Difficult insertion of IUS

New technologies are ready to revolutionize glomerular imaging and significantly improve or replace immunofluorescence and electron microscopy, which have driven research and diagnosis of glomerular diseases for over 50 years. Advanced forms of transmission and scanning electron microscopy have revealed the detailed spatial relationships of the glomerular basement membrane, podocytes, and endothelial cells. These may be overshadowed by super resolution microscopy (SRM), which combines the advantages of immunofluorescence and electron microscopy, offers high resolution identification of specific molecules, and images large, physiologically relevant volumes of the glomerulus. Rapidity, ease of use and low cost with some types of SRM make them potentially suitable for routine diagnosis. SRM visualizes structures below the classical diffraction limit of conventional light microscopy by adding a time variable to either the illumination of the specimen, or to the fluorescence signal emitted by it. Ensemble techniques vary illumination and include Structured Illumination Microscopy (SIM) and Stimulation Emission Depletion Microscopy (STED). Single molecule localization techniques vary the light emission by fluorescence labels in the specimen, and include Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM). Technologies such as expansion microscopy and genetic labeling can also create effective super resolution imaging by non-optical, specialized preparation techniques. All technologies require dark field fluorescence and some require computer image analysis and reconstruction. Replicating successful application in other areas of biology, SIM, STED, and STORM have visualized normal and nephrotic disease podocytes, and have confirmed their appearances to be similar to those seen by electron microscopy, but with added new information on cell configuration and protein localization. STORM has also localized podocyte cytoskeleton and adhesion proteins, and glomerular basement membrane proteins at a resolution never before possible. These pioneering efforts show the promise of super resolution microscopy, and lay the groundwork for future study and new diagnostic tools for glomerular diseases.

Neisseria gonorrhoeae and Chlamydia trachomatis among human immunodeficiency virus-infected women.

<h3></h3> The aim of this study was to evaluate the performance of UCM (Urine Conservation Medium, Novosanis) for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) DNA preservation at room temperature (RT). To compare the stability of CT and NG DNA in unpreserved and UCM-preserved samples, five urine samples were spiked to obtain 99 CT and 75.5 NG DNA copies/µL in the final aliquot. For UCM-preserved samples, UCM was spiked and urine was added in a 2:1 (urine:UCM) ratio to mimic real-life use of Colli-Pee containing UCM. A total of 30 aliquots were analyzed using the Abbott Real-Time CT/NG assay after storage at RT for 1, 8 and 15 days. Mixed-effect regression analysis was used to estimate the effect of UCM on CT and NG Ct values at each timepoint, with urine samples as random effects. After 15 days of storage, all UCM-preserved samples were positive for CT and NG, while only one and three unpreserved samples remained positive for CT and NG respectively. Ct values where samples were negative for CT and/or NG were defined as the cut-off values (cut-off 8d: CT=39.66, NG=39.93; cut-off 15d: CT=39.56, NG=39.92). At each timepoint, Ct-values for CT and NG were significantly lower for preserved compared to unpreserved samples. At 1, 8 and 15 days of storage UCM-preserved samples showed an estimated differences of -6.03, -5.91 and -4.60 cycles for CT, and -6.91, -7.20 and -7.36 for NG respectively. All effects are statistically significant with p&lt;0.01. UCM-preserved urine sampling enables preservation of CT and NG DNA for 15 days of storage at RT. The use of Colli-Pee, whereby the collector tube is prefilled with UCM, allows for immediate mixing of urine and preservative. This offers opportunities for home-based testing where sample quality is maintained during storage and sample shipment to the laboratory by regular postal mail.

Expansion microscopy (ExM) is a powerful high-resolution imaging technique that enhances the spatial resolution of conventional light microscopy by physically enlarging biological specimens by embedding and cross-linking them in a swellable polymer network. This review explores the combination of ExM with commonly used advanced fluorescence imaging modalities, including light sheet fluorescence microscopy (LSFM), stimulated emission depletion (STED), structured illumination microscopy (SIM), single-molecule localisation microscopy (SMLM), and computational super-resolution radial fluctuations (SRRF) to push the boundaries of achievable resolution in biological imaging. By integrating ExM with these optical and analytical approaches, researchers can visualise subcellular structures and molecular complexes with unprecedented clarity, enabling the study of intricate biological processes that are otherwise inaccessible with conventional light microscopy methods. The review covers the theoretical resolutions attainable with each combined technique, example biological questions they can address, and key considerations for optimising their use. Together, these advancements offer novel insights into nanoscale cellular and subcellular structures, opening new avenues for exploration in fields such as neuroscience, cancer research, and developmental biology.

Background Infections with Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV) are among the most prevalent and treatable sexually transmitted diseases in the US. Currently, most available testing methods require patient samples to be split between CT/NG and TV assays. Here, we evaluate performance characteristics of the Seegene Novaplex STI Essential Assay, a real-time multiplex polymerase chain reaction assay for concurrent amplification and detection of multiple pathogens including CT, NG, and TV. Methods To prepare urine samples for extraction, 1 mL of each sample was centrifuged at 13 000 rpm for 15 min. The supernatant was discarded and the pellet was rehydrated in 190 µL of saline. Nucleic acid extraction was performed on a CyBio® FeliX liquid handler (Analytik Jena, Germany) using MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit (Thermo Fisher Scientific, MA). PCR amplification and detection were performed using Seegene Novaplex STI assay (Seegene Inc, South Korea) on a CFX96™ System (Bio-Rad Laboratories, CA). The assay utilizes Seegene MuDT™ technology that allows detection of multiple targets in a single channel without melting curve analysis. For method validation, precision studies were performed within-run and over 5 days. Limits of detection were verified by spiking known negative urine samples with CT/NG standards from Exact Diagnostics (Bio-Rad Laboratories, CA) and TV standards from ZeptoMetrix® (Antylia Scientific, IL). Method comparison was carried out by extracting and analyzing split patient urine samples against cobas® CT/NG assay (Roche Diagnostics, IL) and Solana® Trichomonas assay (Quidel, CA) performed by a reference laboratory. Additional accuracy studies were performed by spiking 20 negative patient samples with variable concentrations of the three pathogens. Results Within-run and between-day precision studies demonstrated consistent and reproducible results for all pathogens. Limit of detection was established at 10 copies/mL for CT and NG and 10 cells/mL for TV. Results of the spike-and-recovery studies were consistent with expected results for all three targets across evaluated concentrations. Patient correlation studies demonstrated 100% agreement with results obtained by the reference laboratory for both NG (32 total samples, 11 positive) and TV (22 total samples, 4 positive), and 96% agreement for CT (29 total samples, 10 positive). Conclusion The Seegene Novaplex STI Essential Assay offers a rapid and reliable method for detecting CT, NG, and TV in urine samples. The capability of the assay to detect multiple targets in a single sample well improves efficiency and turnaround time.

Rectal Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections among men who have sex with men (MSM) are escalating public health concerns. This study aimed to explore (1) the reliability of self-reported sexual positioning as an indicator for rectal CT and NG screening, and (2) factors associated with rectal CT and NG infections in Shenzhen, China. A cross-sectional study was conducted in 2 settings in Shenzhen, China, from April 1, 2021, to March 31, 2022. Data on sociodemographic characteristics, sexual behaviors, and basic CT knowledge were collected. Urine and self-collected rectal swabs were collected for CT and NG testing. In total, 195 MSM participated in the study, and 5.1% tested positive for urogenital CT, 29.2% for rectal CT, 1.0% for urogenital NG, and 8.2% for rectal NG. Among those who reported exclusively insertive anal sex, 69.2% of CT infections and 85.7% of NG infections would have remained undetected with urine testing alone. Risk factors for rectal CT infection included engaging in both insertive and receptive anal sex, with a significant association found for coinfection with rectal NG. Self-reported sexual positioning was found to be an unreliable indicator for CT and NG screening, as a substantial proportion of infections would have remained undetected. The findings suggest that CT and NG screening in China should be offered to all MSM regardless of self-reported sexual positioning, and that the dual CT/NG testing is recommended.

Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) are common etiologic agents of reproductive tract infections (RTIs) with high global prevalence and the potential to cause adverse genital outcomes. This study assessed the positivity rates and co-infection patterns of UU, CT, NG, and HSV among women in Xihu District, Hangzhou, Zhejiang Province, China. We analyzed 5,577 female genital tract specimens collected from January 2018 to December 2024 at Tongde Hospital of Zhejiang Province. Pathogen DNA was detected using real-time PCR. UU susceptibility testing was performed with the Mycoplasma Identification and Susceptibility Test Kit. Among 5,577 specimens, positivity rates were 49.11% for UU, 7.48% for CT, 1.88% for NG, and 10.52% for HSV. A total of 118 mixed infections involving two or more pathogens were identified; the most frequent combination was UU + CT, accounting for 73.73% of mixed infections. UU was the most common organism in co-infections, present in more than 96% of mixed-infection cases. Significant associations were observed between UU and CT (p < 0.001) and between UU and NG (p < 0.05). The overall pathogen positivity rate increased over the seven-year period (p < 0.01). Younger women (≤ 19 years) had the highest positivity for UU, CT, NG, and HSV-1, whereas older women (> 60 years) had the highest HSV-2 positivity. More than 95% of UU isolates were susceptible to doxycycline, minocycline, and josamycin. This study characterizes the epidemiology of UU, CT, NG, and HSV in real-world obstetrics and gynecology outpatient settings in Xihu District, Hangzhou. Given the elevated positivity among younger women, targeted health education and management strategies are warranted.

IntroductionUreaplasma urealyticum (UU) is probably one of the causes of non-gonococcal, non-chlamydial urethritis in men and post-partum endometritis in women. The epidemiology of UU is currently unclear because culture isolation...

ObjectivesThere has been considerable discussion about anorectal Chlamydia trachomatis (CT) in women, with some calling for anorectal CT screening, but little about anorectal Neisseria gonorrhoeae (NG). Given that urogenital NG...

Standard operating procedure in STD clinics commonly has been to test urethral specimens when evaluating males, whether they are heterosexual or men who have sex with men (MSM). Rectal or oropharyngeal specimens may be tested in MSM with symptoms, or in some clinics as screening tests if individuals report sexual practices that would indicate risk of infection at these sites. In a report in this issue of Sexually Transmitted Diseases, Marcus and colleagues studied the prevalence of infection with Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) in the urethra, the oropharynx, and rectum in asymptomatic MSM visiting the San Francisco STD clinic.1 A retrospective analysis of 3398 patient visits found CT or GC at 549 (16.2%) of those visits. The prevalence of infections among these asymptomatic men ranged from a high of 7.8% for rectal CT to a low of 0.4% for urethral GC. Strikingly, 83.8% of CT and GC infections would have been missed if only urethral screening was performed. What was found in the Marcus study, and in others, as well, is that the majority of infections with CT/GC in MSM are not in the urethra. The first major study to make this point was performed by Kent and colleagues, in 2 clinics in San Francisco, where they found that 50% of chlamydial and gonococcal infections were extraurethral.2 That finding has been confirmed by a number of approaches, including retrospective chart reviews and prospectively designed studies (Table 1). While these studies have focused on rectal and pharyngeal CT/GC infections in MSM, it must be noted that such infections are far from rare in women.12,13 In many clinical settings, the infections in women will be seen more often than those in men. These extragenital infections should not be ignored now, and we need more research to better define their potential consequences. Both CT and GC are recognized as causes of proctitis, and even asymptomatic infections have been associated with HIV transmission. 14 Current CDC recommendations call for routine screening of the oropharynx and rectum in MSM who may be exposed to GC or CT at those sites. It is not clear whether pharyngeal infections have a major direct impact on health. But evidence is emerging that infection can be transmitted from the oropharynx to the urethra of sex partners, thus continuing chains of infection.15 All of the cited studies have been made possible by the introduction and application of highly sensitive and specific nucleic acid amplification tests (NAATs) for the diagnosis of CT and GC infections. NAATs are recommended for routine diagnosis of CT/GC infections.15 Where direct comparisons have been done, it has been shown that the increment in sensitivity for NAATs as compared to culture is greater with pharyngeal and rectal specimens than it is with cervical and urethral specimens. An approximate doubling of the number of infections detected has been obtained with NAATs. 4,11 Alas, there remains a gap wherein the best tests are not readily available to the patients and providers who need them. Several NAATs are commercially available and have received FDA clearance for male and female urine specimens, urethral swabs from men, and cervical and vaginal swabs from women. Unfortunately, none have been cleared by the FDA for pharyngeal or rectal specimens. But the CDC, recognizing the superior performance of these tests used on these specimens, has taken an unusual step and recommended the use of NAATs for diagnosis of CT/GC in oropharyngeal and rectal specimens, even though such use has not received FDA clearance. 16 The

We evaluated the prevalence, incidence, and correlates of sexually transmitted infections among pregnant women initiating preexposure prophylaxis. We analyzed data from an ongoing randomized control trial that enrolled pregnant women in Kenya (NCT04472884) who were ≥18 years, between 24 and 32 weeks' gestation, had high HIV risk scores, and initiated oral preexposure prophylaxis within routine antenatal care. A subset of women were sequentially offered Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) testing in pregnancy and at 6 and 9 months after delivery. Directly observed treatment and expedited partner therapy (EPT) were offered to women diagnosed with CT/NG. All pregnant women offered CT and NG testing accepted (n = 221). The median age of women was 26 years (interquartile range 22-30); median gestational age was 27 weeks (interquartile range 25-29). Prevalence of CT and/or NG during pregnancy was 19/221 (8.6%): 4.1% CT, 3.6% NG, and 1% CT and NG coinfection. Women <24 years were 3 times more likely to have prevalent CT/NG infection as older women (adjusted prevalence ratio: 3.12; 95% confidence interval [CI]: 1.24 to 7.85 P = 0.016). Overall, 21 CT/NG infections occurred in 181.2 total person-years of follow-up (incidence 11.6 per 100 person-years, 95% CI: 7.6 to 17.8). Incident CT and/or NG was 7-fold higher among women <24 years compared with older women (adjusted incidence rate ratio = 6.69, 95% CI: 2.55 to 17.54, P < 0.001). Same-day directly observed treatment and EPT acceptance was 95%; at subsequent visits, 79% reported partners accepted EPT, of whom 95% confirmed treatment completion. CT/NG incidence was high among women who initiated preexposure prophylaxis in pregnancy, indicating that CT/NG testing would be high yield in this population.

Infective anoproctitis in men having sex with men: Don’t forget Mycoplasma genitalium

BackgroundThis study aims to explore the infection and age distribution of Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Herpes simplex virus type II (HSV II) among the outpatients of Reproductive Medicine Center in Putian, Fujian Province to provide a clinical basis for the early diagnosis and treatment of various reproductive tract diseases and infertility in this region.MethodsA total of 1736 samples of secretions and exfoliated cervical cells were collected from the outpatients of the Reproductive Medicine Center of the Affiliated Hospital of Putian University from December 2021 to April 2023. The infections of UU, CT, NG and HSVII were detected by real-time fluorescence polymerase chain reaction (PCR), and the infection statuses of the patients with different genders, ages and diagnoses were analysed.ResultsAmong the 1736 patients, 611 were male and 1125 were female. The male patients had higher UU infection rate but lower HSV II infection rate than the female patients. No significant difference in CT and NG infection rates was observed between the genders. The CT infection rate gradually decreased with the increase in the age. The difference in UU, NG and HSV II infection rates among the different age groups was not statistically significant. For UU infection, the male infertile patients had the highest rate of 37.72% (172/456). Meanwhile, the differences in CT, NG and HSV II infection rates among the different diagnosis groups were not statistically significant. Among the male and female infertile patients, the CT infection rate was the highest in the 21–25 years of age group at 11.11% (2/18) and 9.47% (9/95), respectively. No statistically significant difference in UU, CT, NG and HSV II infection rates was observed among the different age groups of patients diagnosed in relation to the family planning guidance and between the male and female patients with other diagnoses results.ConclusionsThis study showed that UU was the most frequently identified pathogen in infertile men in Putian, Fujian Province. The CT infection rate was the highest in people under 20 years old, and the infection showed a tendency toward young individuals. Therefore, the publicity of sexual health knowledge must be strengthened, and the prevention and treatment of venereal diseases among young and middle-aged people must be improved. Moreover, the pathogen infection is related to infertility to a certain extent, which is conducive to clinical diagnosis and treatment.

<h3>Background</h3> Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Trichomonas vaginalis(TV) are among the major causative agents of sexually transmitted infections (STIs). Huge fraction of patients with NG, CT and TV infections are asymptomatic making syndromic case management(SCM) becomes unreliable. Taqman based triplex qPCR assay for simultaneous detection of NG, CT and TV, besides being dependable diagnostic tool, can effectively differentiate between mono-infections and co-infections of these pathogens. <h3>Methods</h3> Endo-cervical dry swabs were procured from non-pregnant females visiting Department of Obstetrics &amp; Gynaecology, VMMC and Safdarjung Hospital, Delhi, India as per ICMR ethical guidelines. DNA was isolated from the samples and presence of the causative agent, if any, was determined using in-house developed Multiplex qPCR assay by employing specific primer-probe sets targeting each pathogen. Analytical sensitivities and specificities were determined for each uniplex assay. In-house developed assay was clinically validated in comparison to published NAAT based assays. <h3>Results</h3> A 108 bp region of orf-1 gene of NG, 133 bp region of pld gene of CT and 147 bp region of pfo-B gene of TV genomic DNA were utilized for identification. Analytical sensitivity of NG uniplex assay was determined to be 10ag of recombinant DNA (3–4 DNA equivalents) and for both CT and TV, it was determined to be 100ag of DNA (30 DNA equivalents). 85 clinical samples have been examined so far and the assay detects NG and CT DNA with 83% and 60% sensitivities, 99% and 100% specificities, 83% and 100% PPV and, 99% and 98% NPV respectively. Two clinical samples among them were co-infected with NG and CT. <h3>Conclusion</h3> In-house developed multiplex qPCR assay provides an opportunity for diagnosis of three highly prevalent and curable STIs in only one assay. The results following validation are motivating enough to project it as a potentially reliable diagnostic tool for their detection.

97: Utility of Wet Prep In Predicting Cervical Infections In Women