Restriction endonuclease-fragment polymorphisms of oral viridans streptococci, compared by conventional and field-inversion gel electrophoresis. Oral streptococci formerly classified as Streptococcus sanguis or Streptococcus mitis have recently been divided into four species. Two additional species have also been proposed for this group. Each species is genetically distinct, but they have many traits in common, which makes it difficult for clinical isolates to be identified by phenotypic tests. Genotypic comparison may provide an alternative approach. This study used DNA fingerprint analysis for comparison of genotypes of 21 reference strains--classified as Streptococcus gordonii, Streptococcus sanguis, Streptococcus oralis, "Streptococcus parasanguis", or "Streptococcus crista" in previous DNA hybridization studies--and 17 clinical and laboratory strains placed in those groups on the basis of phenotypic tests. HinDIII and PvuII digests were run in conventional horizontal agarose gels. SfiI digests of reference strains and two laboratory strains were run in field-inversion gels. Fingerprint patterns were compared by visual examination, cluster analysis of densitometric traces, and lane-matching software. Only two "S. crista" strains and two parent mutant lineages showed fingerprint patterns that were identical by visual examination. Fingerprint patterns of all other strains were unique. Cluster analysis results could not be considered valid, since replicate lanes in different gels were not grouped together. HinDIII and PvuII digests contained too many bands for correct matching by lane-matching software. SfiI digests were correctly matched by computer, with the same results as visual examination. Results indicate that the diversity of strains within these streptococcal species was too great to permit species identification by DNA fingerprint patterns. This genotypic diversity suggests that isolates from unrelated hosts may have been separate for long periods of time.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of drying period and soil moisture on egg hatch of the tadpole shrimp (Notostraca: Triopsidae). Tadpole shrimp (Triops spp.) are potential biological control agents against larval mosquitoes in temporary ponds and flood-irrigated fields. In some rice field situations, however, they may become pests that uproot and eat young rice plants. In cursory observations, it has been reported that tadpole shrimp eggs do not readily hatch on flooding when the soil or substrate containing eggs is moist before flooding. The relationship between drying (moisture content) of soil and tadpole shrimp hatch was determined in studies conducted in mesocosms at the University of California Aquatic and Vector Control Research facilities at Riverside and at Oasis in the Coachella Valley of southern California. In laboratory hatching trials, an increase in hatch of Triops longicaudatus (LeConte) with declining soil moisture content was demonstrated (t = 8.4, P less than 0.001; r2 = 0.76). In field trials in mesocosms at Riverside, egg hatch increased with increased drying period and declining soil moisture content (G = 29.8, P less than 0.01). No hatch of eggs occurred after 3 d of drying, when soil moisture content was high, but a high level of hatching occurred after 7 and 14 d of drying, when soil moisture declined to low levels. At Oasis, soil moisture did not decrease with drying time because of porous soil and capillary action of water from adjacent flooded mesocosms and thick vegetation covering the pond bottoms. Therefore, hatch rates at Oasis were not associated with the length of the drying period (G = 35, P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


The neurohormonal hypothesis: a theory to explain the mechanism of disease progression in heart failure. Because physicians have traditionally considered heart failure to be a hemodynamic disorder, they have described the syndrome of heart failure using hemodynamic concepts and have designed treatment strategies to correct the hemodynamic derangements of the disease. However, although hemodynamic abnormalities may explain the symptoms of heart failure, they are not sufficient to explain the progression of heart failure and, ultimately, the death of the patient. Therapeutic interventions may improve the hemodynamic status of patients but adversely affect their long-term outcome. These findings have raised questions about the validity of the hemodynamic hypothesis and suggest that alternative mechanisms must play a primary role in advancing the disease process. Several lines of evidence suggest that neurohormonal mechanisms play a central role in the progression of heart failure. Activation of the sympathetic nervous system and renin-angiotensin system exerts a direct deleterious effect on the heart that is independent of the hemodynamic actions of these endogenous mechanisms. Therapeutic interventions that block the effects of these neurohormonal systems favorably alter the natural history of heart failure, and such benefits cannot be explained by the effect of these treatments on cardiac contractility and ejection fraction. Conversely, pharmacologic agents that adversely influence neurohormonal systems in heart failure may increase cardiovascular morbidity and mortality, even though they exert favorable hemodynamic effects. These observations support the formulation of a neurohormonal hypothesis of heart failure and provide the basis for the development of novel therapeutic strategies in the next decade.


Protective effect and duration of action of inhaled formoterol and salbutamol on exercise-induced asthma in children. The magnitude and duration of protection against exercise-induced asthma (EIA) afforded by salbutamol and the new, long-acting beta 2-agonist, formoterol, were compared in a double-blind, placebo-controlled crossover study. Twelve children with asthma and EIA (greater than 25% fall from baseline at a pretrial exercise test) were studied on 3 different days receiving, in random order, either formoterol, 12 micrograms, salbutamol, 200 micrograms, or placebo by inhalation. The effect on EIA was evaluated by standardized treadmill-exercise tests repeated at the following times after medication: 1/2 hour (test 1), 3 hours (test 2), and, if the trial drug still demonstrated an effect, 5 1/2 hours (test 3) and 8 hours (test 4). The mean (SD) maximum percent fall in FEV1 at the pretrial test was 45% (14%). Placebo treatment had no effect on EIA. The mean (SD) maximum percent fall in FEV1 was 44% (14%) (test 1) and 39% (13%) (test 2) (not significant). Salbutamol offered good protection against EIA after 1/2 hour (percent fall in FEV1, 18% [18%]; p less than 0.02) but was not significantly different from that of placebo after 3 hours, 39% (13%) fall in FEV1. Formoterol blocked EIA in all the children and demonstrated a significant effect in most children for at least 8 hours. The percent fall in FEV1 after the various tests were 8% (16%) (test 1), 10% (9%) (test 2), 18% (15%) (test 3), and 18% (7%) (test 4; N = 9) (all tests, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


Immunocytochemical analysis of chromaffin cell proliferation in vitro. The bromodeoxyuridine (BrdU) incorporation technique for immunocytochemical labeling of S-phase nuclei was optimized for the study of chromaffin cell proliferation. Sequential fixation in ethanol followed by paraformaldehyde, and the use of DNAse to render incorporated BrdU accessible to antibody, permitted permanent double staining for BrdU and tyrosine hydroxylase. The efficacy of the technique was demonstrated in microcultures of dissociated neonatal rat adrenal glands, in which chromaffin cells exhibited proliferative responses to nerve growth factor and fibroblast growth factor similar to those previously demonstrated by autoradiography. Growth factor responsiveness was observed in both serum-containing and serum-free medium.


Antibiotic-resistant enterococci.

Antibiotic-resistant enterococci. Enterococci have emerged as an important cause of nosocomial infection. Successful antibiotic treatment of serious enterococcal infection usually depends on the synergistic bactericidal effect achieved by the combination of a cell wall-active agent, such as ampicillin or a glycopeptide, and an aminoglycoside. However, the prevalence of enterococci resistant to one or more of these antibiotics is increasing, and has resulted in serious therapeutic difficulties. The mechanisms of antibiotic resistance, and the epidemiology, laboratory diagnosis and management of infection with antibiotic-resistant enterococci are discussed.


Characterization of methicillin-resistant Staphylococcus aureus associated with nosocomial infections in the University Hospital, Kuala Lumpur. Methicillin-resistant Staphylococcus aureus (MRSA) as a hospital pathogen has presented many clinical problems in the University Hospital, Kuala Lumpur, Malaysia since 1978. The need for control of spread of these organisms became evident by 1985 when it was noted that the incidence of MRSA among S. aureus isolated from hospital inpatients had increased from 11.5% in 1979 to 18.8% in 1985. The characteristics of 50 MRSA isolates associated with nosocomial infections in the hospital are described here. The predominant strains produced Type IV coagulase and 84% of isolates studied showed moderate to high resistance to methicillin with MIC values of 25 mg l-1 or higher. All the MRSA isolates that could be phagetyped were susceptible to Group III phages, with 76.6% of the isolates being susceptible to phage 85. At least 10 different patterns were distinguishable by plasmid typing, the majority of isolates harbouring up to four small plasmids.


Microbiology of postoperative wound infection: a prospective study of 1770 wounds. A prospective study of postoperative wound infection was carried out over a 12-month period. Intra-operative swabs from the patients' anterior nares, the opened viscus and parietes were cultured using standard bacteriological techniques. Of the 1770 wounds studied, 167 (9.4%) became infected. Wound infection rates, according to clinical wound types, were clean 5.9%, clean-contaminated 10.7%, contaminated 24.3% and dirty 52.9%. The figures according to microbiological wound types were clean 4.7%, and potentially, lightly and heavily contaminated 15.3%, 22.1% and 30.2% respectively. The commonest causative organisms were Staphylococcus aureus 23.7%, Escherichia coli 16.9%, Staphylococcus epidermidis 13.5% and Pseudomonas aeruginosa 13.0%. When isolated intra-operatively, Enterobacter spp., Proteus spp., Klebsiella spp. and P. aeruginosa appeared to have a high probability of causing postoperative wound infection, but the intra-operative isolation of Bacteroides sp. was a poor predictor of subsequent wound infection.


Bacteriophage phi X-174 as an aerobiological marker for surgical plume generated by the electromagnetic field focusing system. The aerosol of surgical plume could be measured effectively with the use of bacteriophage phi X-174 as a biological marker, in contrast to previous methodologies reported by others. Recovery of virus plaque-forming units was highest from hydrophobic polytetrafluoro-ethylene membranes compared to hydrophobic polycarbonate screen filters or polyvinylidene difluoride depth filters, indicating that the method of virus recovery strongly affects the utility of a virus as an aerobiological marker. With this new method, surgical plume was indeed found in significant amounts when cutting tissue phantoms made with agar containing virus. The Electromagnetic Field Focusing System was used, which is a new thermal surgical device. The nominal power setting did not appear to be a factor in the amount of virus recovered. However, when pulse modulating the power by adjusting the crest factor from 1.4 to 4.3, a measure of the duty cycle for power delivery which adjusted the device from its cutting to haemostatic mode, a nearly five-fold increase in surgical plume, as evidence by the recovery of phi X-174 plaque forming units, was seen. The data indicate that bacteriophage phi X-174 can be used effectively as an aerobiological marker for aerosols generated during clinical procedures, and reinforce the need to use a safety vacuum during aerosol generating procedures. The availability of a safe and economical biological marker for aerosols from clinical procedures, which may lead to acquired infections in hospital personnel, makes evaluation of procedures and containment systems markedly easier. The data also indicates that surgical plume biohazard may be present in other techniques that employ pulse modulation including surgical lasers and electrocauteries.


Bacterial contamination and the effect of filters in anaesthetic circuits in a simulated patient model. In order to investigate bacterial contamination of anaesthetic breathing circuits and means of prevention of this, six different laboratory experiments were performed. These experiments involved the bacterial contamination of Dräger Narkose Spiromat 650 and Dräger AV-1 circle system circuits and of an isolated soda lime carbon dioxide absorber. The effects of anaesthetic gas, gas flow rate and the incorporation of a hydrophobic membrane heat and moisture exchanging bacterial/viral filter (HMEF) at the patient end of these circuits were investigated. It was found that without a HMEF the whole interior of the anaesthetic circuits became contaminated with bacteria. Components closest to the simulated patient showed the highest levels of contamination. Higher gas flows were associated with decreased levels of circuit contamination, presumably because more bacteria were expelled from the system. Halothane (1 volume %) and soda lime were not found to have any demonstrable bactericidal action. The presence of a HMEF between the simulated patient and the Y-piece prevented any detectable contamination from reaching the circuit. Consequently, the presence of a HMEF provides protection of the anaesthetic circuit as well as other patients, healthcare workers and the environment.


Molten metal burn of the foot: a preventable injury. Molten metal burns of the feet remain a common injury to foundry workers. A case is reported of a foundry worker who sustained circumferential molten metal burns of the distal foot and toes necessitating amputation of four toes. This severe injury could easily have been prevented by the use of protective footwear and spats.


Handwashing and disinfection of heavily contaminated hands--effective or ineffective? Hands are among the principal vehicles for transfer of nosocomial pathogens in hospitals. Often, outbreaks of infection are thought to be caused by a lack of compliance with handwashing guidelines, rather than due to the inadequacy of the handwashing agents used. In this study the effectiveness of proper handwashing and the use of three different hand disinfectants: ethanol 70% (E), isopropanol 40% (I) and alcoholic chlorhexidine (70%) (AC) was compared using three volunteers whose fingertips were heavily contaminated with a succession of bacteria including: Enterococcus faecalis, Staphylococcus aureus, Escherichia coli and Enterobacter cloacae. After each contamination, thorough handwashing and application of one disinfectant on the hands were performed three times. Fingerprint-samples were taken before and 1 min after application of the disinfectants. Thorough handwashing with an ordinary liquid soap ('Sterisol') did not reduce the confluent growth of bacteria on fingertips for any of the species used (197 examinations). Only AC had a significant effect on fingers heavily contaminated with S. aureus (126 examinations; AC compared with E and I; P less than 0.0002 and P less than 0.0002 respectively), but did not completely eradicate the bacteria. After contamination with Ent. cloacae (118 examinations), none of the three agents were particularly effective, but E and AC seemed to be somewhat more effective than I (P less than 0.0002 and P less than 0.01 respectively). When successive contamination was performed using all bacterial species, AC was the most effective decontaminant. However, Ent. cloacae was still present on the fingertips after 15 repeated courses of handwashing and applications of disinfectants. Bathing of hands in AC for 20s completely eradicated all bacteria from the hands. The study demonstrates that, when heavily contaminated, an ordinary handwashing followed by disinfectants is not enough to eradicate potentially pathogenic bacteria from the hands.


A randomized prospective study to compare cefotetan with cefuroxime plus metronidazole as prophylaxis in elective colorectal surgery. This study compares the efficacy of cefotetan with the combination of cefuroxime plus metronidazole as antibiotic prophylaxis in elective colorectal surgery when given over the first 24 h postoperatively. There was no significant difference in wound infection rates between the two groups (14.7% for cefotetan and 13.9% for cefuroxime plus metronidazole), or the rates of other infective complications. Adverse reactions occurred with equal frequency in both treatment groups and no serious side effects occurred. Cefotetan is a safe and effective antibiotic for use as prophylaxis in elective colorectal surgery. Its advantages are that it is a single agent with a spectrum covering both aerobic Gram-negative rods and anaerobic organisms and, because of its long half-life, needs only to be given at 12-hourly intervals.


Postoperative wound infection at a university hospital in Jeddah, Saudi Arabia. A prospective study was made of 1418 surgical wounds at the 250-bed King Abdulaziz University Hospital in Jeddah, Saudi Arabia. Daily examinations of wounds, cultures of all suspicious wounds and 28 days outpatient clinic follow-up were performed. The overall infection rate was 9%. The infection rate after clean surgery was 9.5%. High rates of infections were noted after colon resection (19%), caesarean section (19%), abdominal hysterectomy (10%) and cholecystectomy (10%). The infection rates after appendectomy, mastectomy and herniorrhaphy were approximately 7%. A lower rate of infection was seen after thyroidectomy (2%). The incidence of infection was significantly related to pre-operative stay in hospital and to duration of operation.


Simultaneous depletion of CD4+ and CD8+ T lymphocytes is required to reactivate chronic infection with Toxoplasma gondii. C57BL/6 mice chronically infected with an avirulent strain (ME-49) of Toxoplasma gondii were used to study the mechanisms by which T lymphocytes and IFN-gamma prevent reactivation of latent infection. Infected animals were treated with mAb, either anti-CD8, anti-CD4, anti-CD4 plus anti-CD8, anti-IFN-gamma, or anti-CD4 plus anti-IFN-gamma and the mice followed for survival, histopathology, cyst numbers, and spleen cell cytokine responses. In agreement with previously published findings, treatment with anti-IFN-gamma antibodies fully reactivated the asymptomatic infection, inducing massive necrotic areas in the brain with the appearance of free tachyzoites and death of all animals within 2 wk. Mice treated with the combination of anti-CD4 plus anti-CD8 antibodies showed augmented pathology and mortality nearly identical to the anti-IFN-gamma- treated animals. In contrast, treatment with anti-CD4 or anti-CD8 mAb alone failed to result in significantly enhanced brain pathology or mortality. In additional experiments, full reactivation of infection was observed in mice treated with anti-CD4 plus anti-IFN-gamma indicating that CD4+ lymphocytes are not required for the pathology resulting from IFN-gamma neutralization. Cytokine measurements on parasite Ag-stimulated spleen cells from mAb-treated mice indicated that both CD4+ and CD8+ cells produce IFN-gamma whereas only CD4+ cells contribute to parasite Ag-induced IL-2 synthesis. Together, these results suggest that CD4+ and CD8+ lymphocytes act additively or synergistically to prevent reactivation of chronic T. gondii infection probably through the production of IFN-gamma.


Migration of recombinant IL-2-activated T and natural killer cells in the intercellular space of human H-2 glioma spheroids in vitro. A study on adhesion molecules involved. The migration of rIL-2-activated T and NK cells into the intercellular space of glioma tissue was studied using multicellular spheroids grown from the human H-2 glioblastoma cell line as targets. Lymphocytes of all analyzed subtypes migrated into the spheroids, but CD56+ cells were particularly migratory. Lymphocytes and the H-2 tissue expressed adhesion molecule subunits for the following potential cell-cell or cell-matrix interactions: alpha 3 beta 1 (VLA-3) to fibronectin, laminin, and collagen; alpha 4 beta 1 (VLA-4) and alpha 5 beta 1 (VLA-5) to fibronectin; alpha 6 beta 1 (VLA-6) to laminin; alpha 4 beta 1 to VCAM-1; alpha L beta 2 (Leu-CAMa/LFA-1) to CD54 (ICAM-1); CD44 to fibronectin, collagen, laminin, hyaluronate; CD2 to CD58 (LFA-3); and CD56 (N-CAM) to CD56. In the H-2 tissue, CD54 and VCAM-1 were expressed as a gradient. The expression of CD54 was weak in the peripheral zone and the expression was stronger in the quiescent deeper zone, whereas the distribution of VCAM-1 showed an inversed pattern. The low expression of CD54 was up-regulated along the frontier of migrating lymphocytes. The migration was almost totally prevented by the anti-CD18 (beta 2) mAb IB4 and TS1/18, and also strongly inhibited by the anti-CD54 mAb LB-2. Instead, mAb known to inhibit the binding of beta 1 integrins to fibronectin were not significantly inhibitory. However, a combination of the GPEILDVPST and GRGDS peptides, which compete for the binding of alpha 4 beta 1 and alpha 5 beta 1 to fibronectin and may also affect other adhesion systems, partially prevented migration.


Induction and suppression of delayed-type hypersensitivity to non-MHC antigens during lethal graft-versus-host reaction. A delayed-type hypersensitivity reaction (DTH) to non-MHC Ag was detected during a lethal graft-vs-host reaction (GVH) induced by incompatibility for non-MHC Ag alone. In this model, when appropriate doses of B10.D2 bone marrow and lymphoid cells are grafted to irradiated (DBA/2 x B10.D2)F1 recipients, the ensuing GVH, directed against those DBA/2 non-MHC Ag absent from the B10.D2 background, results in virtually 100% mortality in less than 3 mo; donor alloimmunization against the host histocompatibility Ag considerably reduces mortality, and survival rates of 80 to 100% are common. The experiments reported here show that: 1) the cell responsible for DTH induction expresses the CD4+ CD8- phenotype; 2) CD4+ cells likewise seem to play a predominant role in the pathology of lethal GVH in this genetic combination; 3) the alloimmunization protocol that abrogates mortality also abolishes GVH-associated DTH; and 4) this suppressive effect, as shown elsewhere for the protection against mortality, is mediated by CD4- CD8- "double negative" Thy-1+ CD3+ T suppressor cells. Thus, there is a good parallel between lethal GVH and its associated DTH as concerns both induction and suppression of the two phenomena, suggesting that mortality and DTH may represent different manifestations of a common underlying mechanism. Initiation of the effector phase of DTH in the adoptive transfer model seems to be dependent on the presence of a Thy-1+ double-negative cell in the transfer inoculum; the possible relationship of this double-negative cell to the Th-1-type CD4+ DTH-mediating cell recently shown to induce lethal GVH is discussed.


Murine AIDS superantigen reactivity of the T cells bearing V beta 5 T cell antigen receptor. A B cell line, B6-1710, that expresses the defective virus known to induce murine AIDS stimulates a large fraction of nonprimed splenic T cells. Analysis of the T cell population responding to the B6-1710 for TCR V beta-chain usage revealed that, in addition to the previously reported V beta 5-chain-positive T cells, T cells bearing V beta 11 and V beta 12 are also specifically enriched. We have established V beta 5+ T cell lines, clones, and hybridomas expressing identical TCR with different CD4/CD8 phenotypes and demonstrated that T cell reactivity to B6-1710 is, although not absolute, dependent on the presence of CD4 molecules. Further analysis of T cell hybridomas with known J beta-chain usage revealed that D beta- and J beta-chain usage do not play crucial roles in T cell reactivity to B6-1710 B cells. However, T cell hybridomas derived from TCR-V beta gene transgenic mice were found to be heterogeneous for their reactivity to B6-1710, suggesting that the V alpha-chains associating with the transgenic V beta-chain determine T cell responsiveness to B6-1710. These data clearly demonstrate that T cell reactivity to a murine AIDS virus expressing B cell line resembles that previously reported for Mls-like superantigens.


The parasporal inclusion of Bacillus thuringiensis subsp. shandongiensis: characterization and screening for insecticidal activity. The parasporal body of Bacillus thuringiensis subsp. shandongiensis was characterized in terms of its structure, protein composition, and toxicological properties against several types of insects. The crystals of B. thuringiensis shandongiensis appear to consist of a major protein of 144 kDa present in an spherical inclusion, as determined by transmission electron microscopy, titration curve analysis, and SDS-PAGE of the solubilized crystals. A second protein of ca. 60 kDa is present in trace amounts and appears to be associated with a small bar-shaped inclusion. The 144-kDa protein has been characterized by isoelectric point determination, N-terminal amino acid sequence analysis, amino acid analysis, and immunological cross reactivity. Its N-terminal amino acid sequence differed from that of other B. thuringiensis crystal proteins. The 144-kDa protein was not immunologically related to the crystal proteins of two toxic serovars (B. thuringiensis israelensis and B. thuringiensis kurstaki HD-1) and one nontoxic serovar (B. thuringiensis indiana), as shown in immunoblots probed with antiserum raised against the 144-kDa B. thuringiensis shandongiensis protein, the B. thuringiensis israelensis crystal proteins, and the trypsin resistant fragment of B. thuringiensis kurstaki P1 proteins. In contrast to most B. thuringiensis serovars, B. thuringiensis shandongiensis crystals did not dissolve at pH 12. Solubilization was achieved in sodium bicarbonate at pH 8.3 and in the presence of 25 mM dithiothreitol.(ABSTRACT TRUNCATED AT 250 WORDS)


Type I keratinocyte transglutaminase: expression in human skin and psoriasis. A 92-kD transglutaminase (TGase K), expressed in human cultured keratinocytes and stratum corneum, catalyzes a critical step in the formation of the cornified envelope of terminal differentiation. A rabbit polyclonal antibody to TGase K was used to isolate overlapping cDNA clones from a human keratinocyte cDNA expression library. The cDNA clones were sequenced and unequivocally identified as TGase K by comparison to the N-terminal amino acid sequences of two cyanogen bromide fragments from the purified enzyme. The mRNA for Tgase K is expressed in cultured keratinocytes but not in A431 squamous carcinoma cells, in fibroblasts, or in other non-epithelial tissues and cells. Although TGase K protein expression is limited to the upper layers of normal epidermis, the mRNA is generally present throughout the epidermis, suggesting the possibility of post-transcriptional regulation. Precocious expression of TGase K protein occurs in psoriasis, and quantitative Northern blot analysis of TGase K mRNA from normal and involved epidermal biopsies from psoriasis patients suggests that TGase K mRNA levels are increased in psoriatic lesions. By using quantitative laser scanning confocal microscopy (LSCM) and in situ hybridization, the increase of the TGase K mRNA was in the range of 3-7 times in the psoriatic epidermis and was significantly higher compared with normal skin and with paired adjacent skin. Quantitative LSCM provides a powerful and direct method for analysis of gene expression in skin.


ICAM-1 expression in a spontaneously transformed human keratinocyte cell line: characterization by a simple cell-ELISA assay. Interferon gamma (IFN-gamma) is known to induce ICAM-1 on keratinocytes (KC) in vitro, and its expression in vivo is correlated with epidermal T-cell infiltration in various dermatoses. However, the mechanisms for this cytokine-mediated ICAM-1 expression are essentially unknown. We investigated the induction of ICAM-1 by IFN-gamma in HaCaT cells, a spontaneously transformed human KC cell line, using an immunoperoxidase-ELISA with the monoclonal antibody (MoAb) R6.5. HaCaT cells constitutively expressed low levels of ICAM-1, which were upregulated by IFN-gamma. The kinetics and dose response were similar to those published for primary KC, regardless of whether the HaCaT cells were cultured in low- or high-calcium medium. ICAM-1 expression was increased significantly at 4 h with 500 U/ml IFN-gamma, and reached a plateau (approximately 5 x greater than constitutive) by 24 h. At concentrations greater than 10 U/ml for 24 h, IFN-gamma induced ICAM-1 expression in a dose-dependent fashion (half maximal at 100 U/ml). TNF-alpha alone, and in synergistic combination with IFN-gamma, also upregulated the expression of HaCaT ICAM-1. IFN-gamma treatment of HaCaT cells increased the level of ICAM-1 mRNA and enhanced (approximately 3x) the adherence of fluorescently labeled (calcein) human T lymphoblasts, as determined by Northern blotting and an in vitro adhesion assay, respectively. Our findings suggest that HaCaT cells, in conjunction with a simple immunoperoxidase cell-ELISA, provide a reliable system for studying pharmacologic modulation of ICAM-1 on KC.


Effects of ultraviolet radiation on murine epidermal Langerhans cells: doses of ultraviolet radiation that modulate ICAM-1 (CD54) expression and inhibit Langerhans cell function cause delayed cytotoxicity in vitro. Low doses (100 J/m2) of ultraviolet B (UVB) radiation from sunlamp fluorescent FS20 tubes inhibit the ability of freshly isolated murine epidermal Langerhans cells (LC) to support anti-CD3 MoAb-induced T-cell mitogenesis and selectively inhibit the upregulation of ICAM-1 expression by LC without causing appreciable cytotoxicity in short-term (less than or equal to 24 h) incubations (J Immunol 146:3347-3355, 1991). In the present study, epidermal cells (EC) were exposed to UVB radiation or were sham-irradiated and cultured for 24, 48, or 72 h when LC were recovered, enumerated, and assayed for simultaneous expression of I-A antigens and ICAM-1 by flow cytometry. UVB-irradiated LC that had been cultured for 24 h exhibited levels of I-A antigens comparable to those on unirradiated LC but expressed substantially less ICAM-1. After 48 and 72 h, cultured UVB-irradiated LC expressed somewhat lower levels of I-A antigens and markedly less ICAM-1 than unirradiated controls. Although similar numbers of LC were recovered from cultures initiated with UVB-irradiated and unirradiated epidermal cells after 24 h, far fewer identifiable LC were recovered from cultures seeded with irradiated cells at 48 and 72 h (approximately 50 and approximately 10% of control, respectively). The effect of UVB radiation on the survival of LC in vitro was not reversible with exogenous TNF alpha (125 U/ml) alone or granulocyte/macrophage colony-stimulating factor (5 ng/ml) and IL-1 (50 U/ml) in combination, although these cytokines had modest effects on the expression of I-A antigens and ICAM-1 by cultured UVB-irradiated LC. Results of survival studies performed with enriched LC preparations demonstrated that UVB radiation was clearly cytotoxic for LC and did not merely downregulate surface expression of I-A antigens or alter LC buoyant density. Exposure of LC to radiation from blacklight fluorescent (UVA) tubes (0.25 J/cm2) in the presence of 8-methoxypsoralen (1 micrograms/ml; PUVA) or monochromatic UVC radiation (20 J/m2) also inhibited LC accessory cell function. Results of survival studies performed with EC that had been exposed to PUVA or UVC radiation before culture were similar to those of studies performed with UVB-irradiated cells, although PUVA- and UVC-induced LC cytotoxicity was much more pronounced 48 h after culture initiation than UVB-induced cytotoxicity. UVA radiation alone augmented LC recovery at 24 and 48 h, but did not influence I-A antigen or ICAM-1 expression.(ABSTRACT TRUNCATED AT 400 WORDS)


Belgrade virus: a new hantavirus causing severe hemorrhagic fever with renal syndrome in Yugoslavia. Two biologically and genetically distinct hantaviruses were isolated from blood and urine specimens collected from four Yugoslavian patients with clinically severe hemorrhagic fever with renal syndrome (HFRS). Viral isolates from three patients, designated strains Belgrade 1-3, were distinct from Hantaan, Seoul, Puumala, and Prospect Hill viruses as determined by plaque-reduction neutralization tests and restriction analysis of enzymatically amplified M-segment fragments. The fourth isolate, called Kraljevo, was indistinguishable from Hantaan virus. Strains Belgrade 1 and 2, like the Kraljevo strain, caused a fatal meningoencephalitis in newborn mice inoculated with 100 pfu of virus intracerebrally and intraperitoneally. Strain Belgrade 3 was much less neurovirulent, requiring 30,000 pfu of virus to cause fatal disease in mice. These data indicate that two distinct hantaviruses, one of which constitutes a new serotype, cause clinically severe HFRS in Yugoslavia.


Streptococcal pyrogenic exotoxin A and streptolysin O enhance polymorphonuclear leukocyte binding to gelatin matrixes. Autopsy data from cases of streptococcal toxic shock demonstrate accumulation of polymorphonuclear leukocytes (PMNL) within lung and soft tissue microvasculature. Because of the increased prevalence of streptococcal pyrogenic exotoxin A (SPEA)-producing strains associated with streptococcal toxic shock syndrome, experiments were done to determine whether SPEA or streptolysin O (SLO, a thiol-activated cytolysin produced by all group A streptococci) could stimulate PMNL-dependent adherence mechanisms in vitro. SPEA (0.01-10 micrograms/5.5 x 10(6) PMNL) only modestly enhanced PMNL adherence over the entire range of concentrations tested. In contrast, SLO-induced PMNL binding was highly dose dependent (maximal binding, 55.1 +/- 1.6% at 0.5 hemolytic units/5.5 x 10(6) PMNL) and was mediated by CD11/CD18 adherence glycoprotein.


Genetic diversity of penicillin-resistant Neisseria meningitidis. The genetic relatedness of 42 penicillin-resistant Neisseria meningitidis isolates obtained during a 2-year period from a single hospital was studied by multilocus enzyme electrophoresis and by restriction fragment length polymorphism (RFLP) analysis of penicillin-binding protein (PBP) 2 genes. The PBP 2 genes of 7 susceptible strains gave identical RFLP profiles. Sixteen different PBP 2 RFLP profiles were found among the 42 resistant strains, but 4 were found in greater than 1 resistant isolate. Multilocus enzyme electrophoresis revealed a high level of genetic diversity. Four clusters of resistant strains could be distinguished at a genetic distance of 0.75. Resistant strains with the most common PBP 2 RFLP profile were restricted to one of these clusters and may be derived from a common ancestral strain. However, resistant strains with the 3 other common RFLP profiles were distributed in two or more of the clusters.


Immunohistochemical characterization of endocrine cells in experimental exocrine pancreatic cancer in the Syrian golden hamster. Fifty exocrine pancreatic adenocarcinomas and 57 benign tumors induced in Syrian hamsters by N-nitrosobis(2-oxopropyl)amine (BOP) were examined for the presence of argyrophil cells antiinsulin, -glucagon, -somatostatin, -pancreatic polypeptide (PP), -gastrin/CCK, -vasoactive intestinal polypeptide (VIP), and - neuron-specific enolase (NSE) reactive cells. Argyrophil - and antihormone-reactive cells were found in the normal pancreatic ducts and in the acini, as well as in hyperplastic and atypical ducts/ductules, tubular complexes, benign lesions, and in 80% of ductal adenocarcinomas. Insulin and antiNSE-reactive cells were the most common, followed in decreasing frequency by glucagon, somatostatin, and PP cells. Antigastrin-/CCK-and -VIP-reactive cells were found in two cases. Argyrophil cells were present in about 60% of the tumors with Grimelius staining and in 55% of those with Churukian-Schenk staining. Insulin cells were seen in ductal cancer that had grown into a lymph node and in the lymph node metastases of another cancer. A novel finding was the presence of argyrophil and insulin cells within the lumen of some malignant glandular structures. Coexistence of several peptide cells was found in 52% of the cancers. The presence of argyrophil and hormone-producing cells in induced pancreatic ductal/ductular lesions further strengthens the existence of a close developmental relationship between exocrine and endocrine cells of the pancreas.


[Urodynamic study on urinary disturbance after therapy of uterine cancer]. It is well known that urinary disturbance often appears after radical hysterectomy for uterine cancer and is aggravated by additional radiation therapy. The aim of this study is to elucidate the pathogenesis and to establish the treatment method of urinary disturbance after therapy for uterine cancer. Forty-five patients with urinary disturbance and ten normal controls were subjected to this study. Changes in clinical symptoms and findings in the Urodynamic study (UDS) in 12 severe cases were investigated before and after treatment with beta 2-stimulant (Mabuterol HCL). Clinical symptoms in cases treated by radiation therapy alone were rare and mild without any pad exchanges, and appeared 5 years after treatment for uterine cancer. Findings of UDS in these cases were mild low compliance of detrusor at maximum desire to void (Cmdv) and mild low bladder volume at maximum desire to void (Vmdv). In cases treated by radical hysterectomy alone, Cmdv decreased immediatelly after the operation and then maximum urethral closure pressure (cPura) gradually decreased. Concerning the cases treated by radical hysterectomy and radiation, severe low Cmdv and severe low Vmdv appeared 5 years after the treatment for uterine cancer in almost all cases, and low cPura appeared immediately after the operation in half of the cases. Treatment with beta 2-stimulant significantly improved urinary frequency, voided volume and urinary incontinence. In UDS findings, Vmdv and Cmdv were significantly improved by the treatment with beta 2-stimulant. The functional profile length and cPura value were not significantly changed. In uroflowmetry, the maximum flow rate and average flow rate were significantly improved by the treatment with beta 2-stimulant.(ABSTRACT TRUNCATED AT 250 WORDS)


[Establishment and characterization of human ovarian fibrosarcoma cell line and its sensitivity to anticancer agents]. We succeeded in establishing a cell line (KEN-3) for subculture from a fibrosarcoma which originated in the ovary in a girl aged 17 years. Its characteristics and sensitivity to anticancer agents are reported in this paper. 1. Characteristics of established cell line. Lined cells consist of multinucleated giant cells mixed among many spindle-shaped cells. They grow in small colonies and have none of the pavement-like arrangement characteristic of epithelial tumor cells. The number of chromosomes ranged from 45 to 128 (mode: pseudo-triploidy region, 65). The doubling time, cellular density and plating efficiency were 76.9 hours, 5.4 x 10(5)/cm2 and 30.2%, respectively. Concerning tumor markers, CEA and sialyl SSEA-1 were only produced in small quantities. Subculture was possible subcutaneously in the nude mouse with no capacity for the production of ascites. 2. Susceptibility to anticancer agents and GP170 expression. The in vitro susceptibility to about 12 types of anticancer agents was investigated with the MTT assay. IC50/PPC was shown to be less than 1 for Adriamycin only. The sensitivity to CDDP (IC50/PPC: 4.8) was low, and no sensitivity was observed at all to DTIC, which is used frequently for mesenchymal tumors. GP170 (mdr-1 products) was positive in established cells in immunohistochemical stain.


Recent advances in the study of ionizing radiation damage and repair. This workshop, organized under the auspices of the EC Concerted Action Programme on DNA Repair and Cancer, was held at the CRC Gray Laboratory, Northwood, Middlesex, UK, 23-25 October 1991. The 42 participants were drawn mainly from laboratories participating in the EC Concerted Action, with a few visitors from elsewhere. The discussions centred on the increasing convergence of classical radiobiology and biophysics with molecular biology and mammalian cell genetics to study mechanisms of DNA strand break accumulation and repair following exposure to ionizing radiation. There was a strong emphasis on the application of this research both to cancer radiotherapy and to detection of individuals at risk from cancer due to exposure to ionizing radiation. The first two days were organized as six workshop sessions; on the third day we joined forces with Julie Denekamp and dedicated a session to the memory of our late friend and colleague, Nic McNally. The rest of this day was devoted to reviews by his colleagues and collaborators of the fields of research to which he contributed so much. An evening of music and readings, organized by Joanna and Rachel McNally, completed the memorial. Here we review the first seven sessions of the workshop, emphasizing the more recent approaches and the new information they have given us.


A comparison of the chemical repair rates of free radical precursors of DNA damage and cell killing in Chinese hamster V79 cells. One of the important temporal stages of radiation action in cellular systems is the chemical phase, where oxygen fixation reactions compete with chemical repair reactions involving reducing agents such as GSH. Using the gas explosion technique it is possible to follow the kinetics of these fast (greater than 1 ms) reactions in intact cells. We have compared the chemical repair kinetics of the oxygen-dependent free radical precursors leading to DNA single-strand and double-strand breaks, measured using filter elution techniques, with those leading to cell killing in V79 cells. The chemical repair rates for DNA dsb (670s-1 at pH 7.2 and 380s-1 at pH 9.6) and cell killing (530s-1) were similar. This is in agreement with the important role of DNA dsb in radiation induced cell lethality. The rate for DNA ssb precursors was significantly slower (210s-1). The difference in rate between DNA ssb and dsb precursors may be explained on the basis of a dsb free radical precursor consisting of a paired radical, one radical on each strand. The instantaneous probability of one or other of these radicals being chemically repaired and not proceeding to form a dsb will be twice that of ssb radical precursor. This agrees well with the concept of locally multiply damaged sites (LMDS) produced from clusters of ionizations in DNA (Ward 1985).


Characterization of two radiation-induced lesions from DNA: studies using nuclease P1. Exposure of DNA oligomers to ionizing radiation in oxygenated solution reveals that the two lesions formed in highest yield are the 8-hydroxy modification of guanine and the formamido remnant of thymine. The effect of these lesions on the hydrolysis of the phosphodiester bond by nuclease P1 was studied. Whereas the 8-hydroxyguanine lesion does not affect hydrolysis of the adjacent 3' phosphoester bond by nuclease P1, the formamido lesion markedly retards hydrolysis. Consistent with the oligomer results, digestion of irradiated DNA polymer by nuclease P1 plus acid phosphatase yielded the 8-hydroxyguanine lesion, obtained as the modified nucleoside, and the formamido lesion, obtained as a modified dinucleotide.


Constraints on energy deposition and target size of multiply damaged sites associated with DNA double-strand breaks. We suggest that the observed experimental data on relative double-strand break (dsb) yield as a function of radiation quality can act as valuable constraints in defining the type of energy deposition which causes this basic lesion in radiation biology. Both heavy-ion and alpha-particle data show sufficient trends for quantitative comparisons with calculation to be made. We use the technique of track-structure simulation and search for energy-deposition clusters (containing at least a given number of ionizations in a given diameter) whose relative frequencies (compared to sparsely ionizing radiation) correlate with the relative biological effects (RBEs) for dsb induction. We conclude that locally multiply damaged sites (LMDS) which cause dsb are probably energy depositions of at least two to five ionizations localized, respectively, in sites of diameters of 1-4 nm. Although our derived cluster sizes should be viewed in light of the quality of the experimental data and uncertainties in the computer simulations at the nanometre level, it is unlikely that these estimates of cluster sizes would change greatly.


DNA damaging and cell cycle effects of the topoisomerase I poison camptothecin in irradiated human cells. This study addressed the potential radiosensitizing and DNA-damaging actions of the DNA topoisomerase I poison camptothecin (CPT) on SV40 transformed normal (MRC5CVI) and ataxia-telangiectasia (AT5BIVA) fibroblast cell lines. In both cell lines CPT induced a dose-dependent delay of cells in S phase, followed by a dose-dependent trapping in G2/M phase. Acute X-irradiation produced patterns of G2/M arrest and S-phase delay similar to those observed for CPT in the MRC5CVI cell line, but no S phase delay was observed in the AT5BIVA cell line consistent with the ataxiatelangiectasia phenotype of this cell line. X-irradiation of CPT-treated cells resulted in additive prolongation of S phase delay in MRC5CVI cultures and additive effects for cell killing in both cell lines. The potential for topoisomerase I-DNA cross-linking by CPT was not altered by 24h pretreatment with CPT, or by acute X-irradiation. Hypersensitivity of AT5BIVA to CPT was not attributable to elevated levels of complex trapping. These findings suggest that in a rapidly proliferating human tumour there is unlikely to be synergistic therapeutic gain when the two agents are used concurrently, and that previously reported radiosensitization by CPT is restricted to G0 phase cells.


Induction of lacI- mutations in Escherichia coli cells after single and split-dose irradiation. In the lacI system of Escherichia coli, X-ray mutagenesis follows a linear-quadratic curve with suppression; the survival curve is exponential. Dose fractionation leads to nearly complete repair of premutational lesions during an incubation interval of 3.5 h. Repair starts with a delay of 1.5-2 h, suggesting the involvement of an inducible repair/mutation fixation system. The dose-dependence of mutagenesis is described by a simple model assuming two hits being required. A probable explanation might be that the premutagenic lesions consist of two closely spaced lesions on the opposite strands of the DNA molecule.


The effect of fluoride on photodynamic-induced fluorescence changes of aluminium phthalocyanine in Chinese hamster cells. Fluence-dependent changes in the fluorescence of aluminium phthalocyanine (AlPc) were measured in Chinese hamster ovary (CHO) cells using digital fluorescence microscopy of single cells and spectrofluorimetry of cell suspensions. During illumination the fluorescence initially increased and later progressively decreased. In the presence of fluoride, which protects against phototoxicity of AlPc by forming a fluoroaluminium complex, there was no initial increase in fluorescence: it decreased about 10 times faster than in the absence of fluoride. Qualitatively similar results were observed using single-cell fluorescence microscopy, which also showed the dye to be mostly localized in cytoplasmic organelles and membranes. The pattern of localization did not change during illumination. Concomitant assays of dye extracted from cells revealed little photodegradation that could not account for the fluorescence changes. The absorption spectra of AlPc-loaded cells showed some aggregation of the dye prior to light exposure. During illumination the dye was initially monomerized and subsequently progressively reaggregated. In the presence of fluoride no monomerization was seen, and the aggregation proceeded at a much faster rate. It is concluded that the fluorescence changes are not due to major relocalization of AlPc in the cells, but to light-induced monomerization followed by reaggregation. The protective effect of fluoride may be due to the enhanced aggregation rate, because aggregated dye molecules are photochemically inactive. Because D2(0) affects neither the initial enhanced fluorescence in the absence of fluoride nor the rapid decrease in its presence it appears that 1O2 is not involved in the photodynamic reactions leading to these changes.


Study of dose-rate and split-dose effects on the in vitro micronucleus yield in human lymphocytes exposed to X-rays. This paper reports the effects of changes in dose-rate and dose-fractionation on the micronucleus (MN) yield in human lymphocytes exposed to 250 kV X-rays. For the investigation of dose-rate effects whole blood samples of four healthy donors were irradiated with doses ranging from 1 to 4 Gy given at various dose-rates between 0.2 and 40 Gy/h. For the higher doses (3 and 4 Gy) a decline in the MN yield became apparent when the dose-rate was reduced below 1.6 Gy/h. This effect was enhanced systematically by a further lowering of the dose-rate. For lower doses (1 and 2 Gy) the reduction in the MN yield was less pronounced: only a small effect was observed for two donors when a dose of 2 Gy was administered at a dose-rate of 0.2 Gy/h. In the split-dose experiment a dose of 4 Gy was delivered either as a single exposure or in two fractions of 2 Gy, separated by time intervals ranging from 30 min to 10 h. A continuous decrease of the MN yield with increasing interfraction time is observed: after an initial fast decline a further slight reduction in the MN yield occurs. The observed dose-rate and split-dose effects on the MN yield can be attributed to repair of sublethal damage.


Induction of chromosome aberrations by monochromatic X-rays with resonance energy of phosphorus K-shell absorption edge. The induction of chromosome aberrations by monochromatic soft X-rays with energies corresponding to the K-shell absorption edge of phosphorus has been studied in density-inhibited mouse m5S cells. The frequency of dicentrics was markedly enhanced when the cells were irradiated with energy at the K-shell resonance peak (2.153 keV) as compared to those at below (2.146 keV) or above (2.160 keV) the peak. The quantum efficiency was calculated to be 2.7 x 10(-3) for the induction of dicentrics per photoelectric absorption of phosphorus atom in DNA, which was comparable to the known efficiencies of X- or gamma-ray-induced DNA double-strand breaks. However, comparison of the efficiencies based on the absorbed dose indicated that the magnitude of the enhancement was not due solely to the selective photoelectric absorption of the phosphorus atoms in DNA, and suggested the combined contributions of Auger electrons from phosphorus atoms within and outside the DNA molecules.


Membrane oxidative damage induced by ionizing radiation detected by diphenylhexatriene fluorescence lifetime distributions. The sensitivity of the fluorescence lifetime of 1,6-diphenyl- 1,3,5-hexatriene (DPH) to the dielectric constant of its environment has been used to detect oxidative damage to phospholipid membranes induced by ionizing radiation. The DPH fluorescence decay in phospholipid vesicles is described well by a continuous distribution of lifetime values, reflecting the various DPH depths in the bilayer and related to the gradient of the dielectric constant. Ionizing radiation oxidizes unsaturated acyl residues of phospholipids, altering the dielectric constant across the bilayer, sharpening the distribution of DPH lifetimes and increasing the centre of the distribution. Ionizing radiation doses between 22 and 110 Gy were used, and were effective only in the presence of oxygen. A model based on the formation of packing defects in the bilayer describes the phenomenon.


Hypoxia-specific inhibition of recovery from radiation damage by a novel 2-nitroimidazole with a theophylline side chain. A novel 2-nitroimidazole with a theophylline side-chain, 7-(4'-(2-nitroimidazol-1-yl)-butyl)-theophylline, (NITP) was as efficient a hypoxic-cell radiosensitizer as misonidazole. However, if cells were irradiated with NITP under hypoxic conditions and then exposed to the drug under aerobic conditions, a much larger radiosensitizing effect was observed, partly because of a reduction in the size of the shoulder of the survival curve. There was little effect of NITP on the radiosensitivity of well oxygenated cells, even with post-irradiation drug contact. Split-dose survival curves showed that the drug inhibited recovery from radiation damage only when the cells were irradiated under hypoxia but not when irradiations were under oxic conditions. A reduction in the size of the shoulder of the survival curve should allow the hypoxic-cell radiosensitizing efficiency of NITP to be maintained with low doses of radiation used in multifraction cancer radiotherapy. Bifunctional drugs containing both electron-affinic and repair inhibiting groups may represent a new approach to the synthesis of hypoxic-cell targeted adjuncts to radiotherapy.


The inherent cellular sensitivity to 62.5 MeV(p----Be+) neutrons of human cells differing in photon sensitivity. The inherent sensitivity of 20 human cell lines to the 62.5 MeV(p----Be+) clinical neutron beam at Clatterbridge, UK, has been assessed and compared to their sensitivity to 4 MeV photons. The survival curves of the cell lines following neutron irradiation were curvilinear, and the inherent neutron sensitivity varied by 4.5 fold (0.1 survival level) between the extreme values, in the cell lines studied. There was a strong correlation between the sensitivity of these human cells to photon and neutron irradiation. It was concluded that should these in vitro patterns occur in the clinic, the 4-fold variation in RBE and inherent sensitivity to neutrons could result in overall lower local control rates following fast neutron therapy than might be anticipated. It suggests the need for the development of predictive assays as a potential means of selecting tumours most appropriate for neutron therapy.


Inactivation of C3H 10T1/2 cells by monoenergetic high LET alpha-particles. Inactivation of mouse C3H 10T1/2 cells in plateau-phase (7.8 x 10(4) cells/cm2) was studied by using alpha-particles from the irradiation facility installed for radiobiological experiments at the 3 MV Tandem accelerator, University of Naples. Silicon detectors and CR39 plastic track detectors were employed for dosimetric purposes. The cells were exposed to high LET monoenergetic alpha-particles (energy of 1.8 MeV at the centre of the cell nucleus, track-averaged LET of 177 keV/micron and dose-rate of 1.1 Gy/min) and low-LET 80 kVp X-rays. The X-ray survival curve showed a significant shoulder (alpha/beta = 9 Gy) while the survival curve for alpha-particles was close to exponential. The mean lethal dose of alpha-particles was 0.77 +/- 0.02 Gy and the RBE was 5.2 at 80% survival and 3.0 at 5% survival. Survival of exponentially growing cells (2 x 10(4) cells/cm2) following irradiation with the alpha-particle beam is also reported. The nuclear areas of 10T1/2 cells were measured as 299 +/- 9 micron 2 and 250 +/- 8 micron 2 for cells in log phase and plateau phase, respectively. The inactivation cross-section, obtained from the mean lethal dose, was 34 micron 2 and 37 micron 2 for cells in log phase and plateau phase, respectively. These values appear to be the maximum measured values for the inactivation cross-section of 10T1/2 cells as a function of the alpha-particle LET. This saturation cross-section is very similar to the saturation values reported in the literature for other mammalian cell lines.


Strontium-90 induced bone tumours in beagle dogs: effects of route of exposure and dose rate. Bone tumours from beagles exposed by inhalation to 90SrCl2 at the Inhalation Toxicology Research Institute (ITRI), by chronic ingestion of 90Sr at the Laboratory of Energy-Related Health Research (LEHR), and by injection of 90Sr citrate at the University of Utah were analysed to determine if the bone tumour characteristics differed among the three studies. The range of average skeletal doses at which the bone tumours occurred was similar in all three studies, but differences in the skeletal distribution, histological phenotype, and time to death were observed. The differences observed were attributed to the difference in dose-rate pattern obtained in the chronic ingestion study, in contrast to the inhalation and injection studies. In general, however, the differences noted in bone tumour characteristics were subtle, and would be unlikely to make an impact on models developed to assess the risk of human exposure to 90Sr.


Effects of the perfluorochemical emulsion FMIQ on the radiation response of EMT6 tumours. The effects of FMIQ, a perfluorochemical emulsion based on perfluoro-N-methyldecahydroisoquinoline, were examined using BALB/c mice and EMT6 mammary carcinomas. The radiobiological effects of FMIQ were similar to those found previously for Fluosol in the same tumour/host system. Although the perfluorochemical content (20% w/v) and oxygen-carrying capacity of FMIQ are similar to those of Fluosol, the formulation of FMIQ offers some advantages over that of Fluosol. For example, FMIQ has greater stability during storage. FMIQ also is formulated without pluronic F-68 and is based on a perfluorochemical (FMIQ) having a shorter tissue dwell time than the perfluorotripropylamine in Fluosol; it therefore may produce fewer side-effects than Fluosol. The lifetime of the circulating perfluorochemical droplets in BALB/c mice was longer than FMIQ than for Fluosol; this could offer an advantage in fractionated radiotherapy. These findings give reason to expect that FMIQ may prove to be a better emulsion than Fluosol for clinical use as an adjunct to cancer therapy.


Mls: a link between immunology and retrovirology. The nature of the mysterious minor lymphocyte stimulating (Mls) antigens has recently been clarified. These molecules which were key elements for our current understanding of immune tolerance, have a strong influence on the mouse immune system and are encoded by the open reading frame (orf) of endogenous and exogenous mouse mammary tumor viruses (MMTV's). The knowledge that these antigens are encoded by cancerogenic retroviruses opens an interdisciplinary approach for understanding the mechanisms of immune responses and immune tolerance, retroviral carcinogenesis, and retroviral strategies for infection.


Prognostic significance of HER-2 oncoprotein expression in breast cancer: a 30-year follow-up. To study retrospectively the long-term prognostic significance of HER-2 oncoprotein expression in breast cancer (BC). Two hundred nine consecutive female patients with invasive operable BC from a defined urban population were observed for a median of 30 years. Tissue expression of HER-2 oncoprotein was demonstrated by using an immunoperoxidase procedure. Fifty-five (26%) patients had cancer and a positive HER-2 oncoprotein stain reaction. They had significantly worse 10- and 25-year survival rates than those patients who had a negative stain reaction in their cancer (31% v 48% and 31% v 39%, respectively; P = .004). HER-2 expression was also associated with a poorer survival among patients who had axillary nodal metastases (P = .003; n = 104), but not among those patients who did not have metastases. HER-2 expression was related to the ductal histologic type, poor histologic grade, and high mitotic count, but not to tumor size, axillary nodal status, DNA ploidy, or S-phase fraction (SPF). In a multivariate analysis among patients with nodal metastases, HER-2 expression was an independent prognostic factor (P = .04) that predicted poor survival. However, if the entire series was entered onto the analysis, it did not emerge as an independent factor. HER-2 oncoprotein expression has long-term prognostic significance for predicting poor survival in BC, and it has an independent prognostic value among patients who presented with axillary nodal metastases. This result is contradictory to the argument that HER-2 expression is only a marker for drug resistance because the patients were not given adjuvant drug therapy.


Prognostic importance of c-erbB-2 expression in breast cancer. International (Ludwig) Breast Cancer Study Group. To evaluate the prognostic importance of immunocytochemically determined c-erbB-2 overexpression in the primary tumors of patients with breast cancer. Primary tumors from 1,506 breast cancer patients (760 node-negative and 746 node-positive) who were treated in the International (Ludwig) Breast Cancer Study Group Trial V were studied. Node-negative patients were allocated randomly to either a single cycle of perioperative chemotherapy (PeCT) or no adjuvant treatment, and node-positive patients received either a prolonged chemotherapy (with tamoxifen for postmenopausal patients) or a single perioperative cycle. Tumors from 16% of the node-negative patients and 19% of the node-positive patients were found to be c-erbB-2-positive. In both groups c-erbB-2 positivity correlated with negative progesterone receptors (PR), negative estrogen receptors (ER), and high tumor grade. Lobular carcinomas were all negative, and, thus support the view that such tumors represent a defined subtype of breast carcinoma. The expression of c-erbB-2 was prognostically significant for node-positive but not for node-negative patients. However, in both subgroups, the prognostic significance was greater for patients who had received more adjuvant therapy. For node-positive patients, the effect of prolonged-duration therapy on disease-free survival (DFS) was greater for patients without c-erbB-2 overexpression (hazards ratio [HR], = 0.57; 95% confidence interval [CI], 0.46 to 0.72) than for those with c-erbB-2 overexpression (HR, 0.77; 95% CI, 0.51 to 1.16). Similarly, for node-negative patients, the effect of PeCT on DFS was greater for those without c-erbB-2 overexpression (HR, 0.82; 95% CI, 0.61 to 1.09) than for those with c-erbB-2 overexpression (HR, 1.22; 95% CI, 0.66 to 2.25). We conclude that tumors with overexpression of the c-erbB-2 oncogene are less responsive to cyclophosphamide, methotrexate, and fluorouracil (CMF)-containing adjuvant therapy regimens than those with a normal amount of gene product.


Metabotropic glutamate receptors mediate excitatory transmission in the nucleus of the solitary tract. Following microinjection into the nucleus tractus solitarius (NTS), the effects of glutamate on the baroreceptor reflex are poorly antagonized by kynurenic acid and DL-2-amino-5-phosphonovaleric acid, suggesting the possible involvement of metabotropic glutamate receptors in this response. The metabotropic glutamate receptor agonist 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) depolarized neurons located medial to the tractus solitarius (TS) at the level of the area postrema in coronal sections of the rat NTS. This effect was mimicked by glutamate and was not blocked by antagonists at alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA)/kainate or NMDA receptors. 1S,3R-ACPD also produced an inward current under voltage clamp that was not accompanied by a rise in [Ca2+]i, monitored with the Ca(2+)-sensitive dye fura-2. Conversely, the muscarinic agonist carbachol produced an outward current and a rise in [Ca2+]i. 1S,3R-ACPD reduced both the excitatory and the inhibitory postsynaptic current resulting from single electrical stimuli in the region of the TS. High-frequency stimulation of the TS produced an inward current in the presence of AMPA/kainate and NMDA receptor blockers. This current had similar properties to that produced by 1S,3R-ACPD. Thus, metabotropic glutamate receptors may mediate a component of excitatory transmission in the NTS.


Corticotropin-releasing factor: long-lasting facilitation of the acoustic startle reflex. Intracerebroventricular infusion of corticotropin-releasing factor (CRF) (0.1-1.0 micrograms) produced a pronounced, dose-dependent enhancement of the acoustic startle reflex in rats. This excitatory effect began about 20-30 min after infusion, grew steadily over the 2 hr test period, and lasted at least 6 hr. Higher doses of CRF (10 micrograms) often produced marked facilitation and then inhibition of startle that oscillated repeatedly with a period of 10-20 min. CRF-enhanced startle did not result from an increase in sensitization produced by repetition of the startle stimulus or from a blockade of habituation. Peripheral injections of the autonomic ganglionic blockers hexamethonium (10 mg/kg) or chlorisondamine (3 mg/kg) slightly attenuated the magnitude of CRF-enhanced startle, suggesting a partial role of peripheral sympathetic activation. Intracerebroventricular infusion of the CRF antagonist alpha-helical CRF9-41 (alpha hCRF; 25 or 50 micrograms) blocked CRF-enhanced startle when infused 5 min prior to CRF, indicating a central site of action. CRF-enhanced startle also was reversed when alpha hCRF was given 90 min after infusion of CRF. This suggests that exogenously applied CRF remains in the brain for a very long time after administration or that CRF given exogenously initiates a process that results in a long-lasting activation of endogenous CRF. Because the startle reflex is elevated by both conditioned and unconditioned fear, these data lend further support to the idea that CRF infusion produces a behavioral state that resembles fear or anxiety. Because startle is mediated by a well-defined neural pathway, CRF-enhanced startle may provide a useful behavioral assay to analyze the neural systems upon which exogenous CRF acts to produce its behavioral effects.


Single local interneurons in the locust make central synapses with different properties of transmitter release on distinct postsynaptic neurons. Quantal analysis has been applied to the inhibitory synapses made by single spiking local interneurons onto several nonspiking local interneurons (and motorneurons) in the locust CNS. Transmission at these synapses appears to be mediated by GABA. The apparent reversal potential of the IPSP and inhibitory postsynaptic current were -80 to -85 mV, a value similar to that of the potential evoked by pressure-applied GABA. This reversal potential was 25-30 mV more negative than the resting potential of the nonspiking interneurons in the experimental conditions. The statistical properties of release at these synapses were studied by recording simultaneously from pre- and postsynaptic interneurons with intracellular electrodes. The distribution of postsynaptic potential amplitudes could be described by a simple binomial model, implying uniformity of binomial p (probability of release at a single release site) for each synapse. The mean quantal amplitude was 290 +/- 110 microV, and the mean quantal content m of the IPSPs was 6.25 +/- 2.83. The mean values of binomial n (average size of the releasable pool) and p were 13.11 +/- 2.8 and 0.45 +/- 0.16, respectively. Numerical simulations of statistical experiments were performed to test whether the IPSP amplitude distribution histograms might be misleadingly indicative of quantal release. These simulations showed that such a hypothesis was very unlikely. When a spiking local interneuron was impaled, several of its target interneurons could sometimes be successively sampled. Quantal analysis was then performed with the different IPSPs evoked, in identical conditions, by a same presynaptic interneuron, and the quantal parameters were compared between the synapses. It was found that binomials n and p and their product m generally differed between the synapses made by a given spiking interneuron onto different target neurons. These results show that quantal contents can vary for the many synapses made centrally by one interneuron, and suggest that this variability may arise from differences in release probabilities between the sites associated with different synapses.


Clinical implications of autonomic drugs. Drugs that influence autonomic function are used more frequently than surgeons generally imagine. This article summarizes principles of autonomic pharmacology and highlights specific drugs that are useful to the practicing oral and maxillofacial surgeon.


[Interaction between opiates and neurotransmitters/neuromodulators in spinal analgesia]. Systematically administered morphine produces its antinociceptive effects through a mechanism involving synergism of the spinal and supraspinal opiate systems. Descending inhibitory monoaminergic pathways, including noradrenergic and serotonergic descending systems, participate in this production of antinociception. Recent studies indicate a synergistic interaction between opiates and adrenergic agonists in the spinal cord. The important consideration arising from the apparent synergy is that significant antinociceptive effects can be achieved with nonspecific doses of either agonist, which alone produces only minimal effects. Though synergy between opiates and adrenergic drugs were principally considered, there remains the possibility of spinal synergistic interactions of opiates and other neurotransmitters/neuromodulators. Since decreasing the effective dosage of analgesics may lessen or eliminate the complications of spinal opiates or other analgesics, these combined effects will be extremely useful for clinical pain management.


[Can nicardipine potentiate vecuronium induced neuromuscular blockade?]. The effects of intravenous nicardipine on the neuromuscular blockade produced by single bolus injection of vecuronium were studied in surgical patients undergoing tracheal intubation. We measured the mechanical response of the abductor pollicis muscle to stimulation of the ulnar nerve in a train-of-four sequence at 2 Hz and recorded the amplitudes of the first response (T1). Anesthesia was induced with thiopental 5 mg.kg-1 with or without nicardipine 10 micrograms.kg-1 followed by injection of vecuronium in a dose of either 0.1 or 0.15 mg.kg-1. Onset and duration of neuromuscular blockade were judged by percent depression of T1. The time intervals for 90% and 100% depression in T1 seen in patients who had received vecuronium 0.1 mg.kg-1 with nicardipine (n = 10) were 157.0 +/- 30.8 sec and 192.3 +/- 31.2 sec (mean +/- SD), respectively. These values were significantly shorter than those observed in patients without nicardipine administration (n = 10, P less than 0.05), and were not significantly different from the values in patients who had received vecuronium 0.15 mg.kg-1 (n = 10). On the other hand, the time for 25% recovery in T1 was uninfluenced by nicardipine. Present study indicates that nicardipine pretreatment possibly shortens the onset time after minor or moderate dose of vecuronium.


[Effect of beta adrenergic blocking drugs on the prognostic value of ST-segment depression during exercise electrocardiogram testing]. Exercise testing has been shown to be predictive for future cardiac events in patients with established diagnosis of coronary heart disease. Exercise test parameters associated with poor prognosis may be unreliable if patient is receiving beta adrenergic agents. The purpose of this study was: 1) to compare the results of exercise testing performed before and during beta blocking therapy, and 2) to determine the role of beta blockers in the prognostic significance of the ST-segment response recorded during exercise testing. The study population consisted of 518 patients (mean age 52 +/- 7 years) with coronary heart disease. The diagnosis was based on the presence of one of the following three criteria: 1) typical history and significant ST-segment depression on resting or exercise electrocardiogram, 2) history of myocardial infarction, 3) significant coronary angiographic abnormalities. In all patients symptom-limited exercise test was performed before and two weeks after the onset of beta blocker therapy. The data from the first and second tests were estimated for significance of differences between the mean values with following results: maximal heart rate--135 +/- 21 and 123 +/- 19 bpm (p less than 0.001), maximal work load achieved--98 +/- 43 and 109 +/- 44 W (p less than 0.001), maximal systolic blood pressure--171 +/- 28 and 163 +/- 26 mmHg (p less than 0.001). Occurrence of characteristic ST-segment depression was more frequent during the first than during the second test (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


Intravenous magnesium sulphate in suspected acute myocardial infarction: results of the second Leicester Intravenous Magnesium Intervention Trial (LIMIT-2). The cardiovascular actions of the magnesium ion at pharmacological concentrations include coronary and systemic vasodilatation, platelet inhibition, and antiarrhythmic effects. Magnesium has also been reported to protect myocardial tissue in experimental models of ischaemia and reperfusion. Several small clinical trials in suspected acute myocardial infarction have suggested that early mortality can be reduced by intravenous infusion of magnesium salts in the acute phase, but none has been of sufficient size to be conclusive. We therefore conducted a randomised, double blind, placebo controlled study in 2316 patients with suspected acute myocardial infarction who received either intravenous magnesium sulphate (8 mmol over 5 min followed by 65 mmol over 24 h) or physiological saline. The primary outcome measure was 28-day mortality, which was ascertained in 99.3% of patients. The groups were well balanced for prognostic factors. By intention-to-treat analysis mortality from all causes was 7.8% in the magnesium group and 10.3% in the placebo group (2p = 0.04), a relative reduction of 24% (95% confidence interval 1-43%). Within the coronary care unit the incidence of left ventricular failure was reduced by 25% (7-39%) in the magnesium group (2p = 0.009). There was no significant difference between the groups in the incidence of heart block or the use of antiarrhythmic drugs, direct-current cardioversion, or temporary pacing. Myocardial infarction was confirmed in 65% of each group, with closely similar rises in cardiac enzymes. The side-effects of magnesium treatment were transient flushing, related to speed of injection of the loading dose, and an increased incidence of sinus bradycardia (2p = 0.02). Exploratory subgroup analyses of 28-day mortality did not indicate any effect modification by thrombolysis or aspirin, or by previous treatment with beta blockers, calcium antagonists, or diuretics. Intravenous magnesium sulphate is a simple, safe, and widely applicable treatment. Its efficacy in reducing early mortality of myocardial infarction is comparable to, but independent of, that of thrombolytic or antiplatelet therapy.


21-Hydroxylase, a major autoantigen in idiopathic Addison's disease. Sera from patients with idiopathic Addison's disease commonly react with the zona glomerulosa of adrenal cortex. We used high-resolution western blot analysis of an adrenal microsomal fraction to investigate the target of these antibodies. A protein with an apparent molecular weight of 54 kDa was recognised as the common and major component. Sera identifying this autoantigen (from 12 of 16 patients) also showed strong immunofluorescence staining of a steroid-producing human adrenal adrenocortical cell line, NCI-H295. On application of antisera specific for different cytochrome P450 steroidogenic enzymes (side-chain cleavage enzyme, 21-hydroxylase, 17 alpha-hydroxylase, 11 beta-hydroxylase) the mobility of the 54 kDa protein in western blots corresponded to that of 21-hydroxylase. This parallel behaviour was confirmed by immunoprecipitation, electrophoresis, and autoradiography with the various sera and 35S-methionine-labelled NCI-H295 cell lysates. Preabsorptions of 35S-methionine-labelled cell lysates with the antiserum to 21-hydroxylase, but not with the other enzyme antisera, abolished precipitation of the 54 kDa autoantigen with the patient sera. These results indicate that 21-hydroxylase (P450c21), prominent in the zona glomerulosa of the adrenal cortex, is a major autoantigen in idiopathic Addison's disease.


Serum concentrations of vitamins A and E and early outcome after ischaemic stroke. Formation of free radicals and subsequent lipid peroxidation may have an important role in tissue injury and neuronal cell death after cerebral ischaemia. We conducted a prospective, controlled study to determine whether the endogenous antioxidant vitamins A and E had a protective function in acute ischaemic stroke. The study population consisted of 80 patients seen at the Free University Hospital in Brussels. Entry criteria were occurrence of sudden focal neurological deficit lasting more than 3 h; deficit due to acute ischaemia in the territory of the middle cerebral artery; and investigation within 24 h of onset of the episode. Outcome was assessed within the first 21 days. 80 controls matched for age and sex had various neurological disorders other than acute ischaemia. Serum concentrations of vitamins A and E were similar in the study and control groups. In the study population a serum vitamin A concentration higher than the mean of 2.27 mumol/l was associated with a higher frequency of complete recovery within the first 24 h (p less than 0.05), decreased mortality (p = 0.038), and a better outcome as assessed by the Mathew scale of neurological deficit (p less than 0.03) and the Barthel index. There was no significant difference in outcome between patients with vitamin E concentrations above or below the mean of 35.3 mumol/l. Our results suggest a beneficial effect of a high serum vitamin A concentration on early outcome in ischaemic stroke.


Anti-endothelial antibodies and coronary artery disease after cardiac transplantation. Accelerated coronary artery disease is the most serious complication after cardiac transplantation. The disease has a multifactorial aetiology, with little agreement about the relative importance of the various risk factors. We have investigated the frequency of anti-endothelial antibodies against human umbilical vein endothelial cells by one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis and western blotting. Peptide-specific anti-endothelial antibodies were found in 15/21 heart transplant recipients with accelerated coronary artery disease, and 1/20 transplant patients who had not developed the disease. Positive immunofluorescence of patients' serum on frozen sections of coronary vessels confirmed the endothelial specificity of antibodies. These results provide evidence of an immune aetiology for transplant-associated coronary artery disease and could have important implications for its diagnosis and therapy.


Prediction of normal-tissue tolerance to radiotherapy from in-vitro cellular radiation sensitivity. The success of radiotherapy depends on the total radiation dose, which is limited by the tolerance of surrounding normal tissues. Since there is substantial variation among patients in normal-tissue radiosensitivity, we have tested the hypothesis that in-vitro cellular radiosensitivity is correlated with in-vitro normal-tissue responses. We exposed skin fibroblast cell lines from six radiation-treated patients to various doses of radiation and measured the proportions surviving. There was a strong relation between fibroblast sensitivity in vitro and normal-tissue reactions, especially acute effects. Assessment of radiosensitivity could lead to improved tumour cure rates by enabling radiation doses to be tailored to the individual.


Intravascular ultrasound imaging combined with coronary angioplasty. A novel catheter combining ultrasound imaging and coronary balloon angioplasty was used in the treatment of 69 coronary-artery lesions in 51 patients. The ultrasound transducer enables real-time cross-sectional imaging and qualitative and quantitative assessment of the vessel wall before and after angioplasty. The combination catheter successfully dilated 67 lesions. There was a characteristic three-layered appearance, representing intima, media, and adventitia, in 60 cases. Intravascular imaging provided information on the vessel wall unobtainable by standard contrast angiography in 28 cases and influenced our management in 6 cases.


Intermediate-term results of heart-lung transplantation for cystic fibrosis. Between September, 1984, and March, 1991, 79 patients underwent heart-lung transplantation for end-stage cystic fibrosis at the Harefield Hospital. Short-term outcome has already been reported, and we now present intermediate-term results. The overall actuarial patient survival was 69% at 1 year, 52% at 2 years, and 49% at 3 years. 17 patients had diabetes mellitus with a survival of 62% to 1 year and 51% to 2 years. 23 patients had one or more other possible high-risk factors, and survival of these patients was 64% at 1 year and 57% at 2 years, compared with 71% and 49%, respectively, in the low-risk group (n = 56). Pseudomonas aeruginosa infection was the most common respiratory infection encountered postoperatively. 92% of patients had at least one episode of acute rejection during the first 3 postoperative months. Lung function was greatly improved after transplantation, the mean forced expiratory volume in 1 s and forced vital capacity increasing from 22% and 35% predicted, respectively, preoperatively to 68% and 70% predicted, respectively, by the sixth postoperative month. This improvement was maintained at 1, 2, and 3 years after transplantation. Lymphoproliferative disorders (4 patients) were successfully treated. Obliterative bronchiolitis developed in 17 patients and the cumulative probability of getting this complication at 1, 2, and 3 years postoperatively was 17%, 23%, and 48%, respectively. Overall, 7 patients were retransplanted. There was no coronary artery disease in the 37 patients who underwent coronary angiography at 1 year, 14 at 2 years, and 9 at 3 years after surgery. 58 patients donated their hearts for subsequent "domino" heart transplantation. Our 5 1/2-year experience with heart-lung transplantation is encouraging but the shortage of donor organs and the complication of obliterative bronchiolitis are the two main obstacles to be overcome.


Health effects of white-water canoeing.

Health effects of white-water canoeing. There is little quantitative information on the relation between water quality and disease attack rates after recreational activities in fresh water. We conducted a prospective cohort study to measure the health effects of white-water and slalom canoeing in two channels with different degrees of microbial contamination. Site A, fed by a lowland river, showed high enterovirus concentrations (arithmetic mean 198 pfu per 10 litre and moderate faecal coliform concentrations (geometric mean 285/dl); at site B, from an upland impoundment, all samples were free of enteroviruses and the geometric mean faecal coliform concentration was 22/dl. Between 5 and 7 days after exposure canoeists using site A had significantly higher incidences of gastrointestinal and upper respiratory symptoms than canoeists using site B or non-exposed controls (spectators). Like seawater bathers, fresh-water canoeists can be made ill by sewage contamination. The hazard of fresh water may be best measured by counting of viruses rather than bacteria.


Indian medical journals.

Indian medical journals. Although Indian doctors produce half the articles published from the third world, little has been written about Indian medical journals. We examined 75 of the 113 serious English-language journals published in India. Of the 22 included in the Cumulated Index Medicus only 8 were judged by Indian and foreign referees to be of international standard. A survey of Indian authors indicated that foreign journals were chosen for the best papers because of their wider circulation, better refereeing practices, and punctuality. More than 98% of medical articles from India probably go unnoticed by the international medical community.


Leukaemia incidence after iodine-131 exposure. Leukaemia is one of the most prominent late effects of exposure to ionising radiation. We have studied the incidence of leukaemia among 46,988 Swedish patients exposed to iodine-131 (131I) for diagnostic reasons or to treat hyperthyroidism or thyroid cancer. The observed number of leukaemias was compared with that expected based on incidence data from the general population. The mean absorbed dose to the bone marrow was estimated as 14 mGy (range 0.01-2.226). 195 leukaemias occurred more than 2 years after exposure, and the standardised incidence ratio (SIR) was 1.09 (95% confidence interval 0.94-1.25). Similar, but again not significantly, increased risks were seen for chronic lymphocytic leukaemia (CLL) (SIR = 1.08), a malignant condition not found to be increased after irradiation, and for non-CLL (SIR = 1.09). The risk of leukaemia did not vary by sex, age, time, or radiation dose from 131I. One reason for the absence of a radiation effect, other than chance, includes the possible lowering of risk when exposure is protracted over time as occurs with 131I. Excess leukaemia risks of more than 25% could thus be excluded with high assurance in this population of mainly adults. These results should be reassuring to patients exposed to 131I in medical practice and to most individuals exposed to the fall-out from the Chernobyl accident.


Efficacy of traditional Chinese herbal therapy in adult atopic dermatitis. There has been considerable interest in traditional Chinese herbal therapy (TCHT) as a new treatment for atopic dermatitis. To establish the efficacy and safety of this treatment, a daily decoction of a formula containing ten herbs that has been found to be beneficial in open studies was tested in a double-blind placebo-controlled study. 40 adult patients with longstanding, refractory, widespread, atopic dermatitis were randomised into two groups to receive 2 months' treatment of either the active formulation of herbs (TCHT) or placebo herbs, followed by a crossover to the other treatment after a 4-week washout period. The main outcome measures were extent and severity of erythema and surface damage as judged by standardised body scores. The patients' own assessments of the overall response to treatment were also sought. The geometric mean score for erythema at the end of active treatment was 12.6 (95% confidence interval [CI] 5.9 to 22.0) and at the end of the placebo phase was 113 (65 to 180). The geometric mean score for surface damage was 11.3 (5.8 to 21.8) and 111.0 (68 to 182), respectively. The 95% CI for the mean geometric ratio for the two values with active treatment was 0.04 to 0.22 for erythema (p less than 0.0005) and 0.04 to 0.27 for surface damage (p less than 0.0005). Of the 31 patients who completed the study and expressed a preference, 20 preferred that phase of the trial in which they received TCHT whereas 4 patients preferred placebo (p less than 0.02). There was a subjective improvement in itching (p less than 0.001) and sleep (p less than 0.078) during the TCHT treatment phase. No side-effects were reported by the patients although many commented on the unpalatability of the decoction. TCHT seems to benefit patients with atopic dermatitis. Palatability of the treatment needs to be improved and its safety assured.


Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Intracytoplasmic sperm injection (ICSI) is a promising assisted-fertilisation technique that may benefit women who have not become pregnant by in-vitro fertilisation (IVF) or subzonal insemination (SUZI) of oocytes. We have used ICSI to treat couples with infertility because of severely impaired sperm characteristics, and in whom IVF and SUZI had failed. Direct injection of a single spermatozoon into the ooplasm was done in 47 metaphase-II oocytes: 38 oocytes remained intact after injection, 31 became fertilised, and 15 embryos were replaced in utero. Four pregnancies occurred after eight treatment cycles--two singleton and one twin pregnancy, and a preclinical abortion. Two healthy boys have been delivered from the singleton pregnancies and a healthy boy and girl from the twin pregnancy.


Waterborne transmission of epidemic cholera in Trujillo, Peru: lessons for a continent at risk. The epidemic of cholera that began in Peru in January, 1991, marked the first such epidemic in South America this century. Subsequently, over 533,000 cases and 4700 deaths have been reported from nineteen countries in that hemisphere. We investigated the epidemic in Trujillo, the second largest city in Peru. Trujillo's water supply was unchlorinated and water contamination was common. Suspect cholera cases were defined as persons presenting to a health facility with acute diarrhoea between Feb 1, and March 31, 1991. We studied a cohort of 150 patients who had been admitted to hospital and conducted a matched case-control study with 46 cases and 65 symptom-free and serologically uninfected controls; we also carried out a water quality study. By March 31, 1991, 16,400 cases of suspected cholera (attack rate 2.6%), 6673 hospital admissions, and 71 deaths (case-fatality rate 0.4%) had been reported in the province of Trujillo. 79% of stool cultures of patients with diarrhoea presenting to a single hospital yielded Vibrio cholerae O1. In the case-control study, drinking unboiled water (odds ratio [OR] 3.1, 95% confidence interval [CI] 1.3-7.3), drinking water from a household water storage container in which hands had been introduced into the water (4.2, 1.2-14.9), and going to a fiesta (social event) (3.6, 1.1-11.1) were associated with illness. The water quality study showed progressive contamination during distribution and storage in the home: faecal coliform counts were highest in water from household storage containers and lowest in city well water. V cholerae O1, biotype El Tor, serotype Inaba, was isolated from three city water samples. Cholera control measures in Trujillo should focus on treatment of water and prevention of contamination during distribution and in the home. Trujillo's water and sanitation problems are common in South America; similar control measures are needed throughout the continent to prevent spread of epidemic cholera.


Evaluation of endoscopy for early detection of gastric-stump cancer. The value of endoscopic surveillance of postgastrectomy patients for the early detection of gastric-stump cancer is controversial. Using data from an Amsterdam postgastrectomy cohort of 2633 patients, we have done a retrospective analysis of the effect on mortality from gastric cancer of endoscopic surveillance in these patients. Between 1976 and 1982, 504 symptom-free patients from the Amsterdam cohort participated in an endoscopic surveillance programme. All patients were followed up until 1988. Relative to the general Dutch population, risk of death from gastric cancer was less among patients who took part in surveillance than among those who did not participate. However, differences were small and similar differences existed for risk of death from lung and colorectal cancers, suggesting the presence of selection. Our study seems to confirm that large-scale surveillance of postgastrectomy patients is not justified.


Aortic stenosis and bleeding gastrointestinal angiodysplasia: is acquired von Willebrand's disease the link? An association between aortic stenosis and haemorrhage from gastrointestinal angiodysplasia has been recognised for many years, but no explanation for this link has been found. Remarkably, aortic valve replacement, rather than bowel resection, corrects the bleeding. Aortic stenosis can be complicated by acquired von Willebrand's disease type IIA (vWD-IIA), which is corrected after valve replacement, and gastrointestinal angiodysplasia is a common site of bleeding in older patients with acquired or congenital vWD. Could the stenotic aortic valve lead to an acquired, reversible deficiency of the largest multimers of plasma von Willebrand factor (equivalent to vWD-IIA) and thus explain the association with gastrointestinal haemorrhage?


Double-blind study of selective decontamination of the digestive tract in intensive care. Selective decontamination of the digestive tract (SDD), by means of non-absorbable antibiotics, to prevent infection in intensive-care units (ICUs) remains controversial; there is evidence that the regimen reduces the incidence of secondary infection, but no convincing reduction in morbidity or mortality has been shown and the costs and effect on microbial resistance patterns need further study. In a double-blind, placebo-controlled trial, we have tried to find out whether SDD should be used routinely in all ICU patients at high risk of secondary infection. All patients admitted to the ICU who were thought likely to stay in the unit for at least 5 days and to need intubation for longer than 48 h were enrolled and randomly allocated to groups receiving placebo or SDD (amphotericin, colistin, and tobramycin applied to the oropharynx and enterally); all patients received intravenous cefotaxime for 72 h. Of 322 patients randomised, 83 were withdrawn (80 ICU stay or duration of intubation too short, 3 protocol violations). 239 medical, trauma, and surgical patients completed the trial period (114 SDD, 125 placebo). There were no differences between SDD and placebo groups in incidence of infection (30 [26%] vs 43 [34%] patients; p = 0.22), duration of ICU stay (mean 16.2 [14.3] vs 16.8 [12.3] days), hospital stay (29.9 [SD 25.0] vs 31.9 [22.2] days), or mortality (21 [18%] vs 21 [17%]). SDD substantially increased the costs of intensive care. Mechanisms other than bacterial colonisation of the gut may bring about substantial numbers of secondary infections in ICUs. Routine use of SDD in multidisciplinary ICUs cannot be recommended.


Long-term thyroxine treatment and bone mineral density. Studies of the effect of thyroxine replacement therapy on bone mineral density have given conflicting results; the reductions in bone mass reported by some have prompted recommendations that prescribed doses of thyroxine should be reduced. We have examined the effect of long-term thyroxine treatment in a large homogeneous group of patients; all had undergone thyroidectomy for differentiated thyroid cancer but had no history of other thyroid disorders. The 49 patients were matched with controls for age, sex, menopausal status, body mass index, smoking history, and calcium intake score; in all subjects bone mineral density at several femoral and vertebral sites was measured by dual-energy X-ray absorptiometry. Despite long-term thyroxine therapy (mean duration 7.9 [range 1-19] years) at doses (mean 191 [SD 50] micrograms/day) that resulted in higher serum thyroxine and lower serum thyrotropin concentrations than in the controls, the patients showed no evidence of lower bone mineral density than the controls at any site. Nor was bone mineral density correlated with dose, duration of therapy, or cumulative intake, or with tests of thyroid function. There was a decrease in bone density with age in both groups. We suggest that thyroxine alone does not have a significant effect on bone mineral density and hence on risk of osteoporotic fractures.


Neuroendocrine carcinoma in a patient with hairy cell leukemia: a case report. Second malignancies are common in hairy cell leukemia. We report a case of a neuroendocrine carcinoma arising in a patient who had been diagnosed with hairy cell leukemia 6 years earlier. This case is the first report of these two tumors' occurring together. The pathogenetic basis for the presence of these two uncommon tumors in our patient is discussed.


Ammonia-induced alterations in the metabolism of glutamate and aspartate in neuronal perikarya and synaptosomes of rat cerebellum. The effect of subacute and acute doses of ammonium acetate was studied on the production of 14CO2 from 14C-labeled glutamate and aspartate by neuronal perikarya and synaptosomes isolated from rat cerebellum. Studies with inhibitors for aminotransferases (aminooxy acetic acid) and glutamate dehydrogenase (glutamic acid diethyl ester) indicated that transamination reactions play a major role in this process. There was a suppression in this process in hyperammonemic states. Activities of the enzymes, aspartate aminotransferase, alanine aminotransferase, glutamate dehydrogenase and glutaminase were decreased in both preparations in hyperammonemic states. Activity of glutamine synthetase was unaltered.


ATP mediates fast synaptic transmission in mammalian neurons. In addition to its diverse functions inside cells, ATP can act at several types of cell-surface receptor. One of these (P2X-purinoceptor) is believed to be a ligand-gated cation channel. The presence of P2X receptors on autonomic, sensory and central neurons suggests that ATP might be released to act as a fast excitatory synaptic transmitter. Here we record excitatory synaptic potentials and currents from cultured coeliac ganglion neurons which are mimicked by ATP, blocked by the P2-purinoceptor antagonist suramin, desensitized by alpha,beta-methylene-ATP and unaffected by antagonists acting at nicotine, 5-hydroxytryptamine, N-methyl-D-aspartate (NMDA), non-NMDA glutamate, gamma-aminobutyric acid (GABA), noradrenaline or adenosine receptors. We conclude that ATP is the neurotransmitter at this neuroneuronal synapse.