Syrian hamster infected with Leishmania infantum: A new experimental model for inflammatory myopathies

Idiopathic inflammatory myopathies (IIMs) are inflammatory disorders of unknown origin. On the basis of clinical, histopathological, and immunological features, they can be differentiated into three major and distinct subsets: dermatomyositis; polymyositis; and inclusion‐body myositis. Although a few animal models for IIM are currently available, they lack several characteristic aspects of IIMs. The aim of our study was to examine skeletal muscle involvement in an experimental animal model of visceral leishmaniasis, a disseminated infection caused by the protozoan parasite Leishmania infantum, and to compare features of associated inflammation with those of human IIM. Syrian hamsters infected intraperitoneally with amastigotes of L. infantum were killed at 3 or 4 months post‐infection, and the skeletal muscles were studied. Focal inflammation was predominantly observed in the endomysium and, to a lesser extent, in perivascular areas. Degenerating muscle fibers were also found, as well as myonecrosis. Immunofluorescence with confocal laser scanning microscopy was used to characterize the phenotype of inflammatory infiltrates and the distribution of MHC class I and II in muscle biopsies. The infiltrating inflammatory cells consisted mainly of T cells, and CD8⁺ T cells were found in non‐necrotic muscle fibers that expressed MHC class I on the sarcolemma. In addition to T cells, several macrophages were present. The model we are proposing closely resembles polymyositis and may be useful in studying certain aspects of this disease such as the role of T cells in muscle inflammation and myocytotoxicity, while also providing novel therapeutic targets. Muscle Nerve, 2009     

Polymorphisms of the XRCC3 C722T and the RAD51 G135C genes and the risk of head and neck cancer in a Polish population

Genetic variations in DNA repair genes may affect an individual's susceptibility to head and neck cancer. We performed a case–control study to test the association between head and neck cancer risk and two polymorphisms: the C722 T of the XRCC3 and the G135C of the RAD51 —genes of DNA double strand break (DSB) repair by homologous recombination (HRR). Genotypes were determined by PCR-restriction fragment lenght polymorphism (PCR-RFLP). DNA was isolated from peripheral blood lymphocytes of a group of 288 patients consisting of 97 subjects with precancerous hyperplastic laryngeal lesions (PHLL) and 191 subjects with head and neck squamous cell carcinoma (HNSCC) as well as 353 healthy control donors. We found an association between PHLL and the 722CT (OR 6.67; 95% CI 3.02–14.74) as well as 722 TT (OR 4.65; 95% CI 2.30–9.43) variants of the XRCC3 gene. Similar relation was observed between these genotypes and HNSCC (OR 2.59; 95% CI 1.61–4.16 and OR 5.54; 95% CI 3.22–9.52, respectively). Moreover, we also observed an association between PHLL (OR 6.04; 95% CI 3.69–9.90) and HNSCC (OR 6.04; 95% CI 3.69–9.90) and the135GC variant of the RAD51 gene. The gene–gene interaction between XRCC3 and RAD51 polymorphic variants may contribute to higher prevalence of PHLL. The increased risk of this disease was observed in case of the combination of the 722CT/135GC (OR 3.81; 95% CI 1.55–9.75) as well as the 722 TT/135GC genotypes (OR 5.33; 95% CI 1.96–14.47). The presence of the same genes combinations plays a part in higher probability of HNSCC occurrence (OR 2.42; 95% CI 1.22–4.79 for 722CT/135GC and OR 3.63; 95% CI 1.69–7.76 for 722 TT/135GC). We also found an association between these XRCC3 or RAD51 polymorphic variants and smoking status in PHLL (ORs 2.85–10.28 and 1.82–7.35, respectively) and HNSCC patients (ORs 2.94–13.93 and 1.36–3.94, respectively) as well as alcohol intake among PHLL (ORs 3.44–6.12 and 3.52–8.43, respectively) and HNSCC subjects (ORs 2.71–7.01 and 2.33–4.62, respectively). In conclusion our data showed that the C722 T and the G135C polymorphisms of the XRCC3 and the RAD51 genes might be associated with HNSCC. Finally we suggested that these polymorphisms might be used as predictive factor of precancerous lesion for head and neck cancer in a Polish population.     

Recombinant factor VIIa in the management of postpartum bleeds: an audit of clinical use

BACKGROUND: This paper presents an analysis of 25 patient cases in which recombinant factor VIIa was used in the management of postpartum hemorrhage, including severe and/or life-threatening bleeds. Anecdotal experiences in the empirical use of this agent are described and dosing regimens, effects on bleeding, and safety data are presented. METHODS: Data were extracted from the international, internet-based, voluntary registry, haemostasis.com. Search results were manually cross-checked against monthly summary reports and cases of confirmed postpartum hemorrhage were analyzed by the authors. Case providers were contacted individually to approve the use of their cases, supply any missing data, and validate the data already held. RESULTS: Of 43 reported gynecological admissions for hemorrhage, 13 were excluded as they did not relate to childbirth, and 5 due to insufficient data. The remaining 25 records, all associated with postpartum hemorrhage, were submitted by 14 doctors from 5 countries. Following administration of recombinant factor VIIa, bleeding stopped in 18 cases (72%), markedly decreased in 2 (8%), and decreased in 4 (16%). Bleeding increased following recombinant factor VIIa administration in only 1 patient (4%). Requirements for replacement blood products and crystalloids/colloids were also greatly curtailed. A full recovery was achieved by most patients (22/25, 88%) with few complications, even when recombinant factor VIIa was administered as salvage therapy. There were no thrombotic complications associated with recombinant factor VIIa administration. CONCLUSIONS: This review provides the largest aggregate of cases in which recombinant factor VIIa has been used to control obstetrical bleeding. A review of these cases suggests that this agent may be a useful and safe adjunctive therapy in the management of postpartum hemorrhage.     

How do general practitioners recognize the definition of multimorbidity? A European qualitative study

Background: Multimorbidity is a challenging concept for general practice. An EGPRN working group has published a comprehensive definition of the concept of multimorbidity. As multimorbidity could be a way to explore complexity in general practice, it was of importance to explore whether European general practitioners (GPs) recognize this concept and whether they would change it.

Objectives: To investigate whether European GPs recognize the EGPRN concept of multimorbidity and whether they would change it.

Methods: Focus group meetings and semi-structured interviews as data collection techniques with a purposive sample of practicing GPs from every country. Data collection continued until saturation was reached in every country. The analysis was undertaken using a grounded theory based method. In each national team, four independent researchers, working blind and pooling data, carried out the analysis. To ensure the internationalization of the data, an international team of 10 researchers pooled the axial and selective coding of all national teams to check the concept and highlight emerging themes.

Results: The maximal variation and saturation of the sample were reached in all countries with 211 selected GPs. The EGPRN definition was recognized in all countries. Two additional ideas emerged, the use of Wonca’s core competencies of general practice, and the dynamics of the doctor–patient relationship for detecting and managing multimorbidity and patient’s complexity.

Conclusion: European GPs recognized and enhanced the EGPRN concept of multimorbidity. These results open new perspectives regarding the management of complexity using the concept of multimorbidity in general practice.

Key Messages

• European general practitioners recognize the EGPRN enhanced, comprehensive concept of multimorbidity.

• They add the use of Wonca’s core competencies and the patient–doctor relationship dynamics for detecting and managing multimorbidity.

• The EGPRN concept of multimorbidity leads to new perspectives for the management of complexity.

     

Pretreatment serum levels of bFGF and VEGF and its clinical significance in endometrial carcinoma

ObjectiveIn this study, we examined the frequency of serum elevation as well as the prognostic significance of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in endometrial cancer (EC) type I and a biologically aggressive variant of EC type II.Materials and methodsPretreatment serum levels of bFGF and VEGF were evaluated by commercially available enzyme-linked immunosorbent assay (ELISA) for cancer patient samples with type I EC (n = 70) and type II EC (n = 64) and compared to a cohort of normal individuals (n = 64). Values were correlated with clinicopathological characteristics and outcome.ResultsMedian pretreatment VEGF values were 470.4 pg/ml (range, 164.3–598.4 pg/ml) for type I EC, 608.8 pg/ml (range, 354.2–783.6 pg/ml) for type II of EC patients and 215.6 pg/ml (range, 128.3–332.9 pg/ml) for normal healthy subjects (p < 0.001). Elevated serum VEGF concentration correlated significantly with advanced FIGO stage in type II EC (p = 0.011). Median values of bFGF were 10.7 pg/ml (range, 0.5–22.5 pg/ml) for type I EC, 21.2 (range, 0.5–62.4 pg/ml) for type II EC and 1.1 (range, 0–7.2 pg/ml) in controls (p < 0.0001). The pretreatment bFGF levels correlated with advancing tumor stages in types I and II EC (p  < 0.05). Multivariate analysis with Cox proportional hazard regression models revealed that high bFGF serum level correlated with shorter overall survival (OS) in type I EC (HR, 0.39, p < 0.001) and in type II EC (HR, 0.47, p = 0.01) and disease-free survival (DFS) (HR, 0.53, p = 0.03 and HR, 0.51, p = 0.02, respectively).ConclusionHigh preoperative bFGF levels predict a poor prognosis in patients with EC, and the prognostic value is independent of established prognostic parameters. These data suggest that bFGF might potentially serve as a marker in prognosis and offer a possibility to individualize treatment regimen.     

Distribution of collagen type VII in connective tissues of postmenopausal stress-incontinent women

Objective.?This study aimed to disclose the distribution changes of collagen VII-immunoreactive (CVII-IR) structures in the arcus tendineus fasciae pelvis (ATFP) of postmenopausal women with stress urinary incontinence (SUI).

Patients and methods.?Fifty-five postmenopausal patients with a history of incontinence were examined for SUI. In patients with SUI, colposuspension was performed either after intravaginal estrogen therapy (ESTR) or without it (NON-ESTR). Age-matched patients without incontinence served as controls. During the surgery, connective tissue specimens from the ATFP and the rectus muscle external fascia (RMEF) were collected and prepared for immunohistochemistry.

Results.?SUI was diagnosed in 23 patients, 20 of them entered either the ESTR or the NON-ESTR group. No differences were found in the distribution of CVII-IR structures in RMEF specimens obtained from all groups. The organization of CVII-IR fibrils in the ATFP of stress-incontinent women was severely affected by degenerative processes. Within the ESTR group, the degree of CVII-IR fiber disintegration was lower.

Conclusions.?Connective tissue from the urogenital suspensory apparatus of women with SUI demonstrates a degenerative distribution pattern of collagen type VII fibers.     

Integrin expression in the dermis during scar formation in humans

To evaluate changes leading to human wound reorganization we examined by immunohistochemistry the expression of several extracellular matrix (ECM) receptors (alpha2 chain of VLA-2, alpha3 chain of VLA-3, alpha6 chain of VLA-6, alphav, and beta1/beta3 chains of integrins) in a series of biopsies of human skin wounds healing by primary intention. The first time point investigated in this study was day 6 after injury, i.e. when a fibrin clot has been almost completely replaced by the granulation tissue. Gradual changes in integrin expression in granulation tissue and in the dermal scar were observed from the first time point investigated and were characterized by an up-regulation of alpha2beta1 complex, alphav integrin subunit, and beta1 integrin subunit. At day 27, the expression of the alpha2 chain of VLA-2 in the scar decreased. The expression of alphav and beta1 integrin subunits decreased but was still detectable by day 35. Vitronectin expression from day 7 onwards was also increased and colocalized to the area of the wounded dermis, and decreased by day 27. Our data suggests that, during the remodelling of the provisional matrix of the wound, dermal fibroblasts express transiently mainly alpha2 and alphav subunits of integrins associated with up-regulation of the beta1 subunit. It seems that up-regulation of some chains of integrins may be involved in the control of deposition of ECM components associated with wound healing.