mgpB genotyping and genetic diversity for antimicrobial resistance of Mycoplasma genitalium.

Introduction. Antimicrobial resistance (AMR) among Mycoplasma genitalium is a global issue. Understanding the transmission dynamics of infection is an important factor in reducing the occurrence of AMR.Hypothesis/Gap Statement. There is limited information on the genotyping and AMR traits of M. genitalium.Aims. Single-locus sequence-based (SLSB) mgpB sequence typing and genetic diversity analyses of AMR M. genitalium isolated from patients in the Republic of Korea were performed to clarify the transmission dynamics and eludicate proper management.Methodology. Sanger sequencing of mgpB, 23S rRNA, parC and gyrA genes from a total of 103 M. genitalium-positive specimens from 89 patients was carried out.Results. Twenty-seven different mgpB genotypes (GTs) were identified; 12 had been reported previously and 15 had not. GT7 and GT8 occurred frequently (n=38, 36.89 %, and n=16, 15.53 %, respectively). The genetic diversity of the AMR-determining sites was randomly dispersed among the different GTs. However, these GTs were classified into two phylogenetically distinct clusters that were significantly correlated with patient age and genetic diversity at positions 2058 and 2059 in the 23S rRNA gene. The GTs of 20 consecutive samples from 6 patients were compared to investigate temporal changes in GTs. One specimen changed its GT during follow-up, suggesting a new infection.Conclusions. mgpB sequence typing can be a reliable tool for epidemiological studies. Two clusters have different characteristics in terms of genetic diversity. The cluster with genetic diversity in the AMR-determining site may be explained by the high prevalence of the specimens and subsequent antimicrobial exposure during the study period.

Molecular mechanisms of macrolide and fluoroquinolone resistance among Korean isolates of Mycoplasma genitalium over a period of five years 2014-2019.

Antimicrobial resistance in Mycoplasma genitalium has become a global issue, and certain groups have a higher probability of acquiring resistant strains. Little is known about the genetic diversity and characteristics of the antimicrobial resistance-determining sites (ARDSs) of M. genitalium in the Korean population. Therefore, we examined the genetic diversity of the ARDSs of M. genitalium-positive urogenital samples obtained from Korean females (G1) and males (G2) visiting primary care clinics and DNA samples from referred males (G3) with persistent urethritis. From 2014 to 2019, 54 patients from G1, 86 patients from G2, and 68 patients from G3 were included in the study. Sanger sequencing was performed on the 2058/2059 sites in the 23S rRNA gene and quinolone resistance-determining regions (QRDRs) of M. genitalium. The rates of mutation in G1, G2, and G3 were 1.85, 5.81, and 48.53 %, respectively, for A2059G in the 23S rRNA gene (P<0.001); 1.85, 0, and 17.78 %, respectively, for M95R or I in gyrA (P<0.001); 0, 0, and 31.11 %, respectively, for D99N or G in gyrA (P<0.001); and 7.41, 16.28, and 30 %, respectively, for S83R or N or I in parC (P=0.015). A2059G significantly increased the risk of mutations at the gyrA95, gyrA99, and parC83 sites (all P<0.01). In conclusion, although the genetic diversity of the ARDSs of M. genitalium was variable among the groups, it was generally lower in isolates with macrolide resistance and higher in isolates with quinolone resistance in Korea compared with the isolates in other countries. The G3 group demonstrated increased genetic diversity at the A2059G, gyrA95, gyrA99, and parC83 sites.

Roles of Ureaplasma Species in Idiopathic Chronic Prostatitis: A Case-Control Study.

PURPOSE: Because of the inconsistent symptoms associated with Ureaplasma infections, their clinical significances in genitourinary tracts are under debate. Therefore, we evaluated the presence of Ureaplasma urealyticum (UU) and Ureaplasma parvum (UP) in urine samples and examined their associations with chronic prostatitis (CP) through a case and control study. MATERIALS AND METHODS: We included 696 nonchlamydial nongonococcal (NCNG) urine samples from men; 350 were categorized into non-inflammatory CP, 88 in inflammatory CP, and 258 in non-CP group. We amplified a region in the Ureaplasma urease areas from these samples and determined their biovars using the Sanger method. RESULTS: Among the NCNG population, the rates of UU, UP, and non-UU/UP were 3.88%, 6.46%, and 89.66%, respectively. The overall infection rates of non-CP, inflammatory CP, and non-inflammatory CP groups were 4.15%, 6.10%, and 3.65% in UU (p=0.612) and 6.85%, 7.22%, and 6.50% in UP (p=0.968), respectively. UU infection increased the risk of white blood cell (WBC) counts (≥5) in urine (p=0.005). In contrast, UP infections did not increase the risks of urethritis. Re-analysis from the 633 men who were excluded from urethritis effects did not reveal the associations between UU infection and the clinical characteristics of CP. Furthermore, the profiles from the National Institutes of Health-Chronic Prostatitis Symptom Index questionnaire and WBC counts in expressed prostatic secretion were similar among the non-CP and the two CP groups in each Ureaplasma infection. CONCLUSIONS: We found that UU may induce male urethritis. However, Ureapalsma species in urine were not definitively associated with the occurrence of CP.

A case of testicular infarction from the complications of Klebsiella oxytoca induced acute epididymitis.

Herein, we reported a case of testicular infarction in a patient with Klebsiella oxytoca induced acute epididymitis. Acute left epididymitis progressed into testicular infarction requiring orchiectomy in spite of antibiotics treatment. Ordinary urine cultures did not reveal any specific organism, suggesting viable but noncultureable state. We amplified a bacterial 16S ribosomal subunit gene from the urine and orchiectomized samples, and we found K. oxytoca infections from both of them.

The presence of Chlamydia is associated with increased leukocyte counts and pain severity in men with chronic pelvic pain syndrome.

OBJECTIVE: To examine the role of urinary chlamydial infection in patients with chronic prostatitis and/or chronic pelvic pain syndrome (CP-CPPS). METHODS: We recruited men aged 18-55 years over 4 years. We excluded patients with acute urethritis and any acute genitourinary infections. The participants evaluated their CP-CPPS-like symptoms with the Korean version National Institutes of Health-Chronic Prostatitis Symptom Index questionnaires. We measured white blood cell (WBC) counts in expressed prostatic secretion (EPS). In-house nucleic acid amplification test for Chlamydia trachomatis and Neisseria gonorrhoeae detection and WBC counts were performed from the patient's first-voided urine. RESULTS: A total of 765 eligible men were classified into 3 groups: 196 in non-CP-CPPS, 410 in noninflammatory CP-CPPS, and 159 in inflammatory CP-CPPS groups. The chlamydia-infected men showed higher pain, poor quality of life (QOL), and total scores in National Institutes of Health-Chronic Prostatitis Symptom Index questionnaires than the negative men (P = .041 for pain; P = .043 for QOL, and P = .027 for total). Multivariate analysis found that urinary chlamydial infection increased the risk of WBC count ≥16 in EPS (adjusted odds ratio [OR], 2.189; 95% confidence interval [CI], 1.021-4.690; P = .044) and WBC count between 2 and 4 in urine (OR, 5.227; 95% CI, 2.503-10.918; P = .001). In addition, chlamydial infection also increased the risk of inflammatory CP-CPPS than the non-CP-CPPS group (OR, 2.448; 95% CI, 1.010-5.932; P = .044), whereas the patients with noninflammatory CP-CPPS were not affected (OR, 1.6557; 95% CI, 0.738-3.717; P = .221). CONCLUSION: Urinary chlamydial infection increased the pain scores and WBC counts in EPS and worsens the QOL in the patients with CP-CPPS.

Contribution of genetic variation rs266882 to prostate-specific antigen levels in healthy controls with serum PSA below 2.0 ng/ml.

We evaluated the impact of genetic variation in the prostate-specific antigen (PSA) gene (rs266882) on serum PSA levels in healthy men as well as risk factors for benign prostate hypertrophy (BPH) and prostate cancer. The study population comprised 91 men with PSA levels below 2.0 ng/ml as healthy controls, 78 men with PSA 2-10 ng/ml as a BPH group, and 128 prostate cancer patients, all in Korea. DNA was amplified by polymerase chain reaction and the product was sequenced. We found that PSA levels were associated with a G/A polymorphism only in healthy controls. The transition, however, was not associated with PSA levels of BPH and cancer patients, nor was it a risk factor. In conclusion, this genetic factor is important for determining serum PSA levels in the naive group, whereas the disruption of prostatic architecture in BPH or prostate cancer may be a major determining factor for PSA levels.

Modulation of alpha 1 adrenergic receptors on urinary bladder in rat spinal cord injury model.

PURPOSE: Whereas many studies have focused on the vesical changes of the α1 adrenergic receptor (AR) subtypes in partial outlet obstruction, few studies have addressed the modulation of the α1 AR subtypes after spinal cord injury (SCI). Therefore, we studied the modulation of the α1 ARs in urinary bladder in a rat SCI model. METHODS: Four weeks after a SCI, the whole vesical bodies from eight female Sprague-Dawley rats and from eight controls were harvested. The total RNA was extracted from the samples and was used to prepare cDNA. We developed standard plasmid constructs of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and three α1 ARs (α1a, α1b, and α1d) to convert the cycle threshold (Ct) values from real-time polymerase chain reaction (RT-PCR) into subtype mRNA concentrations. The detected Ct values of 16 samples from RT-PCR were interpolated into the standard plasmid curves. RESULTS: All serially diluted standard samples showed very good linearity. The mRNA expression of GAPDH was higher in the SCI group, whereas the mRNA expression of all α1 ARs was lower in the SCI group than in the control animals. The α1a, α1b, and α1d mRNA expression in the controls was 81.7%, 3.3%, and 15.1%, respectively, whereas the α1a, α1b, and α1d mRNA expression in the SCI group was 33.5%, 5.2%, and 60.9%, respectively. CONCLUSIONS: SCI moderates the α1 AR mRNA subtypes in the urinary bladder. The relatively increased α1d or decreased α1a AR mRNA expression may be a therapeutic candidate for controlling the symptoms of neurogenic bladder after SCI.

Effects of CYP4F2 gene polymorphisms on warfarin clearance and sensitivity in Korean patients with mechanical cardiac valves.

BACKGROUND: Recently, a single nucleotide polymorphism of CYP4F2 (rs2108622) was reported to have a significant relationship with the stable warfarin dose. However, the underlying mechanism of CYP4F2 effects on the stable warfarin dose has not been studied. This study aimed to examine the effects of cytochrome P450 (CYP) 4F2 gene on warfarin clearance and sensitivity in Korean patients with mechanical heart valves. METHODS: One hundred ninety-one patients with mechanical heart valves who were on anticoagulation therapy with warfarin and maintained international normalized ratio levels of 2-3 for 3 consecutive times were followed up, retrospectively. Warfarin enantiomer concentrations were determined by a validated high-performance liquid chromatography method. Genotypes of vitamin K epoxide reductase complex subunit 1, CYP2C9, CYP2C19, CYP4F2, human microsomal epoxide hydroxylase, calumenin, and γ-glutamyl carboxylase were determined. RESULTS: From multiple linear regression models, vitamin K epoxide reductase complex subunit 1, CYP2C9, CYP4F2, and age were found to have significant effects on warfarin stable dose. The stable warfarin daily doses of patients with the CC, CT, and TT genotypes in the CYP4F2 gene were 5.34 ± 2.04, 5.33 ± 1.64, and 6.55 ± 2.12 mg, respectively. The higher dose requirements in patients with TT alleles in CYP4F2 were attributable to a low warfarin sensitivity (international normalized ratio/warfarin plasma concentration); the warfarin sensitivity in CC, CT, and TT genotypes was 2.1 ± 1.2, 1.0 ± 0.4, and 0.8 ± 0.6, respectively. The similarity between the dose requirements of patients with CT and CC alleles was explained through the combined result of warfarin sensitivity and clearance outcomes. Apparent plasma (S)- and (R)-warfarin clearances were found to be 37.7% and 34.1% lower in CT genotype patients than in CC genotype patients, respectively. CONCLUSIONS: The dose variability in CYP4F2 genotypes was attributable to both warfarin clearance and sensitivity differences.

Intravesical epinephrine preserves uroplakin II expression in urinary bladder from cyclophosphamide-induced rat cystitis.

We investigated the attenuated effect of intravesical epinephrine (EPI) on uroplakin II (UPII) expression in cyclophosphamide (CYP)-induced rat cystitis. Sixty-eight Sprague-Dawley female rats were divided into one negative control group (GI) and five intraperitoneally CYP (150 mg CYP/kg)-injected groups (GII-VI) consisting of a positive control group (GII), three groups (GIII-V) with retaining intravesically instillated ameliorating agents for 90 min by urethral ligation until sacrifice, and one group (GVI) with freely voiding after intravesical EPI instillation. The retention groups were further classified into null-treated- (GIII), EPI- (GIV), and vehicle group (GV). All rats were euthanized 24 h after CYP injection. The UPII and α1-adrenergic receptors (AR) levels were measured with real-time polymerase chain reaction (RT-PCR) method and the morphological changes were also evaluated. CYP induced severe cystitis and decreased vesical UPII mRNA level. The EPI-treated groups had showed attenuation effects against submucosal edema and hemorrhage, and preserved UPII expression. Concurrently, intravesical EPI resulted in a significant preservation of both subtypes of α1A- and α1B AR expressions, which was well correlated with the hemostatic pattern in the samples. The obstructed and null-treated group (GIII) revealed severe cystitis and maximally decreased UPII levels, and the diluting effect of vehicle (GV) on CYP toxicity was insignificant on UPII preservation. The UPII level of RT-PCR was well correlated with the UPII immunohistological expression and their morphological changes. Intravesical instillation of EPI preserves UPII expression and attenuates the toxic responses in the bladder in CYP-induced rat cystitis.