Clinical Applications of the Gut Microbiome in Genitourinary Cancers.

Recently recognized as one of the hallmarks of cancer, the microbiome consists of symbiotic microorganisms that play pivotal roles in carcinogenesis, the tumor microenvironment, and responses to therapy. With recent advances in microbiome metagenomic sequencing, a growing body of work has demonstrated that changes in gut microbiome composition are associated with differential responses to immune checkpoint inhibitors (ICIs) because of alterations in cytokine signaling and cytotoxic T-cell recruitment. Therefore, strategies to shape the gut microbiome into a more favorable, immunogenic profile may lead to improved responses with ICIs. Immunotherapy is commonly used in genitourinary (GU) cancers such as renal cell carcinoma, urothelial cancer, and to a limited extent, prostate cancer. However, a subset of patients do not derive clinical benefit with ICIs. Gut microbiome-based interventions are of particular interest given the potential to boost responses to ICIs in preclinical and early-phase prospective studies. Novel approaches using probiotic therapy (live bacterial supplementation) and fecal microbiota transplantation in patients with GU cancers are currently under investigation.

[Low frequency of cancer in the urinary organs when macroscopic hematuria is associated with bacteriuria]. / Låg sannolikhet för cancer i urinorganen vid makroskopisk hematuri och samtidig bakteriuri.

Frequency of cancer in the urinary organs was significantly lower in patients with macroscopic hematuria associated with bacteriuria compared to those without bacteriuria. The predictive value of macroscopic hematuria was <1% in patients ≤ 75 years of age with concomitant bacteriuria. CT-urography added no diagnostic value over and above cystoscopy in patients with macroscopic hematuria with bacteriuria.Bacteriuria with other bacteria than E. coli or S. saprophyticus was associated with findings of bladder tumors.

Risk factors for refractory febrile neutropenia in urological chemotherapy.

During chemotherapy, patients are more susceptible to infectious complications as a result of bone marrow suppression, leading to neutropenia. The purpose of this study is to investigate risk factors for refractory febrile neutropenia (FN) during urological chemotherapy. Our method for suppressing FN is to use granulocyte colony-stimulating factor and prevent upper respiratory infection by masking and gargling. We studied 47 episodes of FN in 39 patients that occurred during urological chemotherapy for urothelial cancer, testicular cancer, and prostate cancer. Among our cases, there were 5 patients with refractory FN; we set risk factors for refractory FN and performed statistical analyses. The average age of the 39 patients was 60.6 years (range, 18-80 years). In 47 FN episodes, the chemotherapy regimen before the occurrence of FN included 15 (31.9 %) MVAC (methotrexate, vinblastine, adriamycin, cisplatin) for urothelial cancer, 5 (10.6 %) DE (docetaxel, estramustin) for prostate cancer, and 3 (6.4 %) TIP (paclitaxel, ifosfamide, cisplatin) for testicular cancer. The antibiotics used to treat FN included 17 (36.3 %) meropenem and 23 (49.0 %) cefepime, and the average duration of antibiotics was 4.4 days (range, 1-12). We investigated risk factors for refractory FN and showed a significant relationship between refractory FN and indwelling urinary catheter or smaller Multinational Association for Supportive Care in Cancer score by multivariate analysis. A future prospective study is needed for further evaluation for risk factors and establishing treatment protocols and guidelines for FN.

Aerobic bacterial flora of the vagina and prepuce of California sea lions (Zalophus californianus) and investigation of associations with urogenital carcinoma.

To investigate the association between genital bacterial infection and urogenital carcinoma in California sea lions (Zalophus californianus), vaginal and preputial swabs for bacterial isolation were taken from 148 free-ranging and 51 stranded California sea lions including 16 animals with urogenital carcinoma. Cytological examination of vaginal or preputial smears showed a majority (65.5%, 57/87) of animals examined had mild or no inflammation. Aerobic bacteria were isolated from 116 (78.4%) wild sea lions and 100% of stranded animals. A total of 403 isolates were identified representing 51 unique bacterial species. The median number of isolates per animal increased with age in the wild group, but there was no difference in the number of isolates per animal between wild and stranded adults. The most common bacteria isolated from the wild sea lions were Psychrobacter phenylpyruvicus (39 isolates), non-hemolytic Streptococcus (35 isolates), Corynebacterium spp. (30 isolates), and Escherichia coli (20 isolates). More bacterial species were isolated from stranded animals than wild animals (33 versus 26) and there was significantly less growth of P. phenylpyruvicus, Corynebacterium spp., and Moraxella-like spp. in the stranded animals. Beta-hemolytic Streptococcus was the only bacterium significantly associated with urogenital carcinomas in California sea lions, but only in females.

Absence of human papillomavirus in squamous cell carcinomas of nongenital skin from immunocompromised renal transplant patients.

BACKGROUND AND DESIGN: Human papillomavirus (HPV) is accepted as a factor in the pathogenesis of genital squamous cell carcinomas. The incidences of both HPV infection and squamous cell carcinoma are increased in immunocompromised renal transplantation patients. The purpose of this study was to determine if HPV DNA is present in squamous cell carcinomas of nongenital skin in immunosuppressed patients. Amplification of HPV DNA was performed using the polymerase chain reaction. The sensitivity and specificity of the polymerase chain reaction was assessed on 19 positive and six negative control specimens. Twenty genital squamous cell carcinomas from nonimmunocompromised patients and 28 nongenital squamous cell carcinomas from renal transplantation patients were then analyzed. RESULTS: Human papillomavirus DNA was identified in 18 of 19 positive control specimens and zero of six negative control specimens. Human papillomavirus DNA was identified in four of 20 genital squamous cell carcinoma specimens. In comparison, no HPV DNA was identified in 28 nongenital squamous cell carcinomas from immunosuppressed hosts (Fisher's Exact Test, P < .025). CONCLUSIONS: These findings support a role for HPV in genital skin cancers and suggest that HPV does not play a role in the increased incidence of squamous cell carcinoma in renal transplantation patients.

[HPV in oncogenesis]. / HPV in der Onkogenese.

During recent years, advances have been made in understanding the functions of the viral oncoproteins E6 and E7. E6 und E7 code for proteins forming complexes with cellular proteins, which regulate the cell cycle. This leads to a disturbance of the physiological control mechanism and to increased proliferation of the infected cells. Enhanced expression of viral oncogenes in cells infected by genital "high risk" HPV types results in chromosomal instability and in an accumulation of mutational events. Cutaneous HPV types apparently follow other strategies to transform their host cells.

Human papillomavirus identified by nucleic acid hybridization in concomitant nasal and genital papillomas.

Presence of human papillomavirus (HPV) as the etiologic agent in nearly all upper respiratory tract recurrent papillomas is well-established. The technique of nucleic acid hybridization now allows specific typing of HPV with a high degree of accuracy. This article reports a series of nine consecutive patients treated for nasal papillomas over the past 9 years. Eight of these patients had a personal history of genital papillomas (seven patients) or exposure (one patient). With the use of in situ hybridization and autoradiographic technique on paraffin-embedded tissue sections, HPV RNA type 6/11 was expressed in eight of nine nasal papillomas, and corresponding HPV types were also found in the two cases with which concurrent anogenital papilloma tissue was also available for analysis. Human papillomavirus RNA types 16 and 18 were not detected in any of the specimens. Signals of HPV messenger RNA type 6/11 were stronger in the fungiform areas than in the inverted areas of papillomas.

Triple cancers in the urogenital area of a patient with aplastic anemia.

Three epithelial neoplastic lesions, perineal Bowenoid papulosis, uterine cervical carcinoma, and bladder transitional cell carcinoma, which occurred in a mildly immunosuppressed patient who had aplastic anemia were studied for human papillomavirus (HPV) infection. In the Bowenoid papulosis, HPV type 16 DNA was identified by polymerase chain reaction (PCR) and by in situ hybridization (ISH). In contrast, in the uterine cervical carcinoma, HPV 16 was not detected, although possibly another unidentified type of HPV in the lesion was suggested by the ISH findings. In the bladder transitional cell carcinoma, neither papillomavirus genus-specific (PGS) antigen nor HPV DNA was found.

Isolation of an infectious oncogenic adenovirus strain from the stimulated lymphocytes of a patient with bladder tumour.

In attempts to grow viruses from tumour cells and circulating lymphocytes, we isolated an oncogenic infectious adenovirus type 18 strain from the phytohaemagglutinin-stimulated peripheral T-lymphocytes of a patient with bladder tumour. The virus genome is assumed to play certain role in the development of the tumour in interaction with other DNA viruses and the damaged immune system.

[Studies of human papillomavirus (HPV) in urological tumors].

Urological tumors were examined for the presence of human papillomavirus (HPV) DNA by using Southern blot hybridization. In 20 male patients with condyloma acuminatum, HPV type 6 was found at 85% (17/20), HPV type 11 at 95% (19/20), HPV type 16 at 5% (1/20) and HPV type 18 at 0% (0/20). In 2 female patients with condyloma acuminatum, HPV types 6, 11, 16 and 18 were found at 100% (2/2), 100% (2/2), 50% (1/2) and 0% (0/2), respectively. All 6 of the patients who were positive for HPV type 6, were also positive for HPV type 11. Two patients were positive for HPV types 6, 11 and 16, the last of which was frequently found in penile cancer and uterine cervical cancer. In 6 patients with penile cancer, two patients were positive for HPV type 16 and negative for HPV types 6, 11 and 18. The remaining 4 patients were negative for all these HPV types. One patient who was positive for HPV type 16 had penile cancer after three previous episodes of penile condyloma acuminatum. From this information, a malignant change in the condyloma acuminatum was assumed to indicate the possible association of HPV type 16 with the process of malignant degeneration. HPV types, 6, 11, 16 and 18 were not detected in a female patient with vulvar cancer. Although HPV was thought to participate in the development of urological tumors except for external genital tumors, all patients examined, consisting of 2 with benign prostatic hypertrophy, 5 with prostatic cancer and 24 with bladder cancer, were negative for HPV types 6, 11, 16 and 18. Eight patients with bladder cancer were negative for HPV type 33.

Human papillomavirus types in anogenital warts of children.

Tissue from anogenital warts of 25 children, 10 of whom were suspected of being victims of sexual abuse, was investigated by dot blot and Southern blot techniques for human papillomavirus (HPV) types. HPV DNA was detected in 22 children, two of whom had double infections. The genital HPV types 6 and/or 11 were detected in 20 children, and in three children other HPV types were found. One had HPV 18 (as well as 11); in a second child a possible skin type, HPV 2, was detected; and the third child was infected with an unidentified type. In three cases genital wart material was available from one of the parents, and in all three the HPV type was the same as that of the child. For nine other children one or both parents were reported to have genital warts. The source of infection appeared to be the adult genital tract, but sexual contact might not be the only means of transmission.

[Persistence of herpes simplex virus and adenoviruses in lymphocytes of patients with urogenital tumors]. / Persistentsiia virusa prostogo gerpesa i adenovirusov v limfotsitakh bol'nykh s opukholiami urogenital'nogo trakta.

Certain viruses with neoplastic potential are known to be capable of participating in tumor formation to impair the immune system. In the present study, this association was investigated in patients with urogenital tract tumors. In studies of latent virus carrier state by means of immunofluorescence techniques using specimens from 118 patients with malignant tumors and 70 control persons it was found that in more than 50% of the patients, latent antigens of herpes simplex virus or adenoviruses were present in 1-3% of circulating lymphocytes. In the control group, virus carrier state in lymphocytes was demonstrated in only few patients. Using lymphocyte blastogenesis test, the effect of nonspecific mitogen (phytohemagglutinin) induced transformation of only 10-50% lymphocytes to lymphoblast cells in patients with malignant tumors, the percentage of transformation being also dependent on the stage of the tumor process. In the control group, treatment of lymphocytes with phytohemagglutinin resulted in transformation of 55-85% of lymphocytes into lymphoblasts. The lymphocytes of the majority of patients with tumors became sensitive to specific herpes-virus and adenovirus antigens, mainly the lymphocytes of the patients whose blood cells were also virus carriers.

Sensitive detection of nucleic acids and protein of human papillomavirus type 6 in respiratory and genital tract papillomata.

We have developed a sensitive method to detect and localize HPV-6 viral DNA, mRNA and protein in biopsy specimens of genital and respiratory tract lesions by using in situ hybridization and immunoperoxidase assays on sections of plastic-embedded tissue. This modified in situ hybridization technique, using ultrathin sections and strand-specific 3H-labelled riboprobes, offers the advantages of superior morphological preservation and detection of viral genomes at low copy number with good resolution. This modified immunocytochemistry provides better sensitivity when compared to previous methods using paraffin-embedded materials. In respiratory tract lesions, immunoperoxidase assay detected only a few capsid antigen positive cells, while in the genital tract lesions, there were more capsid antigen positive cells. Southern transfer analyses and in situ hybridizations demonstrated the presence of more viral nucleic acids in genital tract papillomata than respiratory tract papillomata. Epithelial cells throughout the papillomata were infected by HPV-6 as evidenced by positive hybridization, with more viral DNA present in superficial cells. Our results suggest that genital tract epithelium is more permissive for HPV-6 replication than respiratory tract epithelium. Using stand-specific probes synthesized from subgenomic fragments of the HPV-6 genome in conjunction with nuclease digestions, we were able to demonstrate that HPV-6 transcripts specific to open reading frames (ORFs) E6, E7, E1, L1, and L2 occur in maturing superficial cells. In contrast, transcripts specific to ORFs E1, E2, E4, E5a, and E5b could be detected throughout the whole of the epithelium with more signals noted at the basal cell areas. In addition, the distribution of HPV-6 nucleic acids and protein in a carcinoma in situ of the larynx was analyzed. In comparison to benign respiratory tract papillomata, more viral DNA was found in the malignant lesion, but the pattern and distribution of transcription and capsid antigen was similar.

Detection of human papillomavirus in formalin-fixed, invasive squamous carcinomas using the polymerase chain reaction.

We analyzed 88 formalin-fixed, paraffin-embedded invasive squamous carcinomas for human papillomavirus-related DNA sequences (HPV types 16 and 18) following in vitro gene amplification using the polymerase chain reaction. HPV DNA sequences were found in 35 of 50 (70%) carcinomas of the anogenital region, including four of four (100%) anal, six of eight (75%) vulvar, nine of 14 (64%) vaginal, two of five (40%) penile, and 14 of 19 (74%) cervical tumors. Nine of 25 (36%) oropharyngeal squamous carcinomas contained HPV DNA sequences, including four of 10 (40%) laryngeal, three of eight (38%) buccal, and two of seven (29%) glossal tumors. HPV DNA sequences were not found in 13 esophageal carcinomas. Of the 44 cases that contained viral DNA, HPV-16 was detected in 41 cases (93%) and HPV-18 in five cases (11%), while both types were found in two cases (one anal and one vulvar). HPV DNA sequences were found in 43 of 83 (52%) nonverrucous and in one of five (20%) verrucous carcinomas, but this difference was not significant. These findings demonstrate that HPV DNA sequences are more frequently associated with anogenital than oropharyngeal squamous carcinomas and can be readily detected in routinely processed tissues using the polymerase chain reaction.

Shedding of herpes simplex virus type 1 into saliva after surgery for oral and genital or urological cancer patients.

The shedding of herpes simplex virus type-1 (HSV-1) into the saliva was compared in 28 patients with oral cancer and 26 patients with genital or urological cancer. All subjects tested positive for HSV-1 specific antibody. A statistically significant (p less than 0.001) difference was found: infectious viruses were isolated from 12 (39.8%) of the oral cancer patients versus only 2 (7.6%) of the genital or urological patients. This indicates that direct stimulation of peripheral nerves during surgery was responsible for the greater reactivation of HSV-1.

Latent virus carrying lymphocytes of patients with urological tumours.

A considerable wealth of data suggest that a virus with oncogen character can play a part in the development or survival of tumours, by disturbing immune reaction. We studied this situation in urogenital tumours. Examining the latent virus carrier with an immunofluorescent method in 96 cases with malignant tumours and 70 control cases, it was found that more than 50% of the patients had latent adeno- or herpes simplex virus antigens in 1-3% of the circulating lymphocytes, whereas virus carriers occurred in hardly a few percent in the control group. Using a lymphocyte transformation test, the non-specific mitogen phytohaemagglutinin was able to transform only 10-50% of the lymphocytes of patients with malignant tumours into lymphoblast cells (the percentage also depending on the stage of the tumour). On the other hand, under the influence of phytohaemagglutinin 55-85% of the lymphocytes of the control group turned into lymphoblasts. The lymphocytes of the majority of patients with tumours became sensitive for specific adeno- and herpes simplex virus antigens, mainly the lymphocytes of those whose cells were virus carriers.

Complementability of temperature-sensitive adenovirus mutants with extracts of urogenital tumour cells.

Complementation of two temperature-sensitive (ts) mutants of adenovirus type 5 was attempted with tumour extract in HEp-2 cell cultures at permissive and restrictive temperature. Both ts mutants were successfully complemented with adenovirus gene products extracted from bladder and kidney carcinoma cells. The percentage incidence of complementation appeared to be higher with extracts obtained from female patients than with those obtained from males. The same gene sequences may occur in penile carcinoma cells and in bladder carcinoma cells showing the early signs of malignization. Extracts of seminoma cells and of cells from prostatic hypertrophies contained adenovirus genes less frequently; infectious virus was never obtained from them. The same applies to cells from the non-tumourous ("control") patients tested. The old age of the patients and some properties of the tumours examined suggest that hormonal changes may contribute to adenovirus gene expression and, indirectly, to the malignant process and its continuity.