Successful emergency Lichtenberger lateralisation for immediate bilateral laryngeal immobility after total thyroidectomy: a CARE case report.

OBJECTIVE: This case report discusses a successful emergency Lichtenberger lateralisation procedure after immediate bilateral laryngeal immobility, occurring after total thyroidectomy. METHODS: A 63-year-old female with right-sided vocal fold paralysis due to compression by a multinodular thyroid goitre underwent total thyroidectomy, which resulted in immediate post-operative bilateral vocal fold immobility. The patient had acute-onset post-operative dyspnoea, was promptly re-intubated, and an emergency lateralisation Lichtenberger suture was placed over the right vocal fold and fixated on the outer surface of the neck. RESULTS: After two weeks, her right vocal fold recovered first, with the suture still in place. At four weeks, both vocal folds regained function and the suture was extracted. CONCLUSION: The take-away message is that an emergency lateralisation suture may be a viable option in maintaining airway patency, while allowing for normal deglutition, in patients who would otherwise be candidates for prolonged intubation, posterior cordotomy, medial arytenoidectomy or tracheostomy.

Potential of root nodule nonrhizobial endophytic bacteria for growth promotion of Lotus corniculatus L. and Dactylis glomerata L.

AIMS: This research aimed to isolate and characterize nonrhizobial endophytic bacteria from root nodules of Medicago sativa L. and Lotus corniculatus L. with plant growth-promoting characteristics and to test its activity in a pot experiment with acid soil. METHODS AND RESULTS: Out of 44 nonrhizobial isolates, the majority exhibited indole-3-acetic acid (IAA) production; 29 produced siderophores, few isolates performed phosphate solubilization and/or produced lytic enzymes, while 30% of isolates showed notable antifungal activity. The most promising strains were identified as members of Bacillus, Pseudomonas and Serratia genera, based on 16S rRNA. Bacillus megaterium DZK1BH exhibited the overall best attributes for plant growth promotion and positively influenced the growth of L. corniculatus and Dactylis glomerata. CONCLUSIONS: Root nodule endophytic B. megaterium DZK1BH could potentially be used as a biofertilizer for growing L. corniculatus L. and D. glomerata L. in acid soils, while Bacillus mojavensis L3 is a candidate for further antifungal potential investigation. SIGNIFICANCE OF IMPACT OF THE STUDY: The use of root nodule endophytic bacteria with PGP traits may find its future application in organic agriculture, as their utilization could decrease the use of chemical fertilizers and pesticides and simultaneously promote plant growth, especially in soils with low production quality.

Novel insight into Robert's cytoprotection: complex therapeutic effect of cytoprotective pentadecapeptide pentadecapeptide BPC 157 in rats with perforated stomach throughout modulation of nitric oxide-system. Comparison with L-arginine, ranitidine and pantoprazole therapy and L-NG-nitro-L-arginine methyl ester worsening.

Surgically perforated stomach (since direct injury in rats until persisting defect and huge adhesions (day 1, day 7)) fairly represent an unresolved cytoprotection issue, and thereby, we focused resolving of the immediate triad, particular vascular failure (vessels 'disappear'/empty), prolonged bleeding, debilitated defect large widening. Agents (mg/kg) or saline (controls) were given at 1 min post-injury as an abdominal bath (10 ml/rat throughout 2 min). Within 1 - 15 min post-injury period, with cytoprotective BPC 157 (0.01 µg), the rapidly restored vessels 'run' (vessels filled/reappeared) toward the perforated defect, and there is less bleeding, and defect contraction; advanced perforated lesion healing (day 1) to complete healing (day 7), and less adhesions. With pantoprazole (10 mg), early (vessels (worsening), bleeding (prolongation), defect (attenuated widening)) effect means eventual lesions and adhesions severity as in controls. Ranitidine (10 mg) early effect (vessels (improvement), bleeding (less bleeding), defect (eliminated widening, defect not changed)) means final lesions attenuation, but not complete healing, less adhesions. L-NAME (5 mg) early (vessels worsening, less bleeding, attenuated defect widening) and final (lesions aggravation, more adhesions) effect, versus L-arginine (100 mg) early (vessels improvement, more bleeding, attenuated defect widening) and final (lesions attenuation, less adhesions) effect, combined few simultaneously occurring nitric oxide (NO)-system distinct processes. Finally, in the stomach tissue surrounding defect, increased malondialdehyde (MDA)- and decreased NO-values, BPC 157 reversed to the normal healthy values, and mRNA expression studies (Cox2, VEGFa, Nos1, Nos 2, Nos3, Nkap (NF-kappa-B-activating protein gene)), done at that very early post-perforation-time, indicate a way how BPC 157 may act beneficially in the perforated stomach lesion throughout NO- and prostaglandinds-system.

Introduction of dynamic rate-of-force development scaling factor in progressive drop jumps.

Rapid force generation across submaximal levels has been evaluated with the rate of force development scaling factor (RFD-SF) in different isometric tasks, while such measurement was still not verified in dynamic tasks. Our study was designed to evaluate the feasibility of the RFD-SF in dynamic drop jump (DJ) task (RFD-SFDJ). A total of 55 young athletes performed isometric plantarflexion at different submaximal intensities and 60 DJs (6 different drop heights). For each participant we calculated linearity (r2) and slope in isometric task (RFD-SFPF), eccentric part of DJ (RFD-SFDJ-ECC) and concentric part of DJ (RFD-SFDJ-CON), as well as average jump height (DJH) from each drop height. Our results revealed strong linear force-RFD relationship for isometric plantarflexion (r2 = 0.90 ± 0.06), eccentric (r2 = 0.87 ± 0.09) and concentric phase of DJ (r2 = 0.80 ± 0.18). Significant moderate positive correlations were calculated between RFD-SFPF and RFD-SFDJ-ECC (r = 0.311, p < 0.05) and small negative correlations between RFD-SFDJ-CON and RFD-SF (r = -0.276, p < 0.05). Significant positive moderate correlations were seen only between RFD-SFDJ-ECC and DJH from 10 cm (r = 0.459, p < 0.001) and 15 cm (r = 0.423, p < 0.01). This is the first study to introduce and confirm that RFD-SFDJ can be obtained from the multi-joint tasks (60 jumps) and still provide acceptable reliability and linear relationship. Furthermore, RFD-SFDJ may have greater practical application than RFD-SF assessed under the isometric conditions. This verification of RFD-SFDJ opens opportunities for further research regarding its practical application.

Surface Tensions between Active Fluids and Solid Interfaces: Bare vs Dressed.

We analyze the surface tension exerted at the interface between an active fluid and a solid boundary in terms of tangential forces. Focusing on active systems known to possess an equation of state for the pressure, we show that interfacial forces are of a more complex nature. Using a number of macroscopic setups, we show that the surface tension is a combination of an equation-of-state abiding part and of setup-dependent contributions. The latter arise from generic setup-dependent steady currents which "dress" the measurement of the "bare" surface tension. The former shares interesting properties with its equilibrium counterpart, and can be used to generalize the Young-Laplace law to active systems. Finally, we show how a suitably designed probe can directly access this bare surface tension, which can also be computed using a generalized virial formula.

Asymmetry indicates more severe and active disease in Graves' orbitopathy: results from a prospective cross-sectional multicentre study.

PURPOSE: Patients with Graves' orbitopathy can present with asymmetric disease. The aim of this study was to identify clinical characteristics that distinguish asymmetric from unilateral and symmetric Graves' orbitopathy. METHODS: This was a multi-centre study of new referrals to 13 European Group on Graves' Orbitopathy (EUGOGO) tertiary centres. New patients presenting over a 4 month period with a diagnosis of Graves' orbitopathy were included. Patient demographics were collected and a clinical examination was performed based on a previously published protocol. Patients were categorized as having asymmetric, symmetric, and unilateral Graves' orbitopathy. The distribution of clinical characteristics among the three groups was documented. RESULTS: The asymmetric group (n = 83), was older than the symmetric (n = 157) group [mean age 50.9 years (SD 13.9) vs 45.8 (SD 13.5), p = 0.019], had a lower female to male ratio than the symmetric and unilateral (n = 29) groups (1.6 vs 5.0 vs 8.7, p < 0.001), had more active disease than the symmetric and unilateral groups [mean linical Activity Score 3.0 (SD 1.6) vs 1.7 (SD 1.7), p < 0.001 vs 1.3 (SD 1.4), p < 0.001] and significantly more severe disease than the symmetric and unilateral groups, as measured by the Total Eye Score [mean 8.8 (SD 6.6) vs 5.3 (SD 4.4), p < 0.001, vs 2.7 (SD 2.1), p < 0.001]. CONCLUSION: Older age, lower female to male ratio, more severe, and more active disease cluster around asymmetric Graves' orbitopathy. Asymmetry appears to be a marker of more severe and more active disease than other presentations. This simple clinical parameter present at first presentation to tertiary centres may be valuable to clinicians who manage such patients.

Infrared digital thermography of scrotum in early selection of progressive varicocele.

Varicocele is frequent but correctable cause of male infertility. Varicocelectomy is the most commonly performed operative procedure for varicocele. Majority of varicocele patients do not have fertility problem, therefore surgical correction is not recommended in all prevalent cases. On the other hand, varicocele is a progressive condition in some cases and individual with varicocele is at risk for developing impairment which can ultimately lead to semen deterioration and consequent infertility. Selection of patients with varicocele that will progress and cause infertility is beyond our current diagnostic capabilities. Diagnostic assessment of varicocele depends on physical examination and scrotal ultrasound/doppler. Infrared digital thermography of scrotum is a non-invasive and objective diagnostic method for early varicocele detection by means of temperature measurement on the scrotal skin surface. The criteria for diagnostic use of scrotal thermography were recently presented. We hypothesize that the infrared digital thermography of scrotum could be the cornerstone in detection of varicoceles that tend to progress with impairment of semen quality and will require surgical correction, among all prevalent varicocele cases.

Use of platelet-rich plasma in the treatment of bisphosphonate-related osteonecrosis of the jaw.

Platelet-rich plasma is a concentrate of growth factors and osteoconductive proteins, which can play a major role in bone biology by accelerating and enhancing bone repair and regeneration. This paper describes the results of using platelet-rich plasma in the management of bisphosphonate-associated necrosis of the jaw. Eight patients with a diagnosis of bisphosphonate-associated necrosis of the jaw were surgically treated for debridement and removal of necrotic bone, followed by application of autologous platelet concentrate enriched with growth factors and primary suture of the wound. Patients underwent periodic clinical and radiological follow-up examinations. All patients showed clinical improvement and oral lesions resolved 2-4 weeks after treatment. After an average 14-month follow up period, patients remained asymptomatic. Although not conclusive, the combination of necrotic-bone curettage and platelet-rich-plasma to treat refractory osteonecrosis of the jaw yielded promising results.

Scrotal infrared digital thermography in assessment of varicocele--pilot study to assess diagnostic criteria.

The aim of this study was to assess scrotal thermography in diagnostics of varicocele and suggest potential diagnostic criteria. Twelve patients with clinically diagnosed varicocele were examined with scrotal infrared digital thermography, physical examination and ultrasound/doppler. The main outcome measure was evaluation of thermography diagnostic criteria for varicocele. Mean temperature at left pampiniform plexus was ≥ 34 °C in 83%, and at right pampiniform plexus in all cases was ≤ 34 °C. In 92% of patients, temperature at the left testicle was ≥ 32 °C, whereas at the right testicle it was >32 °C in 50% patients. Temperatures between left and right pampiniform plexus and between left and right testicle were significantly different with P < 0.0001 and P < 0.006 respectively. In all patients, temperature difference between pampiniform plexuses was ≥ 0.6 °C. In 92% of patients, temperature at left pampiniform plexus was equal or higher to thigh temperature with the mean temperature difference of 1.1 ± 1.1 °C. Temperature at right pampiniform plexus was colder than the thigh in 92% of patients. This study suggests diagnostic criteria of five thermographic signs to easily diagnose varicocele. Scrotal thermography presents feasible, short and low cost diagnostic method for varicocele. Further study on a larger number of patients and healthy participants is needed to evaluate sensitivity and specificity of this method.

Effects of a platelet gel on early graft revascularization after anterior cruciate ligament reconstruction: a prospective, randomized, double-blind, clinical trial.

BACKGROUND: Slow graft healing in bone tunnels and a slow graft ligamentization process after anterior cruciate ligament (ACL) reconstruction are some of the reasons for prolonged rehabilitation. AIMS: The purpose of this study was to determine if the use of platelet gel (PG) accelerates early graft revascularization after ACL reconstruction. METHODS: PG was produced from autologous platelet-rich plasma and applied locally. We quantitatively evaluated the revascularization process in the osteoligamentous interface zone in the bone tunnels and in the intra-articular part of the graft by means of contrast-enhanced magnetic resonance imaging (MRI). RESULTS: After 4-6 weeks, the PG-treated group demonstrated a significantly higher level of vascularization in the osteoligamentous interface (0.33 ± 0.09) than the control group (0.16 ± 0.09, p < 0.001). In the intra-articular part of the graft, we found no evidence of revascularization in either group. CONCLUSION: Locally applied PG enhanced early revascularization of the graft in the osteoligamentous interface zone after ACL reconstruction.

Interleukin-17 may be a valuable serum tumor marker in patients with colorectal carcinoma.

The promotion of tumor growth is due to a combination of several mechanisms, including angiogenesis and the abundance of cell-derived inflammatory cytokines. The aim of this study was to investigate the serum levels of interleukin 17 (IL-17) and the expression of p53 and Vascular Endothelial Growth Factor (VEGF), in order to determine the relationship between these markers and serum IL-17 levels in patients with colorectal carcinoma. Serum levels of the proinflammatory cytokine IL-17 in patients with colorectal carcinoma (CRC) (n=40) and in a healthy group (n=37) were analysed by ELISA. Surgically resected specimens of 59 colorectal carcinomas were studied by immunohistochemical staining for VEGF and p53. Analyses by ELISA showed significantly higher IL-17 serum levels in patients with colorectal carcinoma than in control subjects (IL-17; mean 128.52+/-47.62 pg/ml vs. mean 101.91+/-22.46 pg/ml; p=0.022). We also found an inverse correlation between p53 expression and the level of IL-17 in the serum of patients with CRC. In fact, the serum concentration of IL-17 was significantly higher in patients who did not express p53 (p=0.023). There was no significant correlation between the expression of p53 and VEGF. However, concomitant expression of VEGF and p53 showed a significant correlation with the histological and nuclear grade of the carcinoma. The data presented in our study indicate that IL-17 might act as a valuable tumor marker in patients with CRC and that combined analysis of p53 and VEGF expression might provide additional information about tumor features.

Evaluation of rabbit auricular chondrocyte isolation and growth parameters in cell culture.

Auricular cartilage is an attractive potential source of cells for many tissue engineering applications. However, there are several requirements that have to be fulfilled in order to develop a suitable tissue engineered implant. Animal experiments serve as important tools for validating novel concepts of cartilage regeneration; therefore rabbit auricular chondrocytes were studied. Various parameters including isolation procedures, passage number, rate of proliferation and gene expression profile for major extracellular matrix components were evaluated in order to assess the potential use of elastic chondrocytes for tissue engineering. Chondrocytes were isolated from rabbit ear cartilage and grown in monolayer cultures over four passages. Yields of harvested cells and proliferation were analysed from the digestion step to the fourth passage, and changes in phenotype were monitored. The proliferation capacity of cell cultures decreased during cultivation and was accompanied by enlargement of cells, this phenomenon being especially evident in the third and fourth passages. The expression of cartilage specific genes for collagen type II, aggrecan and cartilage non-specific collagen type I was determined. The mRNA levels for all three genes were obviously lower in the primo culture than immediately after isolation. During subsequent cultivation the expression of collagen type II decreased further, while there were only slight changes in expression of aggrecan and collagen type I. This study provides a valuable basis for testing of different tissue engineering applications in rabbit model, where auricular chondrocytes are considered as cell source.

Importancia del cepillado antes de acostarse en la prevención de la caries. Estudio en una población infantil / The importance of night tooth brushing before sleeping in the prevention of dental caries. Study on childhood population

En el presente trabajo nos planteamos si sería imprescindible el cepillado tres veces al día y, en caso contrario, en qué momento del día sería más importante la eliminación mecánica de la placa. Para ello, se planteó un estudio transversal correspondiente a 214 escolares de 6-12 años en el periodo comprendido en el curso 2004-5. A cada niño se le practicó el “Examen parcial o tipo III” recomendado por la Federación Dental Internacional, como procedimiento estándar para un estudio de prevalencia de caries. Hemos obtenido los siguientes resultados: 1. El cepillado de dientes antes de acostarse es un factor protector de la presencia de caries, siendo el correspondiente odd-ratio ajustado de 0.339 (intervalo de confianza al 95% [0.159; 0.722]). 2. El consumir azúcar más de una vez una vez al día tiene más riesgo de aparición de caries que si consumen una vez al día, siendo el correspondiente odd-ratio ajustado de 1.998 (intervalo de confianza al 95% [1.090; 3.665]). 3. Los niños que acuden a un colegio público tienen más riesgo de presentar caries que los que asisten a un colegio privado. Siendo el correspondiente odd-ratio ajustado de 2.019 (intervalo de confianza al 95% [1.105; 3.691]). 4. Se cumplen en la población estudiada los objetivos marcados sobre caries dental por la OMS para el año 2000 (AU)

In the present study we are questioning if the three time a day tooth brushing is really essential and if it is the opposite case, what time of the day would be most important for mechanical elimination of dental plaque. For this purpose we have planed a transversal study on 214 pupils aged 6 to 12 years in the period of 2004-05 scheduled school coarse. On each of them was applied the “Partial Exam type III” which is recommended from International Dental Federation and which is considered as standard procedure on prevalence of dental caries study. We obtained following results: 1. Tooth brushing by night before sleeping is a favourable protection factor from dental caries, being correspondent odd-ratio adjusted on 0.339 (interval of confidence is 95% [0.159; 0.722]). 2. Consuming the sugar more than one time a day causes higher risk for dental caries apparition than one time a day consumption of sugar, being correspondent odd-ratio adjusted on 1.998 (interval of confidence is 95% [1.090;3.665]). 3. The children attending public schools have higher risk of dental caries presence than the children attending private schools being correspondent odd-ratio adjusted on 2.019 (interval of confidence is 95% [1.105;3.691]). 4. The objectives on dental caries marked from WHO are carried out on the studied population (AU)

Phagocytic functions of salivary neutrophils in oral mucous membrane diseases.

BACKGROUND AND PURPOSE: Phagocytic functions of salivary polymorphonuclear neutrophils (sPMNs) have not been comprehensively studied in patients with oral mucous membrane diseases, although available data suggest the role of immunity in their pathogenesis. SUBJECTS AND METHODS: Phagocytic functions of sPMN were determined in 15 patients with acute recurrent aphthous ulceration (RAU), 11 patients with oral lichen planus (OLP) and 20 healthy volunteers. In healthy subjects, the same parameters were also determined in peripheral blood polymorphonuclear neutrophils (bPMNs). Phagocytic activity (proportion of ingesting cells, PA), ingestion ability (number of ingested targets per 100 phagocytes, IA) and intracellular microbicidity (proportion of killed targets, IM) of PMNs separated from peripheral blood and the whole unstimulated saliva were determined by acridine orange method with living yeast cells as targets. RESULTS: Salivary PMNs in healthy individuals showed significant reduction in PA (33% vs. 76%; P < 0.009) and IA (0.47% vs. 2.93%; P < 0.009) and significant increase in IM (12.0% vs. 5.5%; P = 0.011) in comparison with bPMNs. In RAU patients, reduced PA (27% vs. 37%; P = 0.035) and IA (0.25% vs. 0.47%; P = 0.05) were detected, while in OLP patients enhanced IM was detected (12% vs. 19%; P = 0.033) in comparison with healthy controls. CONCLUSION: Salivary PMNs present functional features distinct from those in peripheral blood. Some phagocytic functions of sPMNs are reduced in RAU and enhanced in OLP, indicating their role in pathogenesis or reflecting clinical changes in these conditions.

Immunogenicity of dendritic cells pulsed with cartilage derived soluble alloantigens.

Investigation of allo- and autoreactivity of cartilage components is useful not only for the potential allogeneneic transplantation but also for the treatment of tissue specific degenerative autoimmune diseases. We generated dendritic cells (DCs) to study articular cartilage immunogenicity, by using human peripheral blood mononuclear cells. Dendritic cells are extremely efficient antigen-presenting cells that initiate and modulate T-cell dependant immune responses. After isolation by density centrifugation, GM-CSF and IL-4 were added to adherent mononuclear cells. After 7 days of cultivation the cells in cell culture were: HLA-DR+, CD86+, CD14-, CD80-, CD83, CD19-, CD56-, CD3-. Dendritic cells were then pulsed with a supernatant taken from primary monolayer cell cultures of human allogeneic chondrocytes. The stimulating capacity of antigen-pulsed DCs was measured in a classical proliferation assay using tritium labelled thymidine as a marker of mitosis. Proliferative responses were significantly strong but became much stronger when DCs were concomitantly incubated with soluble antigens and human recombinant TNF-alpha and used subsequently as stimulators. Although TNF-alpha positively influenced the stimulating capacity of pulsed DCs, we were unable to detect major differences in expression of specific DCs' phenotype markers assessed by flow cytometry in the absence or presence of this cytokine.

Protein synthesis of human articular chondrocytes cultured in vitro for autologous transplantation.

Chondrocytes in hyaline cartilage produce typical matrix proteins, the most abundant of them being collagen type II and aggrecan. Chondrocytes in monolayer cell culture dedifferentiate and gain fibroblastic phenotype. The cells gradually start to produce collagen type I while the production of collagen type II and aggrecan decreases. Transplantation of autologous chondrocytes cultured in vitro is used for treatment of aseptic articular cartilage lesions. For this purpose, cartilage biopsy is taken and isolated cells are subsequently proliferated in a monolayer cell culture. When implanted, the cells start to produce specific cartilaginous matrix that fills the defect. Prior to surgical procedure the cells can also be cryopreserved for longer periods of time after reaching appropriate numbers. We tested the influence of cultivation time and number of continuous culture passages as well as the influence of cryopreservation on the matrix protein synthesis of human articular chondrocytes. The ability of dedifferentiated chondrocytes to redifferentiate has been monitored by measuring matrix protein synthesis of the cells, re-seeded in agarose suspension culture. The results obtained show progressive dedifferentiation during monolayer cell culture procedures, facilitated by cryopreservation. Successful redifferentiation of cells re-seeded in suspension cultures was observed regardless of the previous level of chondrocyte dediferentiation.

Determination of optimal transport conditions for biopsies of human articular cartilage as well as for suspensions of cultured chondrocytes used in autologous transplantation.

Autologous transplantation of chondrocytes is currently being promoted as a novel approach for the treatment of deep cartilage lesions. Briefly, the method involves enzyme-mediated release of chondrocytes from cartilage biopsies, the expansion of cells by in vitro cultivation and their re-implantation into the defect. The success of this technique depends on many factors including transport conditions for both, cartilage biopsies from the operating hall to the laboratory and the return transport of final suspension of cultured chondrocytes. To determine the extent of cellular damage in biopsies, chondrocytes were enzymatically isolated following a few days of tissue preservation in different tissue culture media. The proportion of dead cells was assessed by Trypan blue staining and counting. The viability was not dependant of the type of the medium used and remained approximately 50% in all samples, even after 72 h. To develop optimal conditions for transport of final chondrocyte suspension, isolated cells were firstly grown in monolayer cultures. Cell suspensions in media with different additives were injected into special glass containers used for the transport and left at 4 degrees C or 25 degrees C for up to 3 days. During this period every 24 h the samples were taken and viability as well as apoptosis levels were assessed. Viability of cells in suspensions at 25 degrees C decreased significantly and became inadequate already after 48 h. In contrast to that, the proportion of viable cells at 4 degrees C remained above 80% even after 48 h. In the majority of the samples, culture medium containing serum and vitamin C provided the best conditions for long-term preservation of chondrocytes.

Kikuchi-Fujimoto disease. A case report.

Histologically, Kikuchi-Fujimoto disease (KFD) is a necrotising, self-limiting lymphadenitis. It typically affects the lymph nodes of the neck and it is often accompanied by a high fever. The present paper documents the case of a 28-year old male carrier of KFD. From the x-ray images, the KFD clinically appeared to be a submandibular salivary gland tumor. An ultrasound study led us to believe that it was more likely a branchial cyst. Malignant lymphoma can be mistaken for KFD, thus the diagnosis must be precise and the differentiation is crucial.

Vitamin C induced apoptosis in human articular chondrocytes.

Chondrocytes present in articular cartilage survive as a resident cell population throughout the lifespan of the individual organism. However, articular chondrocytes as other cells also undergo apoptosis and there is an ever increasing list of diverse stimuli that can induce this phenomenon in vitro. Our main interest was to investigate potential cytotoxic effects of vitamin C (L-ascorbic acid) on human articular chondrocytes. The present study suggests that vitamin C can induce apoptosis in a cell culture of chondrocytes after 18 h of cultivation. Apoptosis-inducing activity of L-ascorbic acid is dose dependent and significantly affected by the presence of serum. The increased number of vitamin C induced apoptotic cells was associated with DNA fragmentation and morphological changes of the cells.

In vitro functional tests for evaluation of stimulating capacity of cultured human dendritic cells.

Basic functional test for evaluation of in vitro cultured human dendritic cells (DC) is primary allogeneic one-way mixed lymphocyte reaction (MLR). In this way, one can evaluate stimulating capacity, which is a basic characteristic of DC. The proliferation of cells is measured through incorporation of 3H-thymidine. Normally proliferation is measured at days 5-7. We studied kinetics of proliferative responses initiated with different stimulating cell suspensions to evaluate differences and possibly reduce time needed to perform this test. Gradual increase in response from days 1 to 7 and a significant difference from controls (peripheral blood mononuclear cells) seen from day 4 was noted if macrophages were used as stimulators. A consistently higher proliferation, compared to controls, was always found already on day 2 when mature DC were used as stimulators. The reaction peaked 2 to 3 days earlier and was also more than two times more intense. This maximal and significantly higher response, consistently seen already after 48 hours, allows us to confirm the presence of mature DC in stimulating suspensions much earlier than previously.