[The preventive effect of HRT on postmenopausal bone loss]. .625mg/day of conjugated estrogen was administered to women after menopause for five years to investigate the preventive effect of HRT on postmenopausal bone loss. Bone mineral content of subjects before and after HRT was measured by the MD (microdensitometry) method. Subjects were: Group I: Those given HRT for the first time (52 cases); Group II: Those who had had HRT for more than 5 years and then were given another 5 years treatment (34 cases); Those with no HRT (26 cases). In the controls, the average level of sigma GS/D decreased significantly (p less than 0.005) from 2.465 to 2.209 in 5 years, while the average sigma GS/D level increased significantly (p less than 0.005) from 2.462 to 2.532 in Group I, and from 2.536 to 2.539 in Group II. Average bone mineral content after 5 years shown by the % based on the initial bone mineral content as 100% decreased to 90.72% (9.28% decrease) in the controls, but increased to 102.91% in Group I, and 100.26% in Group II. The increase ratios were significantly (p less than 0.005) different in Groups I and II. Thus, HRT was seen to be very effective in preventing bone loss after menopause. The effect in Group II was weaker than that in Group I, but much stronger than in the controls.


Unilateral diaphragmatic paralysis: a matter for concern. A dilemma often faces the clinician who is asked to evaluate unilateral elevation of a hemidiaphragm that is identified on routine chest roentgenogram. The possibilities include neoplasm, infection, neuromuscular disease, trauma, or benign etiologies. We present an asymptomatic patient with this finding to provide some guidelines for the nature and extent of further investigation of unilateral diaphragmatic paralysis.


Synaptic vesicle increase correlated to potentiation of transmission at the synapse of the cat superior cervical ganglion in vivo. Changes in the pattern, number and size of synaptic vesicles during transmitter release were examined in the synapses of the cat superior cervical ganglion (SCG) in vivo in relation to alteration in the amplitude of postganglionic compound action potential (PGP). Stimulation of the preganglionic nerve fibers at 10 Hz caused an increase in the mean amplitude of PGP. It became augmented by approximately 30% compared to control 10-30 sec after starting the stimulation, and then gradually declined to reach a plateau after 4-6 min. This level, about 20% higher in value than control, was sustained until the end of 30 min of stimulation. The nerve terminal was divided into two areas to examine topographically the numerical changes in synaptic vesicles (SVs): zone I on the presynaptic membrane encircled with a diameter equivalent to the active zone length, and zone II occupying the remaining area outside zone I. The synaptic vesicle density in zone I (vesicle number/microns 2) was 96.9 +/- 4.8 (mean +/- S.E.M.) in the unstimulated control ganglia and 128.8 +/- 9.4 (mean +/- S.E.M.) in the ganglia stimulated for 10-30 sec, which was 30% higher in value than control. Then, it decreased slightly reaching a plateau, 20% higher in value than control. The diameter distribution of the SVs showed that their diameters in zone I (56.6 nm mean) were larger than those (51.6 nm mean) in zone II, and that prolonged stimulation induced smaller vesicles in both areas. The results showed that the increase in SV number in zone I correlated well with the elevation of PGP.(ABSTRACT TRUNCATED AT 250 WORDS)


Hydrogen peroxide-induced DNA damages in vitro revealed by electron microscopy. A plasmid pSV2neo (5.6 kb) which is a recombinant SV40-based vector was subjected to hydrogen peroxide treatment in presence of iron in vitro (Fenton reaction). Transmission electron microscopy revealed that H2O2 through the Fenton reaction introduced different types of damages in DNA. Besides the expected nicked circular and linear molecules, single-stranded loops and plasmid multimers could be characterized. These observations on DNA damages might be helpful for better understanding of the effect of hydrogen peroxide on living cells in vivo.


Prevalence of human papillomavirus infections among heterosexual men and women with multiple sexual partners. A prospective study of 65 men and 111 women with multiple heterosexual partners was designed to assess the prevalence and potential risk factors of genital human papillomavirus (HPV) infections. In addition, the HPV reservoir in genital, rectal, and oral mucosa was examined. The specimens for the detection of HPV DNA were taken from different sites such as the urethra and coronal sulcus (men), cervix and labia minora (women), anus, rectum, tongue, and buccal mucosa (both men and women). Women underwent speculum examination and colposcopic evaluation of the anogenital region, and a smear for routine cytological classification was also taken. In men, the anogenital region was examined clinically and colposcopically. The polymerase chain reaction (PCR) was used for the detection of HPV types 6/11, 16, 18, and 33. A high prevalence of HPV infection at one or more sites was detected, in 32% of the male and in 23% of the female participants. Seventeen percent of the male distal urethral specimens were positive for HPV DNA. From the female cervical specimens 14% were found positive. Ten proctal specimens (five men and five women) were positive for HPV DNA without any discernible lesion. The persons from whom these samples were taken denied anal insertive intercourse. No oral manifestation of HPV infection was detected. In both men and women a difference between HPV DNA-positive and -negative persons was not found in relation to known risk factors associated HPV infection.(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of HPV-16 in cell lines and cervical lavage specimens by a polymerase chain reaction-enzyme immunoassay assay. A gene amplification method that combines the polymerase chain reaction with detection of amplified DNA in a solution hybridization/enzyme immunoassay (PCR-EIA) was developed for HPV-16 DNA. Samples were amplified with primers for the E7-E1 region of HPV-16. Amplified DNA products were identified and quantitated by hybridization in solution with a biotinylated RNA probe. Labeled DNA/RNA hybrids were measured semiquantitatively in an enzyme immunoassay using solid phase anti-biotin antibody and liquid phase B-d-galactosidase labeled monoclonal antibody against DNA-RNA hybrids. Enzyme bound to the solid phase was quantitated with a fluorogenic substrate. The assay was linear over 2 log10 dilutions of SiHa cells and the detection limit was three copies of HPV-16 genome. The sensitivity of PCR-EIA for detection of PCR amplified products compared favorably with slot and Southern blots using a 32P-labeled RNA probe. The assay was used to assess HPV-16 infection of uterine cervix in women attending a clinic for sexually transmitted diseases. Twenty-one of the 81 specimens (25.9%), obtained by cervicovaginal lavage, were positive for HPV-16 by PCR-EIA. The assay provides a convenient means to objectively measure HPV DNA amplified with PCR.


Detection of EBV DNA in post-nasal space biopsy tissue from asymptomatic EBV-seropositive individuals. The association between EBV and nasopharyngeal carcinoma (NPC) has been well documented although the precise role of the virus in the genesis of the tumour is not understood. We undertook this study to examine the prevalence of EBV infection in nasopharyngeal tissue obtained from 33 healthy individuals not considered to be at risk of developing NPC. Using polymerase chain amplification (PCR) and in situ hybridization we have identified EBV DNA in 70% (23/33) of the tissues examined. Our data demonstrate that EBV is present at the site of tumour development in the low-risk population and by inference that the virus is also present before the onset of disease in the high-risk group. This survey supports the concept of NPC pathogenesis as a multifactorial process.


Detection of multiple Epstein-Barr viral strains in allogeneic bone marrow transplant recipients. We have previously shown in 3 allogeneic bone-marrow transplant (BMT) recipients that complete replacement of recipient marrow was associated with the elimination of the pretransplant Epstein-Barr virus (EBV) strain of the recipient. To study the kinetics of EBV elimination and reinfection in more detail, we have performed a longitudinal study of BMT recipients combining serology, virus isolation from mouthwashes and peripheral blood, and EBV strain characterization. Oropharyngeal EBV excretion was found to persist after the cytoreductive therapy prior to BMT, whereas EBV-carrying cells in the blood were detected only after 5 weeks following BMT. During the first month post-BMT, 2 different EBV strains could be isolated from sequential mouth-washes of 3 patients. The initial strains were found to persist up to 7, 21, and 29 days post-BMT, whereas the subsequent strains appeared at 21, 42, and 34 days post-BMT, respectively. Thus, the original EBV strain may persist only for a limited time after BMT, and the oropharyngeal epithelium may be reinfected by a new EBV strain from the blood within 3 weeks. With respect to the coexistence of multiple EBV strains, 2 patterns were evident. From the day 62 mouthwash of 1 patient, 1 Type A and 1 Type B strain were isolated. From the day 180 mouthwash of a second patient, a dominant Type A strain was recovered, together with 6 "variant" strains that differed from each other by only a single EBNA protein (EBNA 1). This pattern may be explained by viral recombinations during replication, which may form the basis for the vast polymorphism of EBV observed in unrelated individuals.


Infections of the cervix uteri with human papillomavirus and Chlamydia trachomatis. Apart from infection with human papillomavirus (HPV), other microorganisms may be involved in the development of cervical neoplasia. To study concomitant infections with HPV and Chlamydia trachomatis, cervical specimens from 4 groups of women were examined for the presence of these microorganisms by the polymerase chain reaction. The first group consisted of 143 consecutive samples from women with no cytological abnormalities who participated in a triennial screening program to prevent cervical cancer. In this group 2 samples were found positive for HPV and 2 additional samples were found positive for C. trachomatis. In the second group of 46 cytologically abnormal smears, HPV DNA was detected in 71.7% of the samples and C. tra chomatis in 4.3%. In a third group of 94 histological abnormal biopsies, the HPV prevalence ranged from 15% in mild dysplastic lesions up to 92% in invasive cervical carcinomas. Only 2 biopsies of this group (2.1%) were found positive for C. trachomatis. Finally, a group of cervical scrapes was obtained from women attending a clinic for sexually transmitted diseases. In 52 samples positive for C. trachomatis and 60 samples negative for C. trachomatis, no significant (P = 0.57) difference in the frequency of HPV infections was found (11.5% and 8.3%, respectively). The data show that in these study groups HPV and C. trachomatis are independently occurring agents.


Efficacy of (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine and 9-(1,3-dihydroxy-2-propoxymethyl)guanine for the treatment of murine cytomegalovirus infection in severe combined immunodeficiency mice. Mice with severe combined immunodeficiency (SCID) inoculated intraperitoneally with murine cytomegalovirus (MCMV) develop a wasting syndrome at 3-4 days and die at 6-9 days after the infection. 9-(1,3-Dihydroxy-2-propoxymethyl)guanine (DHPG, ganciclovir) and (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC) were compared for their efficacy against MCMV-induced disease and mortality in SCID mice. Under all treatment conditions, i.e., administration of the test compounds for 5 consecutive days starting on the day of infection (day 0), for 5 consecutive days starting on day 4 after the infection, 2 periods of 3 consecutive days starting on day 0 and day 9 after infection, or as a single dose on day 3 before infection, HPMPC proved far superior to ganciclovir in delaying the onset of the disease and increasing the lifespan of the MCMV-infected mice.


Seroepidemiology of cytomegalovirus infection among children between the ages of 4 and 12 years in Taiwan. To determine risk factors responsible for primary cytomegalovirus (CMV) infection in Taiwan, samples of blood for antibody to CMV were obtained from 362 children aged 4 to 12 years: 58% were found to be positive for anti-CMV IgG antibody. Logistic regression analysis showed that seropositivity correlated with age, method of delivery, duration of breast feeding, and younger age of mother. Neither socioeconomic status nor crowded living conditions showed significant correlation with CMV seropositivity. Primary CMV infection in Taiwan appears to be less related to socioeconomic status shown in western countries, and, since the majority of pregnant women were seropositive in Taiwan, two of the major sources of primary CMV transmission are infected breast milk and the infected genital tract.


Detection of enteroviruses in endomyocardial biopsy by molecular approach. Enteroviruses are considered to be the most common agents implicated in myocarditis and cardiomyopathy. Recent studies have suggested persistent enterovirus infection in chronic disease showing the presence of enteroviral RNA in the myocardium. We used gene amplification by PCR which can demonstrate directly the presence of enteroviral sequences in endomyocardial biopsies. The primers were chosen in the 5' non-coding region of the genome representing highly conserved sequences among enteroviruses and therefore allowed the amplification of the majority of enteroviruses. The hybridization of the amplified products was effected with specific general riboprobe derived from 5' non-coding sequences internal of the amplified fragments. The results include 105 patients distributed in 6 groups: 45 idiopathic dilated cardiomyopathies with 66.7%, 17 alcoholic cardiomyopathies with 52.9%, 10 myocarditis with 30%, 5 multifactorial cardiomyopathies with 40%, 5 patients with immunosuppressive therapy with 100%, and 23 control group without viral etiology with 39.1% positive samples. The study suggested a positive link between viral infection and cardiomyopathies, but did not allow a direct relation between enterovirus infection and idiopathic dilated cardiomyopathy to be established.


Comparison of cervical cytology and the polymerase chain reaction for HPV 16 to identify women with cervical disease in a general practice population. A comparison of the ability of cervical cytology and the polymerase chain reaction (PCR) for human papilloma virus type 16 (HPV 16) to identify women with cervical disease has been performed in a general practice population of 249 women, none of whom were believed to have current cervical disease prior to examination. Within this population, 29 women were found by colposcopy and subsequent histopathology to have evidence of cervical disease [5 with cervical intraepithelial neoplasia (CIN) 3; 8 with CIN 2; and 16 with CIN 1]. The prevalence of HPV 16 in this population was 18.9% (CIN 3, 80%; CIN 2, 50%, CIN 1, 12.5%, normal, 16.8%). Women with severe disease (CIN 2 and CIN 3) had a significantly higher incidence of HPV 16 DNA than those with mild cervical disease (CIN 1) or no cervical abnormality (P = 0.001). There was no significant difference in the ability of either PCR for HPV 16 or cytology to identify women with cervical disease. The combination of screening by cytology and the presence of HPV 16 DNA resulted in the identification of a higher proportion of the women with disease, but this observation did not reach statistical significance. Although the failure to detect disease by the two screening methods was similar, HPV 16 DNA positivity was associated with a higher false-positive rate for disease detection than cytology (P less than 0.03). The PCR assay for detecting HPV 16 in this investigation was shown to have a false-positive rate of 2.4% and a false-negative rate of 10.4%. The prospect of screening women for cervical disease using PCR for HPV 16 is discussed.


Metal complexes of phenobarbituric acid. Chelating behavior of the phenobarbiturate ring. Anticonvulsant properties of the K2[Cu(N-methylphenobarbiturato)]4.8H2O. Na2Ni(phenobarbiturato)4.3H2O, Na2Ni3(phenobarbiturato)2(OH)6.4H2O, and NaZn(phenobarbiturato)2(OH).H2O derivatives were prepared from Ni(II) and Zn(II) and phenobarbital. The Na2Ni(phenobarbiturato)4.3H2O complex is diamagnetic and isostructural with the complex previously reported, Na2Cu(phenobarbiturato)4, suggesting a square-planar environment around the Ni(II) ion. The DMF solutions of this complex show the existence of two species. The EPR spectra of the Cu(II) doped complex show the hyperfine and superhyperfine structures. The covalence parameters Alpha2, Beta2, and Delta2 show a strong bonding in the equatorial plane and suggests the formation of a [CuN4] chromophore. The anticonvulsant properties of the K2Cu(N-methylphenobarbiturato)4.8H2O are reported.


Incorporating feminism into the graduate curriculum. Feminist theory and methods are vital catalysts to teaching/learning in graduate education. This article describes how feminism was integrated into a graduate course, specifically regarding nursing activism and health policy. Major themes include a learning model emphasizing the student's individual strands of life experience, reconstruction of nursing history, and small-group process. Class assignments, student responses, and the implications of feminist perspectives for advanced professional practice are discussed.


Alternatives in nursing education funding. Contracting between private educational institutions and state governments or agencies is developing as a cost-effective method for meeting nursing education demands in an era of scarce resources. This article explores relevant literature and current status of this type of contracting.


Effect of pregraduate preceptorship experience on development of adaptive competencies of baccalaureate nursing students. Kolb's (1984) theory of experiential learning was used as a framework to study 50 baccalaureate nursing students' perceptions of the contributions of a senior preceptorship experience to the development of adaptive competencies. Nursing learning environments were thought to contribute most to divergent and convergent competencies, reflecting the importance of both people-oriented and scientific skills in nursing. The preceptorship had a significant impact on most learning competencies. Significant increases occurred in competencies considered not important for nursing by students prior to the experience, e.g., assimilative competencies such as testing theories and ideas, and accommodative competencies such as leading and influencing others. These results support the notion of further research of nursing learning environments from the experiential learning perspective.


New nurse faculty: core competencies for role development. This study originated from the need to define the role competencies for a beginning nurse educator. A survey of full-time faculty and heads of university and community college nursing programs in Ontario showed a high degree of agreement between the two sets of faculty for teacher role, practice, service, personal and professional growth competencies. Significant differences were noted on competencies involving student evaluation, facilitating student's practice, acting as student's advocate, and research. Agreement was noted on the problems created by role ambiguity and role incompetence, particularly for new faculty. This suggests the need for educational preparation and socialization of nurse faculty.


Nursing faculty and students' attitudes regarding HIV. A multidisciplinary group of health professional educators examined the faculty and student attitudes related to AIDS in undergraduate and graduate nursing programs and in a dental hygiene program. Results indicated consistent differences in attitudes toward homosexuality and intravenous drug users, AIDS-phobia, AIDS-related work stress, and willingness to work with HIV, homosexual, or intravenous-drug-using patients among faculty, undergraduate, and graduate nursing students, and certificate-level dental hygiene students. Faculty and master's-level nursing students consistently indicated the most positive attitudes and behavioral intentions. A one-year follow-up of a sample of undergraduate students revealed little change in these attitudes or behavioral intentions. Implications of these findings for nursing educators are discussed.


Scholarship and socialization: reflections on the first year of doctoral study. The first year of study in our PhD program has both broadened and intensified our knowledge. We have come to expect ourselves to attain better understandings of any given issue or topic within its attendant context and within the larger context of our profession. We have begun to appreciate and to strive for the clarity, thoroughness, and precision of thought and analysis that typify the nurse scholar. We are beginning to recognize who the nurse scholar is, why we want to emulate this person, and how we can become this person. As members of the fourth generation, we are now beginning, through our doctoral program, to live the vision of scholars who preceded us. To fulfill this vision, we understand that learning is a lifelong process that does not end with achievement of the doctorate or with postdoctoral study, but that it is continually evolving. We recognize the crucial role of research in nursing, but temper this recognition with the knowledge that research must progress in a planned manner dovetailing with existing research lest it fail to strengthen the web of nursing knowledge.


Is there a place for Phil Donahue in the classroom? The novel approach of using the Phil Donahue television talk show model in the classroom was successful for us. The model worked well with a panel of guests providing content and the faculty member moderating and directing the discussion by targeting questions and limiting and/or expanding responses as dictated by the objectives for the class. We hypothesize that the Johnny Carson format, where a single guest speaker is interviewed by a faculty member, also would produce excellent results. We encourage colleagues to be creative and experiment in the classroom.


Incomplete digestion of legume starches in rats: a study of precooked flours containing retrograded and physically inaccessible starch fractions. The digestibility of starch in precooked flours from green coat lentils (Lens culinaris Medik) and red kidney beans (Phaseolus vulgaris L.) was investigated by balance experiments using rats treated with antibiotics to suppress hind-gut fermentation. The legume preparations were rich in intact cells filled with denaturated starch and contained retrograded amylose. Between 8% (beans) and 11% (lentils) of the total starch ingested appeared in the feces, indicating a relatively low starch digestibility. Red bean flours of two different particle sizes were similarly digested. Sixty percent of the fecal starch in the bean-fed animals and 70% in the lentil-fed group was retrograded amylose. The in vitro indigestible starch content of the flours was evaluated with three different methods that gave rather different values. The retrograded amylose fraction, measured after alkaline treatment of a dietary fiber residue obtained by enzymic digestion, was quantitatively recovered in the feces. None of the procedures gave accurate estimates of the total in vivo indigestible starch. Fecal excretion of starch in rats not treated with antibiotics indicated that the indigestible starch in lentils was less susceptible to fermentation than that in the red bean preparations.


Short-chain fatty acid production and fiber degradation by human colonic bacteria: effects of substrate and cell wall fractionation procedures. Three dietary fiber sources (corn fiber, oat bran, wheat bran) were analyzed for chemical composition and potential fermentation by human colonic bacteria in vitro. Total dietary fiber (TDF) concentration of substrates was 64.3, 11.1 and 50.4 g/100 g dry matter for corn fiber, oat bran and wheat bran, respectively. Original material (ORIG), TDF fractions and simulated (SIM) cell wall fractions (produced by combining cellulose, hemicelluloses and pectic substances in proportions they represented in the cell wall) from each substrate were fermented in vitro for 6, 12, 18, 24 or 48 h using inoculum prepared from freshly voided feces from each of three human volunteers. Substrate dry matter remaining after 48 h of fermentation was 87.8, 39.8 and 73.5% for TDF fractions of corn fiber, oat bran and wheat bran, respectively. Disappearance of ORIG fractions was considerably greater than that of TDF due to fermentation of nonfibrous material. Disruption of cell wall structure during isolation of polysaccharide fractions allowed for dramatically increased fermentability of SIM relative to TDF. Averaged across all treatments, production of the short-chain fatty acids, acetate, propionate and butyrate, occurred in the molar ratio 63:21:16; however, profiles of short-chain fatty acids produced were influenced by both treatment and inoculum source. Extent of substrate fermentation varied among inoculum donors, implying that colonic microbial activities differ among individuals. Potential colonic fermentability of fiber sources was influenced by substrate, method of fiber preparation and inoculum source.


Perinatal protein deprivation enhances the anticonflict effect measured after chronic ethanol administration in adult rats. The anticonflict effects of ethanol, diazepam and pentobarbital were evaluated in adult rats fed a low protein diet during the perinatal period in the plus-maze test, after single injections and following chronic ethanol administration (1 g.kg-1.d-1 for 30 d). Reactivity to the anticonflict effect of these drugs was similar in control and protein-deprived rats after acute treatment. After chronic ethanol administration, control rats showed tolerance to ethanol and cross-tolerance (i.e., lower reactivity) to the anxiolytic effect of diazepam and pentobarbital. Conversely, protein-deprived rats showed greater reactivity to ethanol and lack of cross-tolerance to diazepam and pentobarbital following chronic ethanol treatment. A significantly greater density of cortical gamma-aminobutyric acid receptors subtype A (GABA-A) was detected in protein-deprived rats after chronic ethanol administration compared with the density after chronic saline treatment, whereas no differences were observed in nourished controls. This suggests that the greater anxiolytic activity detected in protein-deprived rats may correlate with higher GABA-A receptor density.


Soluble dietary fiber and cholesterol influence in vivo hepatic and intestinal cholesterol biosynthesis in rats. Ninety-six male Sprague-Dawley rats were randomly assigned to eight dietary treatments. Rats were fed, with ad libitum access, diets containing 10% dietary fiber as cellulose (control), pectin, psyllium or oat bran with or without 0.3% added cholesterol for 3 wk. Among cholesterol-fed rats, liver total cholesterol was significantly lower in rats fed diets supplemented with either pectin or psyllium compared with those fed cellulose. In contrast, rats fed oat bran with cholesterol had significantly higher liver cholesterol concentrations compared with cellulose-fed animals. Liver total lipid concentrations were significantly lower in groups fed pectin and psyllium with or without dietary cholesterol compared with cellulose-fed controls. Pectin feeding with or without dietary cholesterol significantly lowered plasma total cholesterol compared with cellulose feeding. Oat bran had no effect on plasma total cholesterol compared with control diets. Hepatic sterol synthesis was significantly greater for animals fed soluble dietary fibers compared with those fed cellulose, but the effect on intestinal sterol synthesis was less pronounced.


[A basic study on the association of Epstein-Barr virus with nasopharyngeal carcinoma--detection and genotypic analysis of Epstein-Barr virus associated with nasopharyngeal carcinoma in Japanese patients]. Epstein-Barr virus (EBV) is known to be associated with two human malignant diseases, nasopharyngeal carcinoma (NPC) and endemic Burkitt's lymphoma. In this study, the genotypes of EBV in tissues from 13 NPC patients in Japan were analyzed by Southern blot hybridization using EBV genome fragment probes. Ten of the cases contained reiterated sequences (EBV BamHI-H, -B1*, -K fragments), showing that only one genotype was detected in each specimen. One of these had a BamHI fragment containing a fused sequence of BamHI-Y and -H. In all except one case, a single-sized EBV-joined terminus was observed in each NPC specimen, implying evolution of the carcinoma from a single EBV-infected cell. One metastatic lymph node (which was not a primary epipharyngeal tumor) contained EBV with heterogeneous termini suggesting production of linear virion DNA. The type C variant resulting from loss of a BamHI site between the BamHI-W1* and -I1* regions was observed in 7 of the 10 cases, and the other 3 cases had a separated BamHI-I1* fragment. As reported by Lung et al. (Virology, 177: 44-53, 1990), the type C variant appears to be dominant among Japanese strains, as it is in Southern China. In contrast to their findings, however, the "f" variant with an extra BamHI site in the BamHI-F region which they found to be strongly associated with NPC specimens from Southern China, was detected in only one case. The present study, therefore, did not support the specific association of the "f" variant with NPC in Japanese patients. We conclude that the EBV in NPC tissues exists in variants. Further studies along these lines, could help to explain the epidemiology of EBV.


New hybrids of quipazine and trazodone as selective inhibitors of uptake of 5-hydroxytryptamine. Two new congeners, 4-(chloropropyl)-1-(2-quinolyl)piperazine- and 2-[3-[4-[2-(quinolyl)]-1-piperazinyl]propyl]-1,2,4-triazolo] 4,3-a]pyridin-3(2H)-one, of trazodone were synthesized and found to be potent and selective inhibitors of synaptosomal uptake of 5-hydroxytryptamine [5-HT, serotonin; IC50 = norepinephrine greater than 5 microM, 5-HT = 210-890 nM], with minimal effects in antagonizing (-)-apomorphine-induced climbing behavior and suppression of spontaneous locomotor activity in mice (ED50 greater than 50 mg/kg). The two compounds behaved like atypical antidepressants, since they weakly antagonized reserpine-induced hypothermia. The acute toxicity studies have shown that these compounds were less lethal when compared with imipramine or quipazine. Furthermore, chronic treatments (20 mg/kg, daily for 10 and 21 days) significantly decreased the isoprenaline-induced increase in cyclic AMP in the rat brain cortex, suggesting desensitization of beta-adrenoceptors. These findings point to the effects of these compounds as potential antidepressants dealing with specific serotonergic mechanisms.


Abnormal Papanicolaou smears. Comparison of cytology, colposcopy and cervical swab DNA hybridization. Cervical swab sampling for DNA hybridization was performed on 89 women attending a colposcopy clinic. In situ hybridization studies for human papillomavirus (HPV) types 6/11, 16/18 and 31/33/35 were performed on the specimens. Biopsy specimens were obtained from 65 patients. The cytologic, colposcopic and histologic results were classified into five groups and assigned a score of 0-5, depending upon the diagnosis. An increasing score correlated positively with the presence of HPV, especially types 16/18 and 31/33/35. A comparison was made of the sensitivity, specificity, and positive and negative predictive values for the standard method of colposcopic evaluation versus the standard method combined with DNA cervical swab sampling. No significant changes in the parameters were found when the DNA cervical swab was added.


Electrophilic opioid ligands. Oxygen tethered alpha-methylene-gamma-lactone, acrylate, isothiocyanate, and epoxide derivatives of 6 beta-naltrexol. O6-Ether derivatives of 6 beta-naltrexol in which the ether substituent includes various electrophilic groups have been synthesized in an effort to examine structure-activity requirements at the 6 beta-substituent for receptor affinity and irreversibility of binding in opioid receptor preparations. A series of tethered 6 beta-ethers having terminal epoxides, alpha-methylene-gamma-lactones, and an isothiocyanate group were prepared. The stereochemistry of the alpha-methylene-gamma-lactones was established by convergent synthesis of their reduction products from epoxides of known absolute stereochemistry. In general, the tested compounds showed comparable affinity and selectivity for the receptor subtypes. All were found with high affinity for mu-sites. The terminal epoxide ether diastereomers 8 and 9 were not bound irreversibly in the assay for total opioid receptors. The alpha-methylene-gamma-lactone diastereomers 10 and 11, and their O14-acetyl precursors 20 and 21, respectively, varied in their irreversible effects, but where noted these effects were mu-site selective. Methacrylate ether 7 and isothiocyanate ether 12 were bound irreversibly at both mu- and delta-sites.


Insulin-secreting pancreatic (islet cell) carcinoma in a cat. A functional, insulin-secreting pancreatic (islet cell) carcinoma was diagnosed in a 17-year-old male Siamese cat. Diagnosis was made on the basis of clinical signs (i.e., seizures and stupor) that resolved temporarily after correction of hypoglycemia with feeding or intravenous administration of glucose, the finding of an inappropriately increased serum insulin concentration in the face of hypoglycemia, and prolonged resolution of hypoglycemia after surgical removal of the tumor. Primary islet cell tumor of the pancreas was confirmed by biopsy. The cat died 18 months later, and necropsy revealed metastases to regional lymph nodes and liver. Specimens of the tumor and metastatic lesions both stained positively for insulin.


Ring substituted and other conformationally constrained tyrosine analogues of [D-Pen2,D-Pen5]enkephalin with delta opioid receptor selectivity. The conformationally restricted, cyclic disulfide-containing delta opioid receptor selective enkephalin analogue [D-Pen2,D-Pen5] enkephalin (DPDPE) was modified by 2' (CH3) and 3' (I, OCH3, NO2, NH2) ring substitutions and by beta-methyl conformationally constrained beta-methyltyrosine derivatives in the 1 position. The potency and selectivity of these analogues were evaluated by bioassay in the mouse vas deference (MVD, delta receptor assay) and guinea pig ileum (GPI, mu receptor assay) assays and by radioreceptor binding assays in the rat brain using [3H]CTOP (mu ligand) and [3H][p-ClPhe4]DPDPE (delta ligand). The analogues showed highly variable potencies in the binding assays and in the bioassays. Aromatic ring substituents with positive Hammett constants had decreased potency, while substituents with negative Hammett constraints has increased potency for the opioid receptor. The most potent and most selective compound based on the binding was [2'-MeTyr1]DPDPE (IC50 = 0.89 nM and selectivity ratio 1310 in the binding assays). The 6-hydroxy-2-aminotetralin-2-carboxylic acid-containing analogue, [Hat1]DPDPE, also was highly potent and selective in both assays, demonstrating that significant modifications of tyrosine in enkephalins are possible with maintenance of high potency and delta opioid receptor selectivity. Of the beta-methyl-substituted Tyr1 analogues, [(2S,3R)-beta-MeTyr1]DPDPE was the most potent and the delta receptor selective. The results with substitution of beta-MeTyr or Hat instead of Tyr also demonstrate that topographical modification in a conformationally restricted ligand can significantly modulate both potency and receptor selectivity of peptide ligands that have multiple sites of biological activity.


1,3,6-trisubstituted indoles as peptidoleukotriene antagonists: benefits of a second, polar, pyrrole substituent. 1,6-Substituted and 3,5-substituted indoles and indazoles containing acylamino and N-arylsulfonyl amide appendages are potent antagonists of the peptidoleukotrienes LTD4 and LTE4. A compound from the 3,5-substituted indole series, N-[4-[[5-[[(cyclopentyloxy)carbonyl]amino]-1-methylindol- 3-yl]methyl]-3-methoxybenzoyl]-2-methyl-benzenesulfonamide (ICI 204,219), is undergoing clinical evaluation for asthma. Two new elements of structural diversity were introduced to this series of antagonists. An investigation of pyrrole substituents in the 1,6-substituted indoles demonstrated that substitution at C-2 was detrimental to biological activity, but the incorporation of hydrophilic groups at C-3 was beneficial. The introduction of a propionamide moiety at C-3 enhanced activity by 1 order of magnitude; N-[4-[[6-(cyclopentylacetamido)-3-[2-(N- methylcarbamoyl)ethyl]indol-1-yl]methyl]-3-methoxy- benzoyl]benzenesulfonamide (15c) has a pKB of 10.7 at the LTD4 receptor on guinea pig trachea. Modifications of the acylamino portion of the disubstituted antagonists demonstrated that a transposition of the amide CO and NH atoms was viable. N-Cyclopentylmethyl amides in both the 1,6- and 3,5-disubstituted indole series were 1 order of magnitude less potent than the corresponding cyclopentylacetamides. In both series this potency loss could be regained by the incorporation of a propionamide substituent at either C-3 or N-1, respectively. For example, N-[4-[[6-[N-(cyclopentylmethyl)carbamoyl]-3-[2-(pyrrolidin-1 - methylbenzenesulfonamide (39c) has a pKB of 9.5.


Ovarian cancer family and prophylactic choices. A subject from a family with ovarian cancer who has developed bilateral medullary carcinoma of the breast at the age of 40 is presented. The family is consistent with dominant inheritance of ovarian cancer and 12 female family members at 12.5%, 25%, and 50% risk, including our case, have undergone bilateral prophylactic oophorectomy and been given hormone replacement therapy. Despite the risk of further primary tumours of the breast our patient chose to have treatment with wide excision and radiotherapy. The implications for screening, prophylaxis, and hormone replacement therapy for this family are discussed.


delta-Aminolevulinate couples cycA transcription to changes in heme availability in Rhodobacter sphaeroides. In this paper, the response of the transcriptional control region of the Rhodobacter sphaeroides cytochrome c2 gene, cycA, to intermediates in heme biosynthesis was studied. To determine if cycA transcription was regulated by heme availability, several precursors or analogs of tetrapyrroles were tested. Addition of delta-aminolevulinate (ALA), the first committed intermediate in heme biosynthesis, was shown to inhibit cycA transcription initiation at both the upstream and downstream promoter regions. In addition, an ALA auxotroph, which can grow in the presence of high levels of ALA, showed a 5 to 7-fold reduction in steady-state transcription from cycA::lacZYA operon fusions. To identify genetic elements responsible for negative regulation by ALA, trans-acting mutants with increased expression of cycA were isolated that were resistant to growth inhibition by the heme analog cohemin. These cohemin-resistant mutants (Chr) have elevated levels of several cycA transcripts and they contain cycA transcripts that had not previously been detected in wild-type cells. In addition, cycA transcription in the Chr mutants continues after the addition of ALA. Finally, we found that Chr mutants have increased ALA synthase activity, suggesting that synthesis of cytochrome c2 and ALA synthase are controlled by a common gene product whose activity has been modified in these mutants. A model is presented to explain how changes in tetrapyrrole intermediates could provide an effective signal to control both cycA transcription and ALA synthase synthesis in R. sphaeroides.


Recombination between adenovirus type 12 DNA and a hamster preinsertion sequence in a cell-free system. Patch homologies and fractionation of nuclear extracts. We have previously described a cell-free recombination system derived from hamster cell nuclear extracts in which the in vitro recombination between a hamster preinsertion sequence, the cloned 1768 base-pair p7 fragment, and adenovirus type 12 (Ad12) DNA has been demonstrated. The nuclear extracts have now been subfractionated by gel filtration on a Sephacryl S-300 column. The activity promoting cell-free recombination elutes from the Sephacryl S-300 matrix with the shoulder and not the peak fractions of the absorbancy profile. By using these protein subfractions, in vitro recombinants have been generated between the p7 preinsertion sequence and the 60 to 70 map unit fragment of Ad12 DNA, which has previously shown high recombination frequency. In all of the analyzed recombinants thus produced in vitro, striking patchy homologies have been observed between the p7 and Ad12 junction sequences, and between Ad12 DNA or p7 DNA and pBR322 DNA. The patchy homologies are similar to those found earlier during the analyses of some of the junction sequences in integrated Ad12 genomes in Ad12-induced hamster tumor cell lines. Proteins in the shoulder fractions of the gel-filtration experiment can form specific complexes with double-stranded synthetic oligodeoxyribonucleotides corresponding to several p7 and Ad12 DNA sequences. These sequences participate in the recombination reactions catalyzed by the same column fractions in the shoulder of the absorbancy profile. Such proteins have not been found in the peak fractions. Further work will be required to ascertain that the cell-free recombination system mimics certain elements of the mechanisms of integrative recombination and to purify the cellular components essential for recombination.


X-ray crystal structure of canine myeloperoxidase at 3 A resolution. The three-dimensional structure of the enzyme myeloperoxidase has been determined by X-ray crystallography to 3 A resolution. Two heavy atom derivatives were used to phase an initial multiple isomorphous replacement map that was subsequently improved by solvent flattening and non-crystallographic symmetry averaging. Crystallographic refinement gave a final model with an R-factor of 0.257. The root-mean-square deviations from ideality for bond lengths and angles were 0.011 A and 3.8 degrees. Two, apparently identical, halves of the molecule are related by local dyad and covalently linked by a single disulfide bridge. Each half-molecule consists of two polypeptide chains of 108 and 466 amino acid residues, a heme prosthetic group, a bound calcium ion and at least three sites of asparagine-linked glycosylation. There are six additional intra-chain disulfide bonds, five in the large polypeptide and one in the small. A central core region that includes the heme binding site is composed of five alpha-helices. Regions of the larger polypeptide surrounding this core are organized into locally folded domains in which the secondary structure is predominantly alpha-helical with very little organized beta-sheet. A proximal ligand to the heme iron atom has been identified as histidine 336, which is in turn hydrogen-bonded to asparagine 421. On the distal side of the heme, histidine 95 and arginine 239 are likely to participate directly in the catalytic mechanism, in a manner analogous to the distal histidine and arginine of the non-homologous enzyme cytochrome c peroxidase. The site of the covalent linkage to the heme has been tentatively identified as glutamate 242, although the chemical nature of the link remains uncertain. The calcium binding site has been located in a loop comprising residues 168 to 174 together with aspartate 96. Myeloperoxidase is a member of a family of homologous mammalian peroxidases that includes thyroid peroxidase, eosinophil peroxidase and lactoperoxidase. The heme environment, defined by our model for myeloperoxidase, appears to be highly conserved in these four mammalian peroxidases. Furthermore, the conservation of all 12 cysteine residues involved in the six intra-chain disulfide bonds and the calcium binding loop suggests that the three-dimensional structures of members of this gene family are likely to be quite similar.


Phosphorylation of phospholamban in the intact heart. A study on the physiological role of the Ca(2+)-calmodulin-dependent protein kinase system. The aim of the present study was to further elucidate the physiological role of the calcium-calmodulin (Ca(2+)-Cm)-dependent protein kinase system on phospholamban phosphorylation in the intact functioning heart. The effect of increasing extracellular calcium concentration [Ca]o on phospholamban phosphorylation (PHPL) was studied under different experimental conditions: (a) regular twitches and ryanodine induced-tetani both in the presence and in the absence of 3 x 10(-8) M isoproterenol and (b) Post-stimulation potentiation (PSP), i.e. the potentiation of contractility that follows a period of rapid repetitive stimulation. In the regular twitch, the increase in [Ca]o enhanced contractility both, in the absence and in the presence of beta-stimulation without changing basal or isoproterenol stimulated cAMP levels respectively. This increase in contractility was accompanied by a significant enhancement of PHPL-from 90.6 +/- 16.4 to 216 +/- 35.2 pmols 32Pi/mg protein at 0.25 and 3.85 mM [Ca]o respectively-only when isoproterenol was present. The calmodulin antagonist W-7 significantly decreased the isoproterenol-induced phosphorylation of phospholamban at [Ca]o 1.35 mM. Similar results were obtained under tetanic conditions. When myocardial contractility was enhanced by PSP up to ten-times with respect to the regular twitch, no detectable effect in PHPL was observed. Indirect evidence obtained from skinned rat cardiac trabeculae suggested that the failure of the cAMP-independent mechanisms to phosphorylate phospholamban is not related to a deficient increase in intracellular calcium. The results support the notion that the increase in intracellular calcium induces an increase in PHPL only at high intracellular cAMP levels.


Justification for evaluating new anticancer drugs in selected untreated patients with extensive-stage small-cell lung cancer: an Eastern Cooperative Oncology Group randomized study. Studies have shown that response to a given chemotherapy in previously untreated patients with extensive-stage small-cell lung cancer is superior to that in patients previously treated with other regimens. This finding raises the question of whether it is necessary and ethical to study the effects of new anticancer agents in untreated patients. Such studies appear to be the best test for drug development, but there has been no evaluation of whether survival of untreated patients, whose cancer is sensitive to established drugs, is adversely affected in trials of new drugs. This randomized study of untreated patients with extensive-stage small-cell lung cancer was designed (a) to compare the survival of patients treated with either effective standard chemotherapy or an investigational anticancer drug as initial therapy and (b) to evaluate response rates and toxic effects of such therapies. Eighty-six patients were randomly assigned to receive, as initial therapy, either the standard CAV regimen--cyclophosphamide (1000 mg/m2), doxorubicin (50 mg/m2), and vincristine (1.4 mg/m2) every 3 weeks--or the phase II drug menogaril (200 mg/m2) every 4 weeks. Treatment after induction therapy varied, depending on patient response, but nonresponders and those with disease progression received salvage chemotherapy--etoposide (120 mg/m2 on days 1, 2, and 3) and cisplatin (60 mg/m2 on day 1), repeated every 3 weeks. Of the 43 patients on CAV, 42% responded (eight complete responses and 10 partial responses); 5% of the 43 on menogaril responded (two partial responses) (P = .0001). Twelve (22%) of 54 patients responded to salvage chemotherapy (five complete responses and seven partial responses). Within 3 months from start of treatment, twelve patients died--3 patients in the CAV group and nine patients in the menogaril group (P = .12). The estimated median survival was 37 weeks with menogaril and 45 weeks with CAV (P = .28). At 6 months, survival was 76.7% for the CAV group and 67.4% for the menogaril group. At 12 months, survival rates were 24.4% and 27.9%, respectively. Confidence intervals (95%) for the differences between the proportions surviving in the two groups were -9%-28% at 6 months and -25%-14% at 12 months. Use of CAV resulted in significantly higher occurrence of severe and life-threatening treatment-related complications (P = .002). The confidence intervals for the differences in survival are too wide to conclude that evaluation of a new drug in untreated patients with extensive-stage small-cell lung cancer is or is not harmful. The data do suggest, however, that use of this study design may have no adverse effect on survival.


[Carcinomatous autonomic neuropathy and the autoantibodies in paraneoplastic neuropathy]. Carcinomatous autonomic neuropathy is a syndrome of autonomic dysfunction associated with malignant neoplasm. It is most often associated with small-cell lung cancer. Auto-immune mechanism is suspected to underlie this syndrome. Among the symptoms of autonomic dysfunction, intestinal pseudo-obstruction is most frequently seen. The autonomic dysfunction often precedes the discovery of the cancer. Several autoantibodies have been reported in paraneoplastic neuropathies. Anti-Hu antibody, which was originally reported in paraneoplastic sensory neuronopathy, has also been detected in some cases with autonomic failure. Anticarbohydrate antibody activity has been frequently shown in IgM M-proteins associated with peripheral neuropathies. These include anti-MAG, Po, and sulphated glucuronyl glycolipids in demyelinating neuropathy and anti-GM1 in motor dominant neuropathy or motor neuron disease. Further investigation should reveal more glycoconjugate antigens as targets of possible auto-immune attack in paraneoplastic neuropathies.


Extended radical surgery against gastric cancer: low complication and high survival rates. The present study examines survival benefits for 338 gastric cancer patients from radical surgical treatments, including radical lymphadenectomy of group 3 nodes and combined resection of infiltrated adjacent organs. Curative surgery, with no detectable residual tumor, was possible for 236 patients (69.8%). The radical surgical approach was constant in execution, with 330 of the 338 patients (97.6%) being operated upon by the same surgeon. The remaining patients were operated upon in a similarly radical manner which always included a lymphadenectomy of the N-3 region. Anastomotic leakage was observed in seven patients (3.0%). The postoperative mortality rate was 4.8% for total gastrectomy cases and 3.3% for subtotal gastrectomy cases. The actuarial five-year survival rate for patients receiving curative surgery was 50.0%. These low complication and high survival rates support the concept of extended radical surgery for gastric cancer.


Improvements in hepatocellular carcinoma resection by intraoperative ultrasonography and intermittent hepatic inflow blood occlusion. From September, 1989, to December, 1990 (late period), intraoperative ultrasonography (IOU) and intermittent hepatic inflow blood occlusion were introduced in hepatectomy. Compared with the early period from January, 1983, to August, 1989, the resectability of hepatocellular carcinoma (HCC) increased from 12.1 to 62.1% (P less than 0.0001). More resections on cirrhotic patients (P less than 0.05) and more combined resections with other organs (P less than 0.005) were carried out. Although the operation time was longer (P less than 0.01), less blood loss during surgery and fewer perioperative blood transfusions (P less than 0.001) were found during the late period. Since the rate at which classical resections were performed has reduced (P less than 0.001), postoperative morbidity has also decreased (P less than 0.05). Although the surgical mortality did not differ between the two periods, most deaths in the early period were caused by postoperative hepatic failure which was not found in the late period. Since IOU can clarify the intrahepatic vasculature and identify impalpable and invisible tumors, more precise resections can now be carried out. Intermittent hepatic inflow occlusion reduces blood loss during surgery without increasing risk. We suggest both techniques should be mandatory in hepatectomy for HCC in order for the safety range of resections to be broadened.


Efficacy of nonoxynol 9 contraceptive sponge use in preventing heterosexual acquisition of HIV in Nairobi prostitutes. To determine the efficacy of the nonoxynol 9 contraceptive sponge in preventing sexual acquisition of the human immunodeficiency virus (HIV). Prospective, randomized placebo-controlled trial. Research clinic for prostitutes in Nairobi, Kenya. One hundred thirty-eight HIV-seronegative women were enrolled, of whom 74 were assigned to nonoxynol 9 sponge use and 64 to placebo use. These two groups did not significantly differ with respect to demographic characteristics, sexual practices, or prevalence of genital infections at enrollment, except for a lower number of sex partners per week and a higher initial prevalence of genital ulcers among women assigned to nonoxynol 9 sponge use. Among the 116 women who returned for follow-up, the mean durations of follow-up were 14 and 17 months for the two groups, respectively. HIV seroconversion. Nonoxynol 9 sponge use was associated with an increased frequency of genital ulcers (relative risk [RR], 3.3; P less than .0001) and vulvitis (RR, 3.3; P less than .0001) and a reduced risk of gonococcal cervicitis (RR, 0.4; P less than .0001). Twenty-seven (45%) of 60 women in the nonoxynol 9 sponge group and 20 (36%) of 56 women in the placebo group developed HIV antibodies. The hazard ratio for the association between nonoxynol 9 sponge use and HIV seroconversion was 1.7 (95% confidence interval [CI], 0.9 to 3.0). Using multivariate analysis to control for the presence of genital ulcers at enrollment, the adjusted hazard ratio for the association between nonoxynol 9 sponge use and seroconversion was 1.6 (95% CI, 0.8 to 2.8). Genital ulcers and vulvitis occurred with increased frequency in nonoxynol 9 sponge users. We were unable to demonstrate that nonoxynol 9 sponge use was effective in reducing the risk of HIV infection among highly exposed women.


Radiotherapy for bone metastases of hepatocellular carcinoma. Radiotherapeutic effects of 42 bone metastases in 26 hepatocellular carcinoma patients were retrospectively analyzed. Pain relief was observed in 79% of 37 bone metastases treated (29/37). Antitumor effects and improvement of paralysis were also observed in 88% (8/9) and 17% (1/6), respectively. The toxicities experienced were tolerable. Radiotherapy was an effective means of palliation in the treatment of hepatocellular carcinoma bone metastases.


Interactions of lung cancer cells with the human mesothelial cell monolayer: an in vitro model for cancer invasion. An in vitro monolayer model for cancer invasion was developed, comprising cultured layers of human pleura-derived mesothelial cells (PM cells). The cells were isolated from normal human pleura at autopsy and cultured in RPMI-1640 supplemented with 10% fetal calf serum; they were polygonal in shape, forming a pavement-like structure, and preserved the morphologic characteristics of mesothelial cells. When small cell lung cancer cells (OC-10 cells) were seeded on to the monolayer of PM cells, they attached themselves to the monolayer, invaded it and formed flattened cancer cell nests beneath it. By counting the number of cancer cells penetrated, the in vitro invasiveness (invasive capacity) of the cancer cells was assayed. The invasive capacity of poorly invasive cells (10N), which were cloned from OC-10 cells, was five times less than that of the parental OC-10 cells. These results indicate our system to provide a useful model for studying human cancer cell invasion.


Invasive ductal carcinoma of the breast with a predominant intraductal component: cooperative clinicopathologic study in seven Japanese centers. A nationwide multicenter joint study was undertaken to investigate the clinicopathologic characteristics of invasive ductal carcinoma of the breast with a predominant intraductal component, a diagnostic entity proposed in the histologic typing of breast tumors by the World Health Organization in 1981. A total of 368 tumors, of which 218 were studied pathologically between 1983 and 1987 and 150 between 1971 and 1977, were accumulated for the present study from seven medical institutes in Japan. The incidence of such types of tumor was higher in the latter period than in the former, as was that of pure intraductal carcinomas. Seventy of the 368 tumors displayed metastatic nodal involvement and nine tumor deaths were observed from 150 tumors followed up for over 10 years following radical mastectomy, the overall quantity of the extraductal stromal invasion clearly acting on lymph node metastases and patients' outcome. We consider a total stromal invasion of greater than 5 mm to be critical in the prognosis of patients with tumor of this type.


[Plasma endotoxin measured by the combination of new perchloric acid pretreatment method and endotoxin specific assay in liver cirrhosis]. Using with the newly modified perchloric acid pretreatment method and endotoxin specific assay (Endospecy), the plasma endotoxin in the patients with liver cirrhosis was investigated. The mean value of plasma endotoxin in control (n = 20) was below 9.8 pg/ml. The plasma endotoxin level in liver cirrhosis was 5.7 +/- 5.3 pg/ml (n = 70, mean +/- SD), and 20% of the patients (15 cases) showed above 9.8 pg/ml. Endotoxin level significantly correlated with the severity of liver function based on the Child-Turcotte classification (p less than 0.01). Plasma endotoxin positively correlated with total bilirubin (r = 0.417) and ICG clearance test (r = 0.298) and negatively correlated with prothrombin time (%) (r = 0.497) and HDL-cholesterol (r = 0.578) in the patients with liver cirrhosis. There was no correlation between esophageal varices and plasma endotoxin level. Plasma endotoxin was slightly detected in the patients with decompensated cirrhosis, and these data suggest that plasma endotoxin level in cirrhosis is not so elevated.


[Results of patients in stage I and II primary lung cancer with curative resection]. We analyzed 91 patients who had curative resection from 1982 to 1989 for stage I and II primary lung cancer. The 5-year survival rates were 71.4% for stage I and 44.7% for stage II. The 5-year survival rates were 86.6% for T 1 N 0 M 0, 47.3% for T 2 N 0 M 0, 56.3% for T 1 N 1 M 0, and 37.9% for T 2 N 1 M 0. There was no cancer death in patients with squamous cell carcinoma of T 1 N 0 M 0. For stage I, tumor diameter was significant factor to influence survival. Although there was a statistically significant difference between stage I and II in patients with adenocarcinoma, there was no significant difference in those with squamous cell carcinoma. Chemotherapy had no significant improvement in survival for stage I and II lung cancer.


[Prognostic value of the leukogram and the myeloperoxidase activity of neutrophilic granulocytes in infectious complications in children with severe burns]. Clinical course of the burn disease of a severe, or extremely severe degree was studied in 56 children aging from 1 to 5 years. It was established that intensity of a leukocytic reaction and the redistribution reactions of neutrophil granulocytes with different activity of myeloperoxidase objectively reflected the course of a pathologic process, permitted to predict the development of local and general infective complications, and to make up a rational program for treatment with differentiated use of the methods of detoxication and immunotherapy. Activity of acid phosphatase and content of PAS-positive substances in neutrophil granulocytes didn't correlated with severity of the course of the burn disease.


Construction of an expression vector for human papillomavirus type 16 E7-glutathione S transferase fusion protein. An expression vector for human papillomavirus type 16 (HPV 16) E7-glutathione S transferase fusion protein was constructed. The E7 gene was confirmed by southern blot analysis with a digoxygenin 11-dUTP labeled and simultaneously amplified E7 DNA probe by polymerase chain reaction (PCR). The fusion protein was expressed in E. coli, recovered as inclusion bodies, and was confirmed by western blot analysis using a monoclonal antibody against HPV 16-E7 protein. A polyclonal antibody against the HPV 16-E7 protein was raised in the serum of mice hyper-immuned with the fusion protein. This antibody will be available for screening of patients with cervical cancer.


Influenza virus infections and immunity: a review of human and animal models. Studies of the pathogenesis of influenza infection have involved the extensive use of animal models. The development of the current concepts of immunity to influenza and of the contribution the secretory immune system makes toward the protection of mucosal surfaces against influenza infection would have been impossible without this use of animals. The pathology and clinical signs of influenza infection in both natural and experimental hosts, the advantages and disadvantages of the most common experimental influenza infection models, and the contribution of animal models to the understanding of local and systemic immunity to influenza infection are discussed.


An encephalomyocarditis virus epizootic in a baboon colony. Approximately 80 baboon deaths were caused by encephalomyocarditis virus (EMCV) infection in a 3060 member research and production colony. The epizootic extended over a 9-month period and occurred in baboons ranging from 1 day to 22 years of age. Acute death was the most common history. When clinical disease was detected, it was characterized by labored respiration associated with acute congestive heart failure. The salient necropsy findings were pulmonary congestion and edema, hydropericardium, hydrothorax, ascites, lymph node and splenic hypertrophy, and pale white-to-tan mottled hearts. The most significant histologic lesion was nonsuppurative necrotizing myocarditis. Placental infection with fetal loss occurred. Diagnosis was confirmed by light microscopy, transmission electron microscopy, virus culture, and serology. Rarely, EMCV-induced antibody persisted in surviving baboons for more than 24 months. EMCV-infected feral rats were the probable source of the virus and their control stopped the epizootic. No EMCV neutralizing antibody was detected in colony support personnel or chimpanzees.


Enrofloxacin treatment of long-tailed macaques with acute bacillary dysentery due to multiresistant Shigella flexneri IV. Thirty-four cases of acute bacillary dysentery occurred within 90 days among macaques housed at the California Regional Primate Research Center. Cases were identified by depression, diarrhea with blood and leukocytic exudate, and/or leukocytosis with a left shift. Antimicrobial susceptibility testing of enteric isolates and plasmid profile analyses established an etiologic diagnosis of multiple antibiotic resistant Shigella flexneri IV infection. When standard therapies were invalidated by high frequencies of resistance among the isolates, therapy with enrofloxacin, a fluoroquinolone antimicrobial, was initiated to interrupt the epidemic. Serum concentrations of enrofloxacin and its primary metabolite ciprofloxacin were measured in selected cases. A serum concentration-time data analysis was performed to evaluate the oral enrofloxacin dose and dosing interval for nonfasted macaques. Once daily administration of 5 mg/kg enrofloxacin by gastric intubation produced 24-hour serum concentrations above the MICs for the Shigella isolates from this outbreak.


Skeletal lesions and anemia associated with ascorbic acid deficiency in juvenile rhesus macaques. Young rhesus macaques housed in outdoor corn cribs and fed a commercially prepared primate diet became weak, depressed, were reluctant to move, and expressed locomotor abnormalities. Thirteen severely affected animals were hospitalized for evaluation. Physical examination disclosed swellings and instabilities involving the ends of long bones. Radiography confirmed physeal fractures in 11 of 13 animals. Affected bones included the distal femur, proximal humerus, distal tibia/fibula, and distal radius/ulna. Other, less obvious changes were noted on radiographs. Anemia was a consistent finding. Ascorbic acid deficiency was suspected and therapy was initiated that consisted of vitamin supplements, diet change, cage rest, and support bandages. Feed samples were submitted to a laboratory for analysis and were confirmed deficient in vitamin C. Follow-up radiographs showed large calcifying subperiosteal hematomas in epiphyseometaphyseal regions, consistent with a diagnosis of scurvy. Twelve of 13 animals recovered clinically. Subsequent radiographs documented improvement of initially severe angular deformities associated with displaced fractures.


Hypervitaminosis A and reproductive disorders in rabbits. Reproductive abnormalities in New Zealand White rabbits at a large commercial rabbitry were linked to an excess of dietary retinyl acetate. Fetal resorptions, abortions, and stillbirths were common in pregnant does. Examination of aborted and stillborn fetuses disclosed hydrocephalus, microencephaly, and cleft palate. Analysis of the commercially prepared feed disclosed a total vitamin A content of 102,278 IU/kg, of which 97,618 IU was retinyl acetate (recommended total vitamin A concentrations are 6,000 to 12,000 IU/kg). Levels of vitamin A in the plasma of does with reproductive disorders were 517 to 1,667 ng/ml (normal level is 300 ng/ml), and liver levels were 2,070 to 12,854 micrograms/g (normal range is 50 to 300 micrograms/g).


Genetic diversity of laboratory gray short-tailed opossums (Monodelphis domestica): effect of newly introduced wild-caught animals. The colony of gray, short-tailed opossums (Monodelphis domestica) at the Southwest Foundation for Biomedical Research, the primary supplier of this species for research purposes, was founded with nine animals trapped in 1978 in the state of Pernambuco, Brazil. Since 1984, 14 newly acquired founders from the state of Paraiba, Brazil have contributed to the gene pool of the colony. The animals from Paraiba and their descendants are significantly larger than the founders from Pernambuco and their descendants. The two groups also differ significantly in several measurements of morphologic traits. The changes in proportional contribution of each founder to the colony, and changes in inbreeding coefficients during the colony's history, are evaluated. Using previously established markers and three newly identified markers (ACP2, APRT, and DIA1), we show that the Paraiba-derived animals differ significantly from the original founders in allele frequencies and heterozygosity. The genetic diversity of the colony has been substantially increased by acquisition of the new founders from Paraiba. The colony is highly polymorphic, with 22.2% of loci surveyed by protein electrophoresis being variable. We conclude that the genetic differences between populations and among projects within the colony should be considered in future colony management procedures and in selection of experimental subjects.


Mirrors as enrichment for captive chimpanzees (Pan troglodytes). At many facilities, limitations of the physical environment have reduced the opportunity for captive chimpanzees to live in large, naturalistic social groups. Convex mirrors used to increase visual access of neighboring groups may improve the social environment. This was tested in a study of 28 chimpanzees (Pan troglodytes) group-housed in conventional indoor/outdoor runs. A total of 47.8 hours of behavioral observations were conducted and comparisons made across three conditions: no mirror present, a mirror present with visual access to neighboring conspecifics, or a mirror present with visual access to the neighbors' empty run. When the mirror gave subjects visual access to neighboring animals, facial expressions, sexual, and agonistic behaviors increased, whereas affiliative behavior decreased compared with when no mirror was present. When the mirror gave subjects visual access to a neighbors' empty run, facial expressions and sexual behavior increased compared with when no mirror was present. When the mirror gave subjects visual access to a neighbor's empty run, agonism decreased compared with when a mirror gave subjects visual access to neighboring animals. When subjects had visual access to neighbors, they used the mirror 30% of the total data points; while they had visual access to the neighbors' empty run, they looked during 24% of the total data points. Juveniles' use of the mirror increased over time while adults' use remained stable. Adult males used the mirror less than did the other subjects. These findings indicate that a mirror allowing visual access to neighboring conspecifics has potential as an enrichment device that affects social behavior.


Nephrotoxicity of tiletamine in New Zealand white rabbits. Tiletamine and zolazepam, the two constituents of Telazol, were evaluated independently to determine which agent was responsible for the nephrotoxicity caused by Telazol in New Zealand White rabbits. Five rabbits were injected i.m. with 32 mg/kg of tiletamine, four animals received 7.5 mg/kg of tiletamine, and five rabbits received 32 mg/kg of zolazepam. Urinalysis was performed and blood urea nitrogen and serum creatinine were monitored for 7 days postinjection. In all five rabbits injected with the high dose of tiletamine, blood urea nitrogen and creatinine rose by 3 days postinjection and increased steadily throughout the week. By 4 days postinjection, urine protein and glucose were elevated and cellular and protein casts were present. No serum chemistry or urine abnormalities were detected in rabbits receiving low doses of tiletamine, zolazepam, or in the four control rabbits. All animals were euthanized and necropsied at 7 days postinjection. Histopathology showed severe renal tubular necrosis in all five rabbits injected with 32 mg/kg tiletamine. Mild nephrosis was present in three of four rabbits injected with 7.5 mg/kg of tiletamine. No lesions were present in the zolazepam-injected or control rabbits. The results of this study show that tiletamine is the constituent responsible for the nephrotoxicity of Telazol in rabbits. They further demonstrate that doses commonly used for anesthetic induction or restraint can produce renal lesions in rabbits.


Unidirectional distribution of mosaicism in chimeric rats. Experimental rat chimeras were produced by aggregation of eight-cell embryos from two inbred strains, ACI/Hkm and WKAH/Hkm, which differ from each other in their major histocompatibility complexes and coat colors, and their mosaicism was analyzed. The existence of the isozyme Es-1, a serum cholinesterase specifically produced by WKAH-derived cells, and the agouti coat color due to ACI cells, indicated that all of the rats analyzed were unequivocal chimeras. The proportion of ACI cells in the red blood cell populations of the chimeras varied from 45% to 98%, as determined with a fluorescence-activated cell sorter and a monoclonal antibody against class I (RT1) antigen. Digital analysis of the coat color revealed that the proportion of the ACI type of coat color ranged from 72% to 98% in these chimeric rats. Each phenotype expressed in the coat color was complex and varied in size. The ratios of red blood cells and the coat color inclined toward the ACI type of cell population. Conversely, the rate of the WKAH-cell-type population was less than 50%. A breeding test disclosed chimerism of germ cells in two chimeric rats, and there were more pups with agouti coats than with albino coats. Taken together, it was shown in most of the phenotypes analyzed that the ACI type of cells was predominant in all of the chimeric rats. We discuss the possible causes for this unbalanced distribution in the rats.


Cardiovascular responses to intracerebroventricular infusion of artificial cerebrospinal fluid in anesthetized strain 13 guinea pigs. The cardiovascular effects of constant intracerebroventricular infusion in anesthetized strain 13 guinea pigs were studied. Bilateral cerebroventricles of the animals were catheterized stereotaxically with two 20-gauge blunt needles, penetrating 5 to 6 mm into the skull. Baseline cerebroventricular pressure values were 1.3 +/- 0.6 mmHg. After artificial cerebrospinal fluid was infused into the left ventricle at 0.5 ml/h, left cerebroventricular pressure increased to 8.1 +/- 1.6 mmHg (P less than 0.01), while right cerebroventricular pressure reached 5.6 +/- 2.2 mmHg within 20 minutes. No significant changes in mean blood pressure or heart rate were observed. When intracerebroventricular infusion rate increased to 5.0 ml/h, cerebrospinal fluid pressures of left and right cerebroventricles increased to 40.0 +/- 4.8 and 38.4 +/- 4.7 mmHg within 10 minutes from baseline values of 1.5 +/- 0.5 and 1.7 +/- 0.7 mmHg, respectively. Simultaneously, mean blood pressure and heart rate increased from 72 +/- 4 to 101 +/- 9 mmHg and from 195 +/- 11 to 218 +/- 17 beats/min, respectively. However, 30 to 50 minutes later, mean blood pressure, heart rate, and cerebrospinal fluid pressure decreased abruptly, and two of four animals died. We suggest that this technique with a low infusion rate (less than 0.5 ml/h) can be used to deliver certain drugs into the brain ventricles directly without producing undesirable effects on blood pressure or heart rate.


A simple new method for repeated in vivo cerebral cortex flux measurement in rats. A new application of an indicator dilution technique, using nonradioactive para-aminohippuric acid, is described for superior sagittal sinus blood flow determination in rats. Superior sagittal sinus blood flow, mainly representing cerebral cortex blood flow, amounted to 541 +/- 49 microliters/min in ketamine-anesthetized, but otherwise normal, rats breathing room air. Increasing arterial pCO2 enhanced superior sagittal sinus blood flow (P less than 0.01), providing evidence that this method correctly measures cerebral blood flow. The respiratory quotient was 1.1 and the cerebral cortex metabolic rate of oxygen consumption was +/- 1.45 mumol/min. Cerebral cortex ammonia uptake was not significantly different from zero and of the amino acid fluxes, only alanine differed from zero (P less than 0.05). Flux measurements are crucial in studies of healthy or altered organ metabolism in both experimental animals and postoperative surgical patients. We devised a simple and economical method of repeated cerebral cortex flux measurement in rats that is a potentially valuable tool in metabolic studies of the cerebral cortex.


Continuous intravenous infusion and simultaneous pharmacokinetic monitoring in ambulatory rabbits with a programmable micropump and dual subcutaneous Silastic central venous catheters. We describe a newly developed method of continuous intravenous infusion and simultaneous monitoring of plasma levels of investigational compounds in ambulatory untethered rabbits. Continuous infusion was administered by means of a portable programmable external micropump which permitted adjustable dosing. Simultaneous plasma pharmacokinetic monitoring during infusion was accomplished by dual silastic central venous catheters. The potential applications of the micropump infusion system as an alternative to current methods of continuous infusion in other species and in other studies are further discussed. This method provided a safe, reliable, and well-tolerated method of adjustable continuous intravenous infusion and simultaneous sampling of central venous blood in rabbits.


An immunoradiometric assay for squirrel monkey prolactin. Determination of immunoreactive prolactin in squirrel monkeys has been hampered by the lack of specific antibodies. We investigated the adaptability of a commercially available immunoradiometric assay for human prolactin, which employs two separate monoclonal antibodies (MAb I and II) to human prolactin, to determine the presence of squirrel monkey prolactin. We found that immunoreactivity curves for prolactin in squirrel monkey pituitary homogenates and serum were parallel to human prolactin standards, suggesting that the epitopes recognized by these antibodies were common to both human and squirrel monkey prolactin. Both nonglycosylated (23 kD) and glycosylated (26 kD) forms of squirrel monkey prolactin were detected in squirrel monkey pituitary homogenates by Western blot analysis using [125I]-MAb II. Neither sheep nor rat prolactin was recognized by Western blot analysis, indicating that the assay may be specific for primate prolactins. We examined the effect of ketamine HCl, an anesthetic that has been shown to elevate serum prolactin levels in other primates, on prolactin secretion in squirrel monkeys. Serum prolactin levels increased greater than fourfold after the administration of ketamine HCl (30 mg/kg b.w., i.m.) compared with control levels. Serum prolactin levels were unaffected by anesthesia with pentobarbital sodium (15 mg/kg b.w., i.v.). This assay provides a reliable and sensitive method for determining immunoreactive squirrel monkey prolactin.


Development of mouse oocytes superovulated at different ages. The development of oocytes superovulated at 25, 50, or 90 days in four mouse strains (C57BL/6N, DBA/2N, ICR, and B6D2F1) was examined using the techniques of in vitro fertilization, culture, and transfer of two-cell embryos to pseudopregnant recipients. The highest number of ova were obtained from superovulated 25-day-old mice in all strains. Approximately 80% of oocytes developed to the two-cell stage after in vitro fertilization. Of these living oocytes, 60% developed to weanling stage through the recipient. These results suggested that donor age among 25, 50, or 90-day-old mice has no influence on the viability of superovulated oocytes. Consequently, we conclude that superovulated 25-day-old mice offer an economical and efficient source of viable oocytes for the production of transgenic mice.


Effect of bleeding site on clinical laboratory testing of rats: orbital venous plexus versus posterior vena cava. We sought to determine if there were any differences in the results of clinical laboratory tests between blood samples collected from the orbital venous plexus and the posterior vena cava of adult male rats. Thirty healthy adult male Sprague Dawley rats were anesthetized by ether inhalation, and blood samples were collected successively from the orbital venous plexus (OVP) and the posterior vena cava (PVC) for hematologic (n = 10), serum chemistry (n = 10), and coagulation (n = 10) analyses. The prothrombin and partial thromboplastin times of samples from the OVP were prolonged (17% and 288%, respectively) when compared with samples from the PVC. Respective hematologic biases were as follows: red blood cell count (7%), hemoglobin (6%), hematocrit (5%), mean corpuscular volume (-3%), mean corpuscular hemoglobin (-1%), mean corpuscular hemoglobin content (1%), white blood cell count (13%), and platelet count (-7%). Respective serum chemistry biases were as follows: sorbitol dehydrogenase (-7%), glucose (-7%), blood urea nitrogen (-10%), creatinine (-2%), total protein (4%), albumin (2%), globulin (9%), alkaline phosphatase (5%), lactate dehydrogenase (-6%), aspartate aminotransferase (-5%), alanine aminotransferase (-2%), total bilirubin (0%), direct bilirubin (0%), magnesium (-17%), sodium (4%), potassium (0), chloride (4%), calcium (-2%), phosphorous (-17%), cholesterol (3%), triglycerides (24%), creatinine kinase (-8%), 5'nucleotidase (0%), and total bile acids (4%). For hematologic testing, there were no biologically significant differences between samples collected from the OVP and PVC. The coagulation times and serum Mg and P showed biologically significant differences between samples collected from the OVP and PVC.(ABSTRACT TRUNCATED AT 250 WORDS)


Isotype-specific rabbit antibodies against chinchilla immunoglobulins G, M, and A. Chinchillas have become a preferred animal model for studying otitis media, and are also useful in studying insulin release, gastrin physiology, intestinal infection, and hepatocellular pathophysiology. Immunopathologic studies in the model, however, have been limited by absence of specific antibody reagents against chinchilla immunoglobulins. We describe a method for preparing isotype-specific rabbit antibodies against the heavy-chain components of chinchilla immunoglobulins G, M, and A. Chromatographic techniques were used to isolate chinchilla immunoglobulins from serum and breast milk; heavy-chain fractions were isolated and used as antigens to produce isotype-specific antibodies in New Zealand White rabbits. Enzyme-linked immunosorbent assay of these antisera disclosed anti-light chain cross-reactivity, which was removed by affinity chromatography. The isolation and affinity purification techniques were highly reproducible. The availability of these reagents should greatly enhance the utility of the chinchilla in modeling human disease.


Evaluation of a subcutaneously implanted chamber for antibody production in rabbits. Polyclonal antibody production in subcutaneous chambers was compared to traditional antibody production methods in rabbits. The chamber, a sterilized plastic wiffle golf ball that had been surgically implanted in the subcutis of the thoracic region, was immunized via a percutaneous injection of antigen into the core of the ball through one of the perforations in the chamber wall. Rabbits bearing chambers were immunized on the same schedule and with the same concentrations of antigens as were provided the adjuvant injected rabbits. Fluid volumes of 12 to 22 ml could be removed from each chamber at weekly intervals. Chamber antibody to specific microbial antigens was equal to or better than serum antibody produced to the same antigens with Freund's or acrylamide adjuvants. The comfort of the rabbit, the ease in chamber immunization, and the recovery of high titer antibody in large volumes make the subcutaneous chamber an attractive method for polyclonal antibody production.


Improved method for bile collection in unrestrained conscious rats. We describe an improved method for continuous collection of bile from unrestrained rats. Use of an externally accessible, continuous-loop cannula when cannulating the common bile duct allows for full recovery from anesthetic effects and maintenance of a normal bile salt pool until the cannula loop is cut. Bile resulting from the cut cannula is diverted into a surgically implanted glass collection vessel and removed periodically via an externalized sampling port. Bile flow over a 24-hour collection period averaged 0.98 +/- 0.04 ml/hr (Mean +/- SEM, n = 9) with no gross pathological changes noted upon necropsy. This technique offers the capability of reestablishing conditions as close to physiologic as possible postsurgery to minimize potential artifacts during bile collection.


HIV/AIDS education in Indiana public schools, grades 7-12. Indiana secondary school principals were surveyed regarding HIV/AIDS education status in their schools. A questionnaire from the Centers for Disease Control was evaluated then modified for the survey. More than 520 school principals responded to the questionnaire mailed to all 680 school principals, grades 7-12, in Indiana. Most schools offer some form of HIV/AIDS education in grades 7-12. HIV education was taught by teachers with different educational backgrounds and school roles. Almost 60% of school principals believe students are at risk for HIV infection. Tests of statistical significance were conducted with regard to principals' opinions and actual practice of HIV/AIDS education in schools. Most schools have responded to the need for HIV/AIDS education, but with a variety of formats, teaching personnel, and teacher training.


Alteration of neutrophil (PMN) function by heparin, dexamethasone, and enalapril. The aim of this study is to investigate the effect of a seemingly divergent class of pharmacologic agents, each having been reported to suppress intimal hyperplasia, on neutrophil (PMN) function. Human PMNs were isolated and exposed for 30 min to either saline or one of three different pharmacologic agents, each tested at three different concentrations: Group 1, saline (control, n = 14); Groups 2-4, heparin (5000 units, n = 8; 2500 units, n = 6; 1250 units, n = 6) respectively; Groups 5-7, dexamethasone (4 mg, n = 8; 2 mg, n = 6; 1 mg, n = 6), respectively; and Groups 8-10, enalapril (1.25 mg, n = 8; 0.62 mg, n = 6; 0.31 mg, n = 6). Superoxide anion production was measured by the reduction of cytochrome c in a spectrophotometric assay. Chemotaxis was evaluated by the number of PMNs migrating across a filter using a Neuro Probe chamber. Phagocytosis was determined by the ingestion of opsonized zymosan particles by PMNs. Serum obtained from each PMN donor was used both to opsonize the zymosan and as a chemoattractant in the chemotaxis assay. No agent, at any dose, significantly changed superoxide production when compared to control cells. All three agents significantly inhibited PMN chemotaxis at every dose tested (P less than 0.01). In the phagocytosis assay, both heparin (at high and intermediate doses) and enalapril (at all doses) significantly reduced phagocytic activity (P less than 0.01); however, dexamethasone (at high and intermediate doses) produced a marked stimulation (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Investigation of subunit interactions by radiation inactivation: the case of Na+/K+-ATPase. The target size of Na+/K+-ATPase has been determined by radiation inactivation. To interpret the results, we have performed Monte Carlo simulations of the inactivation process. This seems to be a general method for the interpretation of such studies. The simulation revealed that radiation inactivation can distinguish between monoprotomeric and multiprotomeric models of enzyme action only if the measured reaction requires the actual co-operation of, rather than the mere co-existence of, different protomeres.


Hepatocellular carcinoma in porphyria cutanea tarda: frequency and factors related to its occurrence. Thirty-eight patients with porphyria cutanea tarda (PCT) have been seen in the last 18 years. Five of these patients (13%) developed hepatocellular carcinoma (HCC) during follow-up. We analyzed the differences in clinical, laboratory and liver histology findings at presentation, between patients who developed HCC during follow-up (HCC-group, n = 5) and those who did not (PCT-group, n = 33). Of the clinical features the duration of skin-symptoms was longer in the HCC-group (mean: 10.4 +/- 1.1 years) than in the PCT-group (mean: 1.4 +/- 1 years) (p less than 0.001). No differences in routine laboratory findings were found. Although 11/38 (29%) patients had serologic evidence of a past hepatitis B virus infection and 7/38 (18%) patients had antibodies against hepatitis C virus, no differences in these parameters were found between the PCT-group and the HCC-group. In all 34 liver biopsies a variable degree of siderosis was found (PCT-group vs. HCC-group: NS). Only piecemeal necrosis (p less than 0.01) and advanced fibrosis or cirrhosis (p less than 0.001) were more common in liver biopsies in the HCC-group. In conclusion, factors related to an increased risk of HCC in PCT are: a) a long symptomatic period before start of treatment and b) the presence of chronic active hepatitis and/or advanced fibrosis or cirrhosis in liver biopsies.


Space-occupying lesions of the liver detected by ultrasonography and their relation to hepatocellular carcinoma in cirrhosis. Fifty-four patients with cirrhosis, found to have a space-occupying lesion in the liver by ultrasound (US), underwent US-assisted biopsy of the lesion and were then followed prospectively to define outcome and survival. Histologic examination revealed hepatocellular carcinoma in 26 patients, while five had liver cell dysplasia without hepatocellular carcinoma and 23 had no evidence of tumor or of dysplasia. All five patients with an initial diagnosis of dysplasia developed hepatocellular carcinoma during follow-up and their survival curve was similar to that of patients with liver cancer and significantly worse than that of patients without dysplasia or tumor. There were five false-negative cases of hepatocellular carcinoma among the patients with negative histology. Overall, US-assisted liver biopsy diagnosed malignancy with a sensitivity of 72%, which increased to 86% when dysplasia was considered a pre-neoplastic lesion.


Primary hepatocellular carcinoma with severe hypoglycemia: involvement of insulin-like growth factors. We report a case of severe hypoglycemia and hepatic masses suspected to be an insulin-like growth factor-II (IGF-II)-producing hepatocellular carcinoma. A 62-year-old man presented with mental disorder in the night and early morning associated with extremely low blood sugar levels (less than 21 mg/dl). Computerized axial tomography and ultrasonography revealed a massive tumor in the right lobe of the liver with multiple secondary nodules, and a tumor thrombus in the portal vein. At autopsy 107 days after admission, the liver weighed 3070 g, histologically showing an Edmondson type II tumor with liver cirrhosis. IGF-II in plasma (899 ng/ml) and tumor tissue (2.4 micrograms/g) was higher than that in normal plasma (374-804 ng/ml) and non-tumor liver tissue (0.2 micrograms/ml), while IGF-I (14 ng/ml) was significantly reduced. IGF-II, probably produced by the liver tumor, appeared to be involved in the mechanism of hypoglycemia.


Long-term follow-up of anti-hepatitis C virus antibodies in patients with acute nonA nonB hepatitis and different outcome of liver disease. We screened 74 patients with nonA nonB acute hepatitis (37 of post-transfusion PTH, and 37 of non-post-transfusion, NPTH, origin) for the presence of anti-HCV by tests detecting either C100-3 antibodies alone (ELISA I) or C100-3 plus C33c plus C22-3 antibodies (ELISA II). Samples were taken at the onset of disease and then serially for a period of time ranging from 12 to 60 months. An increased number of anti-HCV positive cases (86% vs 69%) and an earlier seroconversion were observed with the second compared to the first generation ELISA. Positive samples were confirmed by recombinant immunoblot assay (RIBA), which was also used to study the kinetics of the antibody response to individual HCV antigens. Anti-C33c and anti-C22-3 antibodies were the first detectable markers of HCV infection in 80% and 20% of the patients, respectively. Thirty-two percent of the patients studied showed complete and persistent biochemical recovery, whereas 68% maintained a chronic elevation of the transaminase values. Among the 20 patients who showed early and persistent normalization of the transaminase values, complete disappearance of all antibody reactivities was limited to five of them, whereas in four cases only anti-C100-3 and anti-5-1-1 became negative.


A combination of the aldose reductase inhibitor, statil, and the prostaglandin E1 analogue, OP1206.alpha CD, completely improves sciatic motor nerve conduction velocity in streptozocin-induced chronically diabetic rats. In view of the possible implication of multifactorial mechanisms in the pathogenesis of diabetic neuropathy, the aldose reductase inhibitor (ARI), Statil, which ameliorates abnormal sorbitol or myo-inositol metabolism in diabetic nerves, and the prostaglandin E1 (PGE1) analogue, OP1206.alpha CD (OP), which improves diabetic vascular derangements, were administered simultaneously for 2 months to streptozocin (STZ)-induced diabetic rats with 5 months' duration of diabetes, and the effects on sciatic motor nerve conduction velocity (MNCV), Na(+)-K(+)-adenosine triphosphatase (ATPase) activity, and morphology of myelinated nerve fibers (MNF) were compared with the effects of a monotherapy with OP. The combination regimen ameliorated abnormal nerve sorbitol and myo-inositol levels and normalized decreased MNCV and enzyme activity. In contrast, neither sorbitol nor myo-inositol metabolism was ameliorated, and only insufficient improvement of MNCV and morphology of MNF was obtained with a monotherapy with OP. In addition, the combination therapy reversed both a decrease in the percent of large MNF and an increase in the percent of small MNF in diabetic rats, whereas a monotherapy with OP reversed only a decrease in the percent of large MNF. The results might suggest that a multiple-drug therapy with different mechanisms of action has greater effects on diabetic neuropathy than a single-drug therapy and is worthy of clinical consideration.


Cyclic adenosine monophosphate accumulation and beta-adrenergic binding in unweighted and denervated rat soleus muscle. Unweighting, but not denervation, of muscle reportedly "spares" insulin receptors, increasing insulin sensitivity. Unweighting also increases beta-adrenergic responses of carbohydrate metabolism. These differential characteristics were studied further by comparing cyclic adenosine monophosphate (cAMP) accumulation and beta-adrenergic binding in normal and 3-day unweighted or denervated soleus muscle. Submaximal amounts of isoproterenol, a beta-agonist, increased cAMP accumulation in vitro and in vivo (by intramuscular [IM] injection) to a greater degree (P less than .05) in unweighted muscles. Forskolin or maximal isoproterenol had similar in vitro effects in all muscles, suggesting increased beta-adrenergic sensitivity following unweighting. Increased sensitivity was confirmed by a greater receptor density (Bmax) for [125I]iodo-(-)-pindolol in particulate preparations of unweighted (420.10(-18) mol/mg muscle) than of control or denervated muscles (285.10(-18) mol/mg muscle). The three dissociation constant (Kd) values were similar (20.3 to 25.8 pmol/L). Total binding capacity (11.4 fmol/muscle) did not change during 3 days of unweighting, but diminished by 30% with denervation. This result illustrates the "sparing" and loss of receptors, respectively, in these two atrophy models. In diabetic animals, IM injection of insulin diminished cAMP accumulation in the presence of theophylline in unweighted muscle (-66% +/- 2%) more than in controls (-42% +/- 6%, P less than .001). These results show that insulin affects cAMP formation in muscle, and support a greater in vivo insulin response following unweighting atrophy. These various data support a role for lysosomal proteolysis in denervation, but not in unweighting, atrophy.


Surgical treatment of tumours of the skull base. To describe the special features of surgical removal of tumours of the skull base and to review a series of patients treated for skull base tumours. A retrospective review of patients followed up for between six months and six years. A unit specialising in surgery of the skull base at a tertiary referral centre. Sixty-six patients with skull base tumours were referred to the unit between February 1984 and December 1989. Fifty-six of the patients underwent complete or partial surgical removal of the tumour. Radiotherapy was used as primary treatment in 10 patients, for tumour recurrence in four patients and as adjuvant postoperative treatment in 10 patients. In the majority of patients the tumours were benign and have been controlled or are considered cured. Of the seven patients with malignant tumours five have died and two are long-term survivors (one has multiple metastases). Three patients with histologically "benign" tumours died of multiple metastases. There have been a total of 10 deaths in the series but only one occurred in the perioperative period. Cranial nerve palsy was the major postoperative complication. Modern microsurgical techniques and modern anaesthesia and intensive care skills lead to a cure in many patients with skull base tumours previously considered inoperable.


[Maxillofacial and dental abnormalities in some multiple abnormality syndromes. "Cri du chat" syndrome, Wilms' tumor-aniridia syndrome; Sotos syndrome; Goldenhar syndrome]. The paper describes the maxillo-facial and dental anomalies observed in some chromosome and non-chromosome poly-malformative syndromes ("Cri du chat" syndrome; Wilms' tumour; Sotos' syndrome; Goldenhar's syndrome). The Authors emphasise the possibility of diagnosing these multiple deformity syndromes from maxillo-facial alterations in early infancy; anomalous tooth position and structure cal also be successfully treated immediately after the first appearance of teeth. This is a particularly promising field of pediatrics and preventive pediatric medicine.


The Missouri Cardiovascular Health Plan: a focus on prevention. The authors describe a framework for planning, implementing, and evaluating cardiovascular health programs in Missouri. Populations targeted for intervention are identified, and the role of the physician in supporting community-based cardiovascular health promotion is explored.