. The cytotoxicity of these derivatives from DHC was assayed on V79 fibroblast cell line. Three independent endpoints for cytotoxicity were evaluated; namely, the nucleic acid content (NAC), tetrazolium reduction (MTT) and neutral red uptake (NRU). IC(50) values of 540 and 350 microM were obtained for compound II in the NRU and NAC tests, respectively. Compound III was less toxic than the other DHC derivatives (IC(50)=1800 microM) on V79 cells based on NAC assay. Compound IV showed an IC(50) ranging from 350 to 600 microM based on the three endpoints evaluated. The three compounds were less toxic on V79 cells than DHC. DHC, compounds II, III and IV did not change the respiration rate of Escherichia coli on the acute toxicity assay.

Cytotoxicity of prodigiosin and benznidazole on V79 cells.

The cytotoxicity of prodigiosin, an antibiotic and potential trypanocide produced by Serratia marcescens, and Benznidazole, a trypanocidal drug, were assayed on V79 fibroblast cell line. Three independent endpoints for cytotoxicity were evaluated; namely, the nucleic acid content (NAC), MTT reduction and neutral red uptake (NRU). IC(50) values of 1-20 microM were obtained for prodigiosin in the NRU, MTT and NAC tests. Prodigiosin had greater trypanocidal activity (IC(50)=5 microM) than Nifurtimox (IC(50)=150 microM) a known trypanocide drug used in Chagas' disease therapy. Benznidazole was less toxic (IC(50)=2000 microM) than prodigiosin (IC(50)=1-20 microM) in V79 cells based on the MTT and NAC assays. Benznidazole stimulated the NRU until 2 mM. Indeed, the cell viability measured with the NRU was higher at all concentrations of benznidazole tested than that measured by MTT reduction and NAC assays.

Synthesis, antimycobacterial activities and cytotoxicity on V79 of 3-[4'-Y-(1,1'-biphenyl)-4-yl]-N,N-dimethyl-3-(4-X-phenyl)-2-propen-1-amine derivatives.

The derivatives of 3-(4'-bromo-[1,1'-biphenyl]-4-yl)-3-(4-X-phenyl)-N,N-dimethyl-2-propen-1-amine (5a-m) were synthesised through a Friedel-Crafts acylation followed by Wittig reaction. The effects of the compounds on standard strains of Mycobacterium sp. (ATCC) and M. tuberculosis isolated from clinical specimens were evaluated. Also the toxicity was determined on V79 cells line using neutral red uptake (NRU), nucleic acid content (NAC) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction to measure the cellular viability.

Violacein cytotoxicity and induction of apoptosis in V79 cells.

Violacein, a pigment produced by Chromobacterium violaceum, is reported to be a potential drug for the treatment of Chagas' disease. Violacein is also effective against leukemia and lymphoma cells in culture (IC50 10(-8) M). Changes in the nuclear acid content, 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide reduction and neutral red uptake in these cells were used to evaluate the cytotoxicity of violacein in V79 Chinese hamster (M-8) fibroblasts. Violacein was highly cytotoxic to V79 fibroblasts (IC50 5-12 microM). Using the TUNEL method and the Feulgen reaction coupled to image analysis, violacein (5 and 10 microM) was found to trigger apoptosis but not necrosis in V79 cells. The morphological changes seen in the nuclei of these cells included chromatin condensation and a decrease in deoxyribonucleic acid content. These results demonstrating that violacein induces apoptosis in V79 cells strengthen its potential as a therapeutic agent.

Effect of growth conditions on production of violacein by Chromobacterium violaceum (BB-78 strain).

1. Chromobacterium violaceum (strain BB-78 isolated in Brazil) produces violacein, a substance potentially useful in phototherapy and with antibiotic and trypanocide activity. Culture conditions were optimized for the production of violacein and changes in nutrients, temperature and pH were correlated to cellular growth. 2. Methionine was the only absolute requirement for growth. Alanine, arginine, tryptophan and vitamin B12 stimulated growth and the microorganism utilized both the D and L forms of methionine. L-Tryptophan and its metabolites were important as a carbon source for violacein production and bacterial growth. 3. The highest yields of violacein were obtained by incubation in liquid medium and in the absence of light at pH 7.0 and 28 degrees C for 24 h.

Comparison of the antiviral activity and toxicity of a protein magnesium ammonium phospholinoleate anhydride polymer with other antiviral drugs.

1. SB-73, a magnesium ammonium phospholinoleate anhydride aggregate, exhibited antiviral action in vitro in the concentration range of 50 to 100 micrograms/ml against herpes simplex type 1, stomatitis vesicular virus, adenovirus type 5, and in vivo in the dose range of 0.7 to 1.3 mg/kg against canine parvovirus and distemper virus. 2. The lethal dose (LD50) was 2.71 +/- 1.55 g/kg body weight in mice inoculated intraperitoneally. Oral ingestion of the aggregate up to 30 g/kg body weight by mice had no lethal effects during the 14 days of observation. 3. In in vitro cytotoxicity experiments with fibroblasts (V-79 Chinese hamster cell line), no toxic effects were observed with SB-73 concentrations (120 micrograms/ml) having antiviral activity. 4. In a cellular proliferation experiment using hamster V-79 cells, we observed 72% proliferation after treatment of the cells with a high concentration (500 micrograms/ml) of SB-73. 5. Compound SB-73 showed no genotoxicity for human lymphocytes at concentrations of 100 micrograms/ml. 6. When the cytotoxicity and genotoxicity of SB-73 were compared with those of acyclovir, idoxuridine and AZT at 500 micrograms/ml concentrations the compound was found to have effects similar to those of acyclovir.

Gastroprotective effect of essential oil from Croton cajucara Benth. (Euphorbiaceae).

The gastroprotective activity of the essential oil from the bark of Croton cajucara Benth (Euphorbiaceae) was assessed in three different models of experimentally induced gastric ulcer in mice. At oral dose of 100 mg/kg the essential oil reduced gastric lesions induced by hypothermic restraint stress and HCl/ethanol significantly. In the HCl/ethanol model a dose-dependent gastroprotective effect was found. Moreover, significant changes in gastric parameters such as pH, secretion rate and total gastric acid were found after intraduodenal administration of essential oil under ligated pylorus (Shay) conditions. The acute toxicity of essential oil was assessed in mice. The LD50 values were 9.3 and 680 mg/kg for oral and intraperitoneal administrations, respectively. The cytotoxicity of essential oil was studied also. A dose-dependent cell viability inhibition was found in V79 fibroblast cell cultures with an IC50 of 22.9 microg/ml. Our results support the pharmacological study of this essential oil.

Densification, crystallization, and electrical properties of lead zirconate titanate glass-ceramics.

Piezoelectric glass-ceramics in the lead zirconate titanato-lead silicate system were developed. SiO(2) was required for glass formability, and excess PbO allowed low temperature processing. The amounts of those constituents were limited by the optimization of the piezoelectric properties. Only a small region of compositions in this system yielded the desired combination of glass formability, crystallization and densification behavior, and resulting piezoelectric properties. Selected compositions were melted and roller quenched to form glass ribbon, then milled into glass powder. Pressed glass powder densified to closed porosity at 850 degrees C with piezoelectric d(33 ) and g(33) coefficients of 26 pC/N and 33x10(-3 ) Vm/N. The low temperature sintering behavior of these ferroelectric glass-ceramics provides the possibility of incorporating a piezoelectric material as a sensor or actuator in thick film circuits or low-fire multilayer packages.

Modeling of the electrostrictive, dielectric, and piezoelectric properties of ceramic PbTiO(3).

The upper and lower limits of the electrostrictive constants, dielectric permittivities, spontaneous polarizations, and piezoelectric coefficients were calculated for ceramic PbTiO(3) from theoretical single-crystal constants. Experimental ceramic data fall between these upper and lower limits. The large piezoelectric anisotropy d(33)/d(31) of ceramic PbTiO(3 ) was shown to be related to the single-crystal PbTiO(3) electrostrictive anisotropies Q(11)/Q(12 ) and Q(44)/Q(12). The possibility of a change in sign of the ceramic d(31) coefficient due to a slight variation in the single-crystal electrostrictive anisotropies was discussed. The single-crystal and predicted ceramic hydrostatic electrostrictive constants were found to be equal. Using this result the ceramic hydrostatic g(h ) coefficient is always smaller than the single-crystal g (h), but the ceramic hydrostatic d(h) coefficient can be either larger or smaller than the single-crystal d(h) depending on the dielectric anisotropy (epsilon (11)/epsilon(33)) of the single-crystal.