Tranexamic acid effects on cartilage and synovial tissue: an in vitro study for a possible safe intra-articular use.

The possible toxic effects of intra-articular tranexamic acid (TA) are still debated. The aim of this study was to evaluate TA effects on human cartilage fragments and synovial biopsies. Explant culture of minced articular cartilage underwent prolonged TA exposure. Histological analysis, immunofluorescence and colorimetric assay for quantification of s-GAG and DNA were performed at the end term. Synoviocytes were cultured for 48h in presence of TA. Light microscopy and flow cytometry analysis were performed at the end of the exposure to TA and one week after the treatment. TA exposure did not influence i) the chondrocyte outgrowth and migration, ii) the expression of chondrogenic and proliferative markers and iii) the s-GAG/DNA ratio. TA treatment did not affect synoviocytes' morphology and treated cells were phenotypically similar to control cells. This study demonstrated that TA does not negatively affect chondrocytes and synoviocytes cultured in vitro. Thus, our findings may be clinically relevant in order to validate the intra-articular TA administration during orthopedic procedures.

Galectin-3 expression in thyroid fine needle cytology (t-FNAC) uncertain cases: validation of molecular markers and technology innovation.

Thyroid cancer is not very common, accounting for 1-2% of all cancers, with a population incidence of about 0.004%. Currently, the ability to discriminate between follicular adenoma and carcinoma represents the major challenge in preclinical diagnosis of thyroid proliferative lesions. Better discrimination between the two would help avoid unnecessary thyroidectomy and save valuable resources. Over the years, galectin-3 (Gal-3) has been proposed as a diagnostic marker with varied success. In this paper, we used Environmental Scanning Electron Microscopy Immunogold Labelling (ESEM-IGL) to investigate the expression of Gal-3 on Thin-Prep fine needle aspiration cytology (FNAC). We optimized the ESEM-IGL method on thyroid cell lines (RO-82 and FTC-133) comparing our membrane Gal-3 labeling data with Western blot. We evaluated 183 thyroid FNAC from Italian patients with a uncertain pre-surgical diagnosis. ESEM-IGL method marker sensitivity is 71.2%, while specificity is 53.3% and diagnostic efficacy is 61.2%. Our results confirmed that Gal-3 expression is associated with situations of hypertrophy and/or cellular hyperproliferation, pathophysiological situations common both to adenomas and to thyroid carcinomas. The innovation of thyroid FNAC Thin-Prep ESEM-IGL shows the levels of Gal-3 immunolabeling clearly, even through the individual cells of a thyroid nodule. However, Gal-3 alone, as a molecular marker of thyroid cancer, can still have a limited application in pre-surgery diagnosis.

Cross-linked hyaluronic acid sub-micron particles: in vitro and in vivo biodistribution study in cancer xenograft model.

This paper focused on the biodistribution of the cross-linked hyaluronic acid (HA-NPs) sub-micron particles in tumor-bearing mice. Solvent-non solvent method followed glutaraldehyde cross-linking utilized for the fabrication of HA-NPs. Size measurement and morphological analysis were performed by dynamic light scattering and electron microscopy, respectively and the size found to be in the range of 200-400 nm. In vitro viability in LNCaP cell line was assessed by water soluble tetrazolium assay after 24 h of exposure to sub-micron particles and no toxicity was found to higher concentration of 3 mg/mL. Internalization of particles in prostate cancer cell LNCaP were studied by confocal microscopy with FITC labeled submicron particles and involvement of hyaluronan receptor mediated uptake/endocytosis was confirmed by competitive assay. Biodistribution studies were performed in xenograft prostate cancer mice model with fluorophore labeled particles and monitored in tumoral parenchyma with strong fluorescence, meanwhile very less signal in liver, kidney and spleen while no fluorescence found in lung after 24 h of systemic administration; that shown ability of this HA based system to recognize cancer tissue. These result fetched that hyaluronic acid based system is selective for tumoral site and can be utilized to deliver bioactives in specific (targeting) and controlled (temporal) manner to cancerous tissue.

Immunogold labelling in environmental scanning electron microscopy: applicative features for complementary cytological interpretation.

We have combined environmental scanning electron microscopy (ESEM) and immunogold labelling (IGL) for the analysis of cell morphology and surface protein detection on human fine needle aspiration, which is processed in thin uniform monolayer (a single layer of cells) on a glass slide by Thin Prep technology. Among scanning electron microscopy techniques, we choose the environmental modality (ESEM) because it allows a slight manipulation of biological samples and an operational time comparable with cytological techniques. Moreover, the Thin Prep technology confirmed a reproducible cell monolayer on glass smear, minimizing problems for the determination of appropriate amount of material per slide. The first experimental data in ESEM-IGL on biological samples with fine needle aspiration Thin Prep, in human thyroid nodules, showed that cells retained their morphology and provided a clear IGL. The optimization of conditions (i.e. vacuum pressure, temperature and relative humidity) confirmed the possibility to observe an immunolabelled biological sample and morphological signal, joined with compositional informations, due to peculiar characteristics of gaseous secondary electron detector in ESEM. The ESEM-IGL and fine needle aspiration Thin Prep could be used in combination for the interpretation of cell morphology and cell surface immunolabelling. Our paper suggests this use as a powerful diagnostic tool in a pre-surgical evaluations, opening a new applicative window for electron microscopy.

Cemented fixed-bearing PFC total knee arthroplasty: survival and failure analysis at 12-17 years.

BACKGROUND: Total knee arthroplasty (TKA) is the appropriate treatment for degenerative pathology of the knee. Implant surveillance is mandatory to improve clinical results. We present the long-term results of a series of consecutive TKA Press Fit Condylar (J&J), cemented fixed bearing with selective patellar resurfacing in nonselected patients. MATERIALS AND METHODS: In this prospective case series, 223 TKA were clinically and radiographically evaluated using the Hospital for Special Surgery (HSS) knee score and the Knee Society Roentgenographic Evaluation and Scoring System. RESULTS: There were 197 patients, with an average age of 68.4 years [95% confidence interval (CI) 52.7-84.1 years]; 49 arthroplasties were implanted in men (21.1%) and 184 (78.9%) in women. The average follow-up was approximately 13.5 years (162.1 months; 95% CI 132.3-191.9), and it was possible to evaluate 179 implants (76.8% of the implanted prosthesis) in 176 patients. The average HSS score increased from 61.5 (95% CI 60.4-62.7) to 89.4 (95% CI 87.7-.93.5) points. The cumulative average survival rate at 15 years (the endpoint being failure with revision) was 90.6%  ± 2% standard deviation. Resurfacing the patella did not make a difference in terms of implant survival. Progressive radiolucent lines were observed around 20 implants (14.3%); all were revised. CONCLUSIONS: The PFC system is an excellent prosthetic solution. Early clinical complications, mechanical axis and patellar resurfacing do not correlate with implant failure, whereas progressive radiolucent lines do.

[Tissue engineering of parathyroid gland]. / L'ingegnerizzazione tissutale delle cellule paratiroidee.

BACKGROUND: The postoperative hypoparathyroidism is a not rare complication after total thyroidectomy and/or total parathyroidectomy. Attempts to transplant parathyroid tissue began in 1975 with the work of Wells, but still today results are disappointing. However, with the development of tissue engineering techniques, some experimental approaches to build artificial parathyroid are been made. Bioengineered device, actively secreting PTH, for transplant in patients with iatrogenic hypoparathyroidism is unavailable. PATIENTS AND METHODS: Parathyroid cells were obtained from three chronic uremic patients in hemodialysis, operated for secondary hyperparathyroidism. Cell cultures in RPMI medium were subsequently seeded on collagen scaffold (three-dimensional matrix with slow biodegradation). Collagen is the major component of the extracellular matrix and thus is a good substrate for cell adhesion and growth. Culture media, with a low calcium concentration, were optimised to physiologically stimulate parathyroid hormone secretion. Cell cultures were morphologically observed in optical and electron (ESEM) microscopy and metabolically assayed by MTT method until the tenth week. Besides, concentration of parathyroid hormone in the culture medium has been measured for several weeks. RESULTS: After 24 hours of culture in RPMI, cells extracted from human parathyroid glands were nearly all adherent and organised in clusters to resemble the glandular organization. The cellular population consisted predominantly of parathyroid cells (90-95%). On collagen scaffolds, cells maintains an epithelial-like morphology also after 10 weeks, colonizing the scaffold surface and keeping a good proliferative rate with a discrete production of parathyroid hormone. CONCLUSION: The use of parathyroid cells extracted from patients with secondary hyperparathyroidism was certainly an appropriate choice that enabled us to achieve these results, that albeit partial bode well for the experimental in vivo animal model. The bioengineered scaffolds when implanted in the subcutaneous can avoid the dispersion of parathyroid cells, assuring also the possibility to easily remove the implant in case of complications. Our research was aimed primarily to the optimisation of PTH secreting human parathyroid cells cultures and then to the in vitro engineering of human parathyroid glands in three-dimensional collagen scaffolds.

Trends in invasive disease due to Candida species following heart and lung transplantation.

Although invasive candidiasis (IC) causes significant morbidity and mortality in patients who undergo heart, lung, or heart-lung transplantation, a systematic study in a large cohort of thoracic organ transplant recipients has not been reported to date. Clinical and microbiological data were reviewed for 1305 patients who underwent thoracic organ transplantation at Stanford University Medical Center between 1980 and 2004. We identified and analyzed 76 episodes of IC in 68 patients (overall incidence 5.2% per patient).The incidence of IC was higher in lung (LTx) and heart-lung transplant (HLTx) recipients as compared with heart transplant (HTx) recipients (risk ratio [RR] 1.7, 95% confidence interval [CI] 1.1-2.7).The incidence of IC decreased over time in all thoracic organ transplant recipients, decreasing from 6.1% in the 1980-1986 time period to 2.1% in the 2001-2004 era in the HTx recipients, and from 20% in the 1980-1986 period to 1.8% in the 2001-2004 period in the LTx and HLTx recipients.The most common site of infection differed between the HTx and LTx cohorts, with bloodstream or disseminated disease in the former and tracheobronchitis in the latter. IC in the first year after transplant was significantly associated with death in both HTx (RR 2.9, 95% CI 1.8-4.6, P=0.001) and LTx and HLTx patients (RR 3.0, 95% CI 1.9-4.6, P<0.001). The attributable mortality from IC decreased during the 25-year period of observation, from 36% to 20% in the HTx recipients and from 39% to 15% in the LTx and HLTx recipients. There were a significant number of cases caused by non-albicans Candida species in all patients, with a trend toward higher mortality in the HTx group. In conclusion, the incidence and attributable mortality of IC in thoracic organ transplant recipients has significantly declined over the past 25 years.The use of newer antifungal agents for prophylaxis and treatment, the decrease in the incidence of cytomegalovirus disease, and the use of more selective immunosuppression, among other factors, may have been responsible for this change.

Effect of chlorhexidine baths on central-line-associated bloodstream infections in a neonatal intensive care unit in a developing country.

Healthcare-associated infections, particularly central-line-associated bloodstream infections (CLABSIs), are worrisome in neonates. This study describes the impact of chlorhexidine baths on CLABSI rates in a neonatal intensive care unit in a developing country, through a quasi-experimental study undertaken over 62 months (January 2012 to February 2017) divided into two periods: before and after the implementation of a protocol for chlorhexidine baths in July 2014. The rate of CLABSIs per 1000 central-line-days decreased from 8.64 to 4.28 after implementation of the protocol. The use of chlorhexidine baths appears to reduce the number of CLABSIs in neonates.

Liver Transplantation in Hepatitis C-Infected Patients: Experience From a South American Transplant Center.

BACKGROUND: Around 2.4% of the world's population is infected with hepatitis C virus (HCV), and it is the most common cause of liver transplantation (LT) in the world. Latin America (LA), with nearly 9% of the world population, has had a continuous increase in the number of LTs per year. Yet, due to the lack of mandatory data collection and a well-developed health-care system, access to transplantation is limited in most LA countries. We report the first LA experience of HCV-infected LT patients. METHODS: We performed a retrospective cohort study by reviewing the medical histories of all HCV-infected LT patients between 1996 and 2016 who acquired HCV before their LT, at the Fundación Valle del Lilí, Cali, Colombia. RESULTS: Between January 1996 and December 2015, a total of 770 LTs were performed, of which 75 had a cirrhotic liver due to HCV infection. With a median follow-up time of 24.4 months (interquartile range [IQR] 4.7-61.2 months), patient survival was 44.9% and 66.9% for the time periods 1996-2006 and 2007-2015, respectively. Hepatocellular carcinoma (HCC) was present in 30.6% of the patients, and overall postoperative complications had an incidence of 80%. CONCLUSIONS: This is the first report of LT in HCV-infected patients in Colombia and in LA. Our results are comparable to those of other transplant centers worldwide with regard to postoperative complications and patient survival. Patients with LT in the 1996-2006 time frame had higher morbidity and mortality. Studies including larger numbers of patients are needed to determine the reason for this finding.

Purification of ethanolaminephosphotransferase from bovine liver microsomes.

CDP-ethanolamine:diacylglycerol ethanolaminephosphotransferase (EC 2. 7.8.1) has been purified to electrophoretic homogeneity and in a catalytically active form from bovine liver microsomes. The purification method is based on the high hydrophobicity of the protein whose charged sites appear to be masked from the interaction with the chromatographic stationary phases when membranes are solubilized with an excess of non-ionic detergent. The isolated protein has a molecular mass of about 38 kDa, as estimated by SDS-PAGE mobility, and exhibits both ethanolaminephosphotransferase and cholinephosphotransferase activities. Evidence is given that both activities are Mn2+-dependent and that the same catalytic site is involved in cholinephosphotransferase and ethanolaminephosphotransferase reactions. Mg2+-dependent CDP-choline:diacylglycerol cholinephosphotransferase (EC 2.7.8.2) is completely inactivated during the solubilization and purification steps.

Antiovary antibodies, repeated attempts, and outcome of in vitro fertilization.

OBJECTIVE: To determine whether the microtrauma induced by repeated puncture of ovarian follicles could result in the production of autoantibodies in women submitted to in vitro fertilization (IVF). DESIGN: Antiovarian antibodies were assayed in serum samples obtained from women engaged in IVF and in serum samples from healthy controls. SETTING: Blood samples were obtained after oocyte collection in the local hospital's IVF unit. PATIENTS: One hundred ten women undergoing IVF entered the study. Oocyte division, numbers of embryos, and pregnancy were recorded for each woman. Control samples were from 40 individuals with no sign of autoimmune disorder. INTERVENTIONS: Antiovarian antibodies were assayed in indirect immunofluorescence and enzyme-linked immunosorbent assay (ELISA) using human ovary as antigen. Specific immunoglobulin (Ig)G, IgA, and IgM were tested separately in an ELISA. MAIN OUTCOME MEASURE: Antiovarian antibody levels were compared between patients and controls and correlated with the subsequent results of IVF. RESULTS: Significantly higher levels of IgG, IgA, and IgM antiovarian antibodies were observed in the group of IVF women (P less than 0.001). IgM antiovarian antibodies correlated with a lower number of embryos (P less than 0.001) and IVF failure (P less than 0.05). CONCLUSIONS: These data suggest that ovarian trauma may lead to the production of autoantibodies.

Importance of semen preparation in avoidance of reduced in vitro fertilization results attributable to bacteria.

Although the physical treatment of semen for IVF and related procedures is sufficient to remove most organisms present in semen, the pathogenic varieties tend to be more resistant. The supplementation of oocyte culture medium with both penicillin and streptomycin was associated with the eradication of 100% of organisms in the current study. In these circumstances, the presence of pathogenic organisms in the untreated semen is not associated with reduced oocyte fertilization in vitro.

Factors affecting human in vitro fertilization: a multifactorial study.

The influence of four factors on the cleavage rate of 705 mature oocytes submitted for in vitro fertilization (IVF) was assessed with the use of a multifactorial method, i.e., logistic regression. The studied factors were (1) the regimen for cycle stimulation, (2) cumulus cell mass appearance, (3) semen quality, and (4) the time of incubation of oocytes before insemination. The logistic function permitted testing the respective influence of each factor on the cleavage rate, with the level of the other factors taken into account. The most important factor was the stimulation treatment, the association of human menopausal gonadotropin with clomiphene citrate (CC) giving better results than CC alone. Time of incubation was demonstrated to have no influence on the cleavage rate, whereas semen quality had an influence. A problem was raised by the existence of an interaction between the stimulation treatment used and the appearance of the cumulus cell mass. It can be concluded that adequate cycle treatment and eventually couple selection with respect to the semen quality of the male partner are to be considered in view of obtaining better results in IVF attempts.

Intensity of nematode infections in cyclic and non-cyclic rock partridge (Alectoris graeca saxatilis) populations.

Populations of rock partridge (Alectoris graeca saxatilis), in the Trentino province of Italy, exhibit cyclic fluctuations in abundance associated with relatively dry habitat. One of the hypothesis to explain these cycles is that survival of some free living parasitic stages and rates of infection are greater in these areas leading to higher parasite burden. This hypothesis was examined by investigating the intensity of parasite infection in cyclic and non cyclic rock partridge populations. Analyses of 87 intestine samples collected from shot rock partridges during 1994 and 1995 identified 8 species of helminths parasites: Ascaridia compar (P = 33.33%; I = 9.28 +/- 1.78), Heterakis tenuicauda (P = 19.54%; I = 10.29 +/- 4.58), Heterakis gallinarum (P = 1.15%; I = 1.0 +/- 0.0), Heterakis altaica (P = 1.15%; I = 17 +/- 0.0), Aonchoteca caudinflata (P = 6.89; I = 2.17 +/- 0.65), Postharmostomum commutatum (P = 5.75; I = 7.0 +/- 3.48), Brachylaema fuscata (P = 1.15; I = 7.0 +/- 0.0), Platynosomum alectoris (P = 2.29; I = 5.5 +/- 1.5). Cestoda, recorded with a prevalence of 5.75, were not identified to species level. A. compar and H. tenuicauda were prevalent in the rock partridge populations and there was no positive association between these species. Intensity of infection in both species was not influenced by host age, sex or year of study but levels of infection with A. compar burdens were significantly greater in cyclic populations than in non cyclic populations and there was a tendency for H. tenuicauda to be greater in cyclic populations. There was no negative relationship between intensity of infection with A. compar or H. tenuicauda and host body mass. These data provide some support for the hypothesis that these parasites may play a role in generating rock partridge population cycles.

A survey to identify the important macroparasites of rock partridge (Alectoris graeca saxatilis) in Trentino, Italy.

A preliminary survey of the intestinal helminth community of rock partridge (Alectoris graeca saxatilis) was undertaken in the Italian province of Trentino where the host population exhibits cyclic fluctuations in abundance. Six helminth species were recorded from 38 rock partridge intestines collected during the hunting season between October and November 1994. Nematoda were the most prevalent (53%) with lower prevalence of Trematoda (11%) and Cestoda (8%). Ascaridia compar (Schrank, 1790) and Heterakis tenuicauda Linstow, 1883, were identified as the component species (each with prevalence > 10%) in the helminth intestinal community of rock partridge in Trentino. Both of these species exhibit an aggregated distribution.

Takayasu arteritis in a 66-year-old caucasian woman.

A 66 year-old Italian woman was admitted to the hospital for fatigue, malaise and fever. Radial pulses were absent. She underwent echo color Doppler examination and angiography, which pointed out tight stenosis of bilateral axillary arteries; particularly, the echo color Doppler showed diffuse arterial wall thickening in the left common carotid artery (1.7 mm). Clinical features and instrumental findings suggested active Takayasu is arteritis. Corticosteroid therapy was administered and bilateral radial pulse appeared again after two months of steroid therapy. This case demonstrates the presence of Takayasu's arteritis in Western Europe and showed how the disease onset can be observed also in geriatric patients.

[Celioscopy under local anesthesia for collecting oocytes for in vitro fertilization]. / Coelioscopie sous anesthésie locale pour recueil d'ovocyte en vue d'une fécondation in vitro.

Since 1960, laparoscopy under local anaesthesia has been gradually abandoned in France and until now oocytes for in vitro fertilization were collected under general anaesthesia, thus increasing the risk of allergic reactions to anaesthesia in these patients who usually undergo several operations. From February to May, 1983, at the Antoine Béclère Maternity, Paris, 50 laparoscopies were performed under local anaesthesia with lidocaine combined with neuroleptanalgesia with diazepam and fentanyl. The procedure was uneventful in 94% of the cases and only 3 women required general anaesthesia. At least one mature oocyte was collected in 46 of these 50 women and 4 pregnancies have already been recorded. Local anaesthesia enables the woman to participate fully in this first stage of in vitro fertilization, increases the out-patient activities of the hospital unit and reduces the cost of in vitro fertilization.

[A biocompatibility study and the effects of slow-release antibiotic materials in the treatment of periodontal disease. I. The biocompatibility of cellulose acetate charged with 25% tetracycline hydrochloride. A clinical and scanning microscopic study of a case]. / Studio sulla biocompatibilità e gli effetti di alcuni materiali a lento rilascio di antibiotico nel trattamento della malattia parodontale. Nota I. Biocompatibilità dell'acetato di cellulosa caricato con il 25% di tetraciclina cloridrato. Studio clinico e al microscopio a scansione di un caso.

A clinical and microscopical (SEM) investigation has been carried out on the biocompatibility of cellulose acetate fiber tetracycline with 25% of tetracycline hydrochloride (Actisite R). A subject with advanced periodontal disease was selected and a pocket of 8 mm of PD was chosen. A segment of fiber was inserted into the pocket for 8 days. After removal, PD and GI clinical parameters were detected and the fiber removed was analyzed at the scanning electronic microscope. The results showed clinical signs of inflammation after removal of fiber. SEM analysis showed macrophagic reaction, a typical sign of inflammatory response to material. The study suggests the need of more biocompatible materials, easier to use as delivery system of antibiotics in the treatment of periodontal disease.

[A biocompatibility study and the effects of slow-release antibiotic materials in the treatment of periodontal disease. II. The biocompatibility and behavior of polyhydroxyethyl methacrylate (pHEMA) as a slow-release material for tetracycline of metronidazole. A study of 2 cases]. / Studio sulla biocompatibilità e gli effetti di alcuni materiali a lento rilascio di antibiotico nel trattamento della malattia parodontale. Nota II. Biocompatibilità e comportamento del poliidrossietilmetacrilato (pHEMA) come materiale a lento rilascio di tetraciclina o di metronidazolo. Studio su due casi.

The study analyses the possibility of using polyhydroxyethyl methacrylate as a material for the slow release of antibiotic in periodontal pockets. The antibiotics examined were tetracycline and metronidazole. The aim of the study was to evaluate the biocompatibility of the material with periodontal tissue and the efficacy of the 2 prepared systems. Two sites were selected in 2 periodontopathic patients who after non-surgical treatment presented pockets measuring 8 and 7 mm. A sheet of pHEMA containing tetracycline was inserted in one and in the other a sheet containing metronidazole: both were left for 8 days in the chosen pockets. At the start and end of treatment PD and GI clinical indices were measured and the DMDx microbiological test was performed to identify Aa of Pg and Pi. The tissue reaction to pHEMA was evaluated using SEM analysis of two samples collected after 8 days of treatment. The microscopic results showed the optimal biocompatibility of both samples. Differences were noted with regard to clinical and microbiological efficacy. It was observed that the sheet of pHEMA containing tetracycline resulted in the disappearance of bleeding and a reduced depth of survey. Moreover, microbiological results showed a significant reduction in Porphyromonas gingivalis. The sheet of pHEMA containing metronidazole showed a lower level of therapeutic efficacy. Although reduced depth was noted, gingival bleeding was persistent and the reduction of bacteria analysed was not significant. In conclusion, the authors confirm the optimal biocompatibility of the material and its easy application, although further research, especially for pHEMA with metronidazole, must be carried out to improve drug kinetics, trying to maintain an effective local concentration throughout treatment.