Muhammad Shahnawaz Soyfoo

Phagocyte-specific S100A8/A9 protein levels during disease exacerbations and infections in systemic lupus erythematosus.

Objective: S100A8 and S100A9 are calcium binding proteins of the S100 family highly expressed in neutrophils and monocytes. S100 proteins are novel ligands of TLR4 important in modulating inflammation. High levels of S100A8/A9 found in human inflammatory diseases are a marker of disease activity in rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). We determined levels of S100A8/A9 in sera of patients with systemic lupus erythematosus (SLE) and analyzed their relation to clinical variables of disease activity. Methods. A group of 93 patients with SLE were studied over a period of 3 years, and 143 serum samples were analyzed. S100A8/A9 serum concentrations were determined by a sandwich ELISA. Sera from 10 primary Sjögren’s syndrome (pSS) patients and 50 healthy volunteers were used as controls. Correlations to SLEDAI, ANA, anti-dsDNA, WBC, CH50, C4, and CRP were made. In addition, infections were recorded in all SLE patients. Results. Serum levels of S100A8/A9 were significantly (p = 0.04) higher in SLE patients (1412 ± 664 ng/ml) versus healthy controls (339 ± 35 ng/ml) and pSS patients (400 ± 85 ng/ml). The only significant correlation (r = 0.219; p = 0.015) was found was between S100A8/A9 and SLEDAI. Further, SLE patients with concomitant infections had higher serum levels of S100A8/A9 (39300 ± 13375 ng/ml) than those without infections (1150 ± 422 ng/ml). Conclusion. Serum levels of S100A8/A9 are significantly raised in SLE versus pSS patients and healthy controls and can be correlated to a disease activity index. S100A8/A9 is a more relevant marker of infection in SLE patients.

Aquaporin-4 Overexpression in Rat ALS Model

Onset of motoneuron death characterizing amyotrophic lateral sclerosis (ALS) is closely linked to modified astrocytic and glial environments. Here, we show that in the spinal cord from transgenic rat overexpressing mutated human SOD1, aquaporin‐4 mRNA and protein are specifically overexpressed in the gray matter at end stage of disease. Immunohistochemistry and double immunofluorescence allowed to detect, in the spinal cord gray matter of the ALS rat, increased aquaporin‐4 surrounding both vessel and motoneuron perikarya. The use of pre‐embedding immunohistochemistry at electron microscopic level confirmed such localization associated with swollen astrocytic processes surrounding the vessels. The AQP4 immunohistochemical labeling surrounding several motoneuron perikarya was only seen in ALS rats. Identification of this AQP4‐positive cellular type remains unclear. Anat Rec 2009.

Potential involvement of the IL-33–ST2 axis in the pathogenesis of primary Sjögren's syndrome

Objectives To investigate the role of the interleukin (IL)-33–ST2 axis in the pathophysiology of primary Sjögrens syndrome (pSS). Methods Serum levels of IL-33 and sST2 were determined by ELISA. The expression of IL-33 and ST2 was investigated in salivary glands (SG) by immunohistochemistry. PBMC were isolated and stimulated with IL-33, IL-12 and IL-23 and the cytokine profile response was examined by flow cytometry. Intracellular cytokine detection of IFNγ and IL-17 was performed by flow cytometry. Results Serum IL-33 and sST2 levels were increased in pSS patients compared with controls and patients with systemic lupus erythematosus. Expression of IL-33 was upregulated in SG with Chisholm scores of 2 and 3 of pSS patients but comparable with controls for SG with Chisholm score of 4. ST2 expression in SG was downregulated in pSS patients. IL-33 at different concentrations did not increase the secretion of pro-inflammatory cytokines but acted synergistically with IL-12 and IL-23 to promote IFNγ production. NK and NKT cells were identified as main producers of IFNγ in vitro and were found in SG of pSS patients. Conclusions IL-33 is released in pSS, and acts with IL-12 and IL-23 to favour the secretion of IFNγ by NK and NKT cells.

Adenosine triphosphate prevents serum deprivation-induced apoptosis in human mesenchymal stem cells via activation of the MAPK signaling pathways.

Human mesenchymal stem cells (hMSC) are multipotent cells derived from various sources including adipose and placental tissues as well as bone marrow. Owing to their regenerative and immunomodulatory properties, their use as a potential therapeutic tool is being extensively tested. However, one of the major hurdles in using cell‐based therapy is the use of fetal bovine serum that can trigger immune responses, viral and prion diseases. The development of a culture medium devoid of serum while preserving cell viability is therefore a major challenge. In this study, we demonstrated that adenosine triphosphate (ATP) restrained serum deprivation‐induced cell death in hMSC by preventing caspases 3/7 activation and modulating ERK1/2 and p38 MAPK signaling pathways. We also showed that serum deprivation conditions triggered dephosphorylation of the proapoptotic protein Bad leading to cell death. Adjunction of ATP restored the phosphorylation state of Bad. Furthermore, ATP significantly modulated the expression of proapoptopic and antiapoptotic genes, in favor of an antiapoptotic profile expression. Finally, we established that hMSC released a high amount of ATP in the extracellular medium when cultured in a serum‐free medium. Collectively, our results demonstrate that ATP favors hMSC viability in serum deprivation conditions. Moreover, they shed light on the cardinal role of the MAPK pathways, ERK1/2 and p38 MAPK, in promoting hMSC survival. Stem Cells 2015;33:211–218

Clinical Significance of Cryofibrinogenemia: Possible Pathophysiological Link with Raynaud's Phenomenon

Objective. To describe the clinical findings and prevalence of patients with cryofibrinogenemia (CF) and to determine whether CF is associated with primary Raynaud’s phenomenon. Methods. Between June 2006 and December 2009, 227 patients were tested for CF in a single university hospital. Forty-five patients with primary Raynaud’s phenomenon were tested for CF. Results. A total of 117 patients with CF without cryoglobulinemia were included. The main clinical manifestations included skin manifestations (50%) and arthralgia (35%). There were 67 patients with primary CF and 50 patients with secondary CF. There was no significant difference in the mean concentration of the cryoprecipitate in primary CF as compared to the secondary form (172 ± 18.6 vs 192 ± 20.9 mg/dl, respectively; p = 0.41). Highest concentrations of cryoprecipitate were observed in those containing fibrinogen only as compared to cryoprecipitates containing fibrinogen and fibronectin (301 ± 43.5 vs 125 ± 10.6 mg/dl; p < 0.001). Patients having skin necrosis (n = 3) had significantly higher values of cryofibrinogen compared to those without necrosis (638 ± 105 vs 160 ± 10.2 mg/dl; p = 0.0046). Among the 45 patients with primary Raynaud’s phenomenon, 36 had associated CF. There was no significant difference in the mean concentration of the cryoprecipitate in these patients compared to those with primary CF. Conclusion. There seems to be a significant correlation between cryofibrinogen concentration and the severity of the clinical signs, particularly when cryoprecipitate is composed of fibrinogen alone. CF might have a possible pathophysiological role in primary Raynaud’s phenomenon.

Pericarditis Revealing Large Vessel Vasculitis

Large vessels vasculitis and more specifically, Giant cell arteritis, is characterized by increased inflammatory markers, headaches and altered clinical status. Diagnosis is confirmed by biopsy of temporal arteries showing the presence of granuloma and vasculitis. We hereby report the case of a patient presenting initially as pericarditis and revealing large vessel vasculitis using FDG-PET.

Cryofibrinogen levels are increased in non-traumatic osteonecrosis: a new pathogenic clue to osteonecrosis?

OBJECTIVE To determine whether levels of cryofibrinogen are increased in non-traumatic osteonecrosis (ON) and could correlate with disease staging. METHODS We prospectively analysed cryofibrinogen levels by immunofixation electrophoresis in 50 patients with non-traumatic ON, 50 healthy volunteers and 8 patients with traumatic ON. Staging of disease involving the femoral heads and the size of necrotic lesions were assessed by the Association Research Circulation Osseous (ARCO) classification system. RESULTS Mean cryofibrinogen levels in patients with non-traumatic ON were significantly increased relative to healthy controls and to patients with traumatic ON (222.1 ± 20.6, 59.9 ± 5.6 and 52.3 ± 14.9 mg/dl, respectively, P < 0.001). In the non-traumatic ON group, mean cryofibrinogen levels were significantly increased in patients with multifocal ON compared with patients with mono/bifocal ON (276.5 ± 56.5 and 149.3 ± 15.4 mg/dl, respectively, P = 0.03). There were no significant differences in cryofibrinogen levels observed with respect to the size of the necrotic lesions involving the femoral heads. Moreover, cryofibrinogen levels in patients with ON of the femoral heads classified according to the stage of disease were not significantly different between patients with stage 1/2 and patients with stage 3 ON (179.2 ± 31.3 vs 204.1 ± 29.0 mg/dl, respectively; P = 0.813). CONCLUSION Cryofibrinogen levels are increased in non-traumatic ON and, more importantly, in multifocal ON. The fact that cryofibrinogen levels are not correlated with the size of lesions and the stage of disease could imply systemic rather than local involvement characterizing the pathogenesis of ON.

Aquaporin-4 as a potential marker of BBB disruption in ALS models

In response to the letter addressed by Svitlana Garbuzova-Davis (1) on the implication of the blood-brain barrier in ALS, we would like to express our interest and bring some further findings supporting this concept. For years, investigators working in the field of ALS focused their research around motor neuronal death and subsequent pathways. Through studies of mutant SOD1 animal models, we know that non-neuronal cells are important players in the disease process that introduced the concept of ‘non-cell autonomous nature of motor neuron death in ALS’ (2). However, the specific susceptibility of motor neurons to death is still elusive. As pointed out by Svitlana GarbuzovaDavis (1), today a third concept involving the bloodbrain barrier (BBB) is emerging (3,4). Recent reports highlighted the perturbation of the blood-brain barrier and the blood-spinal cord barrier (BSCB) occurring at early, presymptomatic and late-phase disease in mice carrying mutant SOD1. The findings of endothelial cell degeneration (3), impaired expression of tight junction proteins, plasma leakage, and IgG deposits (4) in the spinal cord of mutant SOD1 animals may lead to the conclusion that BBB is seriously compromised. Based on these considerations, motor neurons may be exposed to a toxic environment from the plasma and surrounded by oedema with focal hypoxic conditions throughout the spinal cord tissue. We would like to stress here that the overexpression of aquaporin-4 (AQP4) at the gliovascular swollen end-feet we have observed in the transgenic rat SOD1-G93A (5) fits very well with the observation that AQP4 is involved in both BBB integrity and perturbation (6,7). Our original article on aquaporin-4 overexpression in the lumbar spinal cord of SOD1-G93A rats strengthens the evidence of oedema and BSCB disruption (3). Aquaporin-4, the main protein responsible for water movements in the CNS and expressed in astrocytes, may be involved in clearance or formation of oedema. In agreement with Garbuzova-Davis’s ultrastructural analysis of mutant SOD1 mice, we demonstrated intracellular oedema by evidence of swollen astrocyte end-feet around microvasculature in mutant SOD1 rats. In several other models, AQP4 induction in perivascular astrocytes has been correlated to BBB breakdown and could serve as a marker of BBB integrity (6). In addition, we would like to raise here another observation we made showing that this AQP4 overexpression is also observed in stacked glial processes directly in contact with motor neurons (5). This observation, associated with findings that impaired potassium buffering through loss of Kir4.1 channel (8) and decreased glutamate re-uptake by astrocytes (2), should arouse further our curiosity and could help us to understand the paracrine toxicity to motor neurons. Gathering these data corroborates evidences of BSCB disruption in an animal model of ALS. BSCB dysfunction may contribute to a vicious circle of the disease process by spreading inflammation within the CNS and so weakening susceptible neuronal functions. Further confirmations must be obtained from human disease samples and may lead to therapeutic strategies aiming to restore the BBB integrity in ALS.

Diffuse Muscular Pain, Skin Tightening, and Nodular Regenerative Hyperplasia Revealing Paraneoplastic Amyopathic Dermatomyositis due to Testicular Cancer.

Paraneoplastic dermatomyositis (DM) associated with testicular cancer is extremely rare. We report the case of a patient with skin tightening, polymyalgia, hypereosinophilia, and nodular regenerative hyperplasia revealing seminoma and associated paraneoplastic DM.

Parotid Gland Biopsy as an Additional Diagnostic Tool for Supporting the Diagnosis of Sjögren's Syndrome.

Sjögrens syndrome (SS) is an autoimmune disease characterized by keratoconjunctivitis sicca and xerostomia. There are actually no diagnostic criteria for SS, but classification criteria based on the revised American-European criteria have been elaborated. These include subjective criteria: ocular and oral symptoms, and objective criteria: ocular, histopathological, oral, and serological signs. SS is considered if 4 of the 6 criteria are present, when histopathology or serology is positive, or if 3 of any 4 objective criteria are present. A patient presented with both ocular and oral symptoms and signs but did not meet the SS classification criteria. Indeed, no anti-SSA or anti-SSB antibodies were detected, and minor salivary gland biopsy was normal. To further understand the origin of the sicca symptoms, a parotid gland biopsy was performed and showed important lymphocytic infiltrates. This could account for the sicca symptoms and signs since parotid glands are one the major contributors to salivary flow. Therefore, parotid gland biopsy could be a useful asset for the diagnosis of SS.