Molecular Characterisation of Faecal Bacterial Assemblages Among Four Species of Syntopic Odonates.

Factors such as host species, phylogeny, diet, and both timing and location of sampling are thought to influence the composition of gut-associated bacteria in insects. In this study, we compared the faecal-associated bacterial taxa for three Coenagrion and one Enallagma damselfly species. We expected high overlap in representation of bacterial taxa due to the shared ecology and diet of these species. Using metabarcoding based on the 16S rRNA gene, we identified 1513 sequence variants, representing distinct bacterial 'taxa'. Intriguingly, the damselfly species showed somewhat different magnitudes of richness of ZOTUs, ranging from 480 to 914 ZOTUs. In total, 921 (or 60.8% of the 1513) distinct ZOTUs were non-shared, each found only in one species, and then most often in only a single individual. There was a surfeit of these non-shared incidental ZOTUs in the Enallagma species accounting for it showing the highest bacterial richness and accounting for a sample-wide pattern of more single-species ZOTUs than expected, based on comparisons to the null model. Future studies should address the extent to which faecal bacteria represent non-incidental gut bacteria and whether abundant and shared taxa are true gut symbionts. Pictures of odonates adopted from Norske Art databank under Creative Commons License (CC BY 4.0).

Molecular Evidence of Chlamydia-Like Organisms in the Feces of Myotis daubentonii Bats.

Chlamydia-like organisms (CLOs) are recently identified members of the Chlamydiales order. CLOs share intracellular lifestyles and biphasic developmental cycles, and they have been detected in environmental samples as well as in various hosts such as amoebae and arthropods. In this study, we screened bat feces for the presence of CLOs by molecular analysis. Using pan-Chlamydiales PCR targeting the 16S rRNA gene, Chlamydiales DNA was detected in 54% of the specimens. PCR amplification, sequencing, and phylogenetic analysis of the 16S rRNA and 23S rRNA genes were used to classify positive specimens and infer their phylogenetic relationships. Most sequences matched best with Rhabdochlamydia species or uncultured Chlamydia sequences identified in ticks. Another set of sequences matched best with sequences of the Chlamydia genus or uncultured Chlamydiales from snakes. To gain evidence of whether CLOs in bat feces are merely diet borne, we analyzed insects trapped from the same location where the bats foraged. Interestingly, the CLO sequences resembling Rhabdochlamydia spp. were detected in insect material as well, but the other set of CLO sequences was not, suggesting that this set might not originate from prey. Thus, bats represent another potential host for Chlamydiales and could harbor novel, previously unidentified members of this order. IMPORTANCE: Several pathogenic viruses are known to colonize bats, and recent analyses indicate that bats are also reservoir hosts for bacterial genera. Chlamydia-like organisms (CLOs) have been detected in several animal species. CLOs have high 16S rRNA sequence similarity to Chlamydiaceae and exhibit similar intracellular lifestyles and biphasic developmental cycles. Our study describes the frequent occurrence of CLO DNA in bat feces, suggesting an expanding host species spectrum for the Chlamydiales As bats can acquire various infectious agents through their diet, prey insects were also studied. We identified CLO sequences in bats that matched best with sequences in prey insects but also CLO sequences not detected in prey insects. This suggests that a portion of CLO DNA present in bat feces is not prey borne. Furthermore, some sequences from bat droppings not originating from their diet might well represent novel, previously unidentified members of the Chlamydiales order.

Sediment organic tin contamination promotes impoverishment of non-biting midge species communities in the Archipelago Sea, S-W Finland.

Chironomid species are a vital component in many benthic and terrestrial food webs; they have an important role in the detritus cycle, and are an important source of food for many species. We studied how tributyltin (TBT) in brackish water sediments affect the composition of chironomid species communities. Emergence traps were used at selected sites on a TBT gradient in the Archipelago Sea, S-W Finland. Increased sediment TBT concentration was associated with significant chironomid species turnover, which in turn was related to decreased species diversity (number of species and genera). However, the overall number of individuals did not decrease markedly with increasing TBT contamination. This suggests that the ecological role of chironomids in the food web may be preserved even under severe impoverishment of the chironomid community due to organic tin contamination. The increased prevalence of more TBT tolerant species can potentially lead to a transport of organic tin compounds between aquatic and terrestrial food webs. Furthermore, the reduced diversity of an ecologically influential group might lower the resistance of the entire food web to other environmental hazards and perturbations.

Endo- and ectocervical human uterine epithelial cells distinguished by fibronectin production and keratinization in culture.

Human ecto- and endocervical uterine epithelia grow in culture as stratifying and nonstratifying cell layers, patterns reflecting the differentiation of the corresponding cells in vivo. We found that both cultures synthesized and secreted fibronectin but very little other known glycoproteins of basal laminae, laminin, and type IV collagen. Fibronectin production was observed only in the lining marginal cells of the expanding periphery of ectocervical islands, but differentiating and keratinizing cells had loss fibronectin. Most cells in endocervical epithelial cultures grew as a monolayer and produced fibronectin. After stratification of the ectocervical cells, the cytokeratin polypeptide pattern switched to the large-molecular-weight type. In endocervical cultures, the low-molecular-weight keratin polypeptides persisted. Fibronectin production thus appears to be linked to the state of differentiation of the epithelial cells, and fibronectin may function in the attachment of the basal cells to underlying basal lamina matrix.

Explantation and subculture of epithelial cells from human uterine ectocervix.

Human ectocervical explant cultures were grown in medium with D-valine substituted for L-valine. Pure epithelial cell monolayers were obtained with both dialyzed and undialyzed fetal calf serum. Epithelial cell explant colonies grown in D-valine medium supplemented with undialyzed serum could routinely be subcultured if plated at a density of 1.5 x 10(4) cells/sq cm or higher. Such cultures could be passaged at least three times and could yield up to 21 population doublings per culture lifetime. Contaminating fibroblastic colonies were never detected in these cultures, which were free of surface-associated fibronectin as revealed by immunofluorescent tests. Both primary and passaged epithelial colonies retained many characteristic morphological features of ectocervical epithelium when examined by light and electron microscopy. Such cultures may be of use in investigating the action of viral and chemical carcinogenic agents upon epithelial cells in vitro.

Comparison of mass spectrometry and radioimmunoassay to measure medroxyprogesterone acetate in patients with endometrial cancer.

Serum medroxyprogesterone acetate (MPA) was measured by radioimmunoassay (RIA) and gas chromatography-mass spectrometry (GC-MS) in patients with endometrial cancer. Samples were obtained 3, 6 and 24 h after the oral administration of 100 or 200 mg MPA once a day. The levels obtained by GC-MS were lower (median 16-29%) than those obtained by RIA, which is probably attributable to the presence of metabolites interfering with the RIA. Two commercial MPA formulations gave different MPA serum levels by both RIA and GC-MS. The levels obtained by GC-MS were so low that frequently only partial saturation of the endometrial progesterone receptor may be achieved which may explain why high oral doses are needed to produce optimum therapeutic response.

A T-helper cell epitope overlaps a major B-cell epitope in human papillomavirus type 18 E2 protein.

Cultivated CD4+ T-helper cells from two patients with cervical adenocarcinoma showed responses to a peptide EKTGILTVTYHSETQRTK derived from an E2 protein of human papillomavirus type 18 (HPV 18), but not to a corresponding HPV 16 peptide (HKSAIVTLTYDSEWQRDQ). Serum antibodies in the HPV 18 peptide were also demonstrated in these patients. The GILT motif resembles a common pattern present in many T-cell epitopes, and is located at the beginning of an 11-amino acid-long A-helix structure close to the carboxyterminal end of HPV 18 E2. We conclude that two epitopes (a T-helper cell epitope and a B-cell epitope) overlap in the HPV 18 E2.

Cancer incidence among 78,000 asthmatic patients.

The risk of cancer was evaluated among 77,952 asthma patients with bronchial asthma. The series was obtained through linkage of two registers: the Finnish Social Insurance Institution's file of asthma patients and the Finnish Cancer Registry. There was a significant excess risk of lung cancer in both sexes, the standardized incidence ratio (SIR) being 1.32 among men and 1.66 among women. In women, the risk of cancer of the rectum was significantly increased (SIR 1.42), whereas the risks of cancer of the corpus uteri and multiple myeloma were lower than expected (SIR 0.76 and 0.53, respectively). In men, the incidence of cancer of the larynx was significantly reduced (SIR 0.63) and that of the bladder increased (SIR 1.25). When both sexes were combined, cancers of the colon (SIR 1.17) and rectum (SIR 1.28) also showed a significantly elevated risk. A reduction in risk was seen in stomach cancer (SIR 0.88) and lymphatic leukaemia (SIR 0.55). The increased lung cancer risk may be due to local inflammatory changes. It is possible that differences in the immune system, e.g. natural killer cell activity, explain some of the reduced cancer risks.

Human papillomavirus types 16 and 18 in adenocarcinoma of the uterine cervix.

Many reports have shown a link between human papillomavirus (HPV) and cervical squamous neoplasia. However, the association of HPV with cervical adenocarcinoma has been studied less extensively. The authors evaluated the presence of HPV-DNA in 106 patients with adenocarcinoma of the uterine cervix by in situ hybridization, using 35S-labeled probes for HPV 16 DNA and HPV 18 DNA. The overall prevalence of HPV-DNA was 18% (19 of 106). HPV 16 was present in 2 (2%) cases, HPV 18 was observed in 15 (14%) cases, and both HPV 16 and HPV 18 were found in 2 (2%) cases. There was a correlation between HPV-DNA positivity and tumor stage (P less than 0.01) and tumor size (P less than 0.05), but there was no relationship between HPV-DNA positivity and tumor differentiation, proliferation (S-phase fraction), ploidy, lymph node metastases, or five-year survival rate. These results suggest that HPV 18 DNA is associated with cervical adenocarcinoma but the presence of HPV 18 has no influence on overall survival.

Predominance of serum antibodies to synthetic peptide stemming from HPV 18 open reading frame E2 in cervical adenocarcinoma.

AIMS: To determine if there are type specific differences in serum antibody responses to synthetic peptides derived from human papillomavirus (HPV) open reading frame (ORF) E2 in patients with cervical carcinoma. METHODS: Diagnostic phase sera from 88 age-matched women with cervical adenocarcinoma (AC), cervical squamous cell carcinoma (SC), ovarian cancer (OC) or no gynaecological malignancy were available. Serum IgG and IgA antibodies to synthetic peptides corresponding to a residue of HPV 6, 11, 16, and 18 ORF E2 18 amino acids long and a control peptide from mumps virus were determined by ELISA. RESULTS: Both IgA and IgG antibody positivity to the HPV 18 peptide were associated with increased risk (9.0-fold, confidence limits 1.5-199) for AC. IgA positivity to HPV 11, 16, and 18 peptides was associated with an increased risk for SC. However, the association of IgG antibodies to HPV 16 peptide with SC was not significant. IgA or IgG antibodies to HPV 6 or mumps virus peptides were not associated with increased risk for AC, SC, or OC. CONCLUSIONS: These results suggest a specific role for HPV 18 in AC. Differences in antibody responses to HPV peptide in AC and SC suggest immunopathogenetic differences between the two types of cervical carcinoma.

Colposcopic and histologic findings in cervical chlamydial infection.

Colposcopic and histologic findings in 41 patients with Chlamydia trachomatis-associated cervicitis and in age-matched controls with nonspecific cervicitis are presented. Colposcopy showed exophytic follicular cervicitis in 30 patients in the chlamydia group and in none in the control group. Atypical transformation zone occurred significantly more often in the chlamydia group. Directed biopsy specimens revealed cervical intraepithelial neoplasia in 10 cases in the chlamydial group but in only 1 case in the control group. In 4 patients with chlamydial cervicitis lymphoid follicles were observed in histologic specimens.

Topical treatment of flat vaginal condyloma with human leukocyte interferon.

Human leukocyte interferon cream was evaluated in a double blind placebo-controlled trial in 13 patients with widespread vaginal flat condylomatous dysplasia. Patients applied 12 million units of interferon vaginal cream daily into vagina during four two-week treatment courses separated by one-week intervals. Five of eight patients treated with interferon showed clear remissions in colposcopy. Cytologic examinations, however, revealed cells typical of condyloma in all cases. Two responding patients relapsed one and two months after the treatment. Among five patients using placebo, the lesions remained unchanged in three patients and progressed in two. Interferon treatment did not alter the microbiologic or cytologic picture of Papanicolaou smears, but lymphocytic infiltrations in the stroma next to condylomas were seen in histologic samples. No overt side effects were observed during interferon treatment.

Observer variability in the scoring of colpophotographs.

Colposcopy and cervicography are accepted tools for assessing the cervix for an atypical transformation zone. We studied the validity of the colpophotograph as a measurement tool by determining the agreement of experienced colposcopists using colpophotographs of 50 women. Interobserver agreement was generally fair to good (kappa greater than or equal to 0.40) for the presence of the squamocolumnar junction and the area of ectopia but it was poor (kappa less than 0.40) for the area, border, and color characteristics of an atypical transformation zone. Intra-observer agreement was fair to good for the color characteristics of an atypical transformation zone, but it was poor for the area and border characteristics. We conclude that observer agreement studies should play a role in the validation of methods used in the visual diagnosis of cervical intraepithelial neoplasia. Considerable lack of agreement in reporting cytologic findings is a well-known problem, and lack of agreement might be an even bigger problem in reporting colposcopic findings.

Genital Chlamydia trachomatis infections in patients with cervical atypia.

Of 177 gynecologic outpatients with cervical dysplasia studied, 29 (16%) were found to excrete Chlamydia trachomatis from their urogenital tract. Sera collected from 93 of these patients were significantly more often positive for and showed higher levels of antichlamydial micro-complement fixation (CF) and immunofluorescence (IF) antibodies than sera obtained from the controls. However, when the sera were tested for anti-herpesvirus type II (HSV-II) and cytomegalovirus (CMV) antibodies, no difference between the cases and the controls were encountered.

Intrauterine contraceptive device use in patients with acute salpingitis.

The frequency of intrauterine contraceptive device (IUD) use among 144 hospitalized women with acute salpingitis was compared with that of 229 control women, who were symptomless sexual partners of men examined for non-gonococcal urethritis. The relative risk for salpingitis was higher for IUD than in the group of basal level of disease (p < 0.02). This was mostly caused by the high relative risk in the women with no deliveries. Oral contraceptives showed a contrary picture, the relative risk was significantly lower than on the basal level (p < 0.01). Non-gonococcal and gonococcal infections were equally represented in salpingitis patients with IUDs. The severity of the infection in IUD users did not differ from that of non-IUD users.

PIP: The frequency of IUD use among 144 hospitalized women with acute salpingitis was compared with that of 229 control women who were symptomless sexual partners of men examined for nongonococcal urethritis. The relative risk for salpingitis was higher for those with IUDs than in the group of basal level of disease (P 0.02). This was caused largely by the high relative risk in women with no deliveries. Oral contraceptives showed a contrary picture, with the relative risk significantly lower than on the basal level (P 0.01). Nongonococcal and gonococcal infections were equally represented in salpingitis patients with IUDs. The severity of the infection in IUD users did not differ from that of non-IUD users.

Human papillomavirus DNA in unselected pregnant and non-pregnant women.

Human papillomavirus (HPV) DNA is associated with genital squamous cell neoplasia, especially types 16, 18, and 31. Several studies, using histopathology and DNA hybridization, have shown an increased prevalence of cervical HPV DNA in pregnant women. We determined the prevalence of HPV DNA and the distribution of specific HPV DNA types in exfoliated cervical cells from 748 randomly selected pregnant and 503 non-pregnant women, using a dot blot DNA hybridization technique. The prevalence of HPV DNA was 9.6% in pregnant women and 8.9% in non-pregnant women. Thus, we found no evidence of higher prevalence of HPV DNA in pregnant women. HPV DNA types 16/18 and 31/33/35 were more common than types 6/11. The prevalence of HPV DNA was highest (15%) in the youngest age groups suggesting an association between young age and the prevalence of HPV DNA.

Factors predicting abnormal hysterosalpingographic findings in patients treated for acute pelvic inflammatory disease.

One hundred forty-six consecutive inpatients with acute pelvic inflammatory disease (PID) were randomly treated either with the combination of doxycycline and metronidazole or with the combination of penicillin G/amoxicillin and metronidazole. Forty-three women underwent hysterosalpingography (HSG) 6 months after the treatment, and a multivariate statistical analysis was used to study the effect of 11 different variables (parity, history of previous abortion, history of previous PID, use of an intrauterine contraceptive device (IUD), use of oral contraceptives, presence of adnexal mass, results of cervical cultures for Neisseria gonorrhoeae, and Chlamydia trachomatis, presence of serum antichlamydial antibodies, penicillin-metronidazole treatment, and doxycyclin-metronidazole treatment) on the hysterosalpingographic findings. Abnormal HSG was most likely to occur in women who had antichlamydial antibodies present in the serum, an adnexal mass detected on the initial bimanual examination, or had an IUD (positive associations) and least likely to occur in women who had been treated with the combination of doxycycline and metronidazole (negative association).

Cytopathogenicity of cytomegalovirus to human ecto- and endocervical epithelial cells in vitro.

The effect of cytomegalovirus on cell cultures initiated from human uterine endo- and ectocervix was studied. Pure epithelial cultures were obtained which differed in morphology depending on the source. The ectocervical cells grew in mosaic-like regular epithelial patterns, whereas endocervical cultures had a poorer intercellular cohesion, irregular polygonal cell form and curved cytoplasmic processes. Occasional fibroblastic colonies grew in three out of 32 ectocervical cultures and in none of the endocervical cultures. In the latter no cells with fibroblast-specific surface antigens could be seen. Ectocervical cells were resistant to cytomegalovirus infection; only in three out of 20 cultures could individual altered cells be detected. In contrast, endocervical cultures supported the growth of cytomegalovirus and at best 30 per cent of the explanted colonies showed cytologic alterations as well as virus-specific immunofluorescence. The various types of altered cells are described and it si concluded that endocervical epithelial cells may be the site of replication of human cytomegalovirus so commonly found in cervical secretions.

Clinical and virological findings in patients with cytologically diagnosed gynecologic herpes simplex infections.

Amoung 57,117 routinely collected Papanicolaou smears from the female genital tract there were 90 slides (0.16%) from 85 patients on which a cytologic diagnosis of herpes simplex virus (HSV) infection was made. The characteristic cytologic changes occurred predominantly on the ectocervical part of the smears. The changes were transient but occasionally remained detectable on the corresponding histologic specimens up to 2.5 months. Viral isolation was successful in a majority of cases when performed simultaneously with or less than a week after the cytologic diagnosis. The rate of success rapidly decreased thereafter. Positive fluorescent antibody assays of viral antigens on cytologic smears behaved similarly. Cases with apparent primary infection as well as with secondary infection (recurrences) were included in the material but no differences in the morphology of virally altered cells and be found, indicating that primary and secondary infections could not be distinguished on this basis. There were significantly more cytologic dysplastic changes as well as assorted micro-organism infections in the HSV group as compared to the controls.