Pattern Of Proliferation Research Articles

A comparative study of osteopontin expression, Ki67 index and prognosis in squamous cell carcinoma and cysts of the oral cavity.

BackgroundRecent studies have shown a strong relationship between the expression of osteopontin and oral carcinogenesis. Osteopontin (OPN) has been shown to play a major role in regulating the aggressiveness of cancer cells and promote tumor growth. Odontogenic cysts are an essential aspect of oral and maxillofacial pathology. They are relatively frequent lesions with different clinical behavior. Some of them may have a proliferative pattern of growth and neoplastic nature. Evaluation of osteopontin expression with Ki-67 index may help examine clinical behavior and recurrence of oral squamous cell cancer and radicular cyst patients.MethodsA total of 44 oral cavity cancer cases and 21 cysts samples were analyzed by immunohistochemical staining. Data used for analysis were derived from medical records. The following information was obtained from all patients’ medical records: survival, age, sex, lymph node status, tumor size, and location, as well as grade and histologic type of tumor. Expression status of OPN and Ki-67 was statistically assessed.ResultsOur data demonstrated that for summary immunoreactive scores of OPN and Ki-67 expressions in OSCC vs. RC patients statistical significance was found for both markers’ between OSCC and RC groups. Moreover, osteopontin is significantly higher expressed in larger OSCC tumors.ConclusionsIn conclusion, the role of OPN expression both in oral squamous cancer cells and radicular cyst and possible correlation with demographic and clinicopathological features remain undetermined in some aspects, further high-powered studies to develop a more standardized assessment of Ki-67 and osteopontin expression in OSCC and are needed.

In vivo Oxygen Condition of Human Nasal Inferior Turbinate-Derived Stem Cells in Human Nose

Background and Objective: Human nasal inferior turbinate-derived stem cells (hNTSCs) have been considered as a potent and useful source for regenerative medicine. To most effectively mimic the native environment of inferior turbinate could be very effective to hNTSCs biology. Thus, the purpose of this study was to evaluate partial pressure of oxygen (ppO₂) and temperature in inferior turbinate. Methods: Ten patients were enrolled who underwent endoscopic endonasal transsphenoidal skull base tumor surgery between January 2014 and December 2015. The commercially available OxyLab pO2 monitor gauges the ppO₂ and temperature using a fluorescence quenching technique. Also, hNTSCs were isolated from 10 patients and cultivated under hypercapnic condition (5, 10, and 15%) to mimic hypoxic intranasal conditions. Results: The measured oxygen concentration in submucosa tissue was higher than that at the surface of the inferior turbinate and the temperature in submucosa tissue was higher than the value at the surface of inferior turbinate. The patterns of proliferation were significantly different according to hypercapnic cultivation conditions and there were statistically significant decreased proliferation rates after the exposure of higher CO₂ over a period of 5 days. Conclusions: Intranasal turbinate tissue showed the hypoxia state in concordance with the result of the other tissues or organs. However, indirectly induced hypoxia influenced the influence on the hNTSCs proliferation negatively. Further study is needed to mimic the real hypoxic state, but our results could be used to optimize the culture environment of hNTSCs, thereby producing the stem cells for regenerative therapies.

Tubule-derived lactate is required for fibroblast activation in acute kidney injury.

Acute kidney injury (AKI) is a highly prevalent medical syndrome associated with high mortality and morbidity. Several types of cells, including epithelial cells, vascular endothelial cells, pericytes, and macrophages, participate in the development of AKI. Recently, renal fibroblasts were found to play an important role in the regulation of tubular injury, repair, and recovery after AKI. However, the mechanisms underlying fibroblast activation and proliferation during the progression of AKI remain unclear. In the present study, we found many activated myofibroblasts located in the renal interstitium with an abundance of extracellular matrix deposition following folic acid-induced AKI. The proliferative pattern of tubular epithelial cells and interstitial cells following acute injury was different, indicating that the proliferation of fibroblasts followed the proliferation of tubular epithelial cells. Furthermore, we observed that proliferative tubular epithelial cells preferred aerobic glycolysis as the dominating metabolic pathway in the progression of AKI. Lactate generated from injured tubules was taken up by interstitial fibroblasts in the later stages of AKI, which induced fibroblast activation and proliferation in vitro. Early inhibition of lactate production in tubules by glycolytic inhibitors suppressed fibroblast activation after folic acid-induced injury. Collectively, these results support the important role of fibroblasts in the development of AKI and suggest that lactate produced by glycolysis in tubular epithelial cells is a novel regulator of fibroblast activation and proliferation.

Production, Structural Characterization, and In Vitro Assessment of the Prebiotic Potential of Butyl-Fructooligosaccharides.

Short-chain fatty acids (SCFAs), especially butyrate, produced in mammalian intestinal tracts via fermentation of dietary fiber, are known biofunctional compounds in humans. However, the variability of fermentable fiber consumed on a daily basis and the diversity of gut microbiota within individuals often limits the production of short-chain fatty acids in the human gut. In this study, we attempted to enhance the butyrate levels in human fecal samples by utilizing butyl-fructooligosaccharides (B-FOS) as a novel prebiotic substance. Two major types of B-FOS (GF3-1B and GF3-2B), composed of short-chain fructooligosaccharides (FOS) bound to one or two butyric groups by ester bonds, were synthesized. Qualitative analysis of these B-FOS using Fourier transform infrared (FT-IR) spectroscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), nuclear magnetic resonance (NMR) and low-resolution fast-atom bombardment mass spectra (LR-FAB-MS), showed that the chemical structure of GF3-1B and GF3-2B were [O-(1-buty-β-D-fru-(2→1)-O-β-D-fru-(2→1)-O-β-D-fru-O-α-D-glu] and [O-(1-buty)-β-D-fru-(2→1)-O-β-D-fru-(2→1)-O-(4-buty)-β-D-fru-O-α-D-glu], respectively. The ratio of these two compounds was approximately 5:3. To verify their biofunctionality as prebiotic oligosaccharides, proliferation and survival patterns of human fecal microbiota were examined in vitro via 16S rRNA metagenomics analysis compared to a positive FOS control and a negative control without a carbon source. B-FOS treatment showed different enrichment patterns on the fecal microbiota community during fermentation, and especially stimulated the growth of major butyrate producing bacterial consortia and modulated specific butyrate producing pathways with significantly enhanced butyrate levels. Furthermore, the relative abundance of Fusobacterium and ammonia production with related metabolic genes were greatly reduced with B-FOS and FOS treatment compared to the control group. These findings indicate that B-FOS differentially promotes butyrate production through the enhancement of butyrate-producing bacteria and their metabolic genes, and can be applied as a novel prebiotic compound in vivo.

The Impact of Preoperative Breast Magnetic Resonance Imaging on Surgical Management in Symptomatic Patients With Invasive Lobular Carcinoma.

Objective:Due to an insidious proliferative pattern, invasive lobular breast cancer (ILC) often fails to form a defined radiological or palpable lesion and accurate diagnosis remains challenging. This study aimed to determine the value of preoperative magnetic resonance imaging (MRI) for ILC and its impact on surgical outcomes.Methods:Consecutive symptomatic patients diagnosed with ILC in a tertiary centre over a 9-year period were reviewed. The time from diagnosis until surgery, initial type of surgery/index operation (breast-conserving surgery [BCS]/mastectomy) and the rates of reoperation (re-excision/completion mastectomy) were recorded. Patients were grouped into those who received conventional imaging and preoperative MRI (MR+) and those who received conventional imaging alone (MR–).Results:There were 218 cases of ILC, and 32.1% (n = 70) had preoperative MRI. Time from diagnosis to surgery was longer in the MR+ than the MR– group (32.5 vs 21.1 days, P < .001) even when adjusting for age and breast density. Initial BCS was performed on 71.4% (n = 50) of MR+ patients and 72.3% (n = 107) of the MR– group. While the rate of completion mastectomy following initial BCS was higher in the MR+ group (30.0%, n = 15 vs 14.0%, n = 15; χ2 = 5.63; P = .018), this association was not maintained in multivariable analysis. No difference was recorded in overall (initial and completion) mastectomy rate between the MR+ and MR– group (50.0%, n = 35 vs 37.8%, n = 56; χ2 = 2.89; P = .089). Margin re-excision following BCS was comparable between groups (8.0%, n =4, vs 9.3%, n = 10; χ2 = 0.076, P = .783) despite the selection bias for borderline conservable cases in the MR+ group. The rate of usage of MRI for ILC cases declined over the study period.Conclusion:While MRI was associated with minor delays in treatment and did not reduce overall rates of margin re-excision or completion mastectomy, it altered the choice of surgical procedure in almost a quarter of MR+ cases. The benefit of preoperative breast MRI appears to be confined to select (younger, dense breast, borderline conservable) cases in symptomatic ILC.

Supplementation with phytoestrogens and insoluble fibers reduces intestinal carcinogenesis and restores ER-β expression in Apc-driven colorectal carcinogenesis.

Supplementation with phytoestrogens and insoluble fibers has been reported to reduce duodenal polyps in colectomized familial adenomatous polyposis patients, with a mechanism involving, at least in part, upregulation of estrogen receptor-β subtype, whose expression is lowered during intestinal tumorigenesis. These data suggest a protective effect also in the colon, the main target organ for tumorigenesis in familial adenomatous polyposis and a major cancer type in non-familial (sporadic) cancers. Therefore, we tested whether a similar preparation might reduce tumorigenesis in the colon of Pirc rats (F344/NTac-Apc) mutated in the Apc gene and thus, like familial adenomatous polyposis patients, spontaneously developing multiple tumors in the colon. We first demonstrate that estrogen receptor-β expression in Pirc rat colon is significantly down-regulated compared to age-matched wt rats. Then, Pirc rats aged 1 month were treated for 3 months with Adipol (Adi), a patented preparation containing phytoestrogens and insoluble fibers. Colon tumorigenesis was significantly reduced by Adi treatment (colon tumors/rat were 5.3 ± 0.8 and 2.9 ± 0.3, Mucin Depleted Foci/rat 127 ± 6.6 and 97.1 ± 8.6 in Controls and Adi-treated rats, respectively, means ± SE, P < 0.01). The treatment also normalized colon proliferation pattern along the crypt and significantly increased apoptosis in colon tumors. Estrogen receptor-β expression was increased by Adi treatment, especially in the tumors. These positive effects suggest that Adipol may be exploited as a chemopreventive agent to reduce cancer risk in familial adenomatous polyposis patients and to postpone prophylactic colectomy. Moreover, given the similarities between familial adenomatous polyposis and sporadic colorectal cancer, it might also be used as chemopreventive agent in colorectal cancer patients at risk.

Mechanistic anticarcinogenic efficacy of phytofabricated gold nanoparticles on human lung adenocarcinoma cells

The present study deals with Plectranthus amboinicus (Lour.), multiple potential herbs perceived as a medicinal and culinary adjunct has been used to fabricate gold (Au) nanoparticles (NPs) in an eco-benign fashion. We have primly reported the antioxidant and anticarcinogenic effects of phytofabricated Au NPs towards lung adenocarcinoma (LAC) cells (A549 cell line) in vitro. The herbal concentrate, size and dosage of Au NPs were the key variables that determined the anticarcinogenic efficacy. Results indicated that the herbal concentrate encompassing flavonoids, alkaloids and polyphenolics aided reduction and stabilisation of Au NPs supported by spectral (λ max @ 550 nm) and quantitative analyses. The Fourier transform-infrared spectra anticipated distinct phenolics and protein frequencies involved in Au NPs capping in the course of fabrication. The manipulated Au NPs of 15 nm size has significantly influenced the radical scavenging (48.1 μg) and proliferation pattern of LAC cells (A549) associated with cytotoxic characteristics and IC50 (Au NPs concentration linked with 50% mortality) defined at 16.3 μg/mL in 48 h. This nanotoxicological prospect on the use of nanomaterials as an anticancer agent would be an alternative for the emerging intrinsic resistance of cells to the drug; a great challenge pertaining to safer environmental and biological outcomes.

Características Clínico-Patológicas e Tratamento do Ameloblastoma Unicístico: Relato de Caso

The ameloblastoma is an epithelial benign odontogenic tumor which is locally invasive, slow-growing and presents considerable recidive rates. It is considered the most clinically relevant benign odontogenic tumor, currently being classified as “ameloblastoma”, “unicystic ameloblastoma”, “peripheral or extraosseous ameloblastoma” and “metastatic ameloblastoma”. Unicystic type is the second most prevalent ameloblastoma, occurring in approximately 15% of all ameloblastomas, showing specific characteristics, especially in younger patients. The aim of this study is to clarify the characteristics of this type of ameloblastoma, presenting the case report of a young patient affected by the tumor in the jaw, highlighting clinical, imaginological, histopathological, therapeutic and prognostic aspects involved. In this case, the chosen treatment was, initially, the marsupialization of the lesion and posteriorly the enucleation with peripheral ostectomy and tooth extraction involved. This treatment option has shown good results, decreasing the patients damage when properly indicated, for this, an early and assertive diagnosis is essential, associating clinical and histopathological characteristics. The patient keeps on careful follow-up, since the proliferation pattern of the mural type requires greater attention given its greater possibility of recidive.

ORAL CARCINOGENESIS BIOMARKERS IN ORAL CHRONIC MECHANICAL IRRITATION-ASSOCIATED LESIONS

Previous works have shown clinical data suggesting that chronic mechanical irritation (CMI) could be a risk factor for oral cancer (OC). However, there are no known patterns of biomarkers in CMI indicative of cellular malignancy. Objective: The aim of this study was to identify a biomarker pattern of differentiation (Ck19) and cell proliferation (Ki67) in CMI-associated lesions similar to those observed in OC. Study Design: A cross-sectional study with adult patients (n = 61) of both sexes with lesions associated with CMI and with oral cancer were included. CK19 and Ki67 immunohistochemistry were carried out in specimens of both types of lesions, and the patterns were described according to previous publications. The present study was approved by Ethics Committee of the College of Dentistry, Universidad Nacional de Cordoba. Results: Ki67 overexpression and a CK19 focal pattern with strong intensity in basal and parabasal cells were observed in CMI lesions, similar to that observed in OC. Conclusion:There is no scientific literature on biomarker patterns of oral CMI. Therefore, the described pattern, particularly Ck19, is novel and could be indicative of the relation between CMI and oral carcinogenesis.

Does photobiomodulation change the synthesis and secretion of angiogenic proteins by different pulp cell lineages?

This study aimed to compare the synthesis and secretion of VEGF-A, VEGF-C, VEGF-D, VEGFR1, VEGFR2, and FGF-2 between pulp fibroblasts from human primary teeth (HPF) and stem cell from human deciduous teeth (SHED) before and after photobiomodulation. HPF were obtained from explant technique and characterized by immunohistochemistry, while SHED were obtained from digestion technique and characterized by flow cytometry. HPF (control group) and SHED were plated, let to adhere, and put on serum starvation to synchronize the cell cycles prior to photobiomodulation. Then, both cell lineages were irradiated with 660-nm laser according to the following groups: 2.5 and 3.7 J/cm2. MTT and crystal violet assays respectively verified viability and proliferation. ELISA Multiplex Assay assessed the following proteins: VEGF-A, VEGF-C, VEGF-D, VEGFR1, VEGFR2, FGF-2, at 6, 12, and 24 h after photobiomodulation, in supernatant and lysate. Two-way ANOVA/Tukey test evaluated cell viability and proliferation, while angiogenic production and secretion values were analyzed by one-way ANOVA (P < .05). Statistically similar HPF and SHED viability and proliferation patterns occurred before and after photobiomodulation (P > .05). HPF exhibited statistically greater values of all angiogenic proteins than did SHED, at all study periods, except for FGF-2 (supernatant; 12 h); VEGFR1 (lysate; non-irradiated; 12 h); and VEGFR1 (lysate; non-irradiated; 24 h). Photobiomodulation changed the synthesis and secretion of angiogenic proteins by HPF. HPF produced and secreted greater values of all tested angiogenic proteins than did SHED before and after irradiation with both energy densities of 2.5 and 3.7 J/cm2.

Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon.

Understanding the genetic basis for phenotypic differences is fundamental to the study of macroevolutionary patterns of biological diversity. While technological advances in DNA sequencing have made researching genetic variation in wild taxa routine, fully understanding how these variants affect phenotype requires taking the next step to investigate how genetic changes alter cell and tissue interactions that ultimately produce phenotypes. In this article, we investigate a role for cell proliferation as a developmental source of craniofacial diversity in a radiation of 3 species of Cyprinodon from San Salvador Island, Bahamas. Patterns of cell proliferation in the heads of hatching-age fish differ among species of Cyprinodon, and correlate with differences in allometric growth rate among the jaws of 3 distinct species. Regional patterns of cell proliferation in the head are complex, resulting in an unintuitive result in which lower levels of cell proliferation in the posterior head region are associated with the development of relatively larger jaws in one species. We combine these data with previously published morphological and genomic data to show how studying the mechanisms generating phenotype at the cellular and tissue levels of biological organization can help mechanistically link genomic studies with classic morphological studies.

Role of L. plantarum KX519413 as Probiotic and Acacia Gum as Prebiotic in Gastrointestinal Tract Strengthening.

Probiotics, prebiotics, and synbiotics are potential mediators to maintaining healthy intestinal flora and have garnered an area of wide research in the past few years. The current study assesses the in vivo effects of probiotic (Lactobacillus plantarum MBTU-HK1), prebiotic (acacia gum) (either singly or in combination as a synbiotic on growth performance), biochemical, hematological, physiological, and immunological effects and their role in the reduction of procarcinogen enzyme activities in male Balb/c mice. The absence of treatment-related toxicity and a normal physiological range of biochemical and hematological parameters ensure their safe consumption. The synbiotic group was found to possess lowered cholesterol levels and enhanced protein and mineral content. The probiotic and synbiotic groups reinforced immunoglobulin levels and had a modulatory effect on phagocytosis. A lymphocyte proliferation pattern suggested the stimulatory effect of synbiotic combination on splenocyte viability and proliferation. Total antioxidant capability in the liver was determined by a 2,2-diphenyl-1-picrylhydrazyl assay and all the treatment groups were found to possess increased scavenging activity. Synbiotic and prebiotic treatment was observed to lead to reduced tumor necrosis factor α (TNF-α) levels. Bacterial procarcinogenic fecal enzyme activities were found to be decreased, proving their role in the prevention of colon cancer incidence. This study proves the potency and safety of oral administration of L. plantarum MBTU-HK1 and acacia gum either individually or in combination.

14 Effects of long cooling periods of the ear skin on the isolation and cultivation of bovine fibroblasts for posterior use in cloning via nuclear transfer

Many animals with high genetic value have died suddenly without having their germplasm conserved invitro. To simulate tissue transportation from the farm to the laboratory and to determine how long after an animal’s death it is possible to isolate and cultivate fibroblasts to form a cryobank for further use in nuclear transfer, ears of eight bovine females were obtained on the moment of death and preserved for 30 days at 5°C in a refrigerator. On Days 0, 2, 4, 7, 14, 21, and 30 postmortem (named D0 to D30), fibroblast cultivation was performed on Dulbecco’s modification of minimum essential media (DMEM) medium with 10% of fetal bovine serum. When in cellular confluence, cells were cryopreserved in DMEM with 10% of dimethyl sulfoxide. The following analyses were performed: time that allowed cell culture, time until initial cell attachment and proliferation, time until confluence was reached (counting from when the first cells attached and proliferated around the biopsies until they were confluent), contamination rates, cell concentration on freezing moment, and cell viability through membrane integrity via flow cytometry (using annexin and propidium iodide to verify viable, necrotic, and apoptotic cells). Test of means and Tukey’s test were used to compare data at 5%. All time points allowed cell isolation and culture. Contamination was more prevalent on Days 14, 21, and 30. Remaining results are presented in Table 1. While D0 cells took 4 days to initiate attachment and proliferation around the biopsies, and another 24.0±2.0 days to reach confluence, D30 cells needed 33.5±1.5 days to attach and proliferate and another 31 days to reach cellular confluence, totaling ~64.5±1.5 days from the day of cultivation until freezing against a total of 28±2.0 days for D0 cells. Concentration dropped drastically on Days 14, 21, and 30 and so did cell viability on Days 21 and 30. It was concluded that cooling of the bovine ear skin at 5°C is an important strategy for transporting bovine tissue for long distances and obtaining viable cells up to 30 days after an animal’s death. However, the increase in cooling time interferes with the proliferation and viability cell patterns until and after cryopreservation. The first cloned embryos from D30 cells were transferred to recipient cows to investigate any differences in embryo viability between different days. This second phase of the experiment is still ongoing. Table 1.Mean and standard deviation of initial cell attachment/proliferation, time to confluence, concentration, and viability Day A/P (days) Confluence (days) Concentration (cell/mL) Viability (%) D0 4±0a 24±2a 1.334.375±131.375a 85.6±2.2a D2 5.33±1.5a 28.0±3.1a 1.645.050±252.639a 82.19±16.81a D4 4.33±1.03a 36±4b 1.571.656±234.462a 80.80±30.0a D7 10.0±1.78b 36±0b 1.509.400±347.860a 75.02±28.52a D14 19.6±2.19c 34.5±4.04b 698.125±131.203b 79.23±17.73a D21 24.5±3.5c 36±0b 304.166±47.604c 27.46±12.46b D30 33.5±1.5d 31±0b 311.250±0c 28.71±13.9b a-dDifferent letters represent statistically significant differences (P&amp;lt;0.05).

Etosis, rather than apoptosis or cell proliferation, typifies thrombus progression – An immunohistochemical study of coronary aspirates

BackgroundCoronary thrombosis is a process with unpredictable clinical outcome. Changes of thrombus composition overtime influence tissue repair and stabilization. We investigated rates of cell deaths and cell proliferation at different time points after initiation of thrombosis. MethodsThrombectomy aspirates of 55 myocardial infarction patients were selected and histomorphologically classified as fresh (25), lytic (25), partially fibrocellular (10), completely fibrocellular (10). Paraffin sections were immunostained with anti-(cleaved) caspase-3/Casp3 (apoptosis), Citrullinated histone/CitH 3 (etosis), C-reactive protein/CRP and Ki67 (proliferation) in combination with either Feulgen counterstaining (DNA) or cell markers for granulocytes, macrophages, SMCs, platelets and endothelium. Rates of apoptosis, etosis and proliferation were measured as a percentage of total number of immunopositive pixels versus total number of DNA positive pixels, while co-localization with cell markers was assessed by digital image analysis. ResultsPositive staining of CitH3 was observed more frequently (93%) than Casp3 (70%), Ki67 (79%) or CRP (59%) (p < 0.05). Moreover, rate of etosis, found in granulocytes and macrophages, differed significantly among thrombi of different age, being higher in lytic (12.82) than in fresh (8.52) and late-organized (2.75) (p < 0.05). Such differences were not observed for the rates of apoptosis or cell proliferation related to thrombus age. CRP staining was present in fresh, lytic and organized thrombi, but did not reliably identify necrotic areas. ConclusionsDifferent patterns of cell death and cell proliferation are noticed during progression of coronary thrombus overtime, but with significant differences for only etosis. Etosis could potentially serve as a biomarker for thrombus instability with clinical significance.

ZNF802 (JAZF1), a Possible New Therapeutic Target for Treatment of β-Thalassemia

β-Thalassemias are groups of inherited blood disorders caused by reduced or absent production of β-globin chains of adult hemoglobin (HbA; α2β2). The severely affected patients with β-thalassemia require lifelong blood transfusion and chelation therapy. Reactivation of fetal hemoglobin (HbF; α2γ2) could ameliorate the symptoms of patients via increased total hemoglobin production and reduced imbalanced α- to non-α-globin synthesis. Hemoglobin switching during human development requires activities of several transcription factor proteins, including those bind to the globin gene promoters. However, the regulation of fetal to adult hemoglobin switching is not fully understood. Previously, we have carried out the microarray analysis comparing human erythroblasts derived from therapeutic aborted fetal liver and adult peripheral blood. The results showed that ZNF802 mRNA is among the transcripts that up-regulated in adult erythroblast of greater than 2-fold change. ZNF802, also called JAZF1, is a 27 kDa protein containing two zinc finger motifs that acting as transcriptional repressor by binding to the direct repeat (DR) elements on DNA. In this study, quantitative RT-PCR was performed in order to validate the microarray data. We observed 13-fold up-regulation of ZNF802 in adult erythroid cells compared to those cells derived from fetal liver. To further elucidate the role of ZNF802 in reduction of HbF expression in adult erythroids, the lentivirus carrying ZNF802 specific shRNA and non-targeted shRNA were designed and constructed for transduction in erythroblasts. Hematopoietic CD34+ progenitor cells were isolated from β-thalassemia/HbE patients (n=9) and subjected to ex vivo two-phase liquid erythroid culture. The results showed the reduction of greater than 90% of ZNF802 transcript and nearly undetectable level of ZNF802 protein in cells transduced with the ZNF802 shRNA lentivirus. As expected, the level of γ-globin transcript and levels of HbF were increased in ZNF802 knockdown erythroid cells in β-thalassemia/HbE (5-fold changed and HbF 26.9±7.4% vs 18.5±6.8% in non-targeted shRNA (p-value = 0.04), respectively). Interestingly, we found up-regulation of the embryonic ε-globin and ζ-globin transcripts in knockdown ZNF802 cells compared to non-targeted shRNA control (15-fold and 10-fold changed (p-value = 0.01), respectively). To note, the diminishing of ZNF802 effect on the proliferation and differentiation pattern of erythriod cells was not found. Altogether, our findings suggested that ZNF802 may play a role as the transcriptional repressor of γ-globin gene in human adult erythroid cells, thus served as a possible new therapeutic target for treatment of β-thalassemia. The molecular mechanisms underlying the ZNF802 mediated HbF reduction are underway. Disclosures No relevant conflicts of interest to declare.

Differential expression of pro-inflammatory and anti-inflammatory genes of layer chicken bursa after experimental infection with infectious bursal disease virus

Infectious bursal disease (IBD) is one of the most prevalent infectious diseases caused by IBD virus (IBDV), which results in bursal necrosis and immunosuppression that cause severe damage to the immune system in chickens. Cytokines are important mediators and regulators of both types of host responses. In the present study, layer chickens were artificially challenged with IBDV, and the differential expression of inflammatory genes was explored by using quantitative real-time PCR, which offered basic data for further study of IBDV pathogenesis. Data showed that after IBDV infection, the virus load in the bursa of Fabricius (BF) peaked at 96h and then gradually decreased. Compared with those of the negative-infected group, the mRNA expression levels of pro-inflammatory cytokines (interleukin [IL]-1β, IL-6, IL-7, IL-8, tumor necrosis factor [TNF]-α, transforming growth factor [TGF]-β) and anti-inflammatory cytokine IL-10 in the infected group increased to varying degrees at 12 to 192h, respectively. Furthermore, the IL-1β mRNA expression peaked at 48h; the mRNA transcript levels of IL-6, IL-8, and IL-10 were the highest at 96h; TNF-α mRNA expression peaked at 120h; the IL-7 mRNA expression peaked at 144h; and the TGF-β mRNA transcript level was the highest at 192h. Taken together, these observations indicated that along with the change pattern of IBDV proliferation in BF, the mRNA expression of cytokines (IL-1β, IL-6, IL-7, IL-8, IL-10, TNF-α, TGF-β) obviously increased, and the kinetics of each of these cytokines was different. The kinetics of IL-6/IL-10 mRNA expression ratio was significantly positively correlated with that of the virus load. These results suggest that IBDV infection seriously interferes with the natural immune response mediated by inflammatory cytokines in chickens.

Thalidomide Reduces Cell Proliferation in Endometriosis Experimentally Induced in Rats.

To analyze the effect of thalidomide on the progression of endometriotic lesions experimentally induced in rats and to characterize the pattern of cell proliferation by immunohistochemical Proliferating Cell Nuclear Antigen (PCNA) labeling of eutopic and ectopic endometrium. Fifteen female Wistar rats underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium and fixation of a tissue segment to the pelvic peritoneum. Four weeks after, the animals were divided into 3 groups: control (I), 10mg/kg/day (II) and 1mg/kg/day (III) intraperitoneal thalidomide for 10 days. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma analysis. Eutopic and ectopic endometrial tissue was submitted to immunohistochemistry for analysis of cell proliferation by PCNA labeling and the cell proliferation index (CPI) was calculated as the number of labeled cells per 1,000 cells. Group I showed a mean CPI of 0.248 ± 0.0513 in the gland and of 0.178 ± 0.046 in the stroma. In contrast, Groups II and III showed a significantly lower CPI, that is, 0.088 ± 0.009 and 0.080 ± 0.021 for the gland (p < 0.001) and 0.0945 ± 0.0066 and 0.075 ± 0.018 for the stroma (p < 0.001), respectively. Also, the mean lesion area of Group I was 69.2 mm2, a significantly higher value compared with Group II (49.4 mm2, p = 0.023) and Group III (48.6 mm2, p = 0.006). No significant difference was observed between Groups II and III. Thalidomide proved to be effective in reducing the lesion area and CPI of the experimental endometriosis implants both at the dose of 1 mg/kg/day and at the dose of 10 mg/kg/day.

Contribution to characterization of skin field cancerization activity: morphometric, chromatin texture, proliferation, and apoptosis aspects

BackgroundA skin field cancerization is a cutaneous area with subclinical changes resultant from chronic sun exposure, with a higher predisposition to development of pre-neoplastic and neoplastic lesions. So far, there are no well-defined objective parameters that can indicate their degree of activity. ObjectivesTo describe and compare morphometric aspects and expression of factors related to apoptosis and cell proliferation in actinic keratosis (AK), in both photoexposed and photoprotected epidermis. MethodsA cross-sectional study of patients with actinic keratosis in the forearms, biopsied at two points: the actinic keratosis and the axillary region. The biopsies of the actinic keratosis, perilesional area, and axilla were evaluated through keratinocyte intraepithelial neoplasia (KIN), and immunohistochemistry of p53, survivin, and Ki67. Nuclear morphometry of basal layer cells was performed through digital image analysis: entropy, area, perimeter, Ra, fractal dimension, circularity, color intensity, and largest diameter. ResultsThere were 13 patients included and 38 actinic keratosis biopsied. In morphometry, 1039 nuclei were analyzed, of which 228 represented axillary skin, 396 demonstrated actinic keratosis, and 415 represented the perilesional area to the actinic keratosis. There was a significant difference (p<0.05) in all variables tested for the topographies evaluated. A significant correlation was identified between nucellar morphometric elements, KIN, proliferation markers, and apoptosis. Joint patterns of p53, Ki67, and KIN discriminated the topographies sampled. Study limitationsThis was a cross-sectional study with a small number of patients. ConclusionsThere are patterns of proliferation, resistance to apoptosis, and different cellular morphometrics between photoprotected skin and photoexposed skin. The joint expression of p53, Ki67, and KIN can characterize skin field cancerization activity.

Scrutiny of a RCC Beam by Ansys with Non-Identical Collapsed Mechanisms under Assorted Loading Conditions

Scrutiny of reinforced concrete beam by Ansys with non-identical collapsed mechanisms under assorted conditions. As per is code: IS: 456-2000. to evaluation for finite element examination a beam element various kinds of authors are done their research by using software’s like Ansys, abacus, civil FEM, mat-lab from this one they are studied based on various types of work flows regarding fracture proliferation patterns, in this journal i would like to do numerical examination by FEA software as well as experimental work repercussion analogy from data source. Potential of work is based on the design conditions of design of singly reinforced beam doubly reinforced concrete beam.

Building Biological Flashlights: Orthogonal Luciferases and Luciferins for in Vivo Imaging.

Bioluminescence is widely used for real-time imaging in living organisms. This technology features a light-emitting reaction between enzymes (luciferases) and small molecule substrates (luciferins). Photons produced from luciferase-luciferin reactions can penetrate through heterogeneous tissue, enabling readouts of physiological processes. Dozens of bioluminescent probes are now available and many are routinely used to monitor cell proliferation, migration, and gene expression patterns in vivo. Despite the ubiquity of bioluminescence, traditional applications have been largely limited to imaging one biological feature at a time. Only a handful of luciferase-luciferin pairs can be easily used in tandem, and most are poorly resolved in living animals. Efforts to develop spectrally distinct reporters have been successful, but multispectral imaging in large organisms remains a formidable challenge due to interference from surrounding tissue. Consequently, a lack of well-resolved probes has precluded multicomponent tracking. An expanded collection of bioluminescent probes would provide insight into processes where multiple cell types drive physiological tasks, including immune function and organ development. We aimed to expand the bioluminescent toolkit by developing substrate-resolved imaging agents. The goal was to generate multiple orthogonal (i.e., noncross-reactive) luciferases that are responsive to unique scaffolds and could be used concurrently in living animals. We adopted a parallel engineering approach to genetically modify luciferases to accept chemically modified luciferins. When the mutants and analogs are combined, light is produced only when complementary enzyme-substrate partners interact. Thus, the pairs can be distinguished based on substrate selectivity, regardless of the color of light emitted. Sequential administration of the luciferins enables the unique luciferases to be illuminated (and thus resolved) within complex environments, including whole organisms. This Account describes our efforts to develop orthogonal bioluminescent probes, crafting custom luciferases (or "biological flashlights") that can selectively process luciferin analogs (or "batteries") to produce light. In the first section, we describe synthetic methods that were key to accessing diverse luciferin architectures. The second section focuses on identifying complementary luciferase enzymes via a combination of mutagenesis and screening. To expedite the search for orthogonal enzymes and substrates, we developed a computational algorithm to sift through large data sets. The third section features examples of the parallel engineering approach. We identified orthogonal enzyme-substrate pairs comprising two different classes of luciferins. The probes were vetted both in cells and whole organisms. This expanded collection of imaging agents is applicable to studies of immune function and other multicomponent processes. The final section of the Account highlights ongoing work toward building better bioluminescent tools. As ever-brighter and more selective probes are developed, the frontiers of what we can "see" in vivo will continue to expand.