R1441C mutation in LRRK2 impairs dopaminergic neurotransmission in mice

Dominantly inherited mutations in leucine-rich repeat kinase 2 (LRRK2) are a common genetic cause of Parkinson''s disease (PD). The importance of the R1441 residue in the pathogenesis is highlighted by the identification of three distinct missense mutations. To investigate the pathogenic mechanism underlying LRRK2 dysfunction, we generated a knockin (KI) mouse in which the R1441C mutation is expressed under the control of the endogenous regulatory elements. Homozygous R1441C KI mice appear grossly normal and exhibit no dopaminergic (DA) neurodegeneration or alterations in steady-state levels of striatal dopamine up to 2 years of age. However, these KI mice show reductions in amphetamine (AMPH)-induced locomotor activity and stimulated catecholamine release in cultured chromaffin cells. The introduction of the R1441C mutation also impairs dopamine D2 receptor function, as suggested by decreased responses of KI mice in locomotor activity to the inhibitory effect of a D2 receptor agonist, quinpirole. Furthermore, the firing of nigral neurons in R1441C KI mice show reduced sensitivity to suppression induced by quinpirole, dopamine, or AMPH. Together, our data suggest that the R1441C mutation in LRRK2 impairs stimulated dopamine neurotransmission and D2 receptor function, which may represent pathogenic precursors preceding dopaminergic degeneration in PD brains.     

Interleukin-10–592C/A, –819C/T and –1082A/G promoter variants affect the susceptibility to nephropathy in Tunisian type 2 diabetes (T2DM) patients

Background  The Interleukin (IL)-10 polymorphic variants –1082G/A, –819C/T and –592C/A were linked with obesity, metabolic syndrome, and type 2 diabetes (T2DM). We investigated the hypothesis that IL-10 promoter polymorphisms may be associated with the progression of diabetic nephropathy (DN).
Design  Case-controlled study.
Patients  Study subjects comprised of 515 DN patients, and 402 normoalbuminuric (DWN) T2DM patients.
Measurements  IL-10 genotyping was done by PCR-based assays, and the contributions of the IL-10 polymorphic variants to DN were analysed by haplotype analysis and multivariate regression analysis.
Results  Decreased prevalence of (mutant) –819T allele and –819C/T genotype was seen in DN patients; neither the –1082G/A nor the –592C/A polymorphism was associated with DN. Three-loci haplotype (–1082GA/–819CT/–592CA) analysis identified GTC as DN-protective haplotype. Multivariate regression analysis confirmed the association of GTC haplotype ( P =  0·045; O R  = 0·56, 95% CI: 0·31–0·99), and in addition identified GTA haplotype ( P =  0·044; O R  = 0·54, 95% CI: 0·30–0·98) as independent predictors of DN after controlling for a number of covariates (age, sex, BMI; hypertension, glucose, HbA1c, DN duration, total cholesterol, medications).
Conclusion  This study suggests that IL-10 promoter polymorphism influence the risk of nephropathy in Tunisian T2DM patients.     

Outcome After Ipsilateral Breast Tumor Recurrence in Patients With Early-Stage Breast Cancer Treated With Accelerated Partial Breast Irradiation

PurposeLimited outcomes exist in patients who develop an ipsilateral breast tumor recurrence (IBTR) after accelerated partial breast irradiation (APBI). The purpose of this study was to evaluate these outcomes and patterns of failure in our cohort of patients undergoing APBI.Methods and MaterialsA total of 534 patients with early-stage breast cancer were treated with APBI between 1993 and 2010. Clinical, pathologic, and treatment-related variables were analyzed. Clinical outcomes, including further IBTR, regional recurrence, disease-free survival, cause-specific survival, and overall survival were analyzed.ResultsEighteen (3.3%) patients developed an IBTR, for a 5-year actuarial rate of 2.0%; 14 (77.8%) of the recurrences were thought to represent new primary cancers. After IBTR, 13 (72.2%) patients were managed with salvage mastectomy and 4 (22.2%) patients with a second attempt at breast-conserving therapy. Five-year rates of disease-free survival, cause-specific survival, and overall survival after salvage mastectomy for IBTR were 81%, 100%, and 100%, respectively. In the 4 patients treated with a second attempt at breast-conserving therapy, no IBTR, axillary failure, regional recurrence, or distant metastases were noted at 5 years.ConclusionsIBTRs that developed after APBI resulted in excellent clinical outcomes comparable with those observed after whole-breast irradiation.     

Triazenoindazoles and triazenopyrazolopyridines: Design,synthesis, and cytotoxic activity

Several triazenoindazoles 3a–e and triazinopyrazolopyridines 6a–i were prepared through the reaction of the corresponding 3-amino-4-chloroindazole and 3-aminopyrazolopyridine diazonium salts 2 and 5 with a number of secondary amines. All compounds were evaluated for their in vitro cytotoxic activity on three cell lines, HepG2, MCF7, and HeLa. Most compounds inhibited cell growth with IC₅₀ less than 0.1 μM. Compound 6d was the most potent, with an IC₅₀ of 0.03 μM against HepG2 and 0.05 μM against MCF7 and HeLa cells.     

N‐acetylcysteine reduces disease activity by blocking mammalian target of rapamycin in T cells from systemic lupus erythematosus patients: A randomized,double‐blind,placebo‐controlled trial

Objective

Systemic lupus erythematosus (SLE) patients exhibit T cell dysfunction, which can be regulated through mitochondrial transmembrane potential (Δψm) and mammalian target of rapamycin (mTOR) by glutathione (GSH). This randomized, double‐blind, placebo‐controlled study was undertaken to examine the safety, tolerance, and efficacy of the GSH precursor N‐acetylcysteine (NAC).

Methods

A total of 36 SLE patients received either daily placebo or 1.2 gm, 2.4 gm, or 4.8 gm of NAC. Disease activity was evaluated monthly by the British Isles Lupus Assessment Group (BILAG) index, the SLE Disease Activity Index (SLEDAI), and the Fatigue Assessment Scale (FAS) before, during, and after a 3‐month treatment period. Mitochondrial transmembrane potential and mTOR were assessed by flow cytometry. Forty‐two healthy subjects matched to patients for age, sex, and ethnicity were studied as controls.

Results

NAC up to 2.4 gm/day was tolerated by all patients, while 33% of those receiving 4.8 gm/day had reversible nausea. Placebo or NAC 1.2 gm/day did not influence disease activity. Considered together, 2.4 gm and 4.8 gm NAC reduced the SLEDAI score after 1 month (P = 0.0007), 2 months (P = 0.0009), 3 months (P = 0.0030), and 4 months (P = 0.0046); the BILAG score after 1 month (P = 0.029) and 3 months (P = 0.009); and the FAS score after 2 months (P = 0.0006) and 3 months (P = 0.005). NAC increased Δψm (P = 0.0001) in all T cells, profoundly reduced mTOR activity (P = 0.0009), enhanced apoptosis (P = 0.0004), reversed expansion of CD4−CD8− T cells (mean ± SEM 1.35 ± 0.12‐fold change; P = 0.008), stimulated FoxP3 expression in CD4+CD25+ T cells (P = 0.045), and reduced anti‐DNA production (P = 0.049).

Conclusion

This pilot study suggests that NAC safely improves lupus disease activity by blocking mTOR in T lymphocytes.

     

Elevated Cytomegalovirus and Epstein-Barr virus burden in rheumatoid arthritis: A true pathogenic role or just a coincidence

BackgroundCytomegalovirus (CMV) and Epstein-Barr virus (EBV) have created great interest as their immunomodulatory action and latent forms could play a role in the advance of autoimmune diseases.Aim of the workTo investigate the viral load of CMV and EBV in the serum of rheumatoid arthritis (RA) patients’ and study their association with the clinical and laboratory profiles.Patients and methodsThe study included 50 RA patients and 32 healthy controls. Disease activity score (DAS28) was assessed and the medications received reported. Quantitation of CMV and EBV DNA in serum of all subjects was achieved by real-time polymerase chain reaction.ResultsPatients were 38 females and 12 males with mean age of 43.1 ± 12 years, disease duration of 6.5 ± 5.7 years; age of onset 36.6 ± 12.8 years. CMV DNA was detected in serum of 68% (34\50) patients, while EBV DNA was detected in 40% (20\50). Fourteen (28%) patients had both EBV and CMV DNA detected in serum. No viruses were detected in serum of 10/50 (20%) patients or in healthy controls. The mean viral load of CMV and EBV detected were 42005 ± 24805 copies/ml and 18756 ± 24937 copies/ml respectively. A significant increased frequency of anemia (p < 0.0001), Raynaud's (p < 0.0001), oral ulcers (p = 0.014) and arthritis (p < 0.0001) was detected in those infected with CMV versus those infected with EBV.ConclusionsA high incidence of CMV and EBV was detected in RA patients with increased viral load than described previously. Frequencies of RA disease manifestations are significantly higher in CMV infected patients compared to those infected with EBV.     

Prevalence of fibromyalgia in physicians in training: a cross-sectional study

Clinical Rheumatology - The prevalence of fibromyalgia (FM) in physicians in training (PIT) in Saudi Arabia is unknown. The aim of this study is to evaluate the prevalence of FM in PIT using...