Cell-Seeded Tubularized Scaffolds for Reconstruction of Long Urethral Defects: A Preclinical Study

Background

The treatment options for patients requiring repair of a long segment of the urethra are limited by the availability of autologous tissues. We previously reported that acellular collagen-based tubularized constructs seeded with cells are able to repair small urethral defects in a rabbit model.

Objective

We explored the feasibility of engineering clinically relevant long urethras for surgical reconstruction in a canine preclinical model.

Design, setting, and participants

Autologous bladder epithelial and smooth muscle cells from 15 male dogs were grown and seeded onto preconfigured collagen-based tubular matrices (6 cm in length). The perineal urethral segment was removed in 21 male dogs. Urethroplasties were performed with tubularized collagen scaffolds seeded with cells in 15 animals. Tubularized constructs without cells were implanted in six animals. Serial urethrography and three-dimensional computed tomography (CT) scans were performed pre- and postoperatively at 1, 3, 6, and 12 mo. The animals were euthanized at their predetermined time points (three animals at 1 mo, and four at 3, 6, and 12 mo) for analyses.

Outcome measurements and statistical analysis

Statistical analysis of CT imaging and histology was not needed.

Results and limitations

CT urethrograms showed wide-caliber urethras without strictures in animals implanted with cell-seeded matrices. The urethral segments replaced with acellular scaffolds collapsed. Gross examination of the urethral implants seeded with cells showed normal-appearing tissue without evidence of fibrosis. Histologically, an epithelial cell layer surrounded by muscle fiber bundles was observed on the cell-seeded constructs, and cellular organization increased over time. The epithelial and smooth muscle phenotypes were confirmed using antibodies to pancytokeratins AE1/AE3 and smooth muscle–specific desmin. Formation of an epithelial cell layer occurred in the unseeded constructs, but few muscle fibers formed.

Conclusions

Cell-seeded tubularized collagen scaffolds can be used to repair long urethral defects, whereas scaffolds without cells lead to poor tissue development and strictures. This study demonstrates that long tissue-engineered tubularized urethral segments may be used for urethroplasty in patients.     

Relation of interleukin-10 Promoter Polymorphisms to Adult Chronic Immune Thrombocytopenic Purpura in a Cohort of Egyptian Population

Background: Adult chronic immune thrombocytopenic purpura (chronic ITP) is an autoimmune multifactorial bleeding disorder that occurs because of enhanced peripheral platelet destruction. Treatment decisions can be challenging because the goal of treatment is to prevent severe bleeding, but the risk of bleeding can be difficult to estimate for any individual patient.

Objective: This case–control study was planned to investigate the relationship of interleukin (IL)-10 promoter (IL-10-1082, -819 and -592) polymorphisms with the susceptibility, severity and outcome of adult chronic ITP in a cohort of Egyptian population.

Subjects and Methods: Typing of IL-10 promoter polymorphisms was done using restriction fragment length polymorphism for 62 adult patients with chronic ITP and 73 age- and sex-matched healthy controls.

Results: No significant differences were found between ITP patients and controls regarding the frequency of IL-10 promoter genotypes, alleles or haplotypes. IL-10-592 AA genotype and ATA (IL-10-1082, -819 and -592) haplotype were associated with severe ITP (p?=?0.003, 0.043, respectively).

Conclusion: Our findings suggest that the IL-10 promoter polymorphisms are unlikely to affect the development or treatment outcome of chronic adult ITP in Egyptian population, but IL-10-592 AA genotype and IL-10 (-1082, -819 and -592) ATA haplotype may be associated with disease severity. Because ITP is a complex disease, it is recommended that a multicenter study should be done with large sample size and unified typing technique.     

Evaluation and treatment of erectile dysfunction in the aging male: a mini-review

Before the 20th century, individuals often did not live beyond the reproductive years, and sexuality of the elderly was not an issue. However, in the current era it is known that as life expectancy improves, both men and women are seeking to preserve their sexuality into old age. While the appreciation of sexuality persists with aging, a decline in sexual activity is typically seen with, and can be attributed to both general health problems as well as specific sexual dysfunctions. Erectile dysfunction is the most frequently diagnosed sexual dysfunction in the older male population. This mini-review provides an overview of contemporary literature concerning epidemiology, pathophysiology, assessment and treatment of erectile dysfunction in the aging male.     

The interaction between insulin resistance, liver fibrosis and early virological response in Egyptian patients with chronic hepatitis C

BACKGROUND:

Hepatitis C virus (HCV) infection may induce insulin resistance (IR) irrespective of the severity of liver disease, and there is evidence of a central role for IR in failure to achieve sustained virological response (SVR) in HCV patients.

OBJECTIVE:

To assess IR as a predictor of the severity of hepatic fibrosis in Egyptian HCV patients, and its effect on early viral kinetics and virological response to HCV therapy.

METHODS:

A total of 140 chronic HCV patients were divided into two groups according to the homeostasis model assessment-IR (HOMA-IR). Group 1 consisted of 48 chronic HCV patients with HOMA-IR ≥2, and group 2 consisted of 92 chronic HVC patients without IR (HOMA IR <2). All patients were treated with combination therapy (pegylated interferon-alpha 2a plus ribavirin) for 48 weeks and studied for viral kinetics throughout the period of therapy.

RESULTS:

The study revealed that older age, higher body mass index and HOMA-IR≥2 were significantly associated with advanced fibrosis. Rapid virological response, complete early virological response and SVR were significantly lower in the IR-HCV group compared with the non-IR-HCV group. Univariate and multivariate analyses revealed that older age, fibrosis (F≥3), high viral load (>600,000 IU/mL) and HOMA-IR ≥2 were significantly associated with a lack of viral kinetics as well as SVR. However, HOMA-IR ≥2 was the main independent variable associated with lack of SVR. On the other hand, body mass index, plasma insulin level and HOMA-IR decreased significantly compared with starting levels in patients who achieved SVR. This suggests a cause and effect relationship between HCV infection and IR.

CONCLUSION:

IR in chronic HCV patients is associated with progressive fibrosis and slow viral kinetics, and could be a predictor for lack of rapid and early virological response. Therefore, HOMA-IR levels should be measured and improved before starting antiviral treatment.     

Expression of PD-1 (CD279) and FoxP3 in diffuse large B-cell lymphoma

The role of the microenvironment in high-grade lymphoma is not well defined. In this report, we employ immunohistochemistry to characterise programmed death-1 (PD-1/CD279) and FoxP3 expression in 70 cases of diffuse large B-cell lymphoma (DLBCL). PD-1 is a surface marker characteristic of follicular helper T-cells whilst FoxP3 is characteristic of Tregs. We demonstrate variable infiltration with CD4⁺ T-cells (<10 to >50 % of all lymph node cells) and PD-1^hi cells (0.1 to 1.5 % of all cells). CD4⁺ T-cells can be distributed in clusters or more diffusely and PD-1^hi cells, but not FoxP3⁺ cells, are found in rosettes around lymphoma cells. Cases with high CD4⁺ T-cell numbers tended to have higher numbers of both PD-1^hi and FoxP3⁺ cells. Cases with total CD4⁺ T-cell, PD-1^hi and FoxP3⁺ numbers above the median associate with better clinical outcome. Overall, we demonstrate that infiltration by CD4⁺ T-cells, including both FoxP3⁺ and PD-1^hi subsets, correlates with prognosis in DLBCL. In distinction to previous reported series, patients (91 %) were treated with rituximab-containing regimens, suggesting that the effects of CD4+ T-cell infiltration are maintained in the rituximab era. This work suggests that determinants of total CD4⁺ T-cell infiltration, either molecular characteristics of the lymphoma or the patients’ immune system, and not individual T-cell subsets, correlate with clinical outcome.     

Telephone versus office-based management of warfarin: impact on international normalized ratios and outcomes

Studies have concluded that telephone-based management of warfarin is an effective alternative to in-office management. High rates of patient and physician satisfaction have been reported with telephone-based monitoring. Proposed benefits of telephone-based monitoring include time- and cost savings for patients and healthcare providers alike as well as increased access to care for those patients who have difficulty making in-office appointments. This study aimed to evaluate the impact of telephone versus office-based management of warfarin on extreme INR values. A retrospective cohort study was conducted to assess outcomes of patients receiving warfarin managed either by telephone or in-office appointments. The primary endpoint of the study was the frequency of extreme INR values, defined as an INR ≤1.5 or ≥4.5. A total of 110 patients were evaluated; subjects were distributed 2:1 between the in-office and telephone groups. Baseline characteristics were similar between groups. Subjects followed via telephone had a twofold increase in the incidence of extreme INR values compared to the patients followed in-office (15.18 vs. 7.98 %; p < 0.0001). Overall TTR was similar between groups (85.39 vs. 80.38 %, p = 0.1171). There was no difference between the two groups in the incidence of major bleeding events (2.67 vs. 0 %, p = 1.00), thromboembolic events (8 vs. 0 %, p = 0.1740), or hospitalizations related to anticoagulation therapy (6.67 vs. 0 %, p = 0.1758). Patients monitored via telephone had a higher incidence of extreme INR values than patients followed in-office, which may lead to an increased incidence of adverse outcomes in the long-term. Well-designed, prospective studies are needed to confirm such findings.     

Validation of the International Urogynecology Association’s Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire in Arabic

Introduction and Hypothesis

A valid and reliable Arabic version of the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire, IUGA-revised (PISQ-IR) is needed.

Methods

Of 238 Saudi Arabian women who fulfilled the inclusion criteria, 227 were recruited. A detailed history was obtained and pelvic examination was performed in all participants. An Arabic version of the PISQ-IR was administered with three quality of life measures. Reliability was assessed by factor analysis, and internal consistency was measured with Cronbach’s α. Face validity was established with translation, back-translation, and cognitive reviews. Criterion validity was determined by analysing the correlations between the responses and the clinical examinations and by comparing the responses with those to other validated measures of incontinence and prolapse.

Results

Overall, 67 % of the women were categorized as sexually active and 33 % as sexually inactive. The overall frequency of urinary incontinence was highest (85.5 %), followed by prolapse (59 %) and fecal incontinence (53.3 %). The average score in the factor analysis for dimensions of sexual inactivity was 2.7 and Cronbach’s α was 0.61; an item-by-item assessment of the majority of the scales showed an α value of >0.75, suggesting a moderate-to-high internal consistency. Regarding external validity, strong negative correlations were found with the Pelvic Organ Prolapse Distress Inventory, the Colorectal-Anal Distress Inventory (CRADI), and the Pelvic Floor Impact Questionnaire (PFIQ) in the sexually active group, and positive correlations were found with the CRADI and PFIQ in the sexually inactive group.

Conclusions

The Arabic version of the PISQ-IR is reliable and valid for assessing sexual function in Arabic-speaking women with pelvic floor disorders.

     

Antimicrobial sesquiterpene lactones from Artemisia sieberi

Two new sesquiterpene lactones 3R, 8R-dihydroxygermacr-4(15),9(10)-dien-6S,7S,11RH,12,6-olide (1) and 1R, 8S-dihydroxy-11R,13-dihydrobalchanin(2), together with two known compounds 11-epiartapshin (3) and 3′-hydroxygenkwanin (4), were isolated from Artemisia sieberi. Their structures were elucidated by 1D, 2D NMR, MS, and X-ray diffraction. Compound 4 inhibited Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus with Minimal inhibitory concentration values of 50 and 25 μg/disk, respectively. All the isolated compounds exhibited moderate antifungal activities.