Genetic study of the loss and restoration of Mutator transposon activity in maize: evidence against dominant-negative regulator associated with loss of activity. The Mutator system of transposable elements is characterized by a family of transposons called Mu transposons that share common termini and are actively transposing in Robertson's Mutator (Mu) lines of maize. Mu lines lose transposition activity during propagation by either outcrossing or inbreeding. This loss of transposition activity, which can occur at non-Mendelian frequencies, is in the form of loss of forward transposition activity resulting in a decrease in the generation of new mutations, as well as the loss of mutability of Mu transposon induced mutations, and it has been correlated with hypermethylation of the Mu elements. Previous studies have concluded that restoration of Mutator transposon activity by crossing inactive lines back to active lines is incomplete or transient, and depends upon the sex of the inactive parent. Further, it has been proposed that the inactive system is dominant to the active system, with the dominance possibly mediated through a negative regulatory factor that is preferentially transmitted through the female. In this study, we have examined the frequencies of loss and restoration of Mu transposon activity using a Mu line carrying an insertion in the bronze 1 locus. We find that transmission of Mu transposon activity to non-Mu plants can occur at high rates through males and females, but individual cases of decreased transmission through the male were observed. We also find that in crosses between inactive-Mu and active-Mu plants, reactivation was efficient as well as heritable, regardless of the sex of the inactive parent. Similar results were obtained whether the inactivation occurred in an outcross or a self. In all cases examined, loss of Mu transposon activity was correlated with hypermethylation of Mu elements, and reactivation was correlated with their demethylation. Our results indicate that an inactive Mu system does not exhibit dominance over an active Mu system. We conclude that contrary to current models, inactivation and its maintenance is not obligatorily associated with a dominant negative regulatory factor whether nuclear or cytoplasmic, and we propose a revised model to account for these and other observations.


Colorectal leukotriene B4 synthesis in vitro in inflammatory bowel disease: inhibition by the selective 5-lipoxygenase inhibitor BWA4C. The in vitro synthesis of leukotriene B4 (LTB4) was evaluated in colorectal biopsy specimens and resection tissue from patients with inflammatory bowel disease. The in vitro formation of LTB4 from biopsy tissues stimulated with calcium ionophore A23187 correlated with the degree of mucosal inflammation assessed at sigmoidoscopy, and with neutrophil infiltration measured as myeloperoxidase activity. Biopsy specimens from patients taking prednisolone formed less LTB4 than those from patients not on prednisolone, with comparable levels of inflammation seen at sigmoidoscopy. The formation of LTB4 was reduced dose-dependently by the acetohydroxamic acid 5-lipoxygenase inhibitor BWA4C, with no significant inhibition of prostaglandin E2 or thromboxane B2 synthesis. In inflamed colonic resection tissue from colitic patients, the IC50 for inhibition of LTB4 formation by BWA4C was 0.03 mumol/l, compared with an IC50 of 0.8 mumol/l for NDGA. Thus, BWA4C is a potent and selective inhibitor of LTB4 synthesis in colonic tissue from patients with ulcerative colitis. Acetohydroxamic acid 5-lipoxygenase inhibitors, exemplified by BWA4C, may be useful to evaluate the clinical importance of LTB4 in ulcerative colitis, and offer a novel therapy for the disease.


Proliferation in human gastrointestinal epithelium using bromodeoxyuridine in vivo: data for different sites, proximity to a tumour, and polyposis coli. The distribution of DNA synthesising cells in the crypts of the epithelium in human small and large bowel after injection of bromodeoxyuridine into patients has been studied in relation to the position of the cells in the crypt using immunohistochemistry. Different sites of normal epithelium have been studied. The ileum has a shorter crypt and a very significantly smaller total cell population size. However, it has similar peak labelling index (LI) values to the colon, while the rectum has a lower peak LI value. The mean position of the label occurs at the 17th cell position in the ileum and at about the 22nd position in both the colon and rectum. The overall mean LI is significantly higher in the ileum at 17.8%, intermediate in the colon at 10.3%, and lowest in the rectum at 8.5%. There is thus an inverse relation between the likelihood of developing a tumour and the rate of cell proliferation as measured by the LI. Assuming a value of 8.6 hours for the duration of S, the data suggest that the cell cycle time in the mid crypt region is about 30 hours for the ileum and colon and about 37 hours for the rectum. Samples taken adjacent (within 1 cm) to a tumour show a general dampening of proliferative activity at all cell positions compared with samples taken more than 5 cm from a tumour. This is illustrated by the average LI, which is about 5.4% in the colon adjacent to a tumour compared with 10% distant; comparable values for the rectum are 4.6% and 8.5%. Samples taken from two patients with polyposis coli show distributions with a significant difference in skewness compared with normal colon and a general shifting of the distribution to the right, that is to higher cell positions. There is a significant increase in the mean cell position and the position of the peak LI in the polyposis coli samples.


Neuromuscular hyperexcitability features in patients suffering from musculoskeletal pain: a neuroepidemiologic survey. We studied the reported frequencies of clinical complaints of neuromuscular hyperexcitability (muscle cramps and fasciculations) in random samples of 527 Dutch adults, who were and 253 Dutch adults, who were not suffering from musculoskeletal pain and tenderness. Data were collected by telephone-interview and by self-administered questionnaire. Muscle cramps and fasciculations were recorded more frequently in the category that suffered from musculoskeletal pain (p less than 0.001). This association warrants further investigation into the possible intrinsic role of neuromuscular hyperexcitability in musculoskeletal pain and primary fibromyalgia.


A placebo controlled observer blind immunocytochemical and histologic study of epithelium adjacent to anogenital warts in patients treated with systemic interferon alpha in combination with cryotherapy or cryotherapy alone. To examine biopsy specimens of tissue immediately adjacent to anogenital (AG) warts which had been treated with either cryotherapy plus subcutaneous interferon (IFN) alpha 2a or cryotherapy alone, for histological features of (a) human papilloma virus (HPV) infection (b) localised cellular immune responses, to further characterise any cellular immune infiltrates with tissue immunocytochemistry, and to relate any histological, immunocytochemical findings to the treatment response of nearby AG warts. A randomised placebo controlled observer blind study. Genitourinary Medicine clinic, Department of Immunopathology, Royal Victoria Hospital, Belfast, N. Ireland. Thirty patients with AG warts; 16 treated with IFN alpha 2a plus cryotherapy, and 14 treated with cryotherapy alone. (1) Light microscopic features associated with HPV infection and local cellular immune responses. (2) Indirect immunofluorescence detection of the following cell surface markers: HLA DR, alpha one antitrypsin, CD1, CD3, CD4, CD8, CD22. (3) Clinical response of AG warts to treatment. In pre-treatment biopsies only non specific indicators of HPV infection (acanthosis, 29/30 biopsies, and hyperkeratosis, 7/30 biopsies) were seen on light microscopy. Mononuclear cells were seen both throughout the upper dermis and centred around dermal blood vessels in 19/30 (63.3%) biopsies, and infiltrating into the epidermis in 12/30 (40%) biopsies. On indirect immunofluorescence CD3, CD8, CD4 antigen was detected on the surface of cells throughout the upper dermis in 24/29 (82.7%), 15/29 (51.7%), and 3/29 (10.3%), of biopsy specimens respectively. CD3 antigen, CD8 antigen and CD4 antigen was detected on the surface of cells infiltrating into the epidermis in 18/29 (62%), 7/29 (24.1%), and 6/29 (20.7%) of biopsy specimens respectively. CD1 antigen was seen on the surface of dendritic cells throughout the epidermis in all specimens; CD1 positive cells infiltrated into the upper dermis in 5/29 (17.2%). HLA DR was detected on the surface of dendritic cells throughout the epidermis in 22/29 (75.9%) of specimens, and on the surface of cells scattered both diffusely throughout the upper dermis and centred around dermal blood vessels in all specimens. Alpha one antitrypsin (A1AT) antigen was seen on the surface of cells in the upper dermis in 6/29 (20.7%) of biopsy specimens; no cells expressing CD22 surface antigen were seen. The nature of this local cellular immune response was not altered by treatment of nearby warts with either cryotherapy alone or cryotherapy plus systemic IFN alpha 2a, or related to the therapeutic outcome of these warts. (1) No convincing histological evidence of HPV infection was seen in epithelium surrounding AG warts. (2) A predominantly T cell-mediated immune response (the target of which is uncertain) was seen in this perilesional epithelium. (3) In the dosage regimens used in this study, treatment of AG warts with either systemic IFN alpha 2a plus cryotherapy or cryotherapy alone did not appear to augment localised cellular immune responses (against any presumed subclinical HPV infection) in epithelium surrounding AG warts.


Synaptic mechanisms in long-term potentiation. Long-term potentiation (LTP) is a robust form of synaptic enhancement that far outlasts the duration of active stimulus. LTP represents the most intriguing form of synaptic memory, mainly due to its long duration and associative properties, and is held to be involved in learning processes at the behavioural level. Advances in the understanding of central synaptic functions are now extending interest in LTP to the fine detail of its synaptic mechanisms.


The acetic acid test in evaluation of subclinical genital papillomavirus infection: a comparative study on penoscopy, histopathology, virology and scanning electron microscopy findings. To evaluate colposcopic criteria in acetowhite lesions of the penis ("penoscopy") for the diagnosis of subclinical genitoanal papillomavirus infection (GPVI) compared with histopathological criteria of HPV involvement and to various hybridisation assays for HPV DNA detection, and to depict typical lesions by scanning electron microscopy. The study included 101 randomly selected male partners of females with known GPVI, or with penile symptoms such as itching, burning and dyspareunia who did not exhibit overt genital warts but appeared to be afflicted with acetowhite penile lesions after topical application of 5% acqueous acetic acid. Lesions were judged by penoscopy as either typical, conspicuous or nontypical for underlying HPV infection. Biopsy specimens from 91 men were examined by light microscopy and by either Southern blot (SB), polymerase chain reaction (PCR) and/or in situ hybridisation (ISH) assays for the presence of HPV DNA of the HPV types 6, 11, 16, 18, 31, 33 and 42 (Group A). From another ten men lesions clinically typical for GPVI were also examined topographically by scanning electronic microscopy (Group B). The STD out-patient clinic of the Department of Dermatovenereology of Karolinska Hospital, Stockholm, Sweden. Group A Seventy eight (86%) of the biopsied lesions met the penoscopy criteria of being either typical of or conspicuous for GVPI. The agreement between penoscopy and histopathology was fairly good, as HPV diagnosis was made by both methods in 56 (62%) of the cases. The reliability of applying strict colposcopic hallmarks was further substantiated by the finding that 55 (60%) of the biopsy specimens taken from penoscopically typical/conspicuous lesions contained HPV DNA. However, there are diagnostic pitfalls for the acetic acid test. Coexistence of an eczematoid reaction with changes indicative of HPV influence was detected in six (7%) of the cases, while an inflammatory response only occurred in 17 (19%) of the specimens. Additional histopathological diagnoses (normal epithelium, lichen sclerosus et atrophicus, balanitis circinata parakeratotica, verruca plana) were established in another eight (9%) of the cases. Among the HPV DNA positive cases, all of the HPV types tested for were detected with the exception of HPV 18. A severe penile intraepithelial neoplasia (PIN III) was revealed in five (5%) of biopsies; HPV 16 was present in two and HPV 42 in one of these biopsy specimens. Scanning electron microscopy depiction harmonised with the penoscopy findings showing that subclinical GPVI characteristically exhibits a well demarcated, slightly elevated border and that the central area of lesions often displays a "groove" in which the epithelium appears to be thin with protrusions from beneath that probably represent capillaries. Use of the acetic acid test for evaluation of GPVI should be combined with a colposcopic evaluation based on strict topographic hallmarks, followed by a directed biopsy for light microscopic evaluation. We found that the positive predictive value of colposcopy was as high when correlated with histopathological findings (72%) as when virological methods were used, whether HPV DNA hybridisation testing was performed with the well established SB and ISH assays (45%), or by applying the newly introduced and highly sensitive PCR assay as well (71%). False positivity from the acetic acid test occurs and is mainly due to inflammatory conditions but also to the presence of other conditions. Epithelial fissures are evidently associated with some subclinical GPVI lesions and may potentially represent loci minores for infectious stimuli and perhaps facilitate the transmission of some blood-borne STDs. We prose that the term "papillomavirus balanoposthitis" should be used for penile HPV infection associated with inflammatory responses. Our study indicates that PIN III frequently occurs in a subclinical form and may be associated with not only previously identified "high-risk" HPV types such as type 16, but also with the HPV type 42 that has not previously been considered as oncogenic.


Role of histamine in the chemotactic deactivation of polymorphonuclear leukocytes following incubation with formylmethionyl peptides. To clarify the mechanism of chemotactic deactivation of polymorphonuclear neutrophils (PMN) following incubation with the synthetic dipeptide N-formylmethionyl phenylalanine (FMP), we tested the hypothesis that histamine, which is released from leukocytes during incubation with FMP, could explain this inhibition. Human PMN were incubated in the presence or absence of FMP (10(-7) to 10(-5) M) and histamine measured fluorometrically in the supernatant. Washed PMN were then tested in Boyden chambers against FMP (10(-5) M) and other chemoattractants. Incubation of leukocytes with FMP caused a nonpreferential PMN deactivation which was proportionally and kinetically related to FMP-induced histamine release. No histamine release or chemotactic deactivation was observed in the absence of Ca2+ and Mg2+. The inhibitory effect of the peptide was significantly prevented by an H2 blocker or when using basophil-depleted PMN suspensions. Histaminase abolished the capacity of FMP-incubated leukocyte supernatants to decrease PMN chemotaxis. Preincubation of leukocytes with anti-IgE or a sensitizing allergen caused a significant PMN chemotactic deactivation. These results show that histamine which is released during leukocyte incubation with FMP, contributes, at least in part, to the chemotactic deactivation of PMN.


Activation of human eosinophils by platelet-derived growth factor. Activated eosinophils are believed to be major contributors to the chronic inflammatory sequelae of asthma, but the details of the mechanism of eosinophil activation in vivo are unknown. In our search for physiologically important modes of eosinophil activation, we studied the effects of recombinant human platelet-derived growth factor (PDGF) on human peripheral blood eosinophils. We compared two activation end-points: secretion of granule contents, exemplified by the release of eosinophil peroxidase (EPO), and eosinophil-derived neurotoxin (EDN), and the generation of active oxygen metabolites (O2- production). PDGFc-sis dose dependently stimulated the secretion of large amounts of EPO and EDN from eosinophils. Higher concentrations of PDGF induced a dose-dependent O2- production, especially if the cells were first primed with low concentrations of phorbol ester. These activities were not seen with the AA homodimer of PDGF, suggesting that the activation was receptor dependent. However, several attempts to directly demonstrate the existence of such receptors were unsuccessful. The magnitude of the secretory response to PDGF, and the realization that eosinophils could be easily exposed to this substance as they travel towards the lung, suggests the possibility that this growth factor may be a physiologically important activator of eosinophils in the pulmonary inflammation which is associated with asthma.


Augmentation of pulmonary foreign body granulomatous inflammation in mice by lipopolysaccharide: involvement of macrophage activation and tumor necrosis factor-alpha. We examined the effect of lipopolysaccharide (LPS) on dextran bead-induced lung granulomas using LPS responder (C3H/HeN) and nonresponder (C3H/HeJ) mice. LPS augmented granuloma sizes, TNF-alpha and N-acetyl-beta-D-glucosaminidase levels of lung extracts in C3H/HeN, but not in C3H/HeJ mice. Granuloma macrophages of C3H/HeN mice produced higher levels of superoxide anion and TNF-alpha than those of C3H/HeJ mice. Taken together with our previous report that macrophages and macrophage-derived cytokines are essential for the development of lesions, the present results suggest that activation of macrophages plays an important role in the development and augmentation of pulmonary granulomas.


Hymenopteran venom stimulates adenylate cyclase in cultured human cells through a histamine receptor. We have examined the effect of venom sac extract (VSE), prepared from two hymenopteran species, on intracellular cyclic AMP (cAMP) accumulation in a cultured human cell line. VSE prepared from the oriental hornet (Vespa orientalis) and from the paper wasp (Polistes carolina) induced an increase in accumulation of cAMP in human, transformed, non-pigmented, ciliary epithelial cells. The effect of V. orientalis VSE on intracellular cAMP was dose-dependent. Blocking the response of adenylate cyclase-linked beta-adrenergic receptors was without effect on this cellular response to VSE. On the other hand, the response to VSE prepared from both species was suppressed almost totally by the H2-histamine receptor-specific antagonist, cimetidine. The findings are discussed in the light of earlier observations that described a significant reduction in intraocular pressure in vertebrate animals treated with V. orientalis VSE.


Effect of 1,1-dimethylhydrazine (UDMH) on Corynebacterium parvum-associated immunosuppression in mice. These studies further investigate the immunoenhancement properties of UDMH by utilizing Corynebacterium parvum-induced immunosuppressed mice as well as evaluating activated macrophage production of reactive oxygen intermediates or their effects. Forty-eight hour Con A-induced lymphoblastogenic responses from splenocytes isolated from C. parvum and UDMH-treated Balb/C mice were significantly increased compared with C. parvum alone, although less than normal control mice (no treatment). In vitro bioassay of IL-2 production in cell culture supernatant isolated from these same treatment groups exhibited a pattern of stimulation similar to that of lymphocyte blastogenesis. In addition, UDMH did not interfere with H2O2-mediated suppression of either Con A- or LPS-induced lymphocyte blastogenesis and actually enhanced suppression of Con A-induced lymphocyte cultures at 25 micrograms/ml. We also report that production of superoxide anion from TPA-activated peritoneal macrophages exposed to various concentrations of UDMH in vitro was not affected. Although in vivo exposure to UDMH partially reversed C. parvum-induced immunosuppression in mice, the exact mechanism by which UDMH acts to reverse this immune suppression is not clear. UDMH does not appear to interfere with either activated peritoneal macrophage production of superoxide anion or H2O2-induced suppression of lymphocyte blastogenesis to elicit immune enhancement.


Pharmacokinetics of rufloxacin once daily in patients with lower respiratory tract infections. The pharmacokinetic properties of rufloxacin, a new quinolone antibacterial agent, were evaluated in ten patients with lower respiratory tract infections. Patients were given 400 mg of rufloxacin once a day for seven to nine days. Plasma concentrations of the drug were determined by high-performance liquid chromatography and bioassay at regular intervals during treatment. After the first administration, maximal plasma concentrations were 3.17 +/- 0.36 mg/l (mean +/- SEM) and were reached at 4.2 +/- 0.7 h. At the end of treatment peak plasma concentrations increased to 7.26 +/- 0.52 mg/l. Elimination half-life was 38.2 +/- 2.9 h, with a mean extent of accumulation of 2.96 +/- 0.30. Treatment was well tolerated, with no abnormalities noted during routine laboratory examinations. Two days after the last administration, measurable levels of rufloxacin were still observed in plasma, indicating that the long half-life of rufloxacin assures valuable antibacterial activity even after discontinuation of treatment.


Effects of cannabinoids and cocaine on the mitogen-induced transformations of lymphocytes of human and mouse origins. The effects of delta 9-tetrahydrocannabinol (d9THC), delta 6-tetrahydrocannabinol (d6THC), and cocaine on the mitogen-induced transformations of lymphocytes of human and mouse origins were examined in an in vitro system. None of the three compounds is mitogenic in nature. The effects of the two components of marijuana on the mitogen-induced transformation are biphasic. Both compounds stimulated the lymphocyte transformation at low concentrations and inhibited mitogenesis at high concentrations. Cocaine, on the other hand, neither affected mitogen-induced lymphocyte transformations nor altered d9THC's transformation effects when both compounds were added together. Human lymphocytes and mouse splenocytes appeared to respond in similar patterns.


Progression of squamous cell carcinoma of the uterine cervix from cervical intraepithelial neoplasia infected with human papillomavirus: a retrospective follow-up study by in situ hybridization and polymerase chain reaction. A retrospective study was conducted to reveal the natural history of cervical carcinoma infected with human papillomavirus (HPV) and to document latent persistence of HPV infection. Thirty-eight formalin-fixed, paraffin-embedded hysterectomy specimens of cervical carcinoma and cervical intraepithelial neoplasia (CIN) III were examined for the presence of HPV 6, 11, 16, 18, 31, 33, and 35 DNA by in situ hybridization (ISH) with biotinylated DNA probes. The HPV 6 and 11 were not detected, but HPV 16, 18, 31, 33, and 35 were detected in 63% (24 of 38) of CIN III and cervical carcinoma. The HPV-negative specimens by ISH were subjected to polymerase chain reaction (PCR) for detection of HPV 16 and 52b with high sensitivity. In 11 of 14 specimens, specific amplification of HPV 16 was detected and HPV DNA was demonstrated in 92% (35 of 38). A retrospective examination for the presence of HPV DNA by ISH and PCR was performed on sequential biopsy specimens of eight cases whose hysterectomy specimens were judged as HPV 16/18 positive. In the eight cases, HPV 16 was constantly demonstrated in all biopsy specimens throughout the course, from early dysplastic change to carcinoma. This finding indicates long (up to 10 years) persistence of HPV infection in the natural course of progression to carcinoma of the uterine cervix.


Effect of human papillomavirus infection on estrogen receptor and heat shock protein hsp27 phenotype in human cervix and vagina. In this study we have explored whether, as a consequence of human papillomavirus (HPV) infection, there is inappropriate expression of estrogen receptor and/or of a small heat shock protein of 27,000 daltons (hsp27). Estrogen receptor, hsp27, and HPV structural antigens were detected by immunocytochemistry, while HPV DNA (6/11, 16/18, 31/35/51) was determined by in situ hybridization in cervical and vaginal samples from 40 patients. Most of the samples with HPV infection without atypia showed a shift in estrogen receptor expression since this protein appeared mainly in the intermediate and superficial cell layers. In the serial sections, these layers displayed strong estrogen receptor staining, together with high HPV replication and late HPV gene expression. In the samples with HPV infection and atypia, estrogen receptors were also frequently found in the basal and parabasal cells, but almost 20% of these samples did not show estrogen receptors. The presence of high estrogen receptor expression was not dependent on a particular HPV DNA type. On the other hand, interesting modifications in hsp27 expression were observed in the HPV-infected tissues. The cytoplasm of the cells with koilocytotic changes showed very low hsp27 content. In several samples this protein appeared in the nuclei of the superficial cells, and sometimes it was also observed in the cytoplasm of the basal cells. These changes in estrogen receptor and hsp27 expression suggest that these proteins might have a role in virus-host biology.


The usefulness of clinical measurements of cell proliferation in gynecological cancer. This article critically reviews the available methods for the assessment of cell proliferation in clinical practice, and evaluates their applications as diagnostic and prognostic variables in gynecological cancer. The mitotic count is not a standardised method and should not be considered to have inherent value as a measurement of cell proliferation in the clinical situation. In distinguishing benign and malignant uterine smooth muscle tumors, the degree of mitotic activity can only be interpreted in the context of the other pathological and clinical findings. Cytological atypia and the absence of stromal differentiation appear to be more accurate than mitotic activity in delineating high-grade endometrial stromal sarcomas. The potential biological behavior of placental site trophoblastic tumor cannot be reliably predicted on the basis of mitotic counts, and hysterectomy remains the treatment of choice for this condition. Flow cytometry produces accurate cell kinetic data and provides useful prognostic information in ovarian and endometrial tumors. The value of flow cytometry in cervical cancer is questionable and needs clarification. Complete and partial hydatidiform moles are generally distinguishable on the basis of ploidy studies. However, in view of the existence of a diploid partial mole, serial human chorionic gonadotrophin measurement is the only reliable method of establishing or excluding a diagnosis of persistent trophoblastic disease. The other clinical markers of cell proliferation do not have an established role as diagnostic or prognostic parameters in gynecological cancer and merit further critical evaluation. Estimation of mitotic activity is a simple, readily accessible method of assessing cell proliferation in routine histopathological practice and the role of an accurate mitotic index in diagnosis and prognosis must be investigated to determine the true value of counting mitotic figures in histological sections.


Adenosarcoma of the uterus: a Gynecologic Oncology Group clinicopathologic study of 31 cases. We report on the clinical and pathologic findings in 31 cases of adenosarcoma of the uterus subjected to hysterectomy and staging laparotomy. Nine of 30 patients (30%) have had recurrent tumor and six of 30 (20%) have already died of tumor in a relatively short follow-up period (mean, 38.3 months). Seventeen of 31 cases were diagnosed as adenosarcoma with sarcomatous overgrowth (SO). Ten of these 17 with SO contained focal or extensive rhabdomyosarcoma. In six cases, extrauterine spread was identified as follows (two patients had two sites each): vaginal involvement (two cases), pelvic lymph node metastases (two), positive peritoneal cytologic findings (two), parametrial invasion (one), and ovarian metastasis (one). Extrauterine spread (stage III) (p less than 0.001) and myometrial invasion (p = 0.04) were associated with higher rates of recurrence. The presence of lymphatic and/or vascular invasion, SO, and rhabdomyosarcomatous differentiation also indicated poor prognosis but did not attain statistical significance. Based on this experience, staging laparotomy including peritoneal cytology is suggested in cases of clinical stages I and II adenosarcoma. The differential diagnosis of these tumors is also discussed.


The effects of gonadectomy on glucose tolerance of genetically obese (fa/fa) rats: influence of sex and genetic background. In both humans and rodents the occurrence and severity of obesity-associated non-insulin dependent diabetes mellitus (NIDDM) may be influenced by both gonadal hormones and genetic background. Early gonadectomy (at 3-5 days of age) of female and male Wistar diabetic fatty (WDF) rats and of male Zucker rats allowed us to examine these effects in genetically obese rats carrying the fatty (fa) gene. Impairment of glucose tolerance and insulin sensitivity by obesity, and amelioration or exacerbation (in the case of female rats) of this impairment by gonadectomy were assessed by intragastric glucose tolerance tests when the rats reached adulthood. Both glucose tolerance and insulin sensitivity were significantly deranged in obese WDF rats of both sexes and in obese male Zucker rats compared to lean controls of the same sex and strain. Obese male WDF rats were less glucose tolerant and insulin sensitive than were obese male Zucker rats. Glucose intolerance was not ameliorated by castration in lean or obese male WDF or Zucker rats. Insulin sensitivity was significantly improved by castration in obese male rats of both strains, as fasting plasma insulin levels and total areas under the insulin curves were significantly reduced compared to obese sham-operated controls. This effect was greater in the Zucker than in the WDF male rats. Castration significantly decreased the insulin response areas in obese male Zucker rats, but did not alter those of the obese male WDF rats. Ovariectomy did not alter glucose homeostasis of obese female WDF rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Sub-acute underfeeding but not acute starvation reduces noradrenaline induced thermogenesis in the conscious rat. The thermogenic responses to noradrenaline have been studied during food restriction in conscious rats. Acute starvation for 16-18 h reduced the body weight by 3% but not the resting metabolic rate or the increase in metabolic rate induced by noradrenaline, compared to control rats. Sub-acute underfeeding (eight days) with 50% of the control daily intake decreased body weight by 15% and was associated with a significant (P less than 0.05) reduction in resting metabolic rate and a significant (P less than 0.01) suppression of the thermogenic response to noradrenaline. Thus, food restriction reduces the thermogenic responses to noradrenaline, but only after a certain duration of restricted intake.


Body weight and coronary heart disease mortality: an analysis in relation to age and smoking habit. 15 years follow-up data from the Whitehall Study. 18,403 male civil servants aged 40-64 years were examined in London between 1968 and 1970. Mortality from all causes and specifically from coronary heart disease (CHD) over 15 years of follow-up was initially analysed in relation to deciles of body mass index (BMI = weight/height2) at entry into the study. In older men all causes mortality tended to be higher in those with a low BMI, but this was not so for CHD mortality. The latter was further studied after dividing the population into sub-groups according to age and cigarette smoking. With BMI distribution divided into fifths and five year age groups there were significant positive trends of CHD mortality across the BMI distribution in all age groups except the youngest (40-44 years) and oldest (60-64 years). For analysis by smoking category--never, ex- and current cigarette smoker--three age-specific groups were used: 40-49, 50-59 and 60-64 years. In men aged less than 60 years there were significant positive trends of CHD mortality and BMI in five of the six age and smoking categories, the exception being ex-smokers aged 40-49 years. Associations were strongest in the current smokers. By contrast in men aged 60-64 years there was a significant association between BMI and CHD mortality only in ex-smokers and this was of low order (P = 0.04). The data are compatible with some reports of a lesser association of obesity with mortality risk in older persons and in this data set the observation is not confounded by smoking habit.


Fat distribution and blood lipids in a sample of healthy elderly people. The cross-sectional sample consisted of data for 41 white men and 63 white women, 67-92 years of age who were healthy volunteer participants in the Aging Process Study at the University of New Mexico School of Medicine in Albuquerque, New Mexico. The variables consisted of anthropometric measures of body fatness, blood lipids and blood pressures. Correlations were computed between principal component scores, ratios of body circumferences, W/S2, blood lipid values and blood pressures for each sex. In the men, the significant correlations were of the abdomen/hip and abdomen/thigh ratios with W/S2, and the principal component scores with HDL cholesterol, triglyceride and systolic blood pressure. In the women, the abdomen/hip ratio had a low negative correlation with HDL cholesterol but a low positive correlation with triglyceride levels. The principal component scores also had low correlations with blood pressure and triglycerides. Multiple regressions were used to determine further associations between risk factors and fat distribution indices. In the men, the relationships of age and levels of body fatness with HDL cholesterol were much stronger and more complex than those with triglyceride or systolic blood pressure. In the women, only HDL cholesterol and triglyceride were associated with abdomen/hip ratio after removing the effects of overall fatness. The present findings indicate that a large abdominal circumference, implying a correspondingly large internal adipose tissue deposit, produces negative health alterations in blood lipid levels in this sample of elderly individuals. In younger adults, these changes are considered to increase the risk for cardiovascular disease.


Pregnancy, weight cycling and weight gain in obesity. Pregnancy constitutes a biological cause of weight cycling. Since repeated weight variation has been associated with sustained weight increase and subsequent metabolic complications, the role of pregnancy in affecting body weight was analysed retrospectively in a group of 128 severely obese patients at the obesity unit. Their present mean age was 47.8 +/- 10.7 years, the mean age at birth of their first child 24.1 +/- 5.1 years and their mean present BMI was 37.8 +/- 5.5 kg/m2 (s.d.). Seventy-three percent of the women reported a weight retention of more than 10 kg one year after delivery. The median number of other weight cycling events was seven. Such cycling did not correlate to present BMI or weight increase during any pregnancy. Weight increases during subsequent pregnancies were not correlated to each other. The results support the hypothesis that for women who develop severe obesity, their pregnancy-related weight increases contribute substantially to their body weight development.


Children at familial risk for obesity: an examination of dietary intake, physical activity and weight status. A large cohort of children was divided into two groups based on their parents' weight status. The high risk group had one or two overweight parent(s) (n = 92, 4.4 +/- 0.5 years), while the low risk group had no parent overweight (n = 95, 4.5 +/- 0.5 years). Weight of the two groups was similar at the start of the study but the high risk group gained marginally more weight over a one year period (P = 0.05). Although total energy intake was similar, the high risk group was consuming a larger percentage of energy from fat (P = 0.0004) and a smaller percentage from carbohydrate (P = 0.0002). Observed physical activity as similar for most levels but marginally higher for the stationary level and marginally lower in total activity in the high risk group. These results suggest a pattern that may lead to increased weight gain in a high risk group as they grow older.


Fluoxetine increases insulin action in obese nondiabetic and in obese non-insulin-dependent diabetic individuals. Insulin resistance contributes to the metabolic defects in non-insulin-dependent diabetes mellitus (NIDDM). Anorectic agents have been shown to improve insulin action in NIDDM, irrespective of weight reduction. The serotonin-reuptake inhibiting agent fluoxetine has recently been recognized as an anorectic agent. The effect of fluoxetine on insulin action has not yet been determined. In a double blind placebo controlled crossover study, we examined hepatic and peripheral insulin action by the sequential hyperinsulinemic euglycemic clamp technique with infusion of 3-3H-glucose in eight obese NIDDM and in eight obese nondiabetics, matched for age, sex and body mass index. Body weight was kept constant. After 14 days of fluoxetine, 60 mg daily, in NIDDM half-maximal peripheral glucose uptake was achieved at a lower insulin level than after placebo (ED50pgu 180.5 +/- 25.8 vs 225.3 +/- 39.9 mU/l, P less than 0.05), but not in nondiabetics (140 +/- 15.3 vs 135.3 +/- 22.2 mU/l, n.s.). Maximal peripheral glucose uptake (Vmaxpgu) did not change significantly. Basal hepatic glucose production (HGP) was reduced after fluoxetine in both NIDDM (9.45 vs 10.37 mumol/kg/min) and in nondiabetics (8.57 vs 9.16 mumol/kg/min), although the difference was only significant in nondiabetics (P less than 0.05). Multivariate analysis disclosed no differences in the effect of fluoxetine between NIDDM and nondiabetics. When nondiabetics and NIDDM were considered together, only the most insulin-resistant individuals demonstrated a decrease in ED50pgu (P less than 0.001). Likewise, only the individuals with the most outspoken hepatic insulin resistance demonstrated a decrease in insulin level, at which hepatic glucose production is completely suppressed (HGP0) (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


The body shape preferences of Japanese female students. A questionnaire was developed and administered to 433 female college students in 1985 and 973 females in 1990 to assess their body shape preferences. Their ideal weights for their current heights as estimated by them were analyzed using the body mass index. Our samples perceived their 'average' BMI at -0.5 s.d. of their mean current BMI. As a consequence they were very strict in setting the fat-borderline and tolerant in the thin-borderline. The perceived 'ideal' BMI was at -0.5 s.d. of their perceived 'average' and at about -1.0 s.d. of their actual average. Their extreme slim-body preference might have its root in the lack of proper understanding of what the 'average' body weight was, which might also be affected by the fact that they set their ideal shape at the level even lower than their misunderstood 'average'. The tendency was striking among those who considered their ideal body shape to be 'thin' and 'very thin'. These findings have not changed in the last five years in Japan. As for the 'contentment' regarding their weight, the most satisfied group changed from the nationally estimated 'thin' group to 'very thin' group in the last five years.


Bone structure and calcium metabolism in obese Zucker rats. Obesity is associated with altered bone mass. However, reports on bone status in obesity are inconsistent. Increased or normal bone mass was reported in obese adults but decreased bone mineral content was described in obese children. Therefore we evaluated the obese fa/fa rat as a possible model to assist in studies of bone metabolism in obesity. Obese and lean 14-week-old male rats underwent 24 h balance studies for calcium, magnesium and phosphate. Plasma calcium, magnesium, phosphate, immunoreactive parathyroid hormone, urinary cAMP (cyclic adenosine monophosphate) and femur bone histomorphometry were also analysed. Obese rats were heavier and had higher plasma insulin, cholesterol and triglycerides levels (P less than 0.05). A comparable positive balance for calcium, magnesium and phosphate was found in obese and lean rats. Total plasma calcium was higher in the obese, but albumin corrected calcium and plasma magnesium, phosphate and glucose were similar to the lean. In contrast to human obesity, obese rats were hypercalciuric, hypermagnisuric and hyperphosphaturic (P less than 0.05). iPTH and urinary cAMP were higher in the obese. Femora of fa/fa rats were shorter and lighter. Their bone osteoid surface and bone calcium content were similar to controls. Femora metaphysis in the obese had increased number of trabeculae, decreased trabecular width and higher erosion surface/bone surface ratio. Their diaphysis had increased cortical area/bone area and cortical width/bone width ratios and decreased medullary area. In summary, obese rats have higher iPTH, are hypercalciuric and have decreased bone mass. These last two observations differ from what is described in adult human obesity. Therefore, the obese fa/fa rat is of limited assistance in studies of bone status in adult human obesity. It might be of help in studies of bone metabolism in juvenile obesity.


Gelatinolytic metalloproteinase secretion patterns in ocular melanoma. Fifteen posterior uveal melanoma cell lines were analyzed qualitatively for gelatinolytic and caseinolytic proteinase activity after one to five in vitro passages. All 15 cell lines secreted a gelatinolytic metalloproteinase, with an apparent molecular weight of 72 kD, into protein-free culture media; nine of these secreted an additional gelatinolytic metalloproteinase with an apparent molecular weight of 92 kD. Neither species had the ability to degrade casein. This approach may provide insight into the mechanisms of tumor metastasis in uveal melanoma.


A quantitative assay of retrograde transported HSV in the trigeminal ganglion. The relationship between the dose of Herpes simplex virus type 1 (HSV) inoculated in the cornea and the amount of actively replicating virus recovered from mouse trigeminal ganglion cells 5 d after corneal scratch and inoculation was investigated with a tissue culture plaque assay. A dose response curve of productive viral replication was obtained. The estimated dose of HSV that produces half-maximal recovery of virus within the ganglion was 9.15 x 10(3) plaque forming units per eye, and the maximal amount of HSV recovered was 1.34 x 10(4) pfu per ganglion. This definition of infectivity as a function of dose will be useful for studying the effects of potential inhibitors of the binding, uptake, and transport of HSV by productively or latently infected trigeminal neurons.


[Carpal tunnel syndrome. Clinical and electrophysiologic aspects]. After a short pathophysiological review of what is known about the compression of the median nerve in the carpal tunnel, the symptoms and clinical features of carpal tunnel syndrome (CTS) are discussed. Causes and disorders associated with CTS are then reviewed. The usefulness of electrophysiological tests and procedures are discussed in a comprehensive overview and a strategy for the electrodiagnosis of CTS is proposed in mild or atypical cases.


[Prevention of cytomegalovirus infection following heart transplantation]. Cytomegalovirus (CMV) infection after heart transplantation (HTx) is a severe complication, which leads to long treatment and hospital stay. Even if prophylactic therapy with anti-CMV IgG antibodies is performed, there is a high incidence of infection, especially when the heart from a CMV positive donor is transplanted to a CMV negative recipient (high risk constellation). This study evaluates the prophylactic antiviral therapy with ganciclovir in CMV high risk constellation at HTx. Out of 108 HTx, 29 CMV negative recipients (IgG and IgM) received a heart from a CMV positive donor (IgG pos., IgM neg.). The control group (CO) (n = 8) was treated with anti-CMV IgG antibodies (Cytotect 2 ml/kg at day 0, 1, 2, 7, 14, 21,), whereas the study group (GAN) (n = 13) was treated with ganciclovir (7.5 mg/kg single dose n = 8, or 5 mg/kg in twice daily doses n = 5 from day 1 to 14). Urea, creatinine, white blood cell count and platelet count was controlled daily. No side effects on renal and bone marrow function were noted. Therapy was well tolerated. Both groups had similar immunosuppressive protocol (prophylactic cytolysis, prednisone, azathioprine and cyclosporin A) and were similar in age, sex, preoperative diagnosis and NYHA class. Seroconversion for CMV (IgM and IgG) was observed in 75% of CO and 31% of GAN (p less than 0.05). Clinical manifestations of CMV infection started in the second month after HTx with fever in both groups CMV-organ manifestations developed in 50% (or 67% of infected) in CO (enterocolitis 2, pneumonitis 3, tonsillitis 1), and in 15% (or 50% of infected) in GAN (pneumonitis 2, epididymitis 1) NS.(ABSTRACT TRUNCATED AT 250 WORDS)


[Results of surgical therapy of bronchial cancers with reference to adjuvant therapeutic measures in an advanced stage]. The prognosis of bronchial carcinoma especially in advanced tumor stage is poor despite different methods of treatment. The 5-year survival rate after radical operation is about 25-40% for all stages, and between 17-20% in stage III. Without therapy it is 3.5-9.5%. T3-tumors and N2-lymphnodes represent the limit of surgical therapy. From 1980 to 1988 we operated 456 patients because of bronchial carcinoma, 80% men (mean age 59.2 y) and 20% women (mean age 56.9 y). 83.1% had been smokers, 16.9% had been exposed to contaminants. At admission to the hospital 36% were in stage I, 13% in stage II, 47% in stage III, and 4% in stage IV. squamous cell carcinoma 40.9%, adenocarcinoma 26.1%, large cell carcinoma 21.9%, and small cell carcinoma 11.1%. Looking at all cases we mostly performed a lobectomy, in stage III tumors the most common operation was pneumonectomy. In this group only 57.5% of the patients were resectable. In 77 potentially curative operated patients in stage III with non-small cell carcinoma we performed an adjuvant radio-, chemo- or combined therapy. The cumulative survival rates for all patients in the 1st year were 85% in stage I, 67% in stage II, and 36% in stage III and IV. In the 3rd year 57%, 14% and 10%, and in the 5th year 41%, 7%, and 5%. In stage III the survival rates of the potentially curative operated vs. the exploratively operated patients were in the 1st year 50%/36%, in the 3rd year 16%/10%, and in the 5th year 11%/5%.(ABSTRACT TRUNCATED AT 250 WORDS)


Periodic acid incubation can replace hydrochloric acid hydrolysis and trypsin digestion in immunogold--silver staining of bromodeoxyuridine incorporation in plastic sections and allows the PAS reaction. We have examined the possibility of improving the present methods of detecting bromodeoxyuridine (BrdU) and for combining the PAS reaction with the BrdU detection by means of immunogold-silver staining (IGSS). This was done in testes fixed in Carnoy or Bouin, and in parts of the small intestine which were fixed in Carnoy or periodate-lysine-paraformaldehyde (PLP). All tissues were embedded in a mixture of glycol methacrylate and butanediol-monoacrylate. It was found to be impossible to carry out BrdU detection using HCl hydrolysis and trypsin digestion in combination with a PAS reaction. However, incubation of the plastic sections in periodic acid for a period of 30 minutes appeared to make it possible to eliminate the HCl denaturation step and to carry out a specific PAS reaction. Moreover, after incubation in periodic acid, trypsin digestion was no longer required to make the BrdU label accessible in GMA-embedded sections, nor to re-expose the antigenic sites in plastic sections of tissues fixed with cross-linking fixatives. In this way the loss of cell structures, which is inevitable when trypsin is used, can be avoided. Now a BrdU detection with improved morphology can be combined with the PAS reaction in the same plastic section in order to stain tissue carbohydrates. This is important for tumour diagnosis, where the PAS reaction can be very useful.


Serodiagnostic assay of hepatitis C virus infection using viral proteins expressed in Escherichia coli. Infection with hepatitis C virus (HCV) was analyzed by an enzyme-linked immunosorbent assay based on recombinant viral proteins encoded by regions of the putative viral core, NS3, NS4 and NS5, which were expressed in E. coli. Results showed that 106 of 124 cases (85.5%) of non-A, non-B chronic hepatitis and 43 of 45 cases (95.5%) of hepatocellular carcinoma, negative for HBV marker, were positive for antibodies against at least one of these viral proteins. One of 87 healthy individuals with normal alanine aminotransferase activity was positive for antibody against only the viral core, but was negative for HCV RNA. The serum of one patient with chronic hepatitis was positive for one of these proteins, but negative for HCV RNA. These findings in combination with results on detection of HCV RNA in the sera of patients with non-A, non-B chronic hepatitis indicated that 105 of 124 cases (84.6%) were positive for HCV infection. Sera that were negative for HCV antibodies against all these proteins were also negative for HCV RNA assayed by reverse transcription followed by the polymerase chain reaction. Screening of HCV infection by detecting viral antibodies in circulating blood using all these viral proteins is useful for reducing the number of ambiguous results in screening for viral infection. Thus, this assay system may be useful diagnostic purposes.


Influence of exogenous adrenocorticotropic hormone on estrous behavior in cattle. A two-trial experiment was conducted to determine the influence of ACTH on estrous behavior in cattle. In Trial 1, Holstein heifers (n = 20) received an injection of prostaglandin F2 alpha (PGF) during a synchronized diestrus and 30 h later were allotted randomly to receive (i.m.) either 1) 4 mL of gelatin (Veh) or 2) 320 units of ACTH in 4 mL of gelatin (ACTH). Eleven days after the PGF injection, all heifers were again injected with PGF, and they received either Veh or ACTH to complete a cross-over design. Treatment with ACTH decreased (P less than .05) the duration of estrus (12.0 +/- 1.9 vs 18.0 +/- 1.6 h for Veh) and increased (P less than .001) the interval to estrus after PGF injection (62.9 +/- 2.6 vs 43.7 +/- 2.2 h for Veh). Peak serum concentrations of progesterone (P4) and cortisol (C) were elevated (P less than .001) after ACTH compared with Veh. In Trial 2, ovariectomized Holstein cows (n = 12) were injected (i.m.) with .5 mg of estradiol benzoate (EB) and, 10 h later, were allotted randomly to receive (i.m.) either 1) 4 mL of gelatin (Veh) or 2) 320 units of ACTH in 4 mL of gelatin (ACTH). Seven days after the initial EB injection, all cows were again injected with .5 mg of EB and, 10 h later, received either Veh or ACTH to complete a cross-over design. Treatment with ACTH decreased (P less than .01) the proportion of cows in estrus (2/12 vs 11/12 for Veh) and increased (P less than .01) peak serum concentrations of P4 and C.(ABSTRACT TRUNCATED AT 250 WORDS)


In vitro effects of the ionophore lysocellin on ruminal fermentation and microbial populations. Batch and continuous culture techniques were used to evaluate the effect of the ionophore lysocellin on ruminal fermentation and microbial populations. In batch culture, .5 and 1 ppm (of the fluid) lysocellin markedly decreased (P less than .01) the acetate:propionate ratio without affecting fiber digestion, ammonia concentration, or culture pH. Greater concentrations of lysocellin had negative effects (P less than .05) on fiber digestion and increased (P less than .05) culture pH. In continuous culture, a low level of lysocellin (33 ppm of the diet DM or about .7 ppm of the fluid) decreased pH (P less than .05) and methane (P less than .05) production but had no effect on fiber digestion. Lysocellin tended to increase (P less than .05) OM digestion in corn-based diets but decreased OM digestion in barley-based diets (starch source x lysocellin interaction, P less than .05). In addition, the molar proportion of propionate was increased more in barley- than in corn-based diets. Total anaerobes and amylolytic and lactate-utilizing microorganisms were not affected by the ionophore. In continuous culture, cellulolytic and lactate-producing organisms were insensitive to lysocellin, but, in lysocellin-treated media, cellulolytic organisms were resistant, whereas lactic acid producers were sensitive to lysocellin at 4 ppm. In summary, the ionophore lysocellin alters ruminal fermentation by decreasing ruminal methane production and increasing the molar proportion of propionate; however, effects varied depending on whether corn or barley served as the primary starch source.


In vitro and in vivo studies of factors affecting digestion of feeds in synthetic fiber bags. Dry matter disappearance (DMD) and cellulose disappearance (CD) from alfalfa hay were measured simultaneously in synthetic fiber bags incubated in vitro. In general, DMD, CD, and pH were reduced (P less than .05) inside the bags compared with values in the fermentation flasks. Bags became fully expanded with gas soon after the fermentation started, which seemed to inhibit fluid movement into and out of the bag. By creating a vacuum in the flask, fluid exchange was enhanced, and DMD, CD, and pH increased (P less than .01) inside the bags. Sample size, type of fabric, and fabric pore size affected pH, DMD, and CD. Inoculum source (forage or concentrate diet) did not affect DMD in either bags or flasks. In contrast, DMD in bags incubated in vivo was affected (P less than .01) by animal diet. When bags were incubated in vivo inside a rigid plastic container, both pH and DMD were reduced (P less than .01) compared with bags suspended freely in the rumen. Direct physical action of ruminal contents on the bags apparently enhanced exchange of fluid. Bacterial concentrations in bags suspended freely in the rumen were greater than those in bags incubated inside containers. Synthetic fiber bags may be useful in comparing digestibility between diets; however, caution is needed in evaluating such data in absolute terms.


Comparison of larkspur alkaloid extract and lithium chloride in maintaining cattle aversion to larkspur in the field. Lithium chloride (LiCl) and larkspur (Delphinium barbeyi) alkaloid extract were compared in their effect as an emetic to create taste aversions to fresh larkspur. They were further compared in the field to determine whether the indigenous larkspur alkaloids were more effective in maintaining the aversion when conditioned cattle were subjected to the social pressure (social facilitation) of control cattle eating larkspur. Taste aversions were produced in two groups of 1-yr-old cattle by offering fresh larkspur and then gavaging with either LiCl at 200 mg/kg of BW or larkspur alkaloid extract at 1.1 to 1.6 mL/kg of BW. The third group (control) was gavaged with water. The alkaloid group was slower to form an aversion than the lithium group, requiring four doses compared with two doses. All groups were then taken to larkspur-infested rangeland to test the aversion in the field. In the first field trial in which groups grazed separately, both aversion-induced groups generally abstained from eating larkspur. In the second trial in which all groups grazed together, both aversion-induced groups consumed less than half as much larkspur as the controls, but neither group abstained completely. Larkspur alkaloids did not maintain the aversion to larkspur to a greater degree than did LiCl when aversion-induced cattle were subjected to social facilitation.


The effect of supplementary Aspergillus niger phytase in diets for pigs on concentration and apparent digestibility of dry matter, total phosphorus, and phytic acid in different sections of the alimentary tract. Six barrows of approximately 37 kg BW, fitted with two simple T-cannulas in the duodenum (25 cm posterior to the pylorus) and terminal ileum (12 to 15 cm anterior to the ileocecal junction), were fed two diets containing 2.1 g of P/kg in the form of phytic acid and a low intrinsic phytase activity (corn-soybean meal based diet [Diet A] or a typical Dutch diet [Diet B]) without or with supplementary microbial phytase from Aspergillus niger (var. ficuum) equal to 1,500 phytase units per kilogram of diet, in a crossover design. The apparent duodenal, ileal, and total tract (overall) digestibilities of DM, total P, and phytate P (phytic acid x .282) were calculated using both Cr-NDR (neutral detergent residue mordanted with Cr) and Co-EDTA as dual-phase markers. Concentration of total P in the ileal digesta (P less than .01) and feces (P less than .001) of pigs fed microbial phytase was lower than without this enzyme, irrespective of the diet. Ileal digestibility of total P was 18.5 and 29.8 percentage units higher (which was a 1.7- to 2.9-fold increase) due to added Aspergillus niger phytase (P less than .05). Also, total tract (overall) digestibility increased by 27.0 to 29.7 percentage units (P less than .01). Phytic acid concentration in the duodenal and ileal digesta of pigs receiving microbial phytase was lower (P less than .01 or .001), resulting in its higher ileal digestibility (dephosphorylation rate) by 50.1 percentage units for Diet A and by 75.4 percentage units for Diet B. Irrespective of the treatment, no phytase activity could be detected in the ileal digesta of pigs.


Dietary fiber for dogs: III. Effects of beet pulp and oat fiber additions to dog diets on nutrient intake, digestibility, metabolizable energy, and digesta mean retention time. The objective of this experiment was to determine whether alkaline hydrogen peroxide-treated oat hulls (termed oat fiber; OF) are nutritionally efficacious as a source of dietary fiber in meat-based dog foods. Thirty female English Pointers were assigned in a completely randomized design to isonitrogenous diets. Treatments were 1) control diet, 2) 7.5% added beet pulp (BP), and 3) 2.5, 4) 5.0, and 5) 7.5% added OF. Inclusion of 7.5% BP increased (P less than .05) DM intake and decreased (P less than .05) digestibility of DM and OM compared with the control. Dry matter intake increased (P less than .05) with increasing level of OF and digestibility of DM, OM, and total dietary fiber (TDF) decreased (P less than .05). Digestibility of DM, OM, and TDF were higher for dogs fed the 7.5% BP than for those fed the 7.5% OF treatment. Digestible energy, expressed as a percentage of GE, was greater for the control treatment than for the 7.5% BP treatment. A linear decrease in DE (percentage of GE) was noted as the concentration of OF increased, and the DE value (percentage of GE) for the 7.5% BP treatment was greater (P less than .05) than that for the 7.5% OF treatment. A linear decrease (P less than .05) was noted in ME, expressed as a percentage of GE, as the level of OF increased. Frequency of defecation and mean retention time were unaffected (P greater than .05) by treatment. Oat fiber was an effective substitute for BP in dog diets.


Biokinetics and biliary excretion of radiotocopherol administered orally to sheep. Four adult sheep were given .4 microCi/kg of BW of D-alpha[5-Me-3H]tocopherol orally. Plasma alpha-tocopherol specific activities were measured serially during a 72-h period, after which the sheep were killed. The disposition kinetics were best described by the use of a two-compartment model. The radiotocopherol had a slow distribution phase followed by a relatively slow elimination phase. During the 72-h period of serial monitoring the peak plasma specific activities were observed between 32 to 48 h. At slaughter (72 h after dosing) tissue radioactivity distribution indicated a high rate of accumulation in some glandular organs such as the liver and adrenals. Identification of radioactive components excreted in bile, using HPLC or thin-layer chromatography, showed that unchanged radiotocopherol was present only at very low concentrations (less than 3% of the total recovered radioactivity).


Voluntary intake and ingestive behavior of steers grazing Johnstone or endophyte-infected Kentucky-31 tall fescue. Effects of grazing low-endophyte (Acremonium coenophialum Morgan-Jones and Gams, less than 1% infection) Johnstone (J) or high-endophyte (60% infection) Kentucky-31 (K) tall fescue (Festuca arundinacea Schreb.) on grazing behavior and voluntary intake were studied. Six Angus steers (average initial BW = 326 kg) grazed 1.21-ha plots of each forage cultivar (three steers per cultivar) in four 28-d periods beginning May 27. Daytime observations (0630 until 2130) revealed that J steers spent more (P less than .10) time grazing and lying down and took more (P less than .05) prehensile bites than K steers did; conversely, steers grazing K spent more (P less than .10) time standing and idling than J steers did. Idling time showed a forage x period interaction (P less than .10). Mean OM bite size (grams per bite) was not affected (P greater than .10) by forage but differed (P less than .10) among periods. Limited nighttime observations (2130 until 0630) revealed no effects (P greater than .10) of forage on grazing time or number of prehensile bites taken. Voluntary intakes of OM and NDF did not differ (P greater than .10) between steers grazing J and K; however, a forage x period interaction (P less than .10) existed such that, during Period 1, steers grazing J had greater (P less than .01) OM and NDF intakes than did steers grazing K. These data suggest that cattle grazing endophyte-infected tall fescue display altered daytime grazing behavior and that reduction of voluntary intake attributable to endophyte infection may be less severe under free-grazing than has been reported for controlled environmental conditions.


Diet characteristics, digesta kinetics, and dry matter intake of steers grazing eastern gamagrass. Eastern gamagrass (Tripsacum dactyloides [L.] L.) has attracted attention as a forage crop, but information on its use is lacking. This 2-yr study compared diet quality, ingestive mastication, and ADG by steers grazing eastern gamagrass (GG), flaccidgrass (Pennisetum flaccidum Griseb.), and Tifton 44 bermudagrass (Cynodon dactylon [L.] Pers.). The design was a randomized complete block with two agronomic replicates. The diet selected by steers from GG in May did not differ from the diet selected by steers from flaccidgrass (FG) for IVDMD (77.3%), NDF (44.0%), CP (19.5%), and mean and median particle sizes of the ingesta (1.8 and 1.4 mm). In July, GG diets had three percentage units less IVDMD (P less than .05), 8.4 percentage units more NDF (P less than .05), and 4.5 percentage units less CP (P = .07) than the mean of FG and bermudagrass (BG). The canopy (July) of GG had the greatest proportion of its DM as leaf (59 vs 26% for FG and 22% for BG) and the least proportion as stem (25 vs 40% for FG and 59% for BG). Mean particle size (millimeters) of masticates differed (P = .05) among forages with GG greatest (2.2), followed by FG (1.6), and BG particles were smallest (1.2). Proportion of large (greater than or equal to 2.8 mm), medium (less than 2.8 greater than or equal to .5 mm), and small (less than .5 mm) particles of the masticate DM, and their IVDMD and NDF concentration, interacted with species (P less than .05). Gamagrass masticate had the greatest proportion (28%) of large particles and BG the greatest proportion (23%) of small particles. The least IVDMD occurred for large particles of BG (62.5%) and small particles of GG (63.8%). Digesta kinetics did not differ among species. Characteristics of GG yielded steer ADG of .82 vs .67 kg for FG and .30 kg for BG (P = .05).


Isolation and immortalization of rat pre-type II cell lines. The fetal respiratory distress syndrome is due, in part, to the presence of abundant pre-type II alveolar epithelial cells that have not yet differentiated into mature type II cells. Studies of this syndrome have been limited somewhat by the lack of an adequate in vitro model. In the present study we immortalized pre-type II cells by infecting primary isolates obtained from fetal rat lung with a retroviral construct expressing the adenoviral 12S E1A gene product. The immortalized pre-type II cells retained many of the ultrastructural features typical of pre-type II cells in primary culture, most notably lamellar bodies were not detected and the cells contained abundant stores of glycogen, expressed cytokeratin filaments, and bound the lectin Maclura pomifera. Karyotyping revealed that the cells are diploid. Growth studies demonstrate log phase growth in the presence of serum with a markedly decreased growth rate shortly after the cells reach confluence. Exposure of the immortalized pre-type II cells to hydrocortisone and dibutyryl cAMP resulted in the induction of lamellar bodylike organelles; however, these cells did not secrete surfactant or express surfactant protein A. These cells may serve as useful models for some in vitro studies of fetal type II cell maturation or the fetal respiratory distress syndrome, or both.


Cytotoxicity in feline leukemia virus subgroup-C infected fibroblasts is mediated by adherent bone marrow mononuclear cells. The pathogenesis of retrovirus-induced erythroid aplasia in cats is unknown. In studies to define mechanisms of cytotoxicity associated with retroviral infections, bone marrow mononuclear cells (BMMC) from healthy specific pathogen-free cats were co-cultured with uninfected feline embryonic fibroblasts (FEA cells) and FEA cells infected with feline leukemia virus (FeLV) of subgroup A (FEA-A) or subgroup C (FEA-C). Moderate to marked cytotoxicity (CPE) developed in co-cultures of BMMC and FEA-C cells on Days 5 to 7 of incubation but not in co-cultures of BMMC and FEA-A or BMMC and uninfected cells (FEA-CT). Cytotoxicity was associated with adherent cells of light density (1.056) from bone marrow and peripheral blood, which were positive for alpha naphthyl butyrate esterase activity. Stimulation of adherent cells with phorbol ester or addition of recombinant human tumor necrosis factor-alpha (rhTNF-alpha) caused similar CPE in FEA-CT cells. The TNF-alpha concentrations in the culture supernatants of BMMC+FEA-C were higher than those of BMMC+FEA-A or BMMC+FEA-CT, and addition of anti-TNF antibodies to the cultures blocked the CPE. These data support the hypothesis that macrophages exposed to FeLV-C cause CPE in co-cultures of BMMC and FEA cells by a mechanism involving TNF-alpha. It is suggested that TNF-alpha may be involved in the suppression of hematopoiesis in cats which develop FeLV-C induced erythroid aplasia.


Endothelin-induced changes in intracellular calcium concentration in rat vascular smooth muscle cells: effects of extracellular calcium and atrial natriuretic factor. The purpose of this study was to examine whether different mechanisms might underlie the changes in intracellular calcium concentration ([Ca2+]i) stimulated by high and low concentrations of endothelin, and whether atrial natriuretic factor (ANF) has an inhibitory effect on endothelin-induced [Ca2+]i changes in cultured rat vascular smooth muscle cells (VSMCs). In calcium-replete buffer, cultured monolayers of rat VSMCs superfused with endothelin at a high concentration (10 nM) exhibited a marked transient rise in [Ca2+]i, followed by a sustained elevation, whereas a low concentration of endothelin (0.1 nM) induced a sustained monophasic elevation. When calcium-free buffer was used, 10 nM endothelin induced a transient rise in [Ca2+]i of lesser amplitude, whereas 0.1 nM endothelin did not produce a significant rise. Pretreatment of VSMCs with ANF and cosuperfusion with endothelin failed to inhibit either transient or sustained endothelin-induced changes in [Ca2+]i in calcium-replete buffer.


Effects of new intravascular contrast agents on [Ca2+]i transients and contraction in cultured ventricular myocytes. We examined the effects of four kinds of intravascular contrast agents (amidtrizoic acid, iohexol, iopamidol, and ioxaglic acid) on [Ca2+]i transients (indo-1 fluorescence) and cell contraction (video motion analyzer), using cultured chick embryo ventricular myocytes. Exposure of ventricular myocytes to amidtrizoic acid (a conventional contrast agent) reduced the [Ca2+]i transients and the sensitivity of the contractile elements to [Ca2+]i. Ioxaglic acid (a low osmotic contrast agent) also reduced the [Ca2+]i transients, but did not significantly change the sensitivity of the contractile elements to [Ca2+]i. Neither iohexol nor iopamidol (nonionic contrast agents) reduced the [Ca2+]i transients, but both significantly decreased the sensitivity of the contractile elements to [Ca2+]i. A marked negative inotropic effect of amidtrizoic acid was caused by both calcium binding and hypertonicity. The less marked depression of contractility produced by ioxaglic acid is possibly the result of calcium binding, but is not caused by hypertonicity. The negative inotropism produced by nonionic contrast agents (iohexol and iopamidol) was due to hypertonicity, but not due to alterations in the [Ca2+]i transients. Exposure of ventricular myocytes to nonionic contrast agents (iohexol and iopamidol) slowed decay in the [Ca2+]i transients with increased end-diastolic [Ca2+]i. After washing out the nonionic contrast agents, these parameters returned to control levels. On the other hand, exposure to amidtrizoic acid decreased end-diastolic [Ca2+]i without changing decay time in the [Ca2+]i transients. After washing out amidtrizoic acid, there was a prolongation of half decay time in [Ca2+]i transients with a significant increase in end-diastolic [Ca2+]i and cell position. Diastolic dysfunction just after washout of amidtrizoic acid was possibly caused by an increase in [Na+]i due to sodium influx during exposure to the contrast agent.


Similar oxygen cost of myocardial contractility between DPI 201-106 and epinephrine despite different subcellular mechanisms of action in dog hearts. The effects of DPI 201-106 (a novel, cyclic AMP-independent positive inotropic agent with Ca(2+)-sensitizing and Na(+)-channel agonistic mechanisms) on myocardial mechanics and energetics were assessed in the excised cross-circulated dog left ventricle. In the first protocol, the relation between left ventricular oxygen consumption (VO2) and systolic pressure-volume area (PVA) was analyzed before and during administration of DPI 201-106. The reciprocal of the slope of the VO2-PVA relation has been shown to reflect the contractile efficiency, and the VO2-intercept consists of the oxygen cost of contractility-dependent excitation-contraction coupling and basal metabolism. DPI 201-106 increased Emax (contractility index) and elevated the VO2-PVA relation in a parallel manner, i.e., the VO2-intercept increased without a change in the slope. In the second protocol, the increase in the VO2-intercept of the VO2-PVA relation for a unit increase in Emax (i.e., oxygen cost of enhanced contractility) was compared between DPI 201-106 and epinephrine in a paired manner in each heart. Epinephrine significantly abbreviated the time to end systole, whereas DPI 201-106 did not, suggesting that the mechanism of inotropic action differed between the two drugs. However, the oxygen cost of enhanced contractility was the same between the two drugs in each heart. Therefore, DPI 201-106 did not alter the contractile efficiency nor spare the oxygen cost of enhanced contractility as compared to epinephrine under the present experimental conditions. This suggests that the Ca(2+)-sensitizing effect of DPI 201-106, if any, is too small to spare the oxygen cost of contractility in the blood-perfused, non-failing dog heart.


Radioiodinated anti-hepatocellular carcinoma (HCC) ferritin. Targeting therapy, tumor imaging and anti-antibody response in HCC patients with hepatic arterial infusion. Radioimmunoimaging and radioimmunotherapy with radioiodinated anti-(hepatocellular carcinoma ferritin) antibody (131I- or 125I-FtAb) have been applied in patients with primary liver cancer. A total of 41 patients with surgically unresectable hepatocellular carcinoma (HCC) and receiving hepatic artery ligation and cannulation during exploratory laparotomy were treated with this regimen by intrahepatic arterial infusion. Compared with the control group, a decline of serum alpha-fetoprotein (65.7% versus 42.9%) and shrinkage of tumor (68.3% versus 33.9%) were observed in the treated group, and a higher second-look resection rate (31.7% versus 5.1%) and longer survival (1-year: 61.0% versus 37.3%, 3-year: 25.0% versus 6.9%) resulted. The administration of antibody through a hepatic arterial catheter (n = 16) was compared with intravenous injection (n = 17) in terms of the tumor-imaging sensitivity in 33 patients with liver cancer. The results indicated that hepatic arterial infusion was superior to intravenous injection. The sensitivity 7 days after the administration was 100% in the i.a. group and 76.5% in the i.v. group, the uptake ratio of tumor to liver being 1.74 +/- 0.57 in the former and 1.34 +/- 0.29 in the latter. Furthermore, intrahepatic arterial infusion revealed a lower anti-antibody detection rate than intravenous injection (0/14 versus 4/11).


Substrates and signalling complexes: the tortured path to insulin action. In the last few years several potential substrates of the insulin receptor tyrosine kinase have been identified, purified, and their cDNAs isolated. These putative substrates include: 1) pp15, a fatty acid-binding protein; 2) pp120, a plasma membrane ecto-ATPase; 3) pp42, a MAP serine/threonine kinase; 4) pp85, a subunit of the Type 1 phosphatidylinositol kinase; and 5) pp185, a phosphatidylinositol kinase binding protein. Although the tyrosine phosphorylation of several of these substrates correlates with the signalling capabilities of various mutant receptors, the role of these substrates in mediating any one of insulin's many biological responses is still unknown. In addition, recent data indicate that the tyrosine phosphorylation of pp42 may in fact be due to autophosphorylation, thereby removing it from the list of putative substrates of the insulin receptor kinase. Finally, the present review discusses the question of whether signalling occurs as a result of the tyrosine phosphorylation of substrates or via the formation of signalling complexes.


The insulin-binding domain of insulin receptor is encoded by exon 2 and exon 3. Insulin receptors are disulfide-linked oligotetramers composed of two heterodimers each containing a 130-kDa alpha subunit and a 90-kDa beta subunit. Insulin binds to the extracellular alpha subunit, and in the process stimulates the autophosphorylation of the beta subunit and the expression of tyrosine kinase activity. Studies combining the use of photoaffinity labeling and immunoprecipitation with anti-peptide antibody have directly demonstrated that the cysteine-rich domain, encoded by exon 3, in the alpha subunit is part of the insulin-binding site of the receptor. Experiments with chimeric insulin receptors and chimeric insulin-like growth factor I receptors have confirmed that the cysteine-rich domain constitutes a part of the insulin-binding site. In addition, results from these experiments suggest that the N-terminal sequence, encoded by exon 2, in the alpha subunit also participates in insulin binding. In this review it is proposed that, assuming two insulin-binding sites per each holoreceptor oligotetramer, each insulin-binding domain may contain respectively two sub-domains for hydrophobic and charge contact with insulin, and that high-affinity binding would require the interaction of both subunits with the possibility of each subunit reciprocally contributing one of the sub-domains.


Insulin receptor signaling through non-tyrosine kinase pathways: evidence from anti-receptor antibodies and insulin receptor mutants. Although there is general agreement that insulin receptor tyrosine kinase activity mediates many of the actions of insulin, two types of studies suggest that non-tyrosine kinase dependent pathways may also exist. First, both monoclonal and polyclonal antibodies to the receptor have been shown to mediate many of insulin's actions with little or no stimulation of receptor kinase. Second, insulin receptor mutants, with reduced or no tyrosine kinase activity, have been shown to mediate several actions of insulin. Non-tyrosine kinase pathways that could signal insulin effects through the insulin receptor include non-covalent activation of G proteins, phospholipase Cs, or docking proteins such as IRS-1. Further studies on the chemical structures of phospholipids and their hydrolysis products involved in insulin action will be required to sort out the underlying mechanisms of insulin action via non-tyrosine kinase dependent pathways.


Desensitization of the insulin-secreting beta cell. In human diabetes, inherent impaired insulin secretion can be exacerbated by desensitization of the beta cell by chronic hyperglycemia. Interest in this phenomenon has generated extensive studies in genetic or experimentally induced diabetes in animals and in fully in vitro systems, with often conflicting results. In general, although chronic glucose causes decreased beta-cell response to this carbohydrate, basal response and response to alternate stimulating agents are enhanced. Glucose-stimulated insulin synthesis can be increased or decreased depending on the system studied. Using a two-compartment beta-cell model of phasic insulin secretion, a unifying hypothesis is described which can explain some of the apparent conflicting data. This hypothesis suggests that glucose-desensitization is caused by an impairment in stimulation of a hypothetical potentiator singularly responsible for: 1) some of the characteristic phases of insulin secretion; 2) basal release; 3) potentiation of non-glucose stimulators; and 4) apparent "recovery" from desensitization. Review of some of the pathways that regulate insulin secretion suggest that phosphoinositol metabolism and protein kinase-C production are regulated similarly to the theoretical potentiator and their impairment is a major contributor to glucose desensitization in the beta cell.


Protein-tyrosine phosphatases and the regulation of insulin action. Protein-tyrosine phosphatases (PTPases) play an important role in the regulation of insulin action by dephosphorylating the active (autophosphorylated) form of the insulin receptor and attenuating its tyrosine kinase activity. PTPases can also modulate post-receptor signalling by catalyzing the dephosphorylation of cellular substrates of the insulin receptor kinase. Dramatic advances have recently been made in our understanding of PTPases as an extensive family of transmembrane and intracellular proteins that are involved in a number of pathways of cellular signal transduction. Identification of the PTPase(s) which act on various components of the insulin action cascade will not only enhance our understanding of insulin signalling but will also clarify the potential involvement of PTPases in the pathophysiology of insulin-resistant disease states. This brief review provides a summary of reversible tyrosine phosphorylation events in insulin action and available data on candidate PTPases in liver and skeletal muscle that may be involved in the regulation of insulin action.


Insulin/IGF-1 hybrid receptors: implications for the dominant-negative phenotype in syndromes of insulin resistance. Classical insulin and IGF-1 receptors are alpha 2 beta 2 heterotetrameric complexes synthesized from two identical alpha beta half-receptor precursors. Recent data strongly suggests, however, that nonidentical alpha beta half-receptor precursors can assemble to generate hybrid holoreceptor species both in vivo and in vitro. This review focuses primarily on two types of hybrid receptors. The first type is an insulin/IGF-1 hybrid receptor generated by the association of an alpha beta insulin half-receptor with an alpha beta IGF-1 half-receptor. The second type is one formed from a wildtype (kinase-active) insulin or IGF-1 alpha beta half-receptor and a mutant (kinase-inactive) insulin alpha beta half-receptor. Although the functional properties of insulin/IGF-1 hybrid receptors have not yet been completely defined, wildtype/mutant hybrid receptors are essentially substrate kinase inactive. These data indicate that the mutant alpha beta half-receptor exerts a transdominant inhibition upon the wildtype alpha beta half-receptor within the alpha 2 beta 2 holoreceptor complex. This defect in substrate kinase activity may contribute to the molecular defect underlying some syndromes of severe insulin resistance and diabetes. Heterozygous individuals expressing both wildtype and mutant tyrosine kinase-defective insulin receptor precursors demonstrate varying degrees of insulin resistance and diabetes. In addition, cell lines which express both endogenous wildtype and transfected kinase-defective insulin receptors display markedly decreased insulin and IGF-1 sensitivity and responsiveness. Formation of hybrid receptors which results in premature termination of insulin signal transduction may be one mechanism underlying the observation that kinase-inactive receptors inhibit the function of native receptors.


A monoclonal antibody to the T-cell receptor increases IGF-I receptor content in normal T-lymphocytes: comparison with phytohemagglutinin. The biological effects of the IGFs are mediated through interaction with specific cell surface receptors. It has been previously reported that mitogenic activation of T-lymphocytes by phytohemagglutinin (PHA) is associated with increased IGF-I receptor content. However, the mechanisms which regulate IGF-I receptor expression during T-lymphocyte activation are unknown. To explore further the regulation of IGF-I receptor expression in T-cells, we investigated IGF-I receptor content and mRNA abundance in T-lymphocytes after stimulation either by PHA or OKT-3, the latter being a monoclonal antibody directed against the CD-3 antigen of the T-cell receptor. IGF-I binding in T-cells demonstrated increased IGF-I receptor content after stimulation by both PHA and OKT-3. Peak binding was induced after 72 h of treatment with PHA and 48 h of treatment with OKT-3. Affinity cross-linking of 125I-IGF-I to T-cell membranes demonstrated a single approximately 130 kDa band which was increased after treatment with PHA or OKT-3. This band was inhibited by the addition of alpha-IR3, a monoclonal antibody to the IGF-I receptor. Both PHA and OKT-3 increased IGF-I receptor mRNA abundance with peak increases at 20 h and 60 h, respectively. Parallel increases in IGF-I receptor and beta-actin mRNA abundance were observed, consistent with previous studies demonstrating increased actin gene expression after T-cell activation. Thus, the increase in IGF-I receptor mRNA abundance markedly preceded the increase in IGF-I receptor content after PHA stimulation, but the increase in IGF-I receptor mRNA abundance followed the increase in IGF-I receptor content after OKT-3.(ABSTRACT TRUNCATED AT 250 WORDS)


Retention reproducibility of basic drugs in high-performance liquid chromatography on a silica column with a methanol-high-pH buffer eluent. Changes in selectivity with the age of the stationary phase. The long-term reproducibility of the separation of basic drugs by high-performance liquid chromatography using a silica column with a methanol-high-pH eluent was examined. The relative capacity factors of specific tertiary amines changed systematically compared with the other drugs on storage of the columns for 1 year. The changes were enhanced on storing the column in mobile phase and were accelerated by prolonged washing with water. A comparison of freshly packed columns suggested that similar changes also occurred with the dry silica on storage. These effects may be a contributor to previously observed batch-to-batch differences in the silica.


Clinical evaluation of oral fluid samples for diagnosis of viral hepatitis. Oral fluid samples were compared with serum samples as a specimen source for hepatitis A, B, and C virus markers. Oral fluid was obtained with a treated absorbent pad and tested by using existing commercial enzyme immunoassays with only minor modifications. Compared with serum sampling the sensitivity and specificity of oral sampling were 100% (51 of 51 samples) and 98% (46 of 47 samples) for hepatitis A virus immunoglobulin M, 100% (29 of 29 samples) and 100% (29 of 29 samples) for hepatitis B virus surface antigen, and 100% (13 of 13 samples) and 100% (13 of 13 samples) for hepatitis C virus antibody, respectively. The decline of hepatitis A virus immunoglobulin M in oral samples was parallel to, though somewhat more rapid than, that of hepatitis A virus immunoglobulin M in serum samples. It is proposed that oral sampling represents a safer and more convenient procedure for reliable hepatitis virus testing than blood sampling and that it has wide application in patient and outbreak management.


Enhanced shedding of cytomegalovirus in semen of human immunodeficiency virus-seropositive homosexual men. Site-specific shedding of cytomegalovirus (CMV) was assessed in a longitudinal study of homosexual and bisexual men. At initial testing, CMV was cultured from the semen of 33% (19 of 58) of asymptomatic and mildly symptomatic men who were seropositive for human immunodeficiency virus (HIV) at the time of entry into the study, whereas it was cultured from the semen of 17% (10 of 58) of the men who were HIV seronegative. CMV was isolated much more frequently from semen than from urine or throat washing specimens, and it was rarely recovered from stool or blood, regardless of the subject's HIV serostatus. CMV was cultured from the semen of 31% (16 of 52) of the men relatively early after seroconversion to HIV (mean, 12.8 months). CMV was persistently isolated from the semen of a greater proportion of the HIV-seropositive men than from the semen of the HIV-seronegative men during a 4.5-year follow-up period (52 of 110 - [47%] and 15 of 58 [26%] men, respectively). There was an increased relative risk for shedding of CMV in semen in association with decreased CD4+ cell numbers and increased levels of serum immunoglobulin A. However, there was no association of CMV shedding with an increased risk for the development of AIDS.


Interpretive criteria for susceptibility testing of CI-960 (PD127391, AM-1091), fleroxacin, lomefloxacin, and temafloxacin against Neisseria gonorrhoeae, including drug stability in GC agar medium. CI-960, fleroxacin, lomefloxacin, and temafloxacin were tested against over 100 strains of Neisseria gonorrhoeae. Each organism was tested in triplicate by using agar dilution and disk diffusion methods recommended by the National Committee for Clinical Laboratory Standards. CI-960 was the most potent compound, with a MIC against 90% of the strains tested of 0.008 microgram/ml, and the least active was fleroxacin (MIC against 90% of strains, 0.12 microgram/ml). Only the susceptible interpretive category was recommended for the CI-960 tests as follows: 5-micrograms disk, greater than or equal to 39 mm (MIC correlate, less than or equal to 0.12 microgram/ml). Three interpretive categories were proposed for the other fluoroquinolones as follows: fleroxacin, 5-micrograms disk susceptible at greater than or equal to 33 mm (MIC correlate, less than or equal to 0.25 microgram/ml), intermediate at 28 to 32 mm (MIC correlate, 0.5 microgram/ml), and resistant at less than or equal to 27 mm (MIC correlate, greater than 0.5 microgram/ml); lomefloxacin, 10-micrograms disk susceptible at greater than or equal to 35 mm (MIC correlate, less than or equal to 0.12 microgram/ml), intermediate at 28 to 34 mm (MIC correlates, 0.25 to 0.5 microgram/ml), and resistant at less than or equal to 27 mm (MIC correlate, greater than 0.5 microgram/ml); and temafloxacin, 5-micrograms disk susceptible at greater than or equal to 36 mm (MIC correlate, less than or equal to 0.06 microgram/ml), intermediate at 28 to 35 mm (MIC correlates 0.12 to 0.25 microgram/ml), and resistant at less than or equal to 27 mm (greater than 0.25 microgram/ml). Interpretive agreement between disk diffusion results and the MICs was 100% for each agent, with the exception of lomefloxacin, which had a 0.9% minor error. All drugs were stable in GC agar medium for at least 21 days when stored at 2 to 5 degrees C.


Comparison of different immunostaining techniques and monoclonal antibodies to the lower matrix phosphoprotein (pp65) for optimal quantitation of human cytomegalovirus antigenemia. The main parameters of immunostaining techniques, i.e., the type of fixative, immunocytochemical reaction, and quality of monoclonal antibodies (MAbs), for quantitation of human cytomegalovirus (HCMV) antigenemia in peripheral blood polymorphonuclear leukocytes (currently performed by the indirect immunofluorescence or immunoperoxidase reaction by using MAbs to HCMV pp65) were investigated in order to optimize procedural steps and reagents. Significantly better results (in terms of the number of positive cells) were obtained on multiple cytospin preparations from heart transplant recipients with HCMV viremia when we used (i) formalin instead of methanol-acetone fixation and (ii) the indirect immunofluorescence reaction instead of the immunoperoxidase reaction, the avidin-biotin complex method, or the alkaline phosphatase antialkaline phosphatase procedure. In addition, comparison of the staining capabilities of three MAbs to pp65, which were developed in the laboratory and which were reactive to different epitopes of the protein, with a commercially available MAb (Clonab CMV) for determination of HCMV antigenemia showed that, while individual MAbs did not provide better results, the pool of MAbs detected a significantly higher number of positive peripheral blood polymorphonuclear leukocytes than Clonab CMV did. In addition, the sensitivity of the pool in detecting patients with low levels of viremia (less than 5/2 x 10(5) cells inoculated) as antigenemia positive was 100%, whereas the sensitivity of Clonab CMV was 47%. No differences in the specificities between the two MAb preparations were observed.


High correlation in antibody titers between the Sabin-Feldman dye test and an enzyme-linked immunosorbent assay detecting immunoglobulin G antibodies to the nucleoside triphosphate hydrolase of Toxoplasma gondii. An enzyme-linked immunosorbent assay to detect immunoglobulin G antibodies against nucleoside triphosphate hydrolase, which is a specific and dominant antigen of Toxoplasma gondii, was developed, and the sensitivity and specificity of the test were compared with those of the Sabin-Feldman dye test. One hundred percent agreement was observed in comparative study between those tests on 37 positive and 50 negative human sera. Antibody titers in the enzyme-linked immunosorbent assay test, which were expressed as the reciprocal of the highest positive dilution of serum, were just 100 times those in the dye test on 81% (30 of 37) of the positive sera.


Detection of human group C rotaviruses by an enzyme-linked immunosorbent assay using monoclonal antibodies. An enzyme-linked immunosorbent assay using monoclonal antibodies was established for the detection of human group C rotaviruses. Seventeen clinical samples which were found to contain group C rotaviruses were all strongly positive, whereas 9 samples containing group A rotaviruses and 51 samples lacking rotaviruses were all negative with this test.


Comparison of the Western immunoblot assay and a glycoprotein G enzyme immunoassay for detection of serum antibodies to herpes simplex virus type 2 in patients with AIDS. Herpes simplex virus type 2 (HSV-2) seroprevalence in 68 patients with AIDS was 77% by Western blot (immunoblot) and 44% by glycoprotein G-2 immunoassay. Each of 16 patients with culture-proven HSV-2 infection was positive by Western blot versus 8 by glycoprotein G-2 immunoassay. No differences in age, race, duration of AIDS, acyclovir usage, or HSV-1 seroprevalence were found to explain differences in sensitivity.


MIC and disk diffusion quality control guidelines for Neisseria gonorrhoeae susceptibility tests of cefdinir, cefetamet, CI-960, fleroxacin, lomefloxacin, and temafloxacin. Cefdinir (FK482), cefetamet (Ro 15-8074), CI-960, fleroxacin, lomefloxacin, and temafloxacin have potent activities against Neisseria gonorrhoeae. They were tested in a multilaboratory study to establish quality control guidelines. Quality control ranges for N. gonorrhoeae ATCC 49226 were determined by using multiple GC agar lots, three disk lots, and a number of test replicates consistent with the M23-T guidelines of the National Committee for Clinical Laboratory Standards. The MIC ranges included 2 to 4 log2 dilution steps. The recommended inhibition zone diameter ranges were generally 7 to 8 mm and included greater than or equal to 91.3% of all recorded study results.


Peripheral cholangiocarcinoma: sonographic evaluation. One hundred and seven patients who had intrahepatic tumor that demonstrated by sonography, and proved histologically, to be cholangiocarcinoma (CC), were reviewed retrospectively. Two main tumor patterns were found, namely the nodular form (N = 101) and the infiltrative form (N = 6); 33 of 101 patients with nodular lesions had solitary masses and 18 had multiple masses. The echogenicity of primary tumors were hyperechoic (N = 56), hypoechoic (N = 15), isoechoic (N = 10), and mixed-echoic (N = 20). A peripheral hypoechoic rim was present in 35 primary tumors (34.7%). Peripheral bile duct dilatation was seen in 33 patients (30.8%). Extrahepatic extension was found at operation in 46 of 52 patients (88.5%), while it was demonstrable by ultrasonography in only 16 (30.8%). Five of 6 small lesions (3 cm and less) already had an extrahepatic extension.


Ultrasonographic fine-needle aspiration of pathological masses in the head and neck region. Two hundred and eight ultrasonographic fine-needle aspiration biopsies were performed in patients with head and neck masses and examined cytologically. The average minimum diameter of the masses was 1.5 cm. The smallest punctured lymph node located in the perivascular sheath had a diameter of 0.4 cm. In none of the patients were complications observed. Ninety-seven percent of the diagnoses based on cytological examination were confirmed in the histological examination of surgical biopsies or in the further clinical course.


Mathematical modeling of fetal splenic growth: use of the Rossavik growth model. Growth of the fetal spleen has been monitored by measurements of the maximal length (SL), profile circumference (SC), and profile area (SA) of the spleen, from 20 weeks to 41 weeks, menstrual age. Growth curves for these parameters have been determined using a specially developed growth curve model [P = c(t)k + s (t)]. R2 values of 94.3%, 94.9%, and 96.3% were obtained for SL, SC, and SA, respectively. Variability analysis indicated some progressive increase in variability with menstrual age for these three parameters. Variability data were used with the growth curve models to determine standard curves for SL, SC, and SA. These standard curves provide a superior means for evaluating the normal splenic growth in the fetus and for identifying splenic abnormalities in utero.


New simple method for thyroid volume determination by ultrasonography. A new, simple, and accurate ultrasonic method is elaborated for thyroid volume determination with commercially available high-resolution real-time equipment. The method takes into account the special and variable shape of the thyroid. The measurements were taken in three perpendicular sections. Volume calculations were done by applying the corrected ellipsoid model and computer-aided numerical integration, as well as by the conventional simple ellipsoid model. The accuracy of the method was tested by comparing direct measurements of a silicone rubber phantom with ultrasonography of a tissue-equivalent thyroid phantom. Both phantoms were modeled as casts of the same thyroid, which was obtained at autopsy. From the phantom measurements the deviation between the volume calculated by numerical integration or the corrected ellipsoid method and the real volume determined directly with the silicone rubber phantom was less than 5% on average, while volume calculations using the conventional ellipsoid model involved about +20% systematic error. In vivo studies in 40 subjects gave similar results: the ratios of the volumes calculated by simple and corrected ellipsoid method and by numerical integration [V(E), V(CE), and V(NI), respectively] were [V(E)/V(CE)] = 1.19 +/- 0.11 and [V(NI)/V(CE)] = 1.02 +/- 0.11. In conclusion corrected ellipsoid method is suggested for the clinical practice.


Ultrasound findings in perihepatitis associated with pelvic inflammatory disease. The width of the right anterior extrarenal tissue was increased on ultrasound examination in 9 patients with perihepatitis (Fitz-Hugh-Curtis syndrome). Normal values were obtained in 72 women without detectable or known disease and were found to be between 1 mm and 5 mm (mean 1.8 mm). The 9 patients with laparoscopically verified perihepatitis who demonstrated this thickening had values ranging from 6 mm to 12 mm, (mean 7.7 mm). Although the incidence of this inflammatory thickening in perihepatitis is not known, its value is that in its ultrasonic demonstration a careful search should be made for inflammatory disease in structures related to the anterior pararenal space.