Frequency of antibody to a virus-induced tumor-associated antigen (AG-4) in Japanese sera from patients with cervical cancer and controls.

Microquantitative complement fixation was used to detect antibody to AG-4, a tumor-associated herpesvirus type 2-induced antigen, in sera from Japanese patients with cervical neoplasia. The prevalence of antibody to AG-4 in cervical cancer patients is 47% as compared to 7% in control women. That AG-4 antibody is less prevalent in Japanese than in American cancer patients reflects the frequency of genital herpesvirus type 2 infections in Japan. These findings confirm the association of AG-4 with cervical cancer in a proportion of Japanese patients with neoplastic disease; they implicate factors other than herpesvirus type 2 in another proportion of cervical cancer patients. The possibility that herpesvirus type 1 is one of these factors is considered.

Differential effects of sex steroid hormones on the expression of multiple first exons including a novel first exon of prolactin receptor gene in the rat liver.

In addition to the known four alternative first exons E1(1), E1(2), E1(3) and E1(4) of the rat prolactin receptor (PRL-R) gene, a novel first exon, E1(5), was identified by cDNA cloning of the 5'-end region of PRL-R mRNA in the rat liver. Genomic fragments containing E1(5) and its 5'- or 3'-flanking regions were also cloned from rat kidney genomic DNA. A sequence search for E1(5) revealed that E1(5) is located 49 kb upstream of exon 2 of the PRL-R gene in rat chromosome 2q16. RT-PCR analysis revealed that E1(5) was preferentially expressed in the liver, brain and kidney. Expression profiles of E1(2)-, E1(3)- and E1(5)-PRL-R mRNAs in the liver of male and female rats at 5 days of age and those at 8 weeks of age were examined by RT-PCR. The levels of E1(2)-PRL-R mRNA in the female rat increased remarkably in rats at 8 weeks of age compared with those at 5 days of age, and the levels of E1(5)-PRL-R mRNA in the male rat decreased markedly at 8 weeks of age compared with those at 5 days of age. In the female rat, the levels of E1(2)-PRL-R mRNA at 8 weeks of age decreased with ovariectomy performed at 4 weeks of age and recovered with the administration of beta-oestradiol. On the contrary, the levels of E1(5)-PRL-R mRNA increased with ovariectomy and decreased with the oestrogen treatment. In the male rat liver, the levels of E1(2)-PRL-R mRNA at 8 weeks of age increased strikingly with castration performed at 4 weeks of age and became undetectable with the administration of testosterone. The levels of E1(5)-PRL-R mRNA increased slightly with castration and were restored by testosterone treatment. Removal of gonadal tissues and sex steroid hormone treatment had no effect on the expression levels of E1(3)-PRL-R mRNA in both female and male rat livers. These results indicated that the expression of the PRL-R gene in the liver is regulated by the differential effects of sex steroid hormones on the transcription of the multiple first exons including the novel one.

Epstein-Barr virus-activating principle in human semen.

During the screening of natural and physiological products for their Epstein-Barr virus (EBV)-activating potency, we found that a considerable number of human semen specimens obtained from infertility clinics (30 cases) possess a marked capacity to induce EBV early antigen (EA) in non-producer Raji cell system when assayed in combination with n-butyrate. The EBV EA-inducing activity of the semen samples was comparable to that of the most efficient EBV EA inducers, the plant diterpene esters such as 12-O-tetradecanoylphorbol-13-acetate (TPA). Since many, if not all, of such active agents show overlapping with the tumor promoters in their biological reactions, these findings may provide a new insight for assessing the role of semen in the etiology of certain types of human genital malignancies.

In vitro inactivation of herpes simplex virus by a biological response modifier, PSK.

Herpes simplex virus (HSV) causes herpes genitalis, primary gingivostomatitis and recurrent herpes labialis. In order to elucidate in vivo mechanisms by which PSK, a biological response modifier, exerts a protective effect against HSV infection, we used an in vitro system to study whether PSK inactivated infectivity of HSV-type 1 (HSV-1) and HSV-type 2 (HSV-2) isolated from patients with herpes genitalis in addition to a laboratory-cultured strain of HSV type 1 (HSV-1-GC+). It was found that HSV-1-GC+ was inactivated by PSK in a dose dependent fashion of concentrations of PSK and virus titers. Concentrations of PSK as low as 0.31 mg/ml was shown to inactivate the infectivity of HSV-1-GC+. Inactivation required at least 30 min of incubation at 37 degrees C with maximal inactivation observed at 60 min incubation time. Similar to HSV-1-GC+, clinically isolated strains of HSV-2 were inactivated by PSK although clinically isolated strains of HSV-1 were resistant to PSK, compared with HSV-2. It was also shown that PSK-treated HSV retained the ability to adsorb to the cell membrane, but did not synthesize viral protein(s). These data illustrate that there is a biological difference in the sensitivity to PSK between HSV type 1 and type 2, and also suggest that PSK could inactivate HSV in lesions at peripheral sites of recurrent herpes.

Suppression of recurrent genital herpes simplex virus type 2 infection by Rhus javanica in guinea pigs.

Rhus javanica has been shown to exhibit anti-herpes simplex virus (HSV) activity and potentiate the anti-HSV activity of acyclovir in vitro and in vivo. This extract was examined for its suppressive efficacy on recurrent genital infection in guinea pigs. Guinea pigs were primarily infected intravaginally with HSV type 2 (HSV-2). Prophylactic oral administration, at the dose corresponding to human use, of R. javanica significantly reduced the incidence, severity and/or frequency of spontaneous and severe skin lesions as compared with latently infected guinea pigs administered with water. This prophylactic efficacy was confirmed by the crossover administration, for more than 2 months, of R. javanica and water to the infected guinea pigs. Toxicity, such as weight loss, from R. javanica administration was not observed in the guinea pigs. When recurrent HSV-2 disease was induced by ultraviolet irradiation 3 months after primary infection, the prophylaxis with R. javanica was also significantly effective in reducing the severity of ultraviolet-induced skin lesions. Thus, prophylaxis of recurrent genital HSV-2 infection with R. javanica may preserve the efficacy of acyclovir by reducing both the use of acyclovir and the appearance of acyclovir-resistant viruses.

Efficacy of Cafon gel on cutaneous infection with herpes simplex virus (HSV)-2 and acyclovir-resistant HSV in mice.

Caffeine is known to inhibit replication of herpes simplex virus (HSV)-1 and the therapeutic efficacy of caffeine (Cafon) gel has been shown in a mouse model cutaneously infected with HSV-1. In this study we examined the inhibitory effect of caffeine on infection with HSV-2 and acyclovir-resistant HSV-1 strains, thymidine kinase (TK)-deficient and phosphonoacetic acid (PAA)-resistant HSV-1 in vitro and in vivo. Caffeine inhibited plaque formation of HSV-2 and acyclovir-resistant HSV-1 strains and their EC50 values ranged from 0.42 to 1.11 mg/ml. Topical treatment with Cafon gel was significantly effective in retarding the development of skin lesions caused by cutaneous infection with HSV-2 and PAA-resistant HSV-1 and in reducing the virus yield of the skin infected with TK-deficient HSV-1. The results suggested that Cafon gel would be useful for the topical treatment of cutaneous infection with HSV-2 and acyclovir-resistant HSV strains.

Clinical and virologic studies on female genital herpes.

Ninety women with genital herpes were studied clinically and virologically. From a clinical standpoint, these patients could be classified into 3 clinical types--acute, recurrent, and provoked. When herpes simplex virus (HSV) strains isolated from these patients and asymptomatic virus secretors were typed, 36 strains were determined to be type 1 (HSV-1) and the remaining 54 to be type 2 (HSV-2). The acute type was caused by primary infection of HSV-1 or HSV-2 as well as by infection of HSV-2 in the absence of heterologous antibody. In addition, 35 of 40 HSV strains isolated from recurrent, provoked, and asymptomatic virus secretors were identified as HSV-2, suggesting that HSV-2 may play a major role in establishing latent infection in the female genital region.

Acyclovir treatment of skin lesions results in immune deviation in mice infected cutaneously with herpes simplex virus.

Clinical observations indicate that the antibody response to herpes simplex virus (HSV) in patients undergoing acyclovir treatment is reduced and, although the exact mechanism is not clear, some authors interpret it as immunosuppression. In order to clarify the mechanism, we cutaneously infected mice with HSV-1 and treated the resulting skin lesions with acyclovir. The immune response to infection and treatment in these mice was then analysed. Acyclovir treatment was given orally (20 mg/kg, three times daily), starting on day 0 (D0), 2 (D2) or 4 (D4) after infection and continuing until day 10. The serum antibody titre and the severity of skin lesions were significantly higher in the shortest treatment group (D4) than in the longer treatment groups (D0 and D2). In contrast, a skin test analysing delayed-type hypersensitivity (DTH) to HSV antigen showed that the D0 and D2 groups exhibited stronger DTH than the D4 group. Acyclovir treatment failed to cause a dissociation between DTH and antibody production in mice immunized with inactivated HSV antigen. However, acyclovir treatment in infected mice suppressed the development of skin lesions and resulted in a dissociation between DTH response and antibody production, indicating a typical immune deviation. This was supported by a change in the ratio of the isotype antibody IgG2a to IgG1. The treatment of skin lesions with acyclovir reduced the level of antibody response, as observed clinically. This indicates that the reduced antibody response in patients treated with acyclovir may be, at least in part, due to immune deviation and not immunosuppression.

MR of the submandibular gland: normal and pathologic states.

PURPOSE: To evaluate the MR appearance of normal and pathologic states of the submandibular gland. METHODS: MR images of 22 healthy subjects and 21 patients with histopathologically confirmed disorders of the submandibular gland (five pleomorphic adenomas, two hemangiomas, two malignant lymphomas, one adenoid cystic carcinoma, one squamous cell carcinoma, and 10 cases of sialadenitis) were reviewed. RESULTS: All normal submandibular glands showed higher signal intensity than surrounding muscle but lower intensity than fat on T1-weighted and T2-weighted images. Postcontrast images showed moderate enhancement of the gland. All the tumors had lower signal intensity than the normal submandibular gland on T1-weighted images and had intermediate to high (n = 8) or high (n = 3) signal intensity relative to the normal submandibular gland on T2-weighted images. Six of seven benign tumors were well defined, and three of four malignant tumors were poorly defined. In all cases of sialadenitis, the submandibular gland showed diffusely different signal intensities from the normal gland on both T1-weighted and T2-weighted images. Eight cases of chronic sialadenitis showed lower T2-weighted signal intensities than the normal gland, and this can be explained histopathologically by marked fibrosis and cellular infiltration. CONCLUSIONS: MR imaging can show the presence, extent, margins, and signal intensity changes of pathologic conditions of the submandibular gland.

The occurrence of macrophage-like cholera toxin uptake cells in the intestinal villi of suckling rats.

An oral administration of cholera toxin (CT. 10m g) caused diarrhea in infant rats ranging in age from 1 to 14 days. After administration of the toxin a time sequence study was carried out using highly sensitive immunohistochemical procedures. CT was exclusively incorporated into a type of macrophage-like (ML) phagocytic cell. These cells were identified within the intestinal epithelium of rats suffering choleraic diarrhea. After 2 hrs cells taking up the toxin markedly increased in number and were found in both the mucosa and the lamina propria mucosae. After 4 hrs a small number of ML cells containing CT were still present in the mucosal epithelium, but were no longer observed in the lamina propria. Two kinds of monoclonal antibodies against rat macrophages were used to gain a clue as to the cytological characteristics of ML cells. ED1- or ED2-positive macrophages were demonstrable in the lamina propria and submucosa of the small intestines from control rats. In CT-treated rats a considerable number of cells positive for CT and ED1, or CT and ED2 antisera, were found within the epithelial cell layer and the lamina propria of intestinal villi. It is suggested that many ML cells responsive to CT, if not all, are ED1 and ED2 macrophages and are resident in the villous lamina propria where they can migrate to uptake CT in the intestinal lumen. CT B-subunit and heat-labile toxin (LT) B'-subunit from a mutant strain Escherichia coli were given to the rats in order to know the onset mechanism of toxin uptake. It seems likely that the toxin receptor, GM1 ganglioside, participates in the initiation of CT-uptake mechanism. A possible role of the intestinal ML cells was discussed.

Differentiation of Herpes simplex virus types 1 and 2 in sera of patients with HSV central nervous system infections by type-specific enzyme-linked immunosorbent assay.

OBJECTIVE: To differentiate by enzyme-linked immunosorbent assay (ELISA) using type-specific glycoprotein G herpes simplex virus (HSV) type 1 and type 2 in serum collected from patients with HSV central nervous system (CNS)infections. METHODS: HSV 1 and 2 typing in convalescent sera of 17 patients with HSV acute encephalitis, myelitis, or meningitis was determined by the type-specific IgG ELISA kit (Gull Laboratory, Inc.). HSV CNS infections were diagnosed by polymerase chain reaction (PCR) or conventional serologic tests from acute to convalescent stages. RESULTS: In 13 of 17 patients, HSV type 1 and HSV type 2 antibodies were positive; 11 patients with HSV type 1 and 2 patients with HSV type 2 were found. All 10 patients with encephalitis showed equivocal or positive results for HSVtype 1. In two of three cases of myelitis, HSV type 1 was demonstrated. Each case of myelitis and meningitis reacted to both types 1 and 2. CONCLUSION: These data suggest that the kit is useful for type differentiation of HSV CNS infections from convalescent sera, and can play a supplementary role in HSV typing by PCR or previous serologic tests.

The intramural pelvic nerves immunoreactive for calcitonin gene-related peptide in the rectum of normal and aganglionosis rat.

The distribution of calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerves was investigated immunohistochemically in the rectum of normal, capsaicin-treated and congenital aganglionosis rats. The rectum of the normal rat was densely supplied with both extrinsic and intrinsic nerves exhibiting CGRP-like immunoreactivity. Numerous CGRP-LI nerve fibres were seen in both the myenteric and submucous plexuses. Intrinsic CGRP-LI nerve cell bodies were sparsely found in both the ganglionated plexuses, while a large inflow of extrinsic CGRP-LI nerves was characteristically observed in the rat rectum. CGRP-like immunoreactive fibres were abundant in the intramural pelvic nerves which ascend proximally in the intermuscular zone and connect with the myenteric plexus of the rat distal bowel. As compared with CGRP-positive fibres, SP- or SK-positive fibres in the intramural pelvic nerves were far less frequent. The treatment with capsaicin in the neonatal period led to a marked depletion of CGRP-immunoreactivity in these extrinsic nerves as well as in the most terminal varicose fibres seen in the whole layers of the rectal wall. These findings suggest that the vast majority of CGRP-LI fibres in the intramural pelvic nerves are sensory in nature, and that the positive nerve fibres of extrinsic origin directly innervate each layer of the rat rectum. These CGRP-LI sensory fibres associated with the intramural pelvic nerves, may be of importance in the regulation of rectal and colonic function in normal rats. A dense innervation of CGRP-LI nerve fibres, some of which showed the varicose appearance, was also found in the rectum of congenital aganglionosis rats.(ABSTRACT TRUNCATED AT 250 WORDS)

[Establishment and characterization of cell lines derived from nude mice transplanted squamous cell carcinoma of uterine cervix].

Two human cell lines, KIMI-1 and -2 were established from nude mice transplanted tumor originated from a human squamous cell carcinoma of the uterine cervix. These two cell lines have different shapes, chromosome numbers and tumor markers, respectively.

The simple technique of rectal mucosal biopsy for the diagnosis of Hirschsprung's disease.

The new simple technique of rectal mucosal punch biopsy has been used successfully in 170 cases for diagnosing Hirschsprung's disease. The presented punch biopsy method, using a test tube with a side hole and laryngeal biopsy forceps, has proven to be superior to the rectal suction biopsy method associated with several demerits.

Autonomic innervation of the intestine from a baby with megacystis microcolon intestinal hypoperistalsis syndrome: I. Immunohistochemical study.

A typical case of megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is reported. The patient, an infant girl, was fed only by total intravenous nutrition and is now 3 years old. The distribution of several gut peptides was examined in the resected small intestine using an immunohistochemical method. Vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM)-containing nerve fibers were decreased; however, substance P- and leucine enkephalin (Leu-ENK)-containing fibers were increased. The imbalance between several kinds of gut peptides might be one the causes of aperistalsis in MMIHS. This is the first report about the gut peptides of MMIHS.

A case of VACTEL syndrome with antenatally diagnosed duodenal atresia.

A rare case of VACTEL syndrome with antenatally diagnosed duodenal atresia is reported. We performed one-staged radial surgery for the esophageal atresia with tracheoesophageal fistula and duodenal atresia. The prognosis of VACTEL syndrome with duodenal atresia is very poor. This is the first reported case cured operatively.

Distribution and localization of glial fibrillary acidic protein in colons affected by Hirschsprung's disease.

The distribution and localization of glial fibrillary acidic (GFA) protein were examined by means of immunohistochemistry in normoganglionic, oligoganglionic, and aganglionic segments of colons from 25 patients with Hirschsprung's disease, including four cases of long segment aganglionosis. In normoganglionic segments, GFA protein-positive glial cells were densely distributed within the myenteric plexus, but sparse in the submucous plexus. Aganglionic segments were completely devoid of glial cells with GFA protein immunoreactivity, coinciding with the lack of enteric ganglia. Instead, GFA protein was found specifically in association with the hypertrophic nerve fasciculi and their branches, which were mainly located in the intermuscular zone and submucosal connective tissue in the distal aganglionic segment of diseased bowels. However, two types of short and long segment aganglionosis differed in the distribution pattern of GFA protein; the extrinsic nerve fasciculi in short segment disease extended toward the normoganglionic segment, but in long segment disease they did not reach this area. A moderate number of GFA protein-positive fasciculi were observed within the circular muscle layer of proximal aganglionic and oligoganglionic parts in short segment aganglionosis, while no immunoreactive fasciculi were encountered within the circular muscle layer of the corresponding parts in long segment aganglionosis. Immunohistochemistry for GFA protein can be of excellent diagnostic value for the aganglionic colon with Hirschsprung's disease, since GFA protein immunohistochemistry discloses exclusively extrinsic, hypertrophic nerve fasciculi, characteristic of the bowel in cases of Hirschsprung's disease.

Nodular fasciitis of the upper labial fascia: cytometric and ultrastructural studies.

The rare case of nodular fasciitis in the upper labial fascia is reported. Light microscopic, electron microscopic and cytometric studies were performed. The histopathology corresponded to intermediate and reactive types in the Price and Bernstein classification. Cytometric study of evaluating DNA of fibroblasts revealed a high distinct peak in myxoid and inflammatory areas. Ultrastructurally, fibroblasts and myofibroblasts were identified in the lesion. Reported Japanese cases of orofacial nodular fasciitis are reviewed and compared with the European-American literature.

Oral glucose challenge effects on growth and sex steroid hormones in normal women and women with hypothalamic amenorrhea.

OBJECTIVE: To investigate the effect of growth hormone (GH) on the serum levels of ovarian and adrenal sex steroid hormones in women with hypothalamic amenorrhea (HA). METHODS: A standard 75-g oral glucose tolerance test (OGTT) was performed in 12 normal women and 16 women with stress-related or weight-loss-related amenorrhea. RESULTS: Significant progressive declines in GH, testosterone (T), estradiol (E2) and dehydroepiandrosterone sulfate (DHEAS) were observed during the OGTT in the normal and HA groups. Significant positive correlations were observed between GH and T, E2 and DHEAS. The area under the curve (AUC) for the GH response was significantly greater in the HA group than in the normal group. The ratio of the AUC of insulin-like growth factor to that of GH was significantly lower in the HA group. CONCLUSIONS: Growth hormone may modify ovarian and adrenal sex steroidogenic responses to tropic hormones. A significant degree of GH resistance exists in HA patients. This GH resistance may be related to an ovulatory disturbance.

Free testosterone and abortion in early pregnancy.

OBJECTIVES: To investigate the relationship between percentage of free testosterone (% free T) and the abortion rate in early pregnancy. METHODS: Progesterone (P), estradiol (E2), total T, sex hormone binding globulin (SHBG), and % free T were measured in sera obtained from 60 pregnant women with normal pregnancy (n = 38) and missed abortion (n = 22) at between 4 and 12 weeks' gestation. RESULTS: P, E2, total T, and SHBG in 22 patients with missed abortion were significantly lower than those in normal group, whereas % free T was significantly higher. There was a significant negative correlation between % free T and SHBG concentration in the normal group, but not in the missed abortion group. All the subjects in whom % free T was 1.30% and higher subsequently miscarried, but no subject with % free T less than 0.70% had a miscarriage. CONCLUSIONS: The lower the % free T, the lower the rate of subsequent abortion. The value of % free T may be able to predict pregnancy outcome in early pregnancy.