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BACKGROUND Delta variant (B.1.617.2) was considered the most dangerous SARS-CoV-2 strain; however, in-depth studies on its impact on demographic and clinical characteristics of COVID-19 are scarce. OBJECTIVE We aimed to investigate the shift in demographic and clinical characteristics of the COVID-19 pandemic with the emergence of the SARS-CoV-2 delta variant compared to the wild-type (WT) strain (B.1). METHODS A cross-sectional study of COVID-19 cases in the Indian population caused by wild-type (WT) strain (B.1) and delta variant SARS-CoV-2 was performed. The viral genomic sequence metadata containing demographic, vaccination, and patient status details [N =9500, NDelta=6238, NWT=3262] were statistically analyzed. RESULTS With delta variant, in comparison to WT strain, a higher proportion of young individuals ( CONCLUSIONS The increased involvement of young individuals and women, the lower mean age for illness, higher mortality, and frequent post-vaccination infections were the significant epidemiological concerns with the delta variant.

Objective To investigate the role of calcium/calmodulin-dependent protein kinase IV (CaMK4) in the development of joint injury in a mouse model of arthritis and patients with RA. Methods Camk4-deficient, Camk4flox/floxLck-Cre, and mice treated with CaMK4 inhibitor KN-93 or KN-93 encapsulated in nanoparticles tagged with CD4 or CD8 antibodies were subjected to collagen-induced arthritis (CIA). Inflammatory cytokine levels, humoral immune response, synovitis, and T-cell activation were recorded. CAMK4 gene expression was measured in CD4+ T cells from healthy participants and patients with active RA. Micro-CT and histology were used to assess joint pathology. CD4+ and CD14+ cells in patients with RA were subjected to Th17 or osteoclast differentiation, respectively. Results CaMK4-deficient mice subjected to CIA displayed improved clinical scores and decreased numbers of Th17 cells. KN-93 treatment significantly reduced joint destruction by decreasing the production of inflammatory cytokines. Furthermore, Camk4flox/floxLck-Cre mice and mice treated with KN93-loaded CD4 antibody-tagged nanoparticles developed fewer Th17 cells and less severe arthritis. CaMK4 inhibition mitigated IL-17 production by CD4+ cells in patients with RA. The number of in vitro differentiated osteoclasts from CD14+ cells in patients with RA was significantly decreased with CaMK4 inhibitors. Conclusion Using global and CD4-cell-targeted pharmacologic approaches and conditionally deficient mice, we demonstrate that CaMK4 is important in the development of arthritis. Using ex vivo cell cultures from patients with RA, CaMK4 is important for both Th17 generation and osteoclastogenesis. We propose that CaMK4 inhibition represents a new approach to control the development of arthritis.

Infection is one of the major causes of mortality in patients with systemic lupus erythematosus (SLE). We previously found that CD38, an ectoenzyme that regulates the production of NAD+, is up-regulated in CD8+T cells of SLE patients and correlates with the risk of infection. Here, we report that CD38 reduces CD8+T cell function by negatively affecting mitochondrial fitness through the inhibition of multiple steps of mitophagy, a process that is critical for mitochondria quality control. Using a murine lupus model, we found that administration of a CD38 inhibitor in a CD8+T cell–targeted manner reinvigorated their effector function, reversed the defects in autophagy and mitochondria, and improved viral clearance. We conclude that CD38 represents a target to mitigate infection rates in people with SLE.

The complement system is involved in the origin of autoimmunity and systemic lupus erythematosus. Both genetic deficiency of complement components and excessive activation are involved in primary and secondary renal diseases, including lupus nephritis. Among the pathways, the classical pathway has long been accepted as the main pathway of complement activation in systemic lupus erythematosus. However, more recent studies have shown the contribution of factors B and D which implies the involvement of the alternative pathway. While there is evidence on the role of the lectin pathway in systemic lupus erythematosus, it is yet to be demonstrated whether this pathway is protective or harmful in lupus nephritis. Complement is being explored for the development of disease biomarkers and therapeutic targeting. In the current review we discuss the involvement of complement in lupus nephritis.

The proximal insertion beyond coracoid process of pectoralis minor is considered as hidden culprit of rotator cuff disorders. The ectopic insertion is also associated with thoracic outlet syndrome. The current review was conducted to provide a comprehensive evidence-based assessment of the anatomical characteristics of ectopic insertion of pectoralis minor. A through systematic search was conducted on the major electronic database, PubMed, EMBASE, Google Scholar and Journals of Anatomy, orthopedics, plastic surgery, sports medicine. The primary outcome was to measure the prevalence of ectopic insertion of pectoralis minor tendons. The data extraction was conducted for pooled estimation and metanalysis. A total of 25 studies were included for systematic review. The overall pooled estimate of ectopic insertion of Pectoralis Minor was 19.27% (95% CI 15–24%). The prevalence rate in dissected specimen was 21% (CI 15–28%) and in arthroscopic evaluation was 22% (95% CI 5–59%) which was marginally higher with wide confidence interval due small sample size. The prevalence rate in MRI and USG were 15 and 12%, because MRI and USG have almost similar sensitivity in the detection of anomalous insertion of Pectoralis Minor. The distribution of subtypes of anomalous or ectopic insertion based on Le Double classification was 34% for type I, 42 and 9% for Type III. The incidence of ectopic insertion of pectoralis minor was highest in Japanese population. The female and left side have slightly higher incidence at insignificant level. The preoperative MRI or at least USG evaluation of shoulder joint must be conducted for appropriate surgical planning, because the prevalence of ectopic insertion is around 20%. The preoperative detection of anomalous insertion of pectoralis minor can be crucial in minimizing the incidences of iatrogenic injuries of tendon or post-operative complications.

ImportanceHigher risks of contracting infection, developing severe illness and mortality are known facts in aged and male sex if exposed to the wild type SARS-CoV-2 strains (Wuhan and B.1 strains). Now, accumulating evidence suggests greater involvement of lower age and narrowing the age and sex based differences for the severity of symptoms in infections with emerging SARS-CoV-2 variants. Delta variant (B.1.617.2) is now a globally dominant SARS-CoV-2 strain, however, current evidence on demographic characteristics for this variant are limited. Recently, delta variant caused a devastating second wave of COVID-19 in India. We performed a demographic characterization of COVID-19 cases in Indian population diagnosed with SARS-CoV-2 genomic sequencing for delta variant.ObjectiveTo determine demographic characteristics of delta variant in terms of age and sex, severity of the illness and mortality rate, and post-vaccination infections.DesignA cross sectional studySettingDemographic characteristics, including vaccination status (for two complete doses) and severity of the illness and mortality rate, of COVID-19 cases caused by wild type strain (B.1) and delta variant (B.1.617.2) of SARS-CoV-2 in Indian population were studied.ParticipantsCOVID-19 cases for which SARS-CoV-2 genomic sequencing was performed and complete demographic details (age, sex, and location) were available, were included.ExposuresSARS-CoV-2 infection with Delta (B.1.617.2) variant and wild type (B.1) strain.Main Outcomes and MeasuresThe patient metadata containing details for demographic and vaccination status (two complete doses) of the COVID-19 patients with confirmed delta variant and WT (B.1) infections were analyzed [total number of cases (N) =9500, Ndelta=6238, NWT=3262]. Further, severity of the illness and mortality were assessed in subsets of patients. Final data were tabulated and statistically analyzed to determine age and sex based differences in chances of getting infection and the severity of illness, and post-vaccination infections were compared between wild type and delta variant strains. Graphs were plotted to visualize the trends.ResultsWith delta variant, in comparison to wild type (B.1) strain, higher proportion of lower age groups, particularly Conclusions and RelevanceThe increased involvement of young (0-19 year) and women, lower mean age for contracting infection and symptomatic illness/hospitalization, higher mortality, and frequent incidences of post-vaccination infections with delta variant compared to wild type strain raises significant epidemiological concerns.Key PointsQuestionDid SARS-CoV-2 B.1.617.2 (Delta) variant infections show varied demographic characteristics in comparison to wild type strains?FindingsIn this cross sectional study viral genomic sequences of 9500 COVID-19 patients were analyzed. As the key findings, increased involvement of young (0-19 year) and women, lower mean age for contracting infection and symptomatic illness/hospitalization, higher mortality, and frequent incidences of post-vaccination infections with delta variant in comparison to wild type (WT) strain (B.1) were observed.MeaningThe findings of this study suggest that delta variant has varied demographic characteristics reflecting increased involvement of the young and women, and increased lethality in comparison to wild type strains.

Trauma remains a major cause of death throughout the world, especially for patients younger than 45 years. Due to rapid advances in clinical management, both in the acute and prehospital settings, trauma patients survive devastating injuries at unprecedented rates. However, these patients can often face life threatening complications that stem from the robust innate immune response induced by severe hemorrhage, leading to further tissue injury rather than repair. The complement and coagulation cascades are key mediators in this disordered reaction, which includes the development of trauma‐induced coagulopathy. There is increasing evidence that cross‐talk between these two pathways allows rapid amplification of their otherwise targeted responses and contributes to overwhelming and prolonged systemic inflammation. In this article, we summarize the initial steps of innate immune response to trauma and review the complex complement and coagulation cascades, as well as how they interact with each other. Despite progress in understanding these cascades, effective therapeutic targets have yet to be found and further research is needed both to improve survival rates as well as decrease associated morbidity.

Lung inflammation and damage is prominent in people infected with SARS-Cov-2 and a major determinant of morbidity and mortality. We report the deposition of complement components in the lungs of people who succumbed to COVID-19 consistent with the activation of the classical and the alternative pathways. Our study provides strong rationale for the expansion of trials involving the use of complement inhibitors to treat patients with COVID-19.

Current technologies and available scaffold materials do not support long-term cell viability, differentiation and maintenance of podocytes, the ultra-specialized kidney resident cells that are responsible for the filtration of the blood. We developed a new platform which imitates the native kidney microenvironment by decellularizing fibroblasts grown on surfaces with macromolecular crowding. Human immortalized podocytes cultured on this platform displayed superior viability and metabolic activity up to 28 days compared to podocytes cultured on tissue culture plastic surfaces. The new platform displayed a softer surface and an abundance of growth factors and associated molecules. More importantly it enabled podocytes to display molecules responsible for their structure and function and a superior development of intercellular connections/interdigitations, consistent with maturation. The new platform can be used to study podocyte biology, test drug toxicity and determine whether sera from patients with podocytopathies are involved in the expression of glomerular pathology.

Correct localization of the sacral hiatus is essential for administering a successful caudal epidural block. The purpose of this study is to create a statistical model of sacral hiatus from dorsal sacral parameters to improve the location of the hiatus and thus, reduce the failure rate. The aim of this investigation was to examine the relationship of sacral hiatus morphology and dimension with sacral curvature. This study further examines the dorsal sacral parameters that could affect the sacral hiatus dimension. Adult, human, dry sacra and three-dimensionally reconstructed sacra from computed tomography imaging of normal subjects were included in the study and measured using digital Vernier calipers of 0.01 mm accuracy and Geomagic freeform plus software, respectively. The most frequent shape of the sacral hiatus was an inverted V (48%) followed by inverted U shape (32%), an irregular shape (12.3%), an M shape (4.7) and an A shape (2.8%). The data were represented by mean and standard deviation. Sacra with M-shaped hiatus had the lowest hiatal length (14.21 ± 5.44 mm), whereas sacra with an inverted V-shaped hiatus had the highest length (25.41 ± 11.3 mm). The anteroposterior diameter of the sacral hiatus at the base in males and females was found to be 3.46 ± 1.48 mm and 2.79 ± 0.83 mm, respectively (P

Drug repurposing is the only method capable of delivering treatments on the shortened time-scale required for patients afflicted with lung disease arising from SARS-CoV-2 infection. Mucin-1 (MUC1), a membrane-bound molecule expressed on the apical surfaces of most mucosal epithelial cells, is a biochemical marker whose elevated levels predict the development of acute lung injury (ALI) and respiratory distress syndrome (ARDS), and correlate with poor clinical outcomes. In response to the pandemic spread of SARS-CoV-2, we took advantage of a high content screen of 3,713 compounds at different stages of clinical development to identify FDA-approved compounds that reduce MUC1 protein abundance. Our screen identified Fostamatinib (R788), an inhibitor of spleen tyrosine kinase (SYK) approved for the treatment of chronic immune thrombocytopenia, as a repurposing candidate for the treatment of ALI. In vivo, Fostamatinib reduced MUC1 abundance in lung epithelial cells in a mouse model of ALI. In vitro, SYK inhibition by Fostamatinib promoted MUC1 removal from the cell surface. Our work reveals Fostamatinib as a repurposing drug candidate for ALI and provides the rationale for rapidly standing up clinical trials to test Fostamatinib efficacy in patients with COVID-19 lung injury.

Summary Drug repurposing has the advantage of identifying potential treatments on a shortened timescale. In response to the pandemic spread of SARS-CoV-2, we took advantage of a high-content screen of 3,713 compounds at different stages of clinical development to identify FDA-approved compounds that reduce mucin-1 (MUC1) protein abundance. Elevated MUC1 levels predict the development of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) and correlate with poor clinical outcomes. Our screen identifies fostamatinib (R788), an inhibitor of spleen tyrosine kinase (SYK) approved for the treatment of chronic immune thrombocytopenia, as a repurposing candidate for the treatment of ALI. In vivo, fostamatinib reduces MUC1 abundance in lung epithelial cells in a mouse model of ALI. In vitro, SYK inhibition by the active metabolite R406 promotes MUC1 removal from the cell surface. Our work suggests fostamatinib as a repurposing drug candidate for ALI., Graphical Abstract, Highlights Elevated MUC1 levels predict the development of acute lung injury (ALI) A high-content screen of 3,713 compounds identifies repurposing candidates R406 removes MUC1 from the apical surface of epithelial cells Fostamatinib treatment reduces MUC1 in a mouse model of lung injury, In a high-content screen, Kost-Alimova et al. identify R406, the active metabolite of fostamatinib, as an FDA-approved candidate repurposing compound for the reduction of MUC1 protein levels in lung epithelium in the setting of acute lung injury.

Acute and chronic kidney failure is common in hospitalized patients with COVID-19, yet the mechanism of injury and predisposing factors remain poorly understood. We investigated the role of complement activation by determining the levels of deposited complement components (C1q, C3, FH, C5b-9) and immunoglobulin along with the expression levels of the injury-associated molecules spleen tyrosine kinase (Syk), mucin-1 (MUC1) and calcium/calmodulin-dependent protein kinase IV (CaMK4) in the kidney tissues of people who succumbed to COVID-19. We report increased deposition of C1q, C3, C5b-9, total immunoglobulin, and high expression levels of Syk, MUC1 and CaMK4 in the kidneys of COVID-19 patients. Our study provides strong rationale for the expansion of trials involving the use of inhibitors of these molecules, in particular C1q, C3, Syk, MUC1 and CaMK4 to treat patients with COVID-19.

Development of implantable devices based on the principles of in vitro organogenesis has been hindered due to the prolonged time required to develop an implantable device. Herein we assessed the influence of serum concentration (0.5 % and 10 %), oxygen tension (0.5 %, 2 % and 20 %) and macromolecular crowding (75 μg/ml carrageenan) in extracellular matrix deposition in human corneal fibroblast culture (3, 7 and 14 days). The highest extracellular matrix deposition was observed after 14 days in culture at 0.5 % serum, 2 % oxygen tension and 75 μg/ml carrageenan. These data indicate that low oxygen tension coupled with macromolecular crowding significantly accelerate the development of scaffold-free tissue-like modules. This article is protected by copyright. All rights reserved.

Intestinal ischemia followed by reperfusion leads to local and remote organ injury attributed to inflammatory response during the reperfusion phase. The extent to which ischemia contributes to ischemia/reperfusion injury has not been thoroughly studied. After careful evaluation of intestinal tissue following 30 min of ischemia, we noticed significant local mucosal injury in wild-type mice. This injury was drastically reduced in C3-deficient mice, suggesting C3 involvement. Depletion of circulating complement with cobra venom factor eliminated, as expected, injury recorded at the end of the reperfusion phase but failed to eliminate injury that occurred during the ischemic phase. Immunohistochemical studies showed that tissue damage during ischemia was associated with increased expression of C3/C3 fragments primarily in the intestinal epithelial cells, suggesting local involvement of complement. In vitro studies using Caco2 intestinal epithelial cells showed that in the presence of LPS or exposure to hypoxic conditions the cells produce higher C3 mRNA as well as C3a fragment. Caco2 cells were also noted to produce cathepsins B and L, and inhibition of cathepsins suppressed the release of C3a. Finally, we found that mice treated with a cathepsin inhibitor and cathepsin B–deficient mice suffer limited intestinal injury during the ischemic phase. To our knowledge, our findings demonstrate for the first time that significant intestinal injury occurs during ischemia prior to reperfusion and that this is due to activation of C3 within the intestinal epithelial cells in a cathepsin-dependent manner. Modulation of cathepsin activity may prevent injury of organs exposed to ischemia.

Tissue engineering by self-assembly uses the cells' secretome as a regeneration template and biological factory of trophic factors. Despite the several advantages that have been witnessed in preclinical and clinical settings, the major obstacle for wide acceptance of this technology remains the tardy extracellular matrix formation. In this study, we assessed the influence of macromolecular crowding (MMC)/excluding volume effect, a biophysical phenomenon that accelerates thermodynamic activities and biological processes by several orders of magnitude, in human corneal fibroblast (HCF) culture. Our data indicate that the addition of negatively charged galactose derivative (carrageenan) in HCF culture, even at 0.5% serum, increases by 12-fold tissue-specific matrix deposition, while maintaining physiological cell morphology and protein/gene expression. Gene analysis indicates that a glucose derivative (dextran sulfate) may drive corneal fibroblasts toward a myofibroblast lineage. Collectively, these results indicate that MMC may be suitable not only for clinical translation and commercialization of tissue engineering by self-assembly therapies, but also for the development of in vitro pathophysiology models.

Aim: Topographically modified substrates are increasingly used in tissue engineering to enhance biomimicry. The overarching hypothesis is that topographical cues will control cellular response at the cell–substrate interface. Materials & methods: The influence of anisotropically ordered poly(lactic-co-glycolic acid) substrates (constant groove width of ˜1860 nm; constant line width of ˜2220 nm; variable groove depth of ˜35, 306 and 2046 nm) on in vitro and in vivo osteogenesis were assessed. Results & discussion: We demonstrate that substrates with groove depths of approximately 306 and 2046 nm promote osteoblast alignment parallel to underlined topography in vitro. However, none of the topographies assessed promoted directional osteogenesis in vivo. Conclusion: 2D imprinting technologies are useful tools for in vitro cell phenotype maintenance.

C3a is important in the regulation of the immune response as well as in the development of organ inflammation and injury. Furthermore, C3a contributes to liver regeneration but its role in intestinal stem cell function has not been studied. We hypothesized that C3a is important for intestinal repair and regeneration. Intestinal organoid formation, a measure of stem cell capacity, was significantly limited in C3-deficient and C3a receptor (C3aR) 1-deficient mice while C3a promoted the growth of organoids from normal mice by supporting Wnt-signaling but not from C3aR1-deficient mice. Similarly, the presence of C3a in media enhanced the expression of the intestinal stem cell marker leucine-rich repeat G-protein-coupled receptor 5 (Lgr5) and of the cell proliferation marker Ki67 in organoids formed from C3-deficient but not from C3aR1-deficient mice. Using Lgr5.egfp mice we showed significant expression of C3 in Lgr5+ intestinal stem cells whereas C3aR1 was expressed on the surface of various intestinal cells. C3 and C3aR1 expression was induced in intestinal crypts in response to ischemia/reperfusion injury. Finally, C3aR1-deficient mice displayed ischemia/reperfusion injury comparable to control mice. These data suggest that C3a through interaction with C3aR1 enhances stem cell expansion and organoid formation and as such may have a role in intestinal regeneration.

Modular tissue engineering is based on the cells’ innate ability to create bottom-up supramolecular assemblies with efficiency and efficacy still unmatched by man-made devices. Although the regenerative potential of such tissue substitutes has been documented in preclinical and clinical setting, the prolonged culture time required to develop an implantable device is associated with phenotypic drift and/or cell senescence. Herein, we demonstrate that macromolecular crowding significantly enhances extracellular matrix deposition in human bone marrow mesenchymal stem cell culture at both 20% and 2% oxygen tension. Although hypoxia inducible factor - 1α was activated at 2% oxygen tension, increased extracellular matrix synthesis was not observed. The expression of surface markers and transcription factors was not affected as a function of oxygen tension and macromolecular crowding. The multilineage potential was also maintained, albeit adipogenic differentiation was significantly reduced in low oxygen tension cultures, chondrogenic differentiation was significantly increased in macromolecularly crowded cultures and osteogenic differentiation was not affected as a function of oxygen tension and macromolecular crowding. Collectively, these data pave the way for the development of bottom-up tissue equivalents based on physiologically relevant developmental processes.

A key challenge of in vitro organogenesis is the development in timely manner tissue equivalents. Herein, we assessed the simultaneous effect of oxygen tension (0.5%, 2% and 20%), foetal bovine serum concentration (0.5% and 10%) and macromolecular crowding (75 μg/ml carrageenan) in human dermal fibroblast culture. Our data demonstrate that cells cultured at 2% oxygen tension, in the presence of carrageenan and at 0.5% serum concentration deposited within 3 days in culture more extracellular matrix than cells grown for 14 days, at 20% oxygen tension, 10% serum concentration and in the absence of carrageenan. These data suggest that optimal oxygen tension coupled with macromolecular crowding are important in vitro microenvironment modulators for accelerated development of tissue-like modules in vitro. Statement of Significance To enable clinical translation and commercialisation of in vitro organogenesis therapies, we cultured human dermal fibroblast at 2% oxygen tension, under macromolecular crowding conditions (75 μg/ml carrageenan) and at low foetal bovine serum concentration (0.5%). Within 3 days in culture, more extracellular matrix was deposited under these conditions than cells grown for 14 days, at 20% oxygen tension, 10% FBS concentration and in the absence of crowding agents. These data bring us closer to the development of more clinically relevant tissue-like modules.

Polysaccharides are frequently incorporated into scaffolds for tissue engineering applications to improve mechanical and biological properties. We evaluated the influence of a Ficoll® scaffold on collagen films, a scaffold that is extensively used for soft and hard tissue repair. To avoid cytotoxicity issues associated with chemical reagents, the influence of genipin, a naturally occurring crosslinking agent, was assessed. Ultra-structural level collagen films formed with and without Ficoll showed a fine fibrillar structure whereas genipin crosslinked films showed a coarse fibrillar and partially nodular structure. In contrast, glutaraldehyde crosslinked films lost their fibrillar pattern. Crosslinking significantly increased denaturation temperature (p

Development of implantable devices based on the principles of in vitro organogenesis has been hindered due to the prolonged time required to develop an implantable device. Herein we assessed the influence of serum concentration (0.5% and 10%), oxygen tension (0.5%, 2% and 20%) and macromolecular crowding (75 μg/ml carrageenan) in extracellular matrix deposition in human corneal fibroblast culture (3, 7 and 14 days). The highest extracellular matrix deposition was observed after 14 days in culture at 0.5% serum, 2% oxygen tension and 75 μg/ml carrageenan. These data indicate that low oxygen tension coupled with macromolecular crowding significantly accelerate the development of scaffold-free tissue-like modules. Copyright © 2016 John WileySons, Ltd.

Bladder cancer is a common tumor of the urinary tract, accounting for 6% to 8% of all male malignancies and 2% to 3% of all female malignancies. Urothelial carcinoma (UC) of bladder is the second most common urologic malignancy after prostate cancer. Earlier report has elucidated immunologic unreactivity in cancer patients. Cytokines play a pivotal role in the induction of cell mediated and humoral immunity. Quantification of cytokine response in cancer patients can give significant insights about the cellular immunologic potency against the neoplastic cells. In the present study, we aimed to assess alterations of Th1 and Th2 derived cytokines in progression of UC of bladder by determining their circulatory concentration in bladder cancer patients and healthy controls and to correlate the observations with grade and severity of the disease.The study cohort consisted of 122 subjects; 72 patients with bladder UC (28, low grade; 17, high grade; 27, muscle invasive) and 50 healthy controls. The circulatory levels of various cytokines were measured using commercially available sandwich enzyme linked immunosorbent assay (ELISA) kit from BD Biosciences, San Diego, CA, and were statistically correlated according to the grade and the severity of disease.The serum levels of typical Th1 cytokines: IL-2 and IFN-γ were found to be significantly lower (P0.001) while levels of Th2 cytokines i.e., IL-4, IL-5, and IL-10 were significantly higher (P0.001) in patients than in controls. The levels of all the cytokines were correlated with the grade and severity of the disease. There were significant differences between the patients with low grade tumors and muscle invasive tumors for all cytokines (P0.001); except IL-10 (P0.626).The results of our study delineate that in bladder tumor patients a marked polarization exists towards the expression of Th2 type cytokines while Th1 remain suppressed. Furthermore, the levels of all the cytokines alter according to the grades of the tumor. This can give significant insights about the use of Th1 type cytokines for the administration of immunotherapy to bladder cancer patients. Development of new strategies attempting to manipulate the equilibrium between Th1 and Th2 cells would be beneficial in the management of UC of bladder in future.

Parthenium contact dermatitis is a major health problem caused by a cosmopolitan weed Parthenium hysterophorus. It is a T cell-mediated immune injury and disease manifests as itchy erythematous papules, papulovesicular and plaque lesions on exposed areas of the body. We studied the involvement of T(H)1/T(H)2/T(H)17/Treg type responses by assaying various cytokines in Parthenium dermatitis.The study includes 50 patients of Parthenium dermatitis confirmed by patch testing and 50 healthy subjects. The serum levels of T(H)1, T(H)2, T(H)17 and Treg cytokines were estimated by high sensitivity sandwich ELISA and were compared statistically between groups using ANOVA.The mean concentration of T(H)1 cytokines (p0.001) and IL-17 (p0.001) were increased significantly as compared to controls. In contrast, decrease in levels of IL-10 (p0.002) and TGF-β (p0.001) were significant and levels of IL-4 (p0.262) were insignificant whereas no alterations in the total IgE concentrations (p0.976) was observed.The induction of T(H)1 and T(H)17 cytokines reinforce the need of detailed analysis of immune dysregulation in Parthenium dermatitis and might add some insight in the pathogenesis, diagnosis and current treatment modalities of this disease.

Background and Objectives: Host factors are critical in regulating tumor growth, and cytokines that modulate immunological control may be of importance in cervical cancer. To study this, the cytokines were measured in peripheral blood mononuclear cells in women with cervical cancer, before and after neoadjuvant chemoradiation, and assessed their correlation with therapeutic response. Methods: Ninety patients with advanced cervical cancer and 90 healthy controls were enrolled, and human papillomavirus status was determined. Cytokines were estimated by enzyme-linked immunosorbent assay in pretreatment samples after chemotherapy, brachyradiation, and after follow-up. Response to therapy was assessed during and after therapy and after 1 and 3 years of follow-up. Results: Pretreatment levels of type 1 cytokines (IL-2 and IFN-γ) showed significant decrease, whereas type 2 cytokines (IL-4 and IL-10) showed significant increase in patients versus controls. After chemotherapy, a mild increase in type 1 cytokine levels was observed in complete responders versus partial/nonresponders, which became highly significant after completion of therapy and remained significant during follow-up. A slight decrease in type 2 cytokine levels was seen in complete responders versus partial/nonresponders, which remained insignificant for IL-10 even after chemoradiation. Conclusions: This important finding suggests that pretreatment type 1 cytokine levels and the extent of their change during treatment can predict the therapeutic response to neoadjuvant chemoradiation in advanced cervical cancer.

Pemphigus vulgaris (PV) is classical example of antigen-driven severe autoimmune bullous skin disorder. Auto reactive T cells are critical for the induction and regulation of antibody production. With regard to cytokine production profiles, it has been reported that qualitative as well as quantitative alterations in cytokine production can result in activation of inefficacious effector mechanisms and therefore, complex and severe impairment in immune functions. The purpose of this study was to observe the alterations in the levels of T(H)1 [Interleukin-2 (IL-2), Interferon-gamma (IFN-gamma)] and T(H)2 (IL-4 and IL-10) cytokines in the sera from patients affected with PV and compared with Pemphigus foliaceus and healthy subjects. This work is aimed to comprehend the involvement of T(H)1 and T(H)2 cells as inflammatory infiltrate in the modulation of acantholysis and production of pemphigus lesions. Seventy PV, 13 PF and 50 healthy, age-matched individuals without any generalized skin diseases were included in this study. The diagnosis of PV and PF patients was confirmed by histopathology (hematoxylin and eosin) and / direct immunofluorescence. The levels of T(H)1 cytokines (IL-2 and IFN-gamma) and T(H)2 cytokine markers (IL-4 and IL-10) were estimated by high sensitivity ELISA kits. All patients with PV and PF showed significantly (p < 0.000) elevated levels of T(H)2 cytokines (IL-10 and IL-4) as compared with healthy controls. However, the mean concentration of T(H)1 cytokines (IL-2 and IFN-gamma) was significantly decreased in patients as compared to healthy individuals. Both T(H)1 and T(H)2 cytokines did not show any significant difference between PV and PF cases. Current concepts support the idea that PV, induced by autoantibodies against Dsg3, is the consequence of an imbalance between Dsg3-reactive T(H)2 and T(H)1 cells that may be critical for the maintenance of tolerance against Dsg3. Cytokine profile for confirmed PV cases showed direct evidences for involvement of T cell responses. Increase in IL-4 and IL-10 shows induction of T(H)2 cells in the pathogenesis of autoimmune disorders Pemphigus vulgaris. The decreased levels of IL-2 and IFN-gamma might demonstrate the inhibitory effects by IL-4 and IL-10, which suppress the expansion of T(H)1 population.

Problem Despite progress in immunobiology, pre-eclampsia (PE) remains one of the most common reasons for women to die during pregnancy. The widespread pathophysiological mechanisms are endothelial dysfunction, oxidative stress and inflammation. The aim of this study was to assess the alteration in the levels of leptin, interleukin (IL)-10 and inflammatory cytokines [tumour necrosis factor (TNF)-α, IL-6 & IL-8] in pre-eclamptic (severe and mild), healthy pregnant and non-pregnant women and correlate these parameters with disease severity. Method of study The levels of leptin, IL-10 and inflammatory cytokines were measured by high sensitivity enzyme-linked immunoabsorbant assay. The study subjects were 54 pre-eclamptic women (ten severe and 45 milder), compared by age matched 50 healthy pregnant and 27 non-pregnant women. Kruskal–Wallis non-parametric analyses of variance followed by Mann–Whitney U-test were used for statistical analysis. Results The levels of leptin, TNF-α, IL-6 & IL-8 in pre-eclamptic subjects were increased significantly when compared with the healthy control pregnant and non-pregnant (P

Objective: To compare the levels of 3 oxidative stress markers (glutathione peroxidase [GPX], superoxide dismutase [SOD], and malondialdehyde [MDA]) and 2 antioxidants (vitamin C and lycopene) in healthy and pre-eclamptic pregnant women. Methods: Circulating levels of GPX, SOD, MDA, vitamin C and lycopene were measured in 50 healthy pregnant women and 50 women with pre-eclampsia (PE) (41 with mild PE and 9 with severe PE) attending the antenatal clinic or admitted to the maternity ward of the All-India Institute of Medical Sciences, New Delhi, India. Results: The levels of GPX, SOD and MDA were significantly higher in women with PE than in controls, and the increase was higher in women with severe PE (Pb0.001 using analysis of variance and the Kruskal Wallis test). The levels of vitamin C and lycopene were significantly lower in women with PE than in controls, with a greater decrease in women with severe PE. Conclusion: Increased levels of oxidative stress markers and decreased levels of antioxidants in pre-eclamptic women suggest that oxidative stress markers play a significant role in the pathophysiology of preeclampsia, and that supplemental dietary antioxidants may have a beneficial role in the prevention of pre-eclampsia in women at high-risk for this condition. D 2006 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

Tissue engineering by self-assembly hypothesises that optimal repair and regeneration can be achieved best by using the cells’ inherent ability to create organs with proficiency still unmatched by currently available scaffold fabrication technologies. However, the prolonged culture time required to develop an implantable device jeopardises clinical translation and commercialisation of such techniques. Herein, we report that macromolecular crowding, a biophysical in vitro microenvironment modulator, dramatically accelerates extracellular matrix deposition in cultured human corneal, lung and dermal fibroblasts and human bone marrow mesenchymal stem cells. In fact, an almost 5 to 30 fold increase in collagen type I deposition was recorded as early as 48 hours in culture, without any negative effect in cell phenotype and function.

Advancements in molecular and cell biology have led to the development of cell-based therapies to treat injured or degenerated tissues. [1] The rationale of this concept is that functional regeneration can be achieved best by using the innate capacity of cells to create their own tissue-specific extracellular matrix (ECM) avoiding the shortfalls of man-made devices. Although direct cell injections have demonstrated very promising preclinical and clinical outcomes, [2] the mode of administration offers little control over local retention and distribution of the injected cell suspensions[3] leading to scattered therapeutic efficiency. This deficiency has led to the development of living substitutes for skin[4] and blood vessel[5] composed of cells seeded on a collagen scaffold. Notwithstanding the efficacious results in preclinical models and clinical trials, it soon became apparent that the presence of the scaffold hinders tissue remodelling and function. [6] These drawbacks led to the development of the scaffold-free cell-sheet tissue engineering (CSTE)[7] or tissue engineering by self-assembly (TESA), [8] a therapy that offers the fabrication of a contiguous cell sheet that is stabilised by cell-cell contacts and endogenously produced ECM. Despite the documented, in preclinical and clinical setting, positive outcomes for skin, [9] blood vessel, [10, 11] cornea, [12, 13] heart, [14] lung, [15] liver[16] and bone[17] replacement, only Epicel® (Genzyme, USA) for skin and LifeLine™ for blood vessel (Cytograft, USA) have been commercialised so far. This limited technology transfer from bench-top to clinic has been attributed to the substantial long period of time required for ex vivo culture (e.g. 14-35 days for corneal epithelium; [13] 84 days for corneal stromal; [18] 28 days for corneal endothelium;[19] 70 days for lung cell-sheet; [15] and 196 days for blood vessel[11] ) that often leads to loss of native phenotype and cell senescence.[20] Here, we propose a biophysical approach, termed macromolecular crowding (MMC), that increases thermodynamic activities and biological processes by several orders of magnitude,[21] as means to create ECM-rich tissue equivalents. The principle of MMC is derived from the notion that in vivo cells reside in a highly crowded/dense extracellular space and therefore the conversion of the de novo synthesised procollagen to collagen I is rapid. [22] However, in the even substantially more dilute than body fluids (e.g. urine: 36-50g/l; blood: 80g/l) culture conditions (e.g. HAM F10 nutrient medium: 16.55g/l; DMEM/F12 medium: 16.78g/l; DMEM high glucose and L-glutamine medium: 17.22g/l), the rate limiting conversion of procollagen to collagen I is very slow (Figure 1a). We propose that the addition of inert polydispersed macromolecules (presented as spherical objects of variable diameter in Figure 1b) in the culture media will facilitate amplified production of ECM-rich living substitutes. We thank Dr Oliver Carroll for laboratory management; and Mr Maciek Doczyk (http://doczykdesign.com) for his support in the preparation of Figure 1 of this manuscript. This work is supported by Science Foundation Ireland, Research Frontiers Programme, Project Number: SFI‐09‐RFP‐ENM2483 to D.Z.; Science Foundation Ireland, E.T.S. Walton Visitor Awards Programme, Project Number: 08/W.1/B2568 to M.R., A.P. and D.Z.; Health Research Board, Project Number: HRA_POR/2011/84 to D.Z.; and College of Engineering and Informatics, Postgraduate Scholarship Scheme, NUI Galway to P.K. and D.Z. peer-reviewed

Traditional ex vivo culture setups fail to imitate the native tissue niche, leading to cellular senescence, phenotypic drift, growth arrest and loss of stem cell multipotency. Growing evidence suggests that surface topography, substrate stiffness, mechanical stimulation, oxygen tension and localised density influence cellular functions and longevity, enhance tissue-specific extracellular matrix deposition and direct stem cell differentiation. In this review, we discuss how these cues will facilitate engineering of physiological in vitro microenvironments to enable clinical translation of cell based therapies and development of in vitro models for drug discovery applications.

Background: Recent studies have demonstrated an association between insulin growth factor (IGF) and insulin growth factor binding protein-3 (IGFBP-III) serum levels and increased risk for various cancers. However, little information is available on clinical implications of the IGF system in Indian patients with cervical cancer. This study explored associations by analyzing their expression profiles in cervical cancer cases. Materials and Methods: Totals of 50 patients with advanced cervical cancer and 40 healthy controls were enrolled. Human papillomavirus (HPV) and cervical biopsy sample were obtained from all participating women. Circulatory levels were estimated by ELISA and the tissue expression was assessed using RT-PCR and Western blotting. Results: Levels of IGF-I and II showed significant increase whereas IGFBP-III showed significant decline in all patients as compared to controls. Spearman correlation analysis between IGFs and HPV status showed significant correlations. Conclusions: We demonstrated elevated circulating levels and tissue expression of IGF-I and IGF-II in advancer cancer cervix patients, as compared with controls, with a converse trend being apparent for IGFBP-III. In future, associations of the IGF system and clinical outcome of cervical cancer patients in post treatment samples might point to significance in disease mapping as a prognostic marker after validation with a larger patient series.

The objective was to study the alterations in circulating pro-/antioxidants in patients with cancer of the posterior one third of the tongue, before and after concomitant chemoradiation, and to assess variation in their levels to therapeutic response. Sixty patients with newly diagnosed squamous cell carcinoma of the posterior one third of the tongue and 60 healthy controls were enrolled. Blood samples were collected before and after chemoradiation and after 3 months of follow-up. Pro-/antioxidant levels were estimated using standard methods. Response to therapy was assessed during and after therapy and after 3 months of followup. Pretreatment levels of lipid peroxide were significantly elevated while antioxidant levels were lowered in cancer patients compared to controls. After chemotherapy, lipid peroxidation showed a significant decline, while antioxidants showed a significant rise in complete responders, compared with partial/nonresponders, and remained highly significant after therapy and during follow-up. Pretreatment levels of antioxidant-oxidant parameters and the extent of their change during treatment can predict the therapeutic response to concomitant chemoradiation in carcinoma tongue.

Establishing CYFRA 21-1 detection for noninvasive differential diagnosis of urothelial carcinoma (UC) of bladder would help to improve assessment and follow-up of patients, as well as to improve screening of high-risk groups. The study group comprised of 147 subjects including 72 patients with UC of bladder, 75 controls and 17 follow-up cases. The levels of CYFRA 21-1 in serum, urine and urinary cell lysate were estimated by high sensitivity ELISA. Our results indicate that urinary CYFRA 21-1 provides a high value of overall sensitivity for UC of bladder and is also useful even for detection of low grade tumors that might indicate possible earlier detection and treatment administration.

Cervical cancer is the most common cancer in Indian women. Oxidative stress is potentially harmful to cells and ROS are involved in multistage carcinogenesis, in initiation and promotion. The aim was to study the alterations in the circulating pro-/anti-oxidants in advanced cervical cancer patients, before and after neoadjuvant chemoradiation and to assess the relevance of the variation in the levels to therapeutic response. 90 patients with advanced cancer cervix (FIGO IIIa-IVa) and 90 healthy controls were enrolled. Blood samples were collected: before and after chemotherapy, after radiation and after 1 year on follow-up. Pro-/anti-oxidant levels were estimated using standard methods. Response to therapy was assessed during and after therapy and after 1 year of follow-up. The pre-treatment levels of plasma lipid peroxide were significantly elevated; while antioxidant levels were lowered in cancer patients; when compared to controls. After chemotherapy, lipid peroxidation showed a significant decline in complete responders, as compared with partial/non-responders and remained highly significant after therapy and during follow-up. Anti-oxidant enzymes showed a mild increase (P < 0.05), after chemotherapy in complete responders, as compared with partial/non-responders and remained highly significant after therapy and on follow-up. This important finding suggests that pre-treatment levels of antioxidant-oxidant parameters and the extent of their change during treatment can predict the therapeutic response to neoadjuvant chemoradiation in advanced cancer cervix. Oxidant-antioxidant profile merits investigation as markers of response, survival, and recurrence in larger prospective studies, which might throw light on their possible use as predictors of chemoradiosensitivity of cervical tumors.

Vitiligo is an acquired skin disease, characterized by white areas on the skin due to loss of functional melanocytes. The pathogenesis of the disease is still unclear. Published data show the involvement of oxidative stress in the pathophysiology of vitiligo. A total of 30 vitiligo patients and 30 healthy controls were included in this study. We estimated serum levels of malondialdehyde (MDA), vitamins E and C, total antioxidant activity and whole blood levels of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in vitiligo patients and controls. We found significantly higher levels of MDA and significantly lower levels of SOD, GPx, vitamins C and E and total antioxidant activity in vitiligo patients compared with controls. This study is a maiden attempt to report on antioxidant parameters of both generalized/localized-type Indian vitiligo patients. Our results confirmed that oxidative stress may play an important role in the pathogenesis of vitiligo and cause melanocyte damage in vitiligo.

Urothelial carcinoma of bladder is the second most common urological malignancy after prostate cancer. Recently, there has been increased interest in research of the role of free radicals and antioxidant materials in the prevention, treatment, and alleviation of therapy-related side effects of cancer. In the present study, we aimed to assess the alterations in the levels of antioxidant vitamins, activities of defense enzymes, circulating lipid peroxide, and total antioxidant activity (AOA) in patients with urothelial carcinoma of bladder and correlate these changes with the grade and severity of the disease.The study cohort consisted of 90 subjects; 50 patients with bladder UC (25, low grade; 10, high grade; 15, muscle invasive) and 40 healthy controls. Vitamins C and E, malondialdehyde (MDA), and AOA were estimated using standard protocols. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assayed using commercially available kits.The serum levels of vitamins C and E, whole blood levels of SOD and GPx, and serum AOA was significantly lower (P0.001) while serum MDA levels were significantly higher (P0.001) in patients than in controls, indicating presence of oxidative stress in bladder UC patients. The levels of all the biochemical parameters were correlated with the grade and severity of the disease. There were significant differences between the patients with low grade tumors and muscle invasive tumors for all parameters (P0.001); except AOA (P0.279).The observed redox imbalance in UC of bladder in correlation with the grade and stage, as a consequence of decreased levels of antioxidant vitamins, enzymes, and AOA, along with increased MDA levels in circulation, may be important factors in tumor development and growth. Our results suggest that with advancing stage of bladder UC, the levels of oxidative stress increase, while levels of antioxidant molecules decrease. These findings suggest possible use of antioxidant supplementation as prophylactic agents for prevention and treatment of bladder cancer.

Corrigendum to “Oxidative stress markers and antioxidant levels in normal pregnancy and pre-eclampsia” [Int J Gynecol Obstet 94 (2006) 23–27] J.B. Sharma ⁎, A. Sharma , A. Bahadur , N. Vimala , A. Satyam , S. Mittal a a Department of Obstetrics and Gynaecology, All-India Institute of Medical Sciences, New Delhi, India b Department of Biochemistry, All-India Institute of Medical Sciences, New Delhi, India

Background: The diagnosis of pemphigus vulgaris (PV) and pemphigus foliaceous (PF) rests upon clinical, histological and immunofluorescence features. Enzyme-linked immunosorbent assay (ELISA) test and immunoblot (IB) assay have shown variable sensitivity and specificity. Aims: We compared the utility of ELISA and IB in pemphigus patients. Methods: Sixty-six pemphigus cases (PV-54, PF-12) and 72 controls (other vesicobullous disorders and healthy controls) were inducted. ELISA for anti-Dsg 3 and 1 antibodies and IB assay were performed. Results: On ELISA, both mean anti-Dsg 1 and 3 titers were raised in PV and PF. Mean anti-Dsg 1 in mucocutaneous PV was significantly higher than in mucosal PV and mean anti-Dsg 3 was significantly raised in PV than in PF. Anti-Dsg 1 and 3 in the control group were negative. Sensitivity and specificity of ELISA in PV was 98.14% and 90.5% while in PF it was 91.6% and 61.1%, respectively.On IB in PV, 36 cases (66.67%) showed the 130 kDa and 160 kDa antigen bands, 12 (22.2%) only the 130 kDa and six (11.1%) only the 160 kDa band. Eight of the nine pure mucosal cases (88.8%) showed only the 130 kDa. In PF, only the 160 kDa antigen was detected. These antigens were not identified in the control group. Sensitivity and specificity of IB in PV was 88.9% and 100% and in PF it was 100% and 95.2%, respectively. Conclusion: Both tests could differentiate pemphigus from other dermatoses, including other blistering disorders. ELISA could not make a distinction between PV and PF or between the various clinical phenotypes of PV. IB differentiated between PV and PF and the different clinical variants of PV.