Effects and Eradication of Mycoplasma Contamination on Patient-derived Colorectal Cancer Organoid Cultures.

Patient-derived organoids are a useful platform for identification and testing of novel precision oncology approaches. Patient-derived organoids are generated by direct culture of patient samples. However, prior to development into patient-derived organoids, these samples are often processed for clinical use, opening the potential for contamination by Mycoplasma and other microbes. While most microbes can be detected by visual inspection, Mycoplasma can go undetected and have substantial impacts on assay results. Given the increased use of patient-derived organoids, there is a growing need for a standardized protocol to detect and remove Mycoplasma from organoid models. In the current study, we report a procedure for Mycoplasma removal by passaging organoids through mice as patient-derived organoid xenografts. In vivo passage of patient-derived organoids followed by re-establishment was 100% effective at decontaminating colorectal patient-derived organoids (n = 9), based on testing with the Sigma LookOut Mycoplasma PCR Detection Kit. This process can serve as a method to re-establish contaminated patient-derived organoids, which represent precious models to study patient-specific genomic features and treatment responses. SIGNIFICANCE: Organoids are valuable models of cancer. Mycoplasma contamination can alter organoid drug sensitivity, so there is a need for a standardized protocol to detect and remove Mycoplasma from organoids. We report a simple procedure for removing Mycoplasma from organoids via in vivo passaging through mice followed by re-establishment of organoids.

Association of the eukaryotic vaginal virome with prophylactic antibiotic exposure and reproductive outcomes in a subfertile population undergoing in vitro fertilisation: a prospective exploratory study.

OBJECTIVE: The objective of this study was to use high-throughput sequencing to describe the vaginal eukaryotic DNA virome in patients undergoing in vitro fertilisation (IVF) to examine associations between the vaginal virome, antibiotic exposure and IVF outcomes. DESIGN: Prospective exploratory study. SETTING: Single academic fertility centre. POPULATION: Subfertile women age 18-43 years undergoing their first IVF cycle with a fresh embryo transfer. METHODS: The primary exposure was prophylactic azithromycin or no azithromycin before IVF. A mid-vaginal swab was obtained at the time of embryo transfer for virome analysis. MAIN OUTCOME MEASURES: The primary outcomes compared between exposure groups were characteristics of vaginal virome and clinical pregnancy rates. Secondary outcomes were virome associations with number of oocytes retrieved, number of blastocysts and implantation rate. RESULTS: Twenty-six women contributed a vaginal swab before embryo transfer. There were no significant differences in IVF outcomes between azithromycin groups. There was no association between viral diversity and clinical pregnancy overall. A higher diversity of herpesviruses and α-papillomaviruses was observed in samples from the azithromycin-treated group compared with the no azithromycin group (P = 0.04). In women that received azithromycin, viral diversity was higher in the group that did not achieve clinical pregnancy compared with those who did (P = 0.06). CONCLUSIONS: We demonstrate that the vaginal eukaryotic virome in women undergoing IVF is associated with antibiotic exposure. Additionally, we demonstrate an inverse trend between viral diversity and pregnancy, with a higher number of viruses detected associated with failure to achieve clinical pregnancy in the azithromycin group. TWEETABLE ABSTRACT: Higher viral diversity is associated with prophylactic antibiotic exposure in subfertile women undergoing IVF.

High-throughput sequencing of cerebrospinal fluid for diagnosis of chronic Propionibacterium acnes meningitis in an allogeneic stem cell transplant recipient.

BACKGROUND: A 40-year-old man with chronic myelogenous leukemia presented multiple times over a period of 3 years with episodes of confusion, wide-based gait and falls because of recurrent hydrocephalus despite repeated therapeutic lumbar punctures. These problems occurred in the context of persistent cerebrospinal fluid (CSF) pleocytosis and leptomeningeal enhancement. Extensive diagnostic workups and therapeutic trials had failed to identify a clinically plausible cause or produce any significant improvement in the CSF and neuroimaging abnormalities. METHODS: We used high-throughput metagenomic shotgun sequencing to identify microbes in 2 CSF samples collected from the patient during his illness. These results were compared to sequence data from 1 CSF sample collected during treatment and 5 control CSF samples from other patients. RESULTS: We found sequences representing 53% and 67% of the Propionibacterium acnes genome in 2 CSF samples collected from the patient during his illness. Directed antimicrobial therapy was administered for 6 weeks with resolution of CSF and neuroimaging abnormalities. Sequencing of a sample obtained during treatment demonstrated that the P. acnes levels were decreased to background levels. After insertion of a ventriculo-peritoneal shunt, the patient returned to baseline status. CONCLUSIONS: High-throughput metagenomic shotgun sequencing revealed P. acnes as the cause of chronic meningitis that had eluded conventional attempts at diagnosis. Treatment directed at this organism resulted in cure of the infection and clinical improvement.

Erectile dysfunction and lower urinary tract symptoms: a consensus on the importance of co-diagnosis.

Despite differences in design, many large epidemiological studies using well-powered multivariate analyses consistently provide overwhelming evidence of a link between erectile dysfunction (ED) and lower urinary tract symptoms (LUTS). Preclinical evidence suggests that several common pathophysiological mechanisms are involved in the development of both ED and LUTS. We recommend that patients seeking consultation for one condition should always be screened for the other condition. We propose that co-diagnosis would ensure that patient management accounts for all possible co-morbid and associated conditions. Medical, socio-demographic and lifestyle risk factors can help to inform diagnoses and should be taken into consideration during the initial consultation. Awareness of risk factors may alert physicians to patients at risk of ED or LUTS and so allow them to manage patients accordingly; early diagnosis of ED in patients with LUTS, for example, could help reduce the risk of subsequent cardiovascular disease. Prescribing physicians should be aware of the sexual adverse effects of many treatments currently recommended for LUTS; sexual function should be evaluated prior to commencement of treatment, and monitored throughout treatment to ensure that the choice of drug is appropriate.

Sexuality and old age.

Sexuality in older people is a multifaceted field of study. Ageing may offer opportunities for and restrictions on sexuality. Ageing individuals may have increased time for and may wish to explore their sexuality, for example, by considering a same-sex relationship, after a long-term partner dies. However, they may also lack a partner or autonomy. They may develop or have increased symptoms from chronic diseases. Consequent medication may affect their sexual functioning. Their attitudes to sex often change and their capacity for sexual activity may diminish unless they seek help. However, they may lack the confidence to do so and the health professionals consulted may be disinclined or ill-equipped to assist them. They may have concerns regarding how their sexuality or their sexual orientation would be perceived should they require residential care or start to lose their memory. We consider the recent work relating to this broad range of influences on sexuality in old age and conclude that, whilst the challenge for researchers is to accurately clarify the impact of individual factors on sexuality and identify helpful potential interventions, increased academic and educational focus on this area is enhancing the prospects for a sexually enjoyable old age.

An e-mail and telephone helpline for sexual problems - results of a 2-year survey of men's sexual concerns.

INTRODUCTION: Sexual problems are relatively common in the community. The under-reporting of such problems calls for alternative ways of getting a true perspective of the prevalence of sexual dysfunction. AIM: To assess and investigate the concerns, ages and nationality of male users of a electronic helpline using it over 2 years (2009-2010). METHODS: Detailed records were kept of all emails and telephone calls to the helpline which included administrative and clinical queries. The clinical calls and some of the emails were answered by the administrator, while the majority of the emails were answered by a physician. This presentation will deal with the users who emailed for assistance with male sexual dysfunction. (n = 673). RESULTS: The helpline received a total of 6142 calls over the period of investigation, including administrative and press calls. Men accounted for 88% of the callers about sexual dysfunction. Erectile dysfunction accounted for 68% of the complaints by men who used the e-mail service. Premature ejaculation, loss of sex drive, genital problems and masturbation worries were other concerns that were received. A large number of men from the Middle East and the Indian sub-continent used the email service, mostly for premature ejaculation and masturbation worries, related to their arranged marriage. CONCLUSIONS: We found confirmation that the commonest sexual complaints in men were of erectile dysfunction and loss of sex drive. Significant correlations were identified between the types of sexual dysfunction and the age and ethnicity of the men who presented with them.

Therapeutic options for postmenopausal female sexual dysfunction.

BACKGROUND: Female sexual dysfunction (FSD) is a multidimensional problem combining biological, psychological and interpersonal elements of multiple etiologies. Menopause-related sexual dysfunction may not be reversible without therapy. Hormonal deficiency does not usually decrease in severity over time. Many options are available for the successful treatment of postmenopausal FSD. OBJECTIVE: To review the pharmacological and non-pharmacological therapies available for postmenopausal FSD, focusing on practical recommendations for managing postmenopausal women with sexual complaints, through a literature review of the most relevant publications in this field. PSYCHOSOCIAL THERAPY: This type of therapy (basic counselling, physiotherapy and psychosexual intervention) is considered an adaptable step-by-step approach for diagnostic and therapeutic strategies, normally combined with biomedical interventions to provide optimal outcomes. PHARMACOLOGICAL THERAPY: For postmenopausal FSD, many interventional options are now available, including hormonal therapies such as estrogens, testosterone, combined estrogen/testosterone, tibolone and dehydroepiandrosterone. CONCLUSIONS: Menopause and its transition represent significant risk factors for the development of sexual dysfunction. FSD impacts greatly on a patient's quality of life. Consequently, it is receiving more attention thanks to the development of effective treatments. Non-pharmacological approaches should be used first, focusing on lifestyle and psychosexual therapy. If required, proven effective hormonal and non-hormonal therapeutic options are available.

Review of drug treatment for female sexual dysfunction.

Female sexual dysfunction (FSD) incorporates various sexual disorders including hypoactive sexual desire disorder, sexual arousal disorder, orgasmic and sexual pain disorders. Although many strategies have been formulated for the treatment of male sexual problems, FSD remains an area that warrants further research and trial studies to identify the most efficacious treatment options. Research has highlighted numerous pharmacological interventions that have been trialled and found to exhibit positive effects. These include hormonal therapies, prostaglandins, dopaminergic agonists, phosphodiesterase type-5 (PDE-5) inhibitors and melanocortin agonists.

Management of psychosexual and relationship problems in general mental health services by psychiatry trainees.

AIMS AND METHODS: All psychiatry trainees (both Basic and Higher Specialist Trainees in Adult and Old Age Psychiatry) in the Nottingham (Mid-Trent rotation) were invited to complete a questionnaire evaluating the assessment and training of psychosexual and relationship problems in the general mental health services. RESULTS: Only 24% of trainees reported to routinely asked patients about psychosexual history, 65% felt comfortable about taking a detailed psychosexual history. 81% of trainees reported inadequacy whilst dealing with psychosexual disorders during their training in psychiatry. Only 30% reported asking patients for potential sexual side-effects when on psychotropic medication. All the trainees would refer the patient to a local psychosexual disorder clinic (if available). CLINICAL IMPLICATIONS: Taking a sexual and relationship (both past and current) history should be an integral part of any psychiatric assessment. Training in human sexuality should be introduced in medical schools. Training in human sexuality is a core part of psychiatric training curriculum and trainees should be tested for their competency in dealing with sexual dysfunctions.

Double-blind placebo-controlled study of testosterone patch therapy on bone turnover in men with borderline hypogonadism.

Clinical studies suggest there may be a threshold concentration of serum testosterone below which replacement will result in skeletal and psychological benefit. We evaluated the response to testosterone in men with borderline hypogonadism. A randomized double-blind placebo-controlled trial in 39 men over age 40 years presenting with sexual dysfunction and a borderline low testosterone level (total testosterone <10 nmol/L or free androgen index <30%). Patients were randomized to Testoderm TTS body patch (5 mg/day, n = 20) or a placebo patch (n = 19) for 6 months, followed by open-label testosterone replacement for a further 6 months in all patients. During the placebo-controlled phase of the study serum testosterone increased significantly on testosterone vs. placebo treatment (p = 0.004); this was associated with a decrease in total body fat mass (p = 0.019) and increase in haemoglobin level (p = 0.036). There were no significant changes in lean body mass, markers of bone turnover, and measures of bone mineral density (BMD). There was evidence of difference in quality of life according to the Male Erectile Dysfunction Quality of Life questionnaire (MEDQoL score, p = 0.017), mainly accounted for by deterioration in the placebo arm. When the active treatment period was combined for placebo and testosterone groups, the within-patient analysis showed a significant effect of testosterone to decrease markers of bone resorption (uNTX/Cr, p = 0.007; iFDPD/Cr, p = 0.0006) and to increase lean body mass (p = 0.001). There was little convincing evidence from this study that testosterone replacement is likely to have major benefit in men over age 40 years with borderline hypogonadism and sexual dysfunction. However, there was evidence of suppression in bone resorption and hence longer and larger studies are needed to examine its effect on BMD.

A comparison between portable ultrasound (MIDUS) and nocturnal RigiScan when confirming the diagnosis of vascular organic erectile disorder.

In recent years, the use of RigiScan and ultrasound to assess erectile dysfunction has fallen from favour. However, in a small minority of specialist cases, where a vascular, neurogenic or psychogenic aetiology requires confirmation, there remains a need for further investigation. To establish if in a preliminary assessment the use of nocturnal RigiScan or male impotence diagnostic ultrasound system (MIDUS) represents best practice as a diagnostic investigation in patients with a history suggestive of vascular organic erectile disorder. Men attending both urological and psychosexual therapy clinics with erectile dysfunction were assessed using a generic assessment schedule. Patients with a history suggestive of vascular erectile disorder were offered the opportunity of dual investigation of their condition. After screening using a provocative RigiScan using visual stimuli that gleaned inconclusive results, patients were offered the chance to enter a study with both nocturnal RigiScan and MIDUS investigation. These were confined for the purposes of this study to RigiScan events, peak systolic flow velocity (PSV) and end-diastolic flow velocity (EDV) from ultrasound examination where an abnormal EDV is defined as in excess of 4.5 cm/s and a normal PSV is variously defined as being greater than 35 cm/s. In all, 38/43 (88%, 95% CI: 76-95%) of men had a nocturnal event exceeding 3 min on the RigiScan investigation. This compares with 17/43 (40%, 95% CI: 26-54%) of men with a normal EDV blood flow of less than 4.5 cm/s (P<0.017) and 32/43 (74%, CI: 60-85%) of men with a normal PSV flow greater than 35 cm/s (NS). Rigiscan and ultrasonography of the cavernosal vessels are of equal usefulness in suspected arterial penile disease although where veno-occlusive disease is suspected, ultrasonography is more specific.

Sexology as a profession in the United Kingdom.

To identify clinicians working within the field of sexual and psychosexual medicine and to identify and quantify the diversity of problems seen by specialist physicians, nurses and therapists working in the UK and compare these with French clinicians, a prospective 89 item questionnaire was sent to 2352 practitioners in the UK. Eight hundred and fourteen completed questionnaires were returned detailing clinician practice and training and the presenting problems of patients in their clinical practice. The commonest presenting problem amongst men was erectile dysfunction (562/814) followed by emotional and relational problems. The commonest problem for women was loss or absence of sexual desire and was of a similar magnitude to that of erectile dysfunction in men. The substantial majority of clinicians working with patients and clients with sexual problems are non-physicians from a diverse professional training background which contrasts with a high number of physicians in France. The high incidence of emotional and relational problems in both men and women would suggest that clinical services require considerable psychotherapeutic consultation space within the service provision.

Psychological difficulties within a group of patients with vulvodynia.

This study reviews and reports on some of the psychological difficulties seen in a group of women with vulval pain (vulvodynia). The investigation involved 164 women (82 suffering with Vulvodynia and 82 women in a control group with general dermatology conditions) to establish the prevalence of psychological difficulties using validated questionnaires (SCL-90R and the IBQ) and reports on the possible effect these may have on sexual and relationship function and satisfaction. The level of psychological difficulties revealed significantly higher levels of psychological distress in the vulvodynia group within the domains of somatisation, obsessive-compulsive, depression, anxiety & phobic symptoms as well as with interpersonal sensitivity hostility and paranoia.

Returning a referral for erectile dysfunction to the referrer by two different routes.

OBJECTIVE: To compare the prescribing pattern and attitude of general practitioners (GPs) in response to a clinic returning a patient referred for erectile dysfunction (ED) to the referrer by two different methods. METHODS: Referrals on a waiting list for an assessment of ED were reviewed and a subgroup of patients with criteria enabling them to be eligible for a prescription under the National Health Service (NHS) identified. The GP was informed either in writing or by telephone that the clinic had written to the patient, suggesting he make direct contact with his GP. A follow-up questionnaire was sent to each GP one month after the initial letter to the patient and contact with the GP. RESULTS: Of 91 questionnaires sent to GPs, 66 (73%) were completed; an additional five GPs corresponded by letter rather than completing the questionnaire. The long waiting time for assessment had led to 35% of patients having already tried sildenafil, and by the time the questionnaire was completed, 57% of patients had tried sildenafil. Ten times as many referrers indicated that they were happy to initiate a prescription for sildenafil than not to do so, for those men eligible for an NHS prescription. More GPs who had received a letter returned the completed questionnaire (80%) than those who had received a courtesy telephone call (64%). There were no differences between the groups of GPs in their attitude to our contact with their patient and no difference in prescribing pattern. Conclusion The provision of guidelines and advice to GPs either by telephone or letter is acceptable practice in reducing long waiting-list times for ED. Safe, simple and effective treatments are available for GPs to use under NHS guidelines.

A physical map of the human genome.

The human genome is by far the largest genome to be sequenced, and its size and complexity present many challenges for sequence assembly. The International Human Genome Sequencing Consortium constructed a map of the whole genome to enable the selection of clones for sequencing and for the accurate assembly of the genome sequence. Here we report the construction of the whole-genome bacterial artificial chromosome (BAC) map and its integration with previous landmark maps and information from mapping efforts focused on specific chromosomal regions. We also describe the integration of sequence data with the map.

A physical map of the human Y chromosome.

The non-recombining region of the human Y chromosome (NRY), which comprises 95% of the chromosome, does not undergo sexual recombination and is present only in males. An understanding of its biological functions has begun to emerge from DNA studies of individuals with partial Y chromosomes, coupled with molecular characterization of genes implicated in gonadal sex reversal, Turner syndrome, graft rejection and spermatogenic failure. But mapping strategies applied successfully elsewhere in the genome have faltered in the NRY, where there is no meiotic recombination map and intrachromosomal repetitive sequences are abundant. Here we report a high-resolution physical map of the euchromatic, centromeric and heterochromatic regions of the NRY and its construction by unusual methods, including genomic clone subtraction and dissection of sequence family variants. Of the map's 758 DNA markers, 136 have multiple locations in the NRY, reflecting its unusually repetitive sequence composition. The markers anchor 1,038 bacterial artificial chromosome clones, 199 of which form a tiling path for sequencing.