A novel two-step method for the formation of tissue-engineered cartilage by mature bovine chondrocytes: the alginate-recovered-chondrocyte (ARC) method.

Most attempts to tissue-engineer cartilage have involved seeding of cultured cells into a biological or synthetic scaffold. We have developed a novel two-step culture approach that makes possible the in vitro formation of cartilaginous-like tissue by mature adult bovine chondrocytes without the aid of a synthetic matrix. The first step consists of culturing chondrocytes under conditions that maintain their rounded shape and their molecular phenotype as assessed by type II collagen and aggrecan production. This step was accomplished by culturing the isolated chondrocytes in alginate beads until the cells have reestablished a proteoglycan-rich cell-associated matrix (CM). The second step consists of culturing the cells with their CM, after recovery from the beads, on a tissue culture insert with a porous membrane. In this study, young adult bovine articular chondrocytes were cultured in alginate beads in the presence of 10% or 20% fetal bovine serum (FBS). After 7 days of culture, the alginate beads were dissolved by incubating the beads for 20 min in sodium citrate buffer, a calcium chelator. Following a brief centrifugation, the cells with their CM were recovered, resuspended in medium containing 10% or 20% FBS and seeded onto a tissue culture insert. After 1 week of culture on the insert, the individual cells with their CM progressively became incorporated into a mass of cartilaginous tissue. Culture with 20% FBS resulted in the best formation of tissues. These tissues, easily recovered from the insert, were then subjected to biochemical and histological analyses. The biochemical results showed that the chondrocytes remain phenotypically stable in the tissues. The de novo tissue has a relatively high ratio of PG/collagen. Histological examination of the tissue revealed it contained a cartilage-like matrix strongly stained with toluidine blue. This scaffold-free system appears ideal to study, in vitro, the development of transplantable cartilaginous tissue.     

Transvaginal sonography in the assessment of distal ureteral calculi.

OBJECTIVES: To explore the morphological and functional findings of transvaginal sonography (TVS) in the diagnosis and management of distal ureteral calculi. METHODS: We retrospectively reviewed the imaging studies in seven cases of distal ureteral calculi (study group) and 20 cases of female volunteers without urinary tract infection (control group). All 27 patients had undergone TVS for assessment of the lower urinary tract. The variables measured on ultrasound included the presence or absence of distal ureteral calculi, the size and location of a calculus if present, the presence or absence of the ureter jet phenomenon, morphological changes of the ureteral papilla and adjoining tissue, and the height of the ureteral papilla, as well as vascular changes seen on Doppler imaging. RESULTS: In the controls, 80% of the ureteral papillae were triangular and 20% trapezoidal. The average (+/-SD) height of the ureteral papillae was 3.5 +/- 0.7 mm on the right and 3.6 +/- 0.9 mm on the left. There was no statistically significant difference between the heights of the right and left papillae (P = 0.619). In the study group, echogenic stones were identified in all seven patients and a hypoechogenic tubular structure connected to the involved papilla was identified in 6/7 patients. The average height of the involved papilla was 6.7 +/- 1.6 mm. Ureteric jets were visible in all patients in both the study and control groups. CONCLUSION: In patients with distal ureteral calculi, TVS provides a rapid, non-invasive and repeatable means of assessing the morphology and function of the distal ureter.     

What is the place of electroneuromyographic studies in the diagnosis and management of pudendal neuralgia related to entrapment syndrome?

Entrapment of the pudendal nerve may be at the origin of chronic perineal pain. This syndrome must be diagnosed because this can result in the indication of surgical decompression of the entrapped nerve for pain relief. Electroneuromyographic (ENMG) investigation is often performed in this context, based on needle electromyography and the study of sacral reflex and pudendal nerve motor latencies. The limits of ENMG investigation, owing to various pathophysiological and technical considerations, should be known. The employed techniques do not assess directly the pathophysiological mechanisms of pain but rather correlate to structural alterations of the pudendal nerve (demyelination or axonal loss). In addition, only direct or reflex motor innervation is investigated, whereas sensory nerve conduction studies should be more sensitive to detect nerve compression. Finally, ENMG cannot differentiate entrapment from other causes of pudendal nerve lesion (stretch induced by surgical procedures, obstetrical damage, chronic constipation...). Thus, perineal ENMG has a limited sensitivity and specificity in the diagnosis of pudendal nerve entrapment syndrome and does not give direct information about pain mechanisms. Pudendal neuralgia related to nerve entrapment is mainly suspected on specific clinical features and perineal ENMG examination provides additional, but no definitive clues, for the diagnosis or the localization of the site of compression. In fact, the main value of ENMG is to assess objectively pudendal motor innervation when a surgical decompression is considered. Perineal ENMG might predict the outcome of surgery but is of no value for intraoperative monitoring.     

Functional correlates of Doppler flow study of the female urethral vasculature.

OBJECTIVES: To examine the effect of individual patient factors (age, parity, body mass index, menstrual cycle, menopause, hormone replacement therapy, bladder neck position and urethral mobility) on the appearance of Doppler flow in urethral vessels, to investigate the association between the Doppler flow parameters and intrinsic urethral function, storage and voiding, and to explore differences in the urethral vasculature between subjects with and without urodynamic stress incontinence (USI). METHODS: Over a 4-year period we prospectively performed imaging studies in 355 women, including 244 who denied any lower urinary tract symptoms within the previous 3 months (Group A) and 111 who had had lower urinary tract symptoms (Group B). Studies included morphologic assessment and Doppler flow investigation of the lower urinary tract. Vascular flow velocity and vessel density in the urethral vasculature were measured. For women in Group B, multichannel urodynamic studies were also performed. RESULTS: The urethral vasculature has five main branches of vessels. Their appearance was not affected by the menstrual cycle or menopause except for those of the anterior vaginal vessel and anterior branch of the middle urethral vessel. Other than that of the posterior urethral vessel, in which there was a correlation with parity, the resistance index (RI) was not affected by individual patient factors. However, there was a correlation between the vascular index (VI) and individual factors such as age (r = -0.336, P = 0.002), body mass index (r = -0.287, P = 0.028), menopause (r = -0.402, P < 0.001), and hormone replacement therapy (r = 0.392, P = 0.027). Only the VI and RI of the posterior urethral vessel correlated significantly with the urethral pressure profile. In subjects with lower urinary tract symptoms, the appearance of the urethral vasculature on power Doppler imaging and the corresponding RI and VI values were not correlated with objective evidence of USI. CONCLUSION: Patient factors may affect specific Doppler flow parameters of the urethral vasculature, which are related to intrinsic resting urethral closure. There is no difference in the appearance of the urethral vasculature in subjects with or without USI.     

Psychosocial effects of vitiligo

A limited number of studies have paid attention to the psychosocial well-being of patients affected with vitiligo. We review the psychosocial effects of vitiligo, how patients deal with them and the psychiatric morbidity in vitiligo patients. Given the appreciable comorbidity, it is important to consider these observations in the management of patients, for example in patient-physician interaction but also in treatment strategies and evaluation of treatments. The effects of the psychological state on the disease itself together with the potential therapeutic implications are reviewed. Based on these data, we suggest how to further improve patient's management.     

Transvaginal sonography in the morphological and functional assessment of segmental dilation of the distal ureter.

Primary megaureter presents a spectrum of findings ranging from mild, clinically unimportant, stable ureterectasis to severe, progressive obstructive hydroureteronephrosis. We report a patient with a double uterus and ipsilateral renal dystrophy. A previous imaging study had revealed a double uterus and obstructed left hemivagina, into which a single vaginal ectopic ureter inserted. On follow-up intravenous urography 8 years later, the left kidney was non-functioning, and there was segmental dilation of the distal right ureter. Transvaginal sonography with real-time scanning and Doppler were useful in exploring the morphological and functional status of this dilated distal ureter.