Salvatore Arena

Altered 'active' antireflux mechanism in primary vesico-ureteric reflux: a morphological and manometric study

To immunolocate c‐kit‐positive interstitial cells of Cajal (ICCs, known to be responsible for pacemaker activity in human ureters, coordinating ureteric motility) in the intramural ureter of patients with different grades of vesico‐ureteric reflux (VUR), to assess the ureteric histology and correlate these findings with manometric patterns.

Activation of adenosine A2A receptors by polydeoxyribonucleotide increases vascular endothelial growth factor and protects against testicular damage induced by experimental varicocele in rats

In rat experimental varicocele, polydeoxyribonucleotide (PDRN) induces vascular endothelial growth factor (VEGF) production, thereby enhancing testicular function. This may point to a new therapeutic approach in human varicocele.

Aquaporin-9 immunohistochemistry in varicocele testes as a consequence of hypoxia in the sperm production site

Aquaporin‐9 (AQP‐9) regulates tissue hydration by promoting transmembrane exchanges of both water and solutes, such as lactate. The latter is a key metabolite of primary spermatocytes and of maturing haploid germ cells (h‐GCs). The present investigation was aimed at immunolocalising human AQP‐9 in both normal and varicocele testes. Histology and immmunocytochemistry were investigated in archival biopsies from 20 varicocele testes and in eight unaffected ones. AQP‐9 immunostaining was performed using a rabbit antibody, and either focal or diffuse cell membrane labelling was recorded. Varicocele testes showed disarranged tubular compartments, with sloughing h‐GCs, tissue hyperhydration, spermiogenesis failure and fibrosis. AQP‐9 immunohistology of the control testes showed a diffuse cell membrane staining of the primary spermatocytes and h‐GCs, without any positive reaction of spermatogonia and Sertoli cells. AQP‐9 cell expression in the varicocele testes was focal or lacking in both adluminal and sloughing GCs. AQP‐9 expression occurs in normal human testis, at cell membrane of primary spermatocytes and h‐GCs, suggesting a possible role of AQP‐9 in the water and lactate transport from Sertoli cells to GCs. AQP‐9 is focal or lacking in adolescent varicocele testes, and this suggests AQP‐9 to be downregulated in such testicular disorder, leading to lactate deprivation with subsequent hypospermatogenesis.

Dextranomer/Hyaluronic Acid Copolymer Implant for Vesicoureteral Reflux: Role of Myofibroblast Differentiation

PURPOSE Dextranomer/hyaluronic acid implantation is associated with a granulomatous inflammatory reaction, replaced by fibrosis. Appearance of myofibroblasts is considered a crucial event in fibrosis, and CD68 positive cells and other factors are implied in their activation. Mast cells are a source of these factors and tryptase can induce fibroblast to express alpha-smooth muscle actin, which is characteristic of myofibroblasts. We evaluated histological changes in refluxing ureters treated with dextranomer/hyaluronic acid and immunolocalized CD68 positive cells, tryptase mast cells and myofibroblasts. MATERIALS AND METHODS We performed histological, histochemical and immunohistochemical analyses in 22 refluxing ureters treated with dextranomer/hyaluronic acid in comparison with 17 refluxing ureters who underwent ureteral reimplantation but did not receive endoscopic bulking agent. We used CD68 antibody for monocytes/macrophages and epithelioid cells, mast cell tryptase mouse antibody for mast cells, and alpha-smooth muscle actin and vimentin antibodies for myofibroblasts. The area of the ureteral lumen in dextranomer/hyaluronic acid treated and untreated ureteral endings was measured. RESULTS Sirius red documented a major grade of histological lesions in dextranomer/hyaluronic acid treated refluxing ureters. CD68 and tryptase mast cell staining showed a significant enhancement of positive cells in dextranomer/hyaluronic acid treated refluxing ureters. Immunostaining for alpha-smooth muscle actin and vimentin displayed a myofibroblastic invasion in dextranomer/hyaluronic acid. Measurement of surface in treated refluxing ureters was significantly less than in untreated refluxing ureters. CONCLUSIONS Our data documented a recruitment of CD68 and tryptase positive cells, abnormal accumulation of collagenous stroma and successive extracellular matrix remodeling through differentiation of myofibroblasts. Myofibroblasts might provoke tissue contraction, decreasing the ureteral diameter and modifying the ureteral length-to-diameter ratio, preventing urine reflux.

Sarcoglycan Subcomplex Expression in Refluxing Ureteral Endings

PURPOSE Functional and structural lesions of ureteral endings seem to alter the active valve mechanism of the ureterovesical junction, causing vesicoureteral reflux. The interaction of the dystroglycan complex with components of the extracellular matrix may have an important role in force transmission and sarcolemma protection, and the sarcoglycan complex is an essential component of the muscle membrane located dystroglycan complex. We performed immunofluorescence and molecular analysis on the expression of sarcoglycan complex subunits. MATERIALS AND METHODS A total of 21 specimens of refluxing ureteral endings were obtained during ureteral reimplantation. Six ureteral ends obtained during organ explantation were used as controls. Immunohistochemical analysis and reverse transcriptase polymerase chain reaction evaluation were performed for alpha, beta, gamma, delta and epsilon-sarcoglycan complex. RESULTS The Spearman test revealed a significant positive correlation between alpha-sarcoglycan complex immunofluorescence intensity and grade of vesicoureteral reflux, while a negative correlation was recorded between epsilon-sarcoglycan complex immunofluorescence intensity and grade of vesicoureteral reflux. CONCLUSIONS Semiquantitative analysis demonstrated a significant grade related impairment of epsilon-sarcoglycan complex coupled with an increased expression of alpha-sarcoglycan complex. This observation suggests that the structural deficiency of the trigonal ureterovesical junction could cause a passive stretching of refluxing urine on the ureter, deranging the multimodular tensegrity architecture of the sarcoglycan subcomplex, or that the sarcoglycan complex could have a key role in the physiopathology of vesicoureteral reflux. In fact, the defect in any of the sarcoglycan complexes results in degeneration of membrane integrity and muscle fiber. An altered configuration of the sarcoglycan complex could explain the structural and functional changes in refluxing ureteral endings. Our observations underline the assumption that primary vesicoureteral reflux might be regarded as a sarcoglycanopathy with marked quantitative deficiency of epsilon-sarcoglycan complex and over expression of alpha-sarcoglycan complex.

Chronic lymphocytic thyroiditis: could it be influenced by a petrochemical complex? Data from a cytological study in South-Eastern Sicily

INTRODUCTION In genetically predisposed individuals, exogenous factors (including pollution) influence the development of Hashimotos thyroiditis/chronic lymphocytic thyroiditis (CLT). CLT may also be a risk factor for associated thyroid cancer. Few data are available on the role of pollution from petrochemical complexes, one of which is located in the Siracusa province (South-Eastern Sicily), in the pathogenesis of CLT. AIMS i) To study the frequency of CLT in fine-needle aspiration cytology (FNAC)-interrogated thyroid nodules from patients who were stably resident in their zones, comparing it in patients living in the petrochemical complex area (zone A) with that of patients from a control area (zone B). ii) To study the frequency of CLT in the FNAC categories of malignancy risk, comparing the two zones. PATIENTS AND METHODS We retrospectively evaluated cytologically adequate slides of 1323 nodules in 1013 outpatients who underwent ultrasound-guided FNAC from 2006 to 2012. We stratified by area of residence, gender, and FNAC categories of malignancy risk. RESULTS CLT was detected with significantly greater frequency in either patients or nodules from zone A compared with zone B (32.0% vs 23.1%, P=0.002 or 28.2% vs 18.8%, P=0.0001), with a female preponderance (F=35.2% vs M=21.1% or 30.4% vs 20.4%, zone A and F=26.5% vs 12.3% or 21.6% vs 9.5%, zone B). Regardless of zone, CLT was approximately twofold more frequent in the suspiciously malignant+malignant classes (TH4+THY5=47.6%, zone A and 32.4%, zone B) compared with the benign+intermediate classes (THY2+THY3=27.3%, zone A and 18.2%, zone B), but with a clear stepwise THY2 through THY5 increase only in zone A (THY2=25.3%, THY5=66.7%; THY2=18.6%, THY5=28.6% in zone B). CONCLUSIONS The petrochemical complex-related pollution is an environmental factor involved in the development of CLT and, likely, in the CLT association with thyroid neoplasms.

Molecular events involved in the morphogenesis of multicystic dysplastic kidney.

Introduction: Wnt-1 is capable of inducing metanephric mesenchyme to undergo tubulogenesis. A relationship between the degree of cystogenesis and reduced E-cadherin (E-cad) expression was described. Syndecan-1 (Sdc-1) has a critical role in kidney development. Materials and Methods: Ten multicystic dysplastic kidneys (MCDKs) were stained with hematoxylin and eosin and immunohistochemistry was performed using Wnt-1, E-cad and Sdc-1 antibodies. Eight unaffected kidneys were used as controls. Results: Strong Wnt-1 immunostaining occurred inside cystic/tubular epithelial cells and in blastematous foci. An immunoreaction was observed in glomerular epithelial cells. In controls, just weak cytoplasmic Wnt-1 positivity was seen in tubular epithelial cells. E-cad reaction was negative in MCDKs while strong immunostaining was common in tubular cells of controls. A strong Sdc-1 immunoreaction depicted cystic, tubular and glomerular epithelial cells in MCDKs while Sdc-1 expression documented weak positivity in tubular epithelium alone. Conclusions: Our data are in accordance with an involvement of Wnt-1 in normal nephrogenesis and with its role in altered epithelial differentiation of metanephric mesenchyme in MCDKs. Wnt-1 signal may function to suppress E-cad expression, a predisposing event for cystogenesis. High expression of Sdc-1 in tubular/cystic epithelial cells of MCDKs might alter the normal transition of stages of the developmental process and modify the anion charge of the glomerular barrier.

Gender-Specific Correlation Of Intranodular Chronic Lymphocytic Thyroiditis With Thyroid Nodule Size, Echogenicity, And Histologically-Verified Cytological Class Of Malignancy Risk

No data are available on the cytologically and histologically demonstrated presence of intranodular chronic lymphocytic thyroiditis (ICLT) and on the ICLT relationship with thyroid nodule characteristics such as size, echotexture and nature (benign or malignant). We wished to fill this gap by analyzing data in a gender-specific fashion. We studied 408 thyroid nodules from 408 consecutive persons (325 females and 83 males). Nodules were isoechoic (n = 268) or hypoechoic (n = 140), ICLT +ve (n = 113 [27.7%]) or ICLT −ve (n = 295), cytologically low-risk (n = 197) or high-risk (n = 211), histologically benign (n = 263) or malignant (n = 145). ICLT prevailed in females (97/113) and in hypoechoic nodules (58/140 [41.4%] vs 55/268 [20.5%], P < 0.0001). Compared to males, females had (i) smaller nodules (18.5 ± 9.4 vs 23.3 ± 13.4 mm, P = 0.0002), a difference due to the isoechoic nodules (21.1 ± 9.8 vs 26.6 ± 14.1 mm, P = 0.0006), (ii) lower rates of high-risk nodules (161/325 [49.5%] vs 50/83 [60.2%], P = 0.082) and malignant nodules (110/325 [33.8%] vs 35/83 [42.2%] P = 0.16). ICLT +ve nodules were smaller than the ICLT −ve ones (15.4 ± 6.9 vs 20.9 ± 11.2 mm, P < 0.0001), a difference due to the isoechoic nodules (17.5 ± 6.5 vs 23.6 ± 11.7 mm, P = 0.0003). The smallest nodules were hypoechoic, cancerous and ICLT +ve nodules in males (9.5 ± 4.0 mm); the largest were isoechoic, cytologically risky and ICLT −ve in males (29.1 ± 13.2 mm). Compared to ICLT −ve nodules, malignancy prevailed in ICLT +ve nodules (55/113 [48.7%] vs 90/295 [30.5%], P = 0.0006), both in hypoechoic (37/58 [63.8%] vs 41/82 [50.0%]) and isoechoic nodules (18/55 [32.7%] vs 49/213 [23.0%]). ICLT +ve hypoechoic nodules of females and ICLT −ve hypoechoic nodules of males had the greatest rate of malignancy (67% both), while ICLT −ve isoechoic nodules of females had the lowest (19%). In conclusion, presence/absence of ICLT is associated with some sexually dimorphic characteristics of thyroid nodules. Adding the specification of ICLT positivity/negativity in cytological reports may help improving the risk of malignancy at least in some groups of thyroid nodules.