Sperm culture and bacterial susceptibility to antibiotics in a large andrological population: prevalence and impact on seminal parameters.

BACKGROUND: The aim of this study was to evaluate (i) the prevalence of subjects with a positive sperm culture (SC) for bacteria in subjects with or without genitourinary tract inflammation (GTI); (ii) the actual distribution of the species analysed, according to Gram stain; (iii) the impact on sperm parameters; and (iv) the actual bacterial susceptibility to antibiotics. METHODS: A total of 930 subjects (18-55) years, were retrospectively studied. All the patients underwent SC and in the case of positive tests (CFU > 106), a microbiological susceptibility analysis. The subjects studied were subdivided into group A (n = 452), with subjective signs of GTI; group B (n = 478), male partners of infertile couples; and group C, 30 healthy normospermic subjects. In group B and in the control group, a semen analysis was performed. RESULTS: Overall, the prevalence of positive SC was 21.5% (200/930). The prevalence of positive SC in group A (113/200; 56.5%) was significantly higher vs. group B (87/200; 43.5%; p = 0.01) and control group (1/30; 3.3%; p = 0.0001). In subjects with GTI, the prevalence of asthenozoospermic (96/285; 33.7%) and oligo-asthenozoospermic (98/285; 34.4%) was significantly higher vs. normospermic, oligo-astheno-teratozoospermic, oligozoospermic and azoospermic subjects (22/285 (7.7%), 48/285 (16.8%), 15/285 (5.3%) and 6/285 (2.1%), respectively; p = 0.001). Finally, Enterococcus faecalis (Gram-positive) and Escherichia coli (Gram-negative) showed the highest prevalence of antibiotic resistance. CONCLUSIONS: The prevalence of positive SC is higher in GTI subjects; however, the SC could also be positive in subjects without GTI. Commonly used antibiotics have an increasing risk of being useless for the treatment of bacterial infections. Finally, the diagnosis of GTIs is important also for male fertility.

A humanized system to expand in vitro amniotic fluid-derived stem cells intended for clinical application.

BACKGROUND AIMS: The amniotic fluid is a new source of multipotent stem cells with therapeutic potential for human diseases. In agreement with the regulatory requirement to reduce and possibly to avoid animal-derived reagents in the culture of cells intended for cell therapy, bovine serum, the most common supplement in the culture medium, was replaced by human platelet-derived growth factors. METHODS: We tested a new culture medium to expand monolayers of human amniotic fluid stem cells (hAFSC) for clinical use. The AFSC were isolated by c-Kit selection and expanded in media supplemented with either bovine serum or a human platelet lysate (Lyset). RESULTS: We compared proliferation kinetics, colony-forming unit percentage, multilineage differentiation, immunophenotypic characterization and inhibition of peripheral blood mononuclear cell proliferation of the two AFSC cell cultures and we found no significant differences. Moreover, the karyotype analysis of the cells expanded in the presence of the platelet lysate did not present cytogenetic abnormalities and in vitro and in vivo studies revealed no cell tumorigenicity. CONCLUSIONS: Platelet derivatives represent a rich source of growth factors that can play a safety role in the homeostasis, proliferation and remodeling of tissue healing. We propose human platelet extracts as a preferential alternative to animal serum for the expansion of stem cells for clinical applications.

The Flipping-Wedge Osteotomy: How 3D Virtual Surgical Planning (VSP) Suggested a Simple and Promising Type of Osteotomy in Pediatric Post-Traumatic Forearm Deformity.

(1) Background: The application of computer-aided planning in the surgical treatment of post-traumatic forearm deformities has been increasingly widening the range of techniques over the last two decades. We present the "flipping-wedge osteotomy", a promising geometrical approach to correct uniapical deformities defined during our experience with virtual surgical planning (VSP); (2) Methods: a case of post-traumatic distal radius deformity (magnitude 43°) treated with a flipping-wedge osteotomy in an 11-year-old girl is reported, presenting the planning rationale, its geometrical demonstration, and the outcome of the procedure; (3) Results: surgery achieved correction of both the angular and rotational deformities with a neutral ulnar variance; (4) Conclusions: flipping-wedge osteotomy may be a viable option to achieve correction in forearm deformities, and it deserves further clinical investigation.

In Vitro and In Vivo Evaluation of 99mTc-Polymyxin B for Specific Targeting of Gram-Bacteria.

BACKGROUND: Infectious diseases are one of the main causes of morbidity and mortality worldwide. Nuclear molecular imaging would be of great help to non-invasively discriminate between septic and sterile inflammation through available radiopharmaceuticals, as none is currently available for clinical practice. Here, we describe the radiolabeling procedure and in vitro and in vivo studies of 99mTc-polymyxin B sulfate (PMB) as a new single photon emission imaging agent for the characterization of infections due to Gram-negative bacteria. RESULTS: Labeling efficiency was 97 ± 2% with an average molar activity of 29.5 ± 0.6 MBq/nmol. The product was highly stable in saline and serum up to 6 h. In vitro binding assay showed significant displaceable binding to Gram-negative bacteria but not to Gram-positive controls. In mice, 99mTc-HYNIC-PMB was mainly taken up by liver and kidneys. Targeting studies confirmed the specificity of 99mTc-HYNIC-PMB obtained in vitro, showing significantly higher T/B ratios for Gram-negative bacteria than Gram-positive controls. CONCLUSIONS: In vitro and in vivo results suggest that 99mTc-HYNIC-PMB has a potential for in vivo identification of Gram-negative bacteria in patients with infections of unknown etiology. However, further investigations are needed to deeply understand the mechanism of action and behavior of 99mTc-HYNIC-PMB in other animal models and in humans.

Validation of QB stress test as a useful tool in the detection of native arteriovenous fistula stenosis: results after 22 months of follow-up.

BACKGROUND: Access flow (QA) surveillance is the best method recommended for early stenosis detection, but in native arteriovenous fistula (AVF), the literature is conflicting about the real need for monthly monitoring of QA, as suggested by the K-DOQI Guidelines. METHODS: From 1 January 2006 to 31 October 2007 (mean 18.0 +/- 4.9 months), we prospectively followed up 224 patients with monthly AVF monitoring by means of clinical examination and QB stress test (QBST). Suspected malfunctioning AVFs were referred to ultrasound dilution technique (UDT) and imaging techniques (Doppler ultrasonography, angiography), with eventually further percutaneous angioplasty (PTA) or surgical revision. RESULTS: We observed a good correlation between QBST and QA measurement obtained by the UDT. Patients with positive QBST had a lower QA than negative QBST subjects (433 +/- 203 vs 1168 +/- 681 ml/min, P < 0.0001). Fifty-four out of 224 (24%) patients were selected for possibly malfunctioning AVF. We found no stenosis in 13 out of 54 (24%) patients, inflow stenosis in 29 out of 54 (54%) patients and outflow stenosis in 12 out of 54 (22%) patients. The QBST positive predictive value for inflow stenosis was 76.3%. The interventional radiologist performed 38 PTA procedures in 33 patients (11 PTA per 100 patient-years) and we surgically created 13 new AVF (3.7 per 100 patient-years). Only five thrombosis episodes occurred in five patients during the follow-up (1.5 thromboses per 100 patient-years). CONCLUSIONS: QBST is a simple, low-cost, not time-consuming test, able to select, together with clinical evaluation, malfunctioning AVF with stenosis located specifically in the inflow tract. Our follow-up data demonstrated that it is possible to achieve a low AVF thrombosis rate by adding QBST in an AVF monitoring program, thus reducing the surveillance burden.

Progenitor Cells Activated by Platelet Lysate in Human Articular Cartilage as a Tool for Future Cartilage Engineering and Reparative Strategies.

Regenerative strategies for human articular cartilage are still challenging despite the presence of resident progenitor cell population. Today, many efforts in the field of regenerative medicine focus on the use of platelet derivatives due to their ability to reactivate endogenous mechanisms supporting tissue repair. While their use in orthopedics continues, mechanisms of action and efficacy need further characterization. We describe that the platelet lysate (PL) is able to activate chondro-progenitor cells in a terminally differentiated cartilage tissue. Primary cultures of human articular chondrocytes (ACs) and cartilage explants were set up from donor hip joint biopsies and were treated in vitro with PL. PL recruited a chondro-progenitors (CPCs)-enriched population from ex vivo cartilage culture, that showed high proliferation rate, clonogenicity and nestin expression. CPCs were positive for in vitro tri-lineage differentiation and formed hyaline cartilage-like tissue in vivo without hypertrophic fate. Moreover, the secretory profile of CPCs was analyzed, together with their migratory capabilities. Some CPC-features were also induced in PL-treated ACs compared to fetal bovine serum (FBS)-control ACs. PL treatment of human articular cartilage activates a stem cell niche responsive to injury. These facts can improve the PL therapeutic efficacy in cartilage applications.

Does Intraoperative Platelet-rich Plasma Improve Clinical and Structural Outcomes after Arthroscopic Repair of Isolated Tears of the Supraspinatus Tendon?

BACKGROUND: Arthroscopic cuff repair is a highly successful technique, but postoperative rehabilitation is complex and the rate of tear recurrence is not negligible. Biological augmentations have been proposed to overcome these drawbacks. The platelet-rich plasma (PRP) is a platelet-rich blood fraction that is applied on the repair site to enhance tendon healing. This study evaluates the effectiveness of PRP application in arthroscopic cuff repair. MATERIALS AND METHODS: A prospective nonrandomized study was carried out on 22 patients undergoing arthroscopic rotator cuff repair. Only patients with isolated and repairable supraspinatus tears were included and divided into two groups: 11 patients (Group A) received intraoperative PRP and 11 patients (Group B) did not. All patients had the same rehabilitation and followup protocol. Clinical-functional parameters (visual analog score, active range of motion, University of California at Los Angeles - UCLA, Constant) were recorded at predefined intervals, and magnetic resonance imaging (MRI) was performed 1 year postoperative. RESULTS: Only one patient of Group B did not complete the study protocol. No intraoperative or postoperative complications were observed. No differences were found in the clinical-functional parameters during the entire study. At 1 year, MRI showed 1 retear in Group A and 2 retears in Group B, but the difference was not significant. CONCLUSIONS: The role of PRP as an adjuvant for surgical repair of rotator cuff tears is controversial. In this study, we could not demonstrate significant advantages of PRP for arthroscopic repair of isolated supraspinatus tears. The potential improvement in the structural outcome should be evaluated in the long term to justify the additional costs related to PRP application.

Subtrochanteric fractures: two case reports of non-union treatment.

Proximal femoral fractures are a very common disease which affect elderly patients after low energy trauma, but can also affect the young population as a consequence of high energy trauma. Classification of sub trochanteric fractures has not yet been clearly defined representing a major issue in comparing different treatment techniques among the scientific literature.These fractures are well known by orthopaedic surgeons for intraoperative difficulties in reduction and post-operative high rate of non-unions and malunions.We present two clinical cases, one treated with an intramedullary device and one with an extramedullary device, both failed and revised with open surgery using a condylar angled bladeplate, following AO principles of anatomical reduction and stable fixation.

Human Articular Chondrocytes Regulate Immune Response by Affecting Directly T Cell Proliferation and Indirectly Inhibiting Monocyte Differentiation to Professional Antigen-Presenting Cells.

Autologous chondrocyte implantation is the current gold standard cell therapy for cartilage lesions. However, in some instances, the heavily compromised health of the patient can either impair or limit the recovery of the autologous chondrocytes and a satisfactory outcome of the implant. Allogeneic human articular chondrocytes (hAC) could be a good alternative, but the possible immunological incompatibility between recipient and hAC donor should be considered. Herein, we report that allogeneic hAC inhibited T lymphocyte response to antigen-dependent and -independent proliferative stimuli. This effect was maximal when T cells and hAC were in contact and it was not relieved by the addition of exogenous lymphocyte growth factor interleukin (IL)-2. More important, hAC impaired the differentiation of peripheral blood monocytes induced with granulocyte monocyte colony-stimulating factor and IL-4 (Mo) to professional antigen-presenting cells, such as dendritic cells (DC). Indeed, a marked inhibition of the onset of the CD1a expression and an ineffective downregulation of CD14 antigens was observed in Mo-hAC co-cultures. Furthermore, compared to immature or mature DC, Mo from Mo-hAC co-cultures did not trigger an efficacious allo-response. The prostaglandin (PG) E2 present in the Mo-hAC co-culture conditioned media is a putative candidate of the hAC-mediated inhibition of Mo maturation. Altogether, these findings indicate that allogeneic hAC inhibit, rather than trigger, immune response and strongly suggest that an efficient chondrocyte implantation could be possible also in an allogeneic setting.

Klebsiella pneumoniae Is Able to Trigger Epithelial-Mesenchymal Transition Process in Cultured Airway Epithelial Cells.

The ability of some bacterial pathogens to activate Epithelial-Mesenchymal Transition normally is a consequence of the persistence of a local chronic inflammatory response or depends on a direct interaction of the pathogens with the host epithelial cells. In this study we monitored the abilities of the K. pneumoniae to activate the expression of genes related to EMT-like processes and the occurrence of phenotypic changes in airway epithelial cells during the early steps of cell infection. We describe changes in the production of intracellular reactive oxygen species and increased HIF-1α mRNA expression in cells exposed to K. pneumoniae infection. We also describe the upregulation of a set of transcription factors implicated in the EMT processes, such as Twist, Snail and ZEB, indicating that the morphological changes of epithelial cells already appreciable after few hours from the K. pneumoniae infection are tightly regulated by the activation of transcriptional pathways, driving epithelial cells to EMT. These effects appear to be effectively counteracted by resveratrol, an antioxidant that is able to exert a sustained scavenging of the intracellular ROS. This is the first report indicating that strains of K. pneumoniae may promote EMT-like programs through direct interaction with epithelial cells without the involvement of inflammatory cells.

Carbapenem resistance confers to Klebsiella pneumoniae strains an enhanced ability to induce infection and cell death in epithelial tissue-specific in vitro models.

BACKGROUND: Carbapenem-resistant Klebsiella pneumoniae strains (KPC-Kp) are emerging worldwide causing different nosocomial infections including those of the urinary tract, lung or skin wounds. For these strains, the antibiotic treatment is limited to only few choices including colistin, whose continuous use led to the emergence of carbapenem-resistant KPC-Kp strains resistant also to this treatment (KPC-Kp Col-R). AIM: Very little is known about the capacity of the different strains of KPC-Kp to invade the epithelial cells in vitro. To verify if the acquisition of carbapenem-resistant and the colistin-resistant phenotypes are correlated with a different ability to infect a series of epithelial cell lines of various tissutal origin and with a different capacity to induce cellular death. MATERIALS & METHODS: We used Klebsiella pneumoniae (KP), KPC-Kp and KPC-Kp Col-R strains, isolated from different patients carrying various tissue-specific infections, to infect a series of epithelial cell lines of different tissutal origin. The invasive capacity of the strains and the extent and characteristics of the cell damage and death induced by the bacteria were evaluated and compared. CONCLUSION: Our results show that both KPC-Kp and KPC-Kp Col-R display a greater ability to infect the epithelial cells, with respect to KP, and that the bacterial cell invasion results in a nonprogrammed cell death.

Total lymphocyte count and in-hospital mortality in older persons with multimorbidity.

BACKGROUND AND AIMS: Low total lymphocyte count (TLC) has been found to be a poor prognostic factor in adults affected by heart diseases, malignancy, and renal failure. The aims of this study were to verify if a low TLC was associated with in-hospital mortality in older persons and to evaluate whether this association was independent of the presence of multiple co-existing diseases (multimorbidity). METHODS: The authors carried out a cross-section analysis of data of 65+ years old patients (n=596) admitted to a Geriatric Unit in Northern Italy. TLC, total white blood cell count (WBC) and serum albumin were assayed the day after admission. The presence and severity of diseases were evaluated with the Geriatric Index of Comorbidity (GIC). Other covariates included age, gender, cigarette smoking, cognition (Mini-Mental State Examination) and function (Activities of Daily Living). Logistic regression models were created to study factors affecting in-hospital death. RESULTS: TLC was inversely correlated with both age and multimorbidity. Patients in the lowest tertile of TLC had the highest association with death during hospitalization (OR 6.1, 95% CI 1.1-33.6) independently of multimorbidity and all the other covariates. Stratifying the sample by degree of multimorbidity, this association was clearest in patients with the least severe multimorbidity (GIC

Avaliação ex vivo da microinfiltração marginal coronária de restauradores provisórios usados em endodontia / Ex vivo evaluation of coronal marginal microleakage of temporary restoratives used in endodontics

O objetivo deste trabalho foi avaliar ex vivo a microinfiltração marginal coronária de restauradores provisóriosusados em endodontia. Foram utilizados quarenta e seis dentes caninos unirradiculares humanos obtidos do estoque do Banco de Dentes da FORP-USP. Após a realização do acesso endodôntico e do preparo químico-mecânico, foi introduzido em seu canal radicular um cone de papel absorvente e, na câmara pulpar, uma mecha de algodão impregnada com solução alcoólica de dimetilglioxima 1 por cento. Os espécimes foram restaurados com os materiais seguindo as orientações dos respectivos fabricantes, sendo cada grupo assim disposto: G1 - Vidrion R®; G2 - Cavit W®; G3 - Villevie®; G4 Bioplic®; controle positivo - não recebeu nenhum tipo de restaurador provisório; controle negativo - restaurado com Cavit W® e completamente impermeabilizadopor duas camadas de esmalte cosmético. Posteriormente, todos os espécimes foram imersos em solução de sulfato de níquel 5 por cento e submetidos à ciclagem térmica por 72h. Os dados foram submetidos ao teste estatísticonão paramétrico de Kruskal-Wallis e mostraram que os restauradores provisórios Villevie® e Bioplic® sofreram menor infiltração, seguidos pelo Cavit W® e, por último, o Vidrion R®. Pode-se concluir que, apesar de osrestauradores provisórios Villevie® e Bioplic® sofrerem menor infiltração, nenhum material impediu totalmente a infiltração marginal coronária