Production of Soluble and Functional Anti-TNF-α Fab' Fragment in Cytoplasm of E. coli: Investigating the Effect of Process Conditions on Cellular Biomass and Protein Yield Using Response Surface Methodology.

With the increasing dominance of monoclonal antibodies (mAbs) in the biopharmaceutical industry and smaller antibody fragments bringing notable advantages over full-length antibodies, it is of considerable significance to choose the most suitable production system. Although mammalian expression system has been the preferred choice in recent years for mAbs production, E. coli could be the favorable host for non-glycosylated small antibody fragments due to the emergence of new engineered E. coli strains capable of forming disulfide-bonds in their cytoplasm.In this study, non-glycosylated anti-TNF-α Fab' moiety of Certolizumab pegol, produced by periplasmic expression in E. coli in previous studies, was produced in the cytoplasm of E. coli SHuffle strain. The results indicated that it is biologically functional by testing the antigen-binding activity via indirect ELISA and inhibition of TNF-α induced cytotoxicity using MTT test. Major factors affecting protein production and, optimized culture conditions were examined by analyzing growth characteristics and patterns of expression in 24 h of post-induction cultivation and, optimization of culture conditions by response surface methodology considering temperature, time of induction and concentration of inducer in small (tube) and shake-flask scale. Based on the results, temperature had the most significant influence on functional protein yield while exerting different impacts in small and shake-flask scales, which indicated that cultivation volume is also an important factor that should be taken into account in optimization process. Furthermore, richness of medium and slower cellular growth rate improved specific cellular yield of functional protein by having a positive effect on the solubility of Fab' antibody.

A Review on Stem Cell Therapy for Neuropathic Pain.

Neuropathic pain is a complex, chronic pain state that is heterogeneous in nature and caused by the consequence of a lesion or disease affecting the somatosensory system. Current medications give a long-lasting pain relief only in a limited percentage of patients also associated with numerous side effects. Stem cell transplantation is one of the attractive therapeutic platforms for the treatment of a variety of diseases, such as neuropathic pain. Here, the authors review the therapeutic effects of stem cell transplantation of different origin and species in different models of neuropathic pain disorders. Stem cell transplantation could alleviate the neuropathic pain; indeed, stem cells are the source of cells, which differentiate into a variety of cell types and lead trophic factors to migrate to the lesion site opposing the effects of damage. In conclusion, this review suggests that stem cell therapy can be a novel approach for the treatment of neuropathic pain.

Combining Growth Factor and Stem Cell Therapy for Stroke Rehabilitation, A Review.

Stroke is a serious, life-threatening condition demanding vigorous search for new therapies. Recent research has focused on stem cell-based therapies as a viable choice following ischemic stroke, based on studies displaying that stem cells transplanted to the brain not only survive but also cause functional recovery. Growth factors defined as polypeptides that regulate the growth and differentiation of many cell types. Many studies have demonstrated that combined use of growth factors may increase results by the stimulation of endogenous neurogenesis, anti-inflammatory, neuroprotection properties, and enhancement of stem cell survival rates and so may be more effective than a single stem cell therapy. This paper reviews and discusses the most promising new stroke recovery research, including combination treatment.

Association between IFN-γ +874 T/A (Rs2430561) Polymorphisms and Bipolar 1 Disorder: A Study in an Ethnic Iranian Population.

BACKGROUND: The pathophysiology of bipolar 1 disorder (B1D), a major psychiatric disorder with inflammatory origins and structural changes in the brain, is of great interest to researchers. Pro-inflammatory biomarkers and specific gene expression play pivotal roles in B1D development, and IFN-γ has emerged as an important inflammatory marker. The aim of this research was to determine whether the INF-γ +874 T/A polymorphism is associated with B1D susceptibility in an ethnic Iranian population. METHODS: The IFN-γ +874 T/A (rs2430561) gene polymorphism was studied in 106 B1D patients and 109 control subjects using sequence specific primers (SSPs) and amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). RESULTS: Significant statistical differences in IFN-γ +874 T/A polymorphism genotype distribution were found between the patients and control subjects (P = 0.0006). Decreased risk of B1D was detected in the codominant model (T/T vs T/A and A/A, OR = 0.19, 95% CI = 0.07-0.49 for T/A, OR = 0.38, 95% CI = 0.12-1.24 for A/A, P value=0.0006), and in the dominant model (T/T vs T/A-A/A, OR = 0.21, 95% CI = 0.08-0.54, P = 0.0005). However, no significant difference in the IFN-γ polymorphism allele distribution was found between the two groups (P = 0.25). CONCLUSION: The IFN-γ +874 T/A polymorphism may have a significant role in BID development.

A Study on the Association of Interleukin-1 Cluster with Genetic Risk in Bipolar I Disorder in Iranian Patients: A Case-control Study.

The pathogenesis of Bipolar I Disorder (BP-I) involves immune-mediated mechanisms, especially an imbalance in pro-inflammatory/anti-inflammatory cytokines in plasma or cerebrospinal fluid. Interleukin-1 (IL-1) gene cluster, coding some of these pro-inflammatory cytokines, might play a role in various neuropathologies related to neuron inflammation. The aim of the present study was to investigate the possible role of IL-1 gene cluster polymorphisms in determining the susceptibility to BP-I in Iranian population. 48 patients with BP-I in Mashhad (in north-eastern Iran), diagnosed by two psychiatrists using SCID (structured clinical interview for DSM disorders) were selected through convenient sampling and were compared with 47 healthy controls, voluntarily enrolled in the study. Patients with non-Persian ethnicity, history of immunoallergic disorders, endocrinopathies, neurologic disorders, and substance-induced mood disorders were excluded from both case and control groups. Genotyping of IL-1 gene cluster polymorphisms, including IL-1a-889, IL-1b +3954, IL-1b-511, and IL-1RN (VNTR) were carried out using Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and compared by SPSS using Fisher's exact and chi-square tests. The frequency of IL-1b-511 CC genotype and C/T allelic frequency were significantly different between BMD patients and healthy controls (p=0.04 and p=0.02, respectively). Among patients, -511 T allele was significantly more frequent in those with a positive history of major depression. Moreover, +3954 T allele was significantly more frequent in early onset BMD patients. The results suggest a positive association between IL-1 gene cluster variation and BP-I. This polymorphism may contribute to genetic vulnerability through its possible role in neuron inflammation.

The relationship between meteorological conditions and homicide, suicide, rage, and psychiatric hospitalization.

This study focuses on the relationship between the incidence of homicide, rage, suicide, and psychiatric hospitalization as violent behaviors with temperature, humidity, and air pressure as specific meteorological variables in the city of Mashhad, in the northeast of Iran. The data were obtained from Iran Meteorological Organization, official registry of Legal Medicine Organization and the local psychiatric hospital, March 2009 to Feb 2010 daily and were analyzed with SPSS-14 using Pearson correlation coefficient, ANOVA, and post hoc analysis tests. The rates of rage and psychiatric admission had a significant relationship with the daily mean air temperature, minimum relative humidity, maximum relative humidity, minimum daily pressure, and maximum daily air pressure (p < 0.0001). There was no significant correlation between homicide and suicide rates with any meteorological variables (p > 0.05). We concluded that, the possibility of nonfatal violence and psychiatric hospitalization would increase in hot and arid weather with low air pressure.