Success factors for implementing and sustaining a mature electronic medical record in a low-resource setting: a case study of iSanté in Haiti.

Electronic health information systems, including electronic medical records (EMRs), have the potential to improve access to information and quality of care, among other things. Success factors and challenges for novel EMR implementations in low-resource settings have increasingly been studied, although less is known about maturing systems and sustainability. One systematic review identified seven categories of implementation success factors: ethical, financial, functionality, organizational, political, technical and training. This case study applies this framework to iSanté, Haiti's national EMR in use in more than 100 sites and housing records for more than 750 000 patients. The author group, consisting of representatives of different agencies within the Haitian Ministry of Health (MSPP), funding partner the Centers for Disease Control and Prevention (CDC) Haiti, and implementing partner the International Training and Education Center for Health (I-TECH), identify successes and lessons learned according to the seven identified categories, and propose an additional cross-cutting category, sustainability. Factors important for long-term implementation success of complex information systems are balancing investments in hardware and software infrastructure upkeep, user capacity and data quality control; designing and building a system within the context of the greater eHealth ecosystem with a plan for interoperability and data exchange; establishing system governance and strong leadership to support local system ownership and planning for system financing to ensure sustainability. Lessons learned from 10 years of implementation of the iSanté EMR system are relevant to sustainability of a full range of increasingly interrelated information systems (e.g. for laboratory, supply chain, pharmacy and human resources) in the health sector in low-resource settings.

Hsp90{beta} and p130(cas): novel regulatory factors of MMP-13 expression in human osteoarthritic chondrocytes.

BACKGROUND: Human osteoarthritic (OA) chondrocytes were previously classified into L (low)- and H (high)-OA according to matrix metalloproteinase-13 (MMP-13) basal levels and interleukin 1beta (IL1beta) inducibility. In H-OA chondrocytes, the regulatory proteins p130(cas) and nuclear matrix protein 4 (NMP4) acting on the MMP-13 promoter were identified. OBJECTIVE: To identify regulators of MMP-13 expression/production in human L-OA chondrocytes, to determine their effect on the expression of other MMPs and the effect of IL1beta on these molecules. METHODS: The identification of the L-OA chondrocyte proteins interacting specifically with the AGRE site of the MMP-13 promoter was performed by mass spectrometry. Heat shock protein 90beta (Hsp90beta), p130(cas) and NMP4 small interfering RNAs (siRNAs) were transfected into L-OA chondrocytes and incubated with or without IL1beta. Gene expression was determined by real-time PCR, MMP-1 and MMP-13 production by ELISA, and signalling pathway activation by western blotting and ELISA. RESULTS: Hsp90beta was identified as a protein of the L-OA/AGRE-specific complex. Silencing p130(cas) and Hsp90beta significantly increased MMP-13 expression (about four- and twofold, respectively) and production. sip130(cas) affected to a lesser extent MMP-1 expression (twofold) and production. siNMP4 showed no effect. Expression of MMP-2, -3, -9 and -14 was unaffected. Silencing both Hsp90beta and p130(cas) had a significant additive effect on MMP-13, but not on MMP-1 expression, the level of which was similar to that with sip130(cas) alone. IL1beta decreased p130(cas) and Hsp90beta expression/production, indicating another pathway by which this cytokine upregulates MMP expression. The IL1beta-triggered signalling pathways responsible for MMP upregulation were unaffected in the silenced cells. CONCLUSION: This study illustrates the complex regulation of MMP-13 by showing the inhibitory effect of the two cytoplasmic molecules, p130(cas) and Hsp90beta, in L-OA chondrocytes.

[Interest of peri-operative immunonutrition in head and neck cancers]. / Intérêt de l'immunonutrition péri-opératoire en carcinologie cervico-faciale.

OBJECTIVE: Authors reported the results of a study on the application of immunonutrion in peri-operative (pre and postoperative) in head and neck cancer for all patients malnourished or not. In preoperative we used an oral treatmentand in postoperative an enteral one. MATERIALS AND METHODS: Prospective study concerning 78 patients (47 malnourished versus 31 not) having had heavy head and neck curative cancerology surgery. The mean follow up was of 10 months (from 7 to 16 month). They peri-operative immuno-enriched diet consisted, in pre-operative of 1000 kcal/j during 7 days of oral immunonutrition (Impact), and in post-operative, 1500 kcal/j during 10 days of enteral immunonutition (Crucial). The nutritional state was evaluated in pre-operative by simple clinical and biological parameters (size, weight, CMI "Corporal Mass Index", albumin, NRI "Nutritional Risk Index"), and in post-operative by the evolution of the weight and the CMI. The palatability of the product used in pre-operative and the patients' compliance to the treatment are studied using the satisfaction's multiple choice question paper. RESULTS: The study showed an improvement of the patients' nutritional and general state (regain appetite, less marked asthenia) and of the quality of life. The product used in preoperative was well tolerated, this oral supplementation led to the same beneficial effects of the enteral's. At eight days in preoperative, the average weight was 62.35 kg, the average CMI was 20.93, and the average NRI was 94.12. In post-operative the patients' nutritional state improved: at eight days, the average loss of weight was 2.82 kg, the average CMI was 22.2. At one and six months after respectively the average gain of weight was 2.17 kg and 6.11 kg, the average CMI was 23.71 and 25.16. The application of this protocol decreased the post-operative complications (13% reduction of the infectious complications and 6% diminution of the fistulas). The time of hospitalization is then reduced (1.7 days), and the life's longevity is improved. CONCLUSION: The results produced by this study, demonstrate the necessity to apply a peri-operative immuno-enriched diet systematically for all the patients with and without a degraded nutritional state, undergoing a heavy head and neck curative cancerology surgery.

Histone deacetylase inhibitors suppress interleukin-1beta-induced nitric oxide and prostaglandin E2 production in human chondrocytes.

OBJECTIVE: Overproduction of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) plays an important role in the pathogenesis of osteoarthritis (OA). In the present study, we determined the effect of trichostatin A (TSA) and butyric acid (BA), two histone deacetylase (HDAC) inhibitors, on NO and PGE(2) synthesis, inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 expression, and nuclear factor (NF)-kappaB DNA-binding activity, in interleukin-1beta (IL-1)-stimulated human OA chondrocytes, and on IL-1-induced proteoglycan degradation in cartilage explants. METHODS: Chondrocytes were stimulated with IL-1 in the absence or presence of increasing concentrations of TSA or BA. The production of NO and PGE(2) was evaluated using Griess reagent and an enzyme immunoassay, respectively. The expression of iNOS and COX-2 proteins and mRNAs was evaluated using Western blotting and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. Proteoglycan degradation was measured with dimethymethylene blue assay. Electrophoretic mobility shift assay (EMSA) was utilized to analyze the DNA-binding activity of NF-kappaB. RESULTS: HDAC inhibition with TSA or BA resulted in a dose-dependent inhibition of IL-1-induced NO and PGE(2) production. IL-17- and tumor necrosis factor-alpha (TNF-alpha)-induced NO and PGE(2) production was also inhibited by TSA and BA. This inhibition correlated with the suppression of iNOS and COX-2 protein and mRNA expression. TSA and BA also prevented IL-1-induced proteoglycan release from cartilage explants. Finally, we demonstrate that the DNA-binding activity of NF-kappaB, was induced by IL-1, but was not affected by treatment with HDAC inhibitors. CONCLUSIONS: These data indicate that HDAC inhibitors suppressed IL-1-induced NO and PGE(2) synthesis, iNOS and COX-2 expression, as well as proteoglycan degradation. The suppressive effect of HDAC inhibitors is not due to impaired DNA-binding activity of NF-kappaB. These findings also suggest that HDAC inhibitors may be of potential therapeutic value in the treatment of OA.

Intracellular localisation of galectin-3 has a protective role in chondrocyte survival.

OBJECTIVE: Although galectin-3 (gal-3) is expressed during arthritic disorders, the part it plays has never been described. The aim of the study was to determine the intracellular roles of gal-3 in chondrocytes and cartilage. METHODS: Following treatment with sodium nitroprusside, a cell death inducer, intracellular levels of total and phosphorylated gal-3 were measured by immunoblots in human osteoarthritic (OA) chondrocytes. Cell viability was also assessed by the lactate dehydrogenase activity in conditioned media from OA chondrocytes or from ATDC5 cells transfected with a gal-3-expressing vector. After generating an OA model by intra-articular injection of 0.5% mono-iodoacetate (MIA), histological evaluation of articular cartilage and subchondral bone was performed in wild-type (WT) and gal-3 knockout (KO) mice aged 6 weeks and 4 months. RESULTS: In vitro experiments demonstrated that intracellular gal-3 had a protective role in chondrocyte survival, which involved its phosphorylation. In contrast to 6-week-old mice, 4-month-old gal-3 KO mice, compared with WT mice, presented OA-like cartilage modifications. OA induction via MIA injection in WT mice generated cartilage lesions similar to those found in gal-3 KO animals. Moreover, OA induction showed a significant decrease in subchondral bone surface in the gal-3 KO mice in contrast to the WT group. CONCLUSIONS: Altogether these findings indicate that intracellular gal-3 has a beneficial effect in articular cells, as its absence in KO mice led to cartilage lesions.

Randomised controlled trial comparing two methods of acetabular cup positioning during total hip arthroplasty.

BACKGROUND: Acetabular cup positioning is an important technical aspect in total hip arthroplasty. Most surgeons estimate cup abduction angle during surgery with the insertion rod position according to the patient's body anatomical landmarks or other reference points in the operating room. High acetabular component abduction angle is associated with an increased risk of dislocation, premature polyethylene wear and osteolysis. METHOD: To evaluate the potential benefits of a new technique for vertical acetabular cup positioning, 100 acetabular cups were randomised to be inserted with or without an inclinometer. Abduction angles were measured on postoperative radiographs by 2 evaluators blind to the treatment group. RESULTS: Of the cups, 57% (27/47) were positioned within the desirable abduction angle range of 40-49 with the inclinometer, compared with 50% (27/53) by visuospatial perception (p=0.454). The proportion of cups positioned outside a safe angle range of 30-55 was low in both groups: 6% (3/47) for the inclinometer group versus 4% (2/53) for the visuospatial perception group (p=0.536). CONCLUSION: The use of an inclinometer did not significantly improve the acetabular cup abduction angle obtained by our group of surgeons when compared with visuospatial perception. Newer techniques such as navigation may be useful in further optimising cup positioning and reducing the outliers.

Understanding the reasons for decline of HIV prevalence in Haiti.

OBJECTIVES: HIV sero-surveillance rounds and projection estimates suggest a decline of HIV prevalence among pregnant women and the general population in Haiti. This study aimed to evaluate the decline of HIV prevalence and understand the reasons for the decline. METHODS: Following an epidemiological analysis, three mathematical models were used to re-create the national epidemic, calculate HIV incidence, and confirm the decline of HIV prevalence. Declining trends in prevalence data were compared with observed trends in behavioural data. RESULTS: HIV progressed rapidly from initial infection to AIDS and death, with people dying twice as fast as in developed countries. With the rapid progression of the disease and the early intervention efforts in securing the blood supply, prevalence among commercial sex workers and blood donors peaked in the late 1980s followed by a decline in the mid-1990s in the general population. The observed decline among pregnant women and in the general population was confirmed after controlling for confounding variables. The Haitians are well informed: there is an increase in condom use with occasional partners at last contact and in abstinence and fidelity, and a decrease in the number of occasional partners. However, the age of sexual debut is lower and the proportion of sexually active youth has increased. CONCLUSIONS: There is evidence of decline in HIV prevalence among pregnant women, specifically among pregnant women living in urban areas and pregnant women 25 years and older, but not among pregnant women living in rural areas and pregnant women 24 years and younger. Although many factors have acted in synergy to halt the AIDS epidemic in Haiti, the main reasons for decline seem to point to mortality and blood safety intervention efforts in the early stages of the epidemic.

Differential regulation of the bone morphogenic protein antagonist chordin in human normal and osteoarthritic chondrocytes.

OBJECTIVE: To investigate the distribution of the bone morphogenic protein (BMP) antagonist chordin in normal and osteoarthritic cartilage and synovial membranes, and its regulation in chondrocytes and synovial fibroblasts by inflammatory and growth factors. METHODS: Localisation of chordin in tissues was undertaken by immunohistochemistry and gene regulation was determined by real time polymerase chain reaction. RESULTS: In normal cartilage, chordin was found at low levels (mean (SD), 7.6 (1.3)%), mainly in the very superficial layers. In osteoarthritis, chordin was also found in the superficial layers (8.9 (1.1)%), though at a significantly higher level (24.7 (1.5)%) in the last two thirds of the cartilage. In contrast to normal cells, chordin mRNA and protein levels were significantly downregulated (p<0.01) in osteoarthritic chondrocytes by all the growth factors tested. Interferon gamma stimulated chordin expression in normal but not in osteoarthritic chondrocytes (p<0.0002), while interleukin 1 beta and tumour necrosis factor alpha did not affect the expression level. However, no difference was found in either the distribution or regulation of chordin in normal and osteoarthritic synovial membranes or synovial fibroblasts. CONCLUSIONS: The differential distribution and regulation of chordin in normal and osteoarthritic cartilage and chondrocytes suggests an involvement of this antagonist in the osteoarthritic process.

Interjoint coordination in lower limbs in patients with a rupture of the anterior cruciate ligament of the knee joint.

Previous studies of movement kinematics in patients with a ruptured anterior cruciate ligament (ACL) have focused on changes in angular displacement in a single joint, usually flexion/extension of the knee. In the present study, we investigated the effect of an ACL injury on the overall limb interjoint coordination. We asked healthy and chronic ACL-deficient male subjects to perform eight types of movements: forward squats, backward squats, sideways squats, squats on one leg, going up a step, going down a step, walking three steps, and stepping in place. Depending on the movement concerned, we applied principal component (PC) analysis to 3 or 4 degrees of freedom (DFs): thigh flexion/extension, knee flexion/extension, ankle flexion/extension, thigh abduction/adduction. The first three DFs were investigated in all movements. PC analysis identifies linear combinations of DFs. Movements with a fixed ratio between DFs are thus described by only one PC or synergy. PCs were computed for the entire movement as well as for the period of time when the foot was in contact with the ground. For both the control and the injured groups, two synergies (PC vectors) usually accounted for more than 95% of the DFs' angular excursions. It was possible to describe 95-99% of some movements using only one synergy. Compared to control subjects, injured subjects employed different synergies for going up a step, walking three steps, squatting sideways, and squatting forward, both in the injured and uninjured legs. Those movements may thus be more indicative of injury than other movements. Although ACL-deficiency did not increase asymmetry (angle between the PCs of the same movement performed on the right and the left sides), this result is not conclusive because of the comparatively low number of subjects who participated in the study. However, the finding that synergies in both legs of patients were different from those in control subjects for going up a step and walking three steps suggests that interjoint coordination was affected for both legs, so that the asymmetry index might have been preserved despite the injury. There was also a relationship between the asymmetry index for squatting on one leg, squatting forward, walking three steps and some of the outcomes of the knee injury and osteoarthritis outcome score (pain, symptoms, activities of daily living, sport and recreation function, and knee-related quality of life). This suggests that significant differences in the asymmetry index could be obtained if more severely-injured patients participated in this study. It is possible that subjects compensated for their mechanical deficiencies by modifying muscle activation patterns. Synergies were not only modified in injured subjects, but also rearranged: the percentage of movement explained by the first PC was different for the injured and/or uninjured legs of patients, as compared to the legs of the control group, for going up a step, going down a step, walking three steps, and squatting forward. We concluded that the analysis of interjoint coordination may be efficient in characterizing motor deficits in people with knee injuries.

Computer based method for the three-dimensional kinematic analysis of combined posterior cruciate ligament and postero-lateral complex reconstructions on cadaver knees.

The aim of this study is to evaluate the effect of combined posterior cruciate ligament (PCL) and postero-lateral corner (PLC) reconstruction on laxity and three-dimensional kinematics of cadaver knees. We performed anatomical double bundle PCL reconstruction, and functional one bundle 'over-the-bottom' PCL reconstruction combined with one type of PLC reconstruction, running from the postero-lateral tibia to an isometric point near the lateral epicondyle of the femur. Our results showed that combined reconstruction was necessary to restore rotatory laxity. PLC reconstruction, according to the technique described, invariably created a shift towards internal rotation of the kinematic curves, compared to the intact knee.

Loss-of-function and residual channel activity of connexin26 mutations associated with non-syndromic deafness.

Connexins are the protein subunits of gap junction channels that allow a direct signaling pathway between networks of cells. The specific role of connexin channels in the homeostasis of different organs has been validated by the association of mutations in several human connexins with a variety of genetic diseases. Several connexins are present in the mammalian cochlea and at least four of them have been proposed as genes causing sensorineural hearing loss. We have started our functional analysis by selecting nine mutations in Cx26 that are associated with non-syndromic recessive deafness (DFNB1). We have observed that both human Cx26 wild-type (HCx26wt) and the F83L polymorphism, found in unaffected controls, generated electrical conductance between paired Xenopus oocytes, which was several orders of magnitude greater than that measured in water-injected controls. In contrast, most recessive Cx26 mutations (identified in DFNB1 patients) resulted in a simple loss of channel activity. In addition, the V37I mutation, originally identified as a polymorphism in heterozygous unaffected individuals, was devoid of function and thus may be pathologically significant. Unexpectedly, we have found that the recessive mutation V84L retained functional activity in both paired Xenopus oocytes and transfected HeLa cells. Furthermore, both the magnitude of macroscopic junctional conductance and its voltage-gating properties were indistinguishable from those of HCx26wt. The identification of functional differences of disease causing mutations may lead to define which permeation or gating properties of Cx26 are necessary for normal auditory function in humans and will be instrumental in identifying the molecular steps leading to DFNB1.

Comparison of two methods for reconstruction of the posterior cruciate ligament using a computer based method: quantitative evaluation of laxity, three-dimensional kinematics and ligament deformation measurement in cadaver knees.

The aim of this paper is to present a biomechanical comparison of two different methods for reconstruction of the posterior cruciate ligament in cadaver knees. We used an original computer-based method allowing precise calculation of three-dimensional (3D) knee kinematic parameters as well as the estimation of combined graft deformation (elongation-flexion-torsion). After isolated posterior cruciate ligament (PCL) dissection, double bundle and 'over-the-bottom' methods were performed successively on each knee using synthetic polyester ligaments. The effect of pre-tensioning was tested with the 'over-the-bottom' method. antero-posterior (A-P) and rotational laxity as well as 3D kinematics were recorded and analysed. Our computer based method allowed us to show that both reconstruction methods were equivalent in restoring A-P and rotational laxity as well as kinematic curves. Combined deformation of the prostheses was equivalent for both ligaments.

Computer-based method for the 3-D kinematic analysis of posterior cruciate ligament and postero-lateral corner lesions.

Posterior cruciate ligament (PCL) rupture, whether or not combined with postero-lateral corner (PLC) tears, are more often diagnosed today thanks to improved imaging techniques. However, due to the lack of reliable instrumentation to quantitatively evaluate the knee, much is still unknown about the function of these ligamentous structures. The aim of this paper is to present results on the effect of progressive resection of the PCL and PLC on knee laxity and 3-D knee kinematics. The results show that 3-D movement analysis is important and complements laxity measurements by helping to interpret the complex alteration of knee function.

Can altered production of interleukin-1beta, interleukin-6, transforming growth factor-beta and prostaglandin E(2) by isolated human subchondral osteoblasts identify two subgroups of osteoarthritic patients.

OBJECTIVE: To determine the capacity of human subchondral osteoarthritic osteoblasts (Ob) to produce interleukin (IL)-1beta, IL-6, transforming growth factor-beta (TGF-beta) and prostaglandin E(2) (PGE(2)), and determine if a relationship exists between IL-1beta, TGF-beta, PGE(2) and IL-6 production. METHODS: We measured the abundance of IL-1beta, IL-6, TGF-beta and PGE(2) using very sensitive ELISA in conditioned-media of human primary subchondral Ob from normal individuals and osteoarthritic patients. Selective inhibition of IL-6 or IL-6 receptor signaling was performed to determine its effect on PGE(2) production whereas the inhibiton of PGE(2) production was performed to determine its effect on IL-6 production. The expression of bone cell markers and urokinase plasminogen activator (uPA) activity was also determined. RESULTS: Osteoarthritic Ob produced all these factors with greater variability than normal cells. Interestingly, the production of IL-6 and PGE(2) by osteoarthritic Ob separated patients into two subgroups, those whose Ob produced levels comparable to normal (low producers) and those whose Ob produced higher levels (high producers). In those cells classified as high osteoarthritic Ob, PGE(2) and IL-6 levels were increased two- to three-fold and five- to six-fold, respectively, compared with normal. In contrast, while using their IL-6 and PGE(2) production to separate osteoarthritic Ob into low and high producers, we found that IL-1beta levels were similar in normal and all osteoarthritic Ob. Using the same criteria, TGF-beta levels were increased in all osteoarthritic Ob compared with normal. Reducing PGE(2) synthesis by Indomethacin [a cyclo-oxygenase (COX) -1 and -2 inhibitor] reduced IL-6 levels in all osteoarthritic Ob, whereas Naproxen (a more selective COX-2 inhbitor) reduced PGE(2) and IL-6 levels only in the high osteoarthritic group. Conversely, PGE(2) addition to osteoarthritic Ob enhanced IL-6 production in both groups. Moreover, the addition of parathyroid hormone also stimulated IL-6 production to similar normal levels in both osteoarthritic groups. In contrast, using an antibody against IL-6 or IL-6 receptors did not reduce PGE(2) levels in either group. The evaluation of alkaline phosphatase activity, osteocalcin release, collagen type I and uPA activity in osteoarthritic Ob failed to show any differences between these cells regardless to which subgroup they were assigned. CONCLUSIONS: These results indicate that IL-6 and PGE(2) production by subchondral Ob can discriminate two subgroups of osteoarthritic patients that cannot otherwise be separated by their expression of cell markers, and that endogenous PGE(2) levels influence IL-6 synthesis in osteoarthritic Ob.

A new generation of artificial ligaments in reconstruction of the anterior cruciate ligament. Two-year follow-up of a randomised trial.

We have undertaken a randomised clinical trial comparing two methods of reconstruction of the anterior cruciate ligament in patients with chronic instability. We used an ipsilateral bone-patellar-tendon-bone autograft in 27 patients and the Ligament Advancement Reinforcement System (LARS) artificial ligament in 26. Assessment before and at two, six, 12 and 24 months after surgery, included the history, physical examination, a modified International Knee Documentation Committee (IKDC) score, the Tegner score, the Knee Injury and Osteoarthritis Outcome Score (KOOS) and instrumented laxity testing. There were no cases of reactive synovitis or of infection of the knee, and there was no difference regarding the failure rate between the two groups. The IKDC showed no significant differences between the two groups at any stage of the follow-up. The KOOS evaluation showed consistently better results in all subscales for the LARS group during the first year of follow-up. After 24 months these differences were no longer evident. Instrument-tested laxity was greater in the LARS group at all stages of follow-up, but the differences were not significant at 24 months. Our findings suggest that at follow-up at 24 months the LARS ligament seems to be a satisfactory treatment option, especially when an early return to high levels of activity is demanded.

[Standards, options and recommendations: Good clinical practice in the dietetic management of cancer patients: hospital catering]. / Standards, options et recommandations: Bonnes practiques de diététique en cancérologie: la prestation alimentaire.

CONTEXT: The "Standards, Options and Recommendations" (SOR) project, started in 1993, involves a collaboration between the Federation of the French Cancer Centres (FNCLCC), the 20 French Regional Cancer Centres, some French public university and general hospitals and private Clinics and medical scientific societies. Its main objective is the development of clinical practice guidelines to improve the quality of health care and outcome for cancer patients. The methodology is based on a literature review followed by a critical appraisal by a multidisciplinary group of experts to produce the draft guidelines which are then validated by specialists in cancer care delivery. OBJECTIVES: To develop clinical practice guidelines for hospital catering for cancer patient using the methodology developed by the Standards, Options and Recommendations project. METHODS: Data were identified by a literature search of Medline and the reference lists of experts in the groups. After the guidelines were drafted, they were validated by independent reviewers. RESULTS: The main recommendations are: 1) While taking into consideration the specific needs of cancer patients, the dietician is responsible for the hygiene, the sanitary quality of alimentation, the equilibrium and nutritional quality of the hospital catering. 2) Ordering and distribution of meals, and clearing up afterwards contribute to the quality of hospital catering and the personnel who do this should have time and be willing to listen to the patients. 3) The ordering of meals should be adapted to individual patient's requirements and must take into account the patient's medication. 4) The method of transporting the food chosen by the institution (cold or warm method) should be respected. The personnel responsible should receive regular and specific training to use the method correctly. 5) The intake of patients with nutritional follow-up should be reliably and reproducibly evaluated by the personnel after every meal. 6) Patient satisfaction should be assessed once a year and the results of this assessment used to improve the quality of hospital catering. 7) The dietician is the interface between the accounts department, the medical wards, the hospital catering department and the patients.

Transforming growth factor-beta induced collagenase-3 production in human osteoarthritic chondrocytes is triggered by Smad proteins: cooperation between activator protein-1 and PEA-3 binding sites.

OBJECTIVE: To examine the signaling pathways leading to transforming growth factor-beta (TGF-beta) induced collagenase-3 production in human osteoarthritic (OA) chondrocytes, as well as the transcription factors and their binding sites involved in the transcriptional control of collagenase-3 gene. METHODS: Identification of the TGF-beta signaling pathway was by Western immunoblotting using specific antibodies for the phosphorylated forms of p44/42 and p38 MAPK, SAPK/JNK, and the Smad2 protein. Electromobility shift assays (EMSA) were carried out for activator protein- (AP-1), polyomavirus enhancer A (PEA-3), activin-response-element-like, Smad-binding-element-like, and TGF-beta inhibitory element oligonucleotides. Supershift assays using antibodies to the Jun, Fos, and Smad families of proteins were used for identification of transcription factors. Chondrocyte transfections were also performed using the -133CAT collagenase-3 promoter plasmid (containing PEA-3, AP-1, and TATA sites) and mutated AP-1 and PEA-3 sites. RESULTS: The primary target of TGF-beta induced collagenase-3 in OA chondrocytes was the Smad2 protein, with significant phosphorylation within 5 min. Contrasting with the Smad2, the untreated OA chondrocytes already had detectable levels of the phosphorylated forms of p38 and p44/42 MAPK. Of the oligonucleotides tested, EMSA revealed that TGF-beta treated OA chondrocyte proteins bound only to the AP-1 and PEA-3. Supershifts with the AP-1 oligonucleotide showed the presence of the Jun (c-Jun, JunB, JunD) and Fos (c-Fos, FosB, Fra-1, Fra-2) proteins in the untreated and TGF-beta treated OA chondrocytes, whereas only Smad proteins (Smad2, 3, 4) were present in the AP-1 binding proteins from the TGF-beta treated chondrocytes. The AP-1 mutation decreased both basal (95%) and TGF-beta induced (99%) collagenase-3 production, whereas the PEA-3 mutation decreased the basal (15%) but more significantly (50%) the TGF-beta induced transcription. CONCLUSION: Smad proteins are the main cytoplasmic signaling pathways in TGF-beta stimulated collagenase-3 in OA chondrocytes. The AP-1 site appears critical for upregulation of collagenase-3 production, but TGF-beta stimulation requires both AP-1 and PEA-3 sites for optimal response.

Antiplatelet and antithrombotic activity of SL65.0472, a mixed 5-HT1B/5-HT2A receptor antagonist.

The antiplatelet and antithrombotic activity of SL65.0472 (7-fluoro-2-oxo-4-[2-[4-(thieno [3,2-c]pyrin-4-yl) piperazin-1-yl]ethyl]-1,2-di-hydroquinoline-acetamide), a mixed 5-HT1B/5-HT2A receptor antagonist was investigated on 5HT-induced human platelet activation in vitro, and in rat, rabbit and canine platelet dependent thrombosis models. SL65.0472 inhibited 5-HT-induced platelet shape change in the presence of EDTA (IC50 values = 35, 69 and 225 nM in rabbit, rat and human platelet rich plasma (PRP)), and also inhibited aggregation induced in human PRP by 3-5 microM 5-HT + threshold concentrations of ADP (0.5-1 microM) or collagen (0.3 microg/ml) with mean IC50 values of 49 +/- 13 and 48 +/- 6 nM respectively. SL65.0472 inhibited thrombus formation when given both intravenously 5 min and orally 2 h prior to assembly of an arterio-venous (A-V) shunt in rats as from 0.1 and 0.3 mg/kg respectively. It was active in a rabbit A-V shunt model with significant decreases in thrombus weight as from 0.1 mg/kg i. v. and at 10 mg/kg p.o. The delay to occlusion in an electric current-induced rabbit femoral artery thrombosis model was increased by 251% (p <0.05) after 20 mg/kg p.o. SL65.0472 (30 microg/kg i.v.) virtually abolished coronary cyclic flow variations (7.2 +/- 1.0/h to 0.6 +/- 0.6/h, p <0.05) and increased minimum coronary blood flow (1.2 +/- 0.8 ml/min to 31.8 +/- 8.4 ml/min, p <0.05) in a coronary artery thrombosis model in the anaesthetised dog. Finally, SL65.0472 significantly increased the amount of blood lost after rat tail transection at 3 mg/kg p.o. Thus the anti-5-HT2A component of SL65.0472 is reflected by its ability to inhibit 5-HT-induced platelet activation, and platelet-rich thrombus formation.

Correlation between patients' satisfaction and objective measurement of knee stability after ACL reconstruction using a patellar tendon autograft.

We evaluated the relationship between patients' satisfaction and objective measurements of knee stability after reconstruction of the ACL using a patellar tendon autograft. An examination of 59 patients 2-7 years after surgery was carried out. Assessment was made by the Knee and Osteoarthritis Outcome Score for patient satisfaction, a modified International Knee Documentation Committee form for clinical knee stability and a Telos stress radiography for PA stability. The results show that patients' satisfaction was much greater than the objective evaluation would suggest. We conclude that documenting mechanical knee stability alone is inadequate for follow-up studies and a questionnaire assessing patient satisfaction should be added.

Functional analysis of a dominant mutation of human connexin26 associated with nonsyndromic deafness.

Cx26 has been implicated in dominant (DFNA3) and recessive (DFNB1) forms of nonsyndromic sensorineural deafness. While most homozygous DFNB1 Cx26 mutations result in a simple loss of channel activity, it is less clear how heterozygous mutations in Cx26 linked to DFNA3 cause hearing loss. We have tested the ability of one dominant mutation (W44C) to interfere with wild-type human Cx26 (HCx26wt). HCx26wt induced robust electrical conductance between paired oocytes, and facilitated dye transfer between transfected HeLa cells. In contrast, oocyte pairs injected with only W44C were not electrically coupled above background levels, and W44C failed to dye couple transfected HeLa cells. Moreover, W44C dramatically inhibited intercellular conductance of HCx26wt when co-expressed in an equal ratio, and the low levels of residual conductance displayed altered gating properties. A nonfunctional recessive mutation (W77R) did not inhibit the ability of HCx26wt to form functional channels when co-injected in the same oocyte pairs, nor did it alter HCx26wt gating. These results provide evidence for a functional dominant negative effect of the W44C mutant on HCx26wt and explain how heterozygous Cx26 mutations could contribute to autosomal dominant deafness, by resulting in a net loss, and/or alteration, of Cx26 function.