Overcoming effector T cell exhaustion in ovarian cancer ascites with a novel adenovirus encoding for a MUC1 bispecific antibody engager and IL-2 cytokine.

T cell-focused cancer immunotherapy including checkpoint inhibitors and cell therapies has been rapidly evolving over the past decade. Nevertheless, there remains a major unmet medical need in oncology generally and immuno-oncology specifically. We have constructed an oncolytic adenovirus, Ad5/3-E2F-d24-aMUC1aCD3-IL-2 (TILT-322), which is armed with a human aMUC1aCD3 T cell engager and IL-2. TILT-322 treatment stimulated T cell cytotoxicity through the increased presence of granzyme B, perforin, and interferon-gamma. Additional immune profiling indicated TILT-322 increased gamma delta T cell activation and impacted other cell types such as natural killer cells and natural killer-like T cells that are traditionally involved in cancer immunotherapy. TILT-322 treatment also decreased the proportion of exhausted CD8+ T cells as demarked by immune checkpoint expression in ovarian ascites samples. Overall, our data showed that TILT-322 treatment led to an enhanced T cell activation and reversed T cell exhaustion translating into high antitumor efficacy when given locally or intravenously. The analysis of blood and tumors isolated from an in vivo patient-derived ovarian cancer xenograft model suggested TILT-322 mediated tumor control through improved T cell functions. Therefore, TILT-322 is a promising novel anti-tumor agent for clinical translation.

GNAS mutation inhibits growth and induces phosphodiesterase 4D expression in colorectal cancer cell lines.

Approximately 5% of colorectal cancers (CRCs) have a gain-of-function mutation in the GNAS gene, which leads to the activation of cAMP-dependent signaling pathways and associates with poor prognosis. We investigated the effect of an activating GNAS mutation in CRC cell lines on gene expression and cell proliferation in vitro, and tumor growth in vivo. GNAS-mutated (GNASmt) HCT116 cells showed stimulated synthesis of cAMP as compared to parental (Par) cells. The most upregulated gene in the GNASmt cells was cAMP-hydrolyzing phosphodiesterase 4D (PDE4D) as detected by RNA sequencing. To further validate our finding, we analyzed PDE4D expression in a set of human CRC tumors (n = 35) and demonstrated overexpression in GNAS mutant CRC tumors as compared to GNAS wild-type tumors. The GNASmt HCT116 cells proliferated more slowly than the Par cells. PDE4 inhibitor Ro 20-1724 and PDE4D subtype selective inhibitor GEBR-7b further suppressed the proliferation of GNASmt cells without an effect on Par cells. The growth inhibitory effect of these inhibitors was also seen in the intrinsically GNAS-mutated SK-CO-1 CRC cell line having high levels of cAMP synthesis and PDE4D expression. In vivo, GNASmt HCT116 cells formed smaller tumors than the Par cells in nude mice. In conclusion, our findings demonstrate that GNAS mutation results in the growth suppression of CRC cells. Moreover, the GNAS mutation-induced overexpression of PDE4D provides a potential avenue to impede the proliferation of CRC cells through the use of PDE4 inhibitors.

Modern perspectives of heavy metals alleviation from oil contaminated soil: A review.

Heavy metal poisoning of soil from oil spills causes serious environmental problems worldwide. Various causes and effects of heavy metal pollution in the soil environment are discussed in this article. In addition, this study explores new approaches to cleaning up soil that has been contaminated with heavy metals as a result of oil spills. Furthermore, it provides a thorough analysis of recent developments in remediation methods, such as novel nano-based approaches, chemical amendments, bioremediation, and phytoremediation. The objective of this review is to provide a comprehensive overview of the removal of heavy metals from oil-contaminated soils. This review emphasizes on the integration of various approaches and the development of hybrid approaches that combine various remediation techniques in a synergistic way to improve sustainability and efficacy. The study places a strong emphasis on each remediation strategy that can be applied in the real-world circumstances while critically evaluating its effectiveness, drawbacks, and environmental repercussions. Additionally, it discusses the processes that reduce heavy metal toxicity and improve soil health, taking into account elements like interactions between plants and microbes, bioavailability, and pollutant uptake pathways. Furthermore, the current study suggests that more research and development is needed in this area, particularly to overcome current barriers, improve our understanding of underlying mechanisms, and investigate cutting-edge ideas that have the potential to completely transform the heavy metal clean up industry.

Menadione sodium bisulfite neutralizes chromium phytotoxic effects in okra by regulating cytosolutes, lipid peroxidation, antioxidant system and metal uptake.

Chromium (Cr) is a major abiotic stress for plant species that significantly impacted plant development and impeded agricultural production. Menadione sodium bisulfite (MSB) has recently manifested a remarkable role in modulating plant defense responses. In the present experiment, Cr caused a significant decrease in growth, relative water contents, and chlorophyll in okra cultivars (Shabnam 786 and Arka Anamika). Cr produced an increase in proline, total soluble proteins, total free amino acids, phenolics, flavonoids, ascorbic acid, hydrogen peroxide (H2O2), malondialdehyde (MDA), and Cr accumulation. Besides, activities of antioxidant enzymes were also higher in Cr-stressed plants. MSB application (50, 100, 150, and 200 µM) profoundly impacted growth and important physio-biochemical characteristics in okra under Cr stress. Better growth in MSB treated plants was associated with lower oxidative damage and better oxidative defense system reflected in the form of higher antioxidant enzyme activities alongside the concentrations of non-enzymatic antioxidant compounds. In this background, cv. Shabnam-786 exhibited greater Cr tolerance over Arka Anamika. The degree of oxidative damage measured in the form of H2O2 and MDA was greater in cv. Arka Anamika. Lower MSB levels (50 and 100 µM) circumvented inhibitory Cr effects in okra, while higher doses proved lethal for plant growth and development.

Frequency and assessment of Nutritional Status of school going children in rural areas of Islamabad.

BACKGROUND AND OBJECTIVES: Nutritional status is considered a significant and positive health indicator. It determines anthropometric measurements of preschool children, the height of children at the time of school entry and prevalence of low birth weight. The objective of the study was to determine the frequency of nutritional status and socio-demographic factors influencing under nutrition among school children of rural Islamabad. METHODS: A cross-sectional study was conducted among school children of the age (4-16 years) from January 2017 to September 2019. The Sample size was 1710. Schools were selected through convenient sampling technique. Frequency and percentages were calculated and inferential statistics were computed to analyze the association of health status with categorical variables by using chi-square by keeping the level of significance <0.05 through SPSS version 20. RESULTS: The mean age of the sample was 9.38 ± 4.14 with the maximum number of children (49.1%) in the age bracket of 5-9. Out of 1710 children, 54.4% had normal weight for age, 25.3% were underweight, 7.5% overweight and 12.8% were found to be obese. Stunting was found to be 26%. Prevalence of being underweight was higher than overweight /obesity particularly in younger and higher age groups as indicated by p-value of 0.000. Comparing with females, male students had significantly higher frequency of being underweight and stunted as reflected by p-value of 0.004 and 0.000 respectively. Univariate analysis also showed a strong association between age and nutritional status as mean weight increased from 39.22 ±5.21 to 63.50± 4.66 and height from 35.67±5.76 to 113.73± 29.22 with advancing age. CONCLUSIONS: Undernutrition remains an ongoing health problem in school going children of rural Islamabad; particularly in male students of younger age groups. School health programs and nutritional interventions need to be strengthened particularly in rural areas of Islamabad.

Therapeutic potential of medicinal plants for the management of scabies.

Sarcoptes scabiei (S. scabiei), a parasite mite which causes scabies disease resulting in serious public health concern. The long-term scabies disease can lead to complications such as septicemia, acute post-streptococcal glomerulonephritis, heart disease, and secondary infections. Timely treatment to the affected patients is required to control the disease and get rid of the causative agent. Delayed diagnosis and inappropriate treatment can lead to serious consequences. The most common treatment strategy is the use of allopathic medicines which can immediately relieve the patient but have the drawback of side effects. The safe and cost-effective alternative treatment strategy is the use of medicinal plants which have beneficial therapeutic potential against variety of diseases due to the presence of many bioactive phytoconstituents with no or minimal side effects. For the present review, the published articles describing scabies disease and its phytotherapeutic modalities were searched through different data bases including Google Scholar, PubMed, Medline, and ScienceDirect using the keywords like S. scabiei, prevalence of scabies disease, and phytotherapy of scabies. A large number of medicinal plants, such as Melaleuca alternifolia, Curcuma longa, Azadirachta indica, Rosmarinus officinalis, Capsicum annuum, Cinnamomum camphor, Solanum nigrum, and Eupatorium perfoliatum, have been reviewed for the promising future treatments of scabies. All the studied plants have many bioactive compounds with potential therapeutic effects against scabies and can be utilized for therapeutic purposes for this disease. This literature study has limitations because of the lack of sufficient data due to limited pre-clinical trials in this particular area. This review provides a baseline to explore the therapeutic potential of these medicinal plants against skin diseases. However, extensive studies are required to identify, authenticate, and characterize the bioactive compounds present in these plants which may lead to value addition in pharmaceutical industries providing the cost-effective way of treatment with minimal side effects.

Molecular Dynamic Simulations to Probe Stereoselectivity of Tiagabine Binding with Human GAT1.

The human gamma aminobutyric acid transporter subtype 1 (hGAT1) located in the nerve terminals is known to catalyze the neuronal function by the electrogenic reuptake of γ-aminobutyric acid (GABA) with the co-transport of Na+ and Cl- ions. In the past, there has been a major research drive focused on the dysfunction of hGAT1 in several neurological disorders. Thus, hGAT1 of the GABAergic system has been well established as an attractive target for such diseased conditions. Till date, there are various reports about stereo selectivity of -COOH group of tiagabine, a Food and Drug Administration (FDA)-approved hGAT1-selective antiepileptic drug. However, the effect of the stereochemistry of the protonated -NH group of tiagabine has never been scrutinized. Therefore, in this study, tiagabine has been used to explore the binding hypothesis of different enantiomers of tiagabine. In addition, the impact of axial and equatorial configuration of the-COOH group attached at the meta position of the piperidine ring of tiagabine enantiomers was also investigated. Further, the stability of the finally selected four hGAT1-tiagabine enantiomers namely entries 3, 4, 6, and 9 was evaluated through 100 ns molecular dynamics (MD) simulations for the selection of the best probable tiagabine enantiomer. The results indicate that the protonated -NH group in the R-conformation and the -COOH group of Tiagabine in the equatorial configuration of entry 4 provide maximum strength in terms of interaction within the hGAT1 binding pocket to prevent the change in hGAT1 conformational state, i.e., from open-to-out to open-to-in as compared to other selected tiagabine enantiomers 3, 6, and 9.

Adenovirus Coding for Interleukin-2 and Tumor Necrosis Factor Alpha Replaces Lymphodepleting Chemotherapy in Adoptive T Cell Therapy.

Lymphodepleting preconditioning with high-dose chemotherapy is commonly used to increase the clinical efficacy of adoptive T cell therapy (ACT) strategies, however, with severe toxicity for patients. Conversely, oncolytic adenoviruses are safe and, when engineered to express interleukin-2 (IL-2) and tumor necrosis factor alpha (TNF-α), they can achieve antitumor immunomodulatory effects similar to lymphodepletion. Therefore, we compare the safety and efficacy of such adenoviruses with a cyclophosphamide- and fludarabine-containing lymphodepleting regimen in the setting of ACT. Human adenovirus (Ad5/3-E2F-D24-hTNF-α-IRES-hIL-2; TILT-123) replication was studied using a Syrian hamster pancreatic tumor model (HapT1) infused with tumor-infiltrating lymphocytes (TILs). Using the oncolytic virus instead of lymphodepletion resulted in superior efficacy and survival. Immune cells responsive to TNF-α IL-2 were studied using an immunocompetent mouse melanoma model (B16.OVA) infused with ovalbumin-specific T (OT-I) cells. Here, the adenovirus approach improved tumor control together with increased intratumoral Th1 cytokine levels and infiltration of CD8+ T cells and CD86+ dendritic cells. Similar to humans, lymphodepleting preconditioning caused severe cytopenias, systemic inflammation, and damage to vital organs. Toxicity was minimal in adenovirus- and OT-I-treated mice. These findings demonstrate that ACT can be effectively facilitated by cytokine-coding adenovirus without requiring lymphodepletion, a rationale being clinically investigated.

Ex-vivo abortifacient activity of Androsace foliosa n-hexane leaves extract on isolated rabbit uterus.

Androsace foliosa is a medicinal herb utilized in different areas of Pakistan for abortifacient, diabetic and liver complications. In the current research, the possible action of the n-hexane leaves extract of the Androsace foliosa on isolated rabbit uterus was examined. Abortifacient activity was examined in the existence of standard antagonist e.g. atropine and salbutamol and a uterine tonic like oxytocin. The isolated rabbit uterus is initially treated with 1mg/kg stilboesterol for 1 complete day. The consequence of oxytocin as uterine contraction agonist was observed. Additionally, antagonists e.g. salbutamol (2µg) and atropine (1-2mg) on the uterine contractile action of the extract were also examined. The A. foliosa n-hexane leaves extract fashion dose correlated amplification in the force of uterine contraction comparable to oxytocin. The drug oxytocin was pragmatic to amplify the uterine contractile action of the extract. Meanwhile pre-treating the tissue with either atropine or salbutamol earlier than administrating the extract indicates the inhibitory action of the drugs on the action of the extract.

Evidence of cognitive flexibility deficits and impaired emotion regulation in childhood survivors of acute lymphoblastic leukaemia: Findings from task switching study.

The objective of the current study was to compare cognitive flexibility and emotion regulation between childhood survivors of Acute Lymphoblastic Leukaemia (ALL) and healthy control subjects. Twenty-five childhood survivors of ALL treated with intrathecal chemotherapy between 2013 to 2016 from Shaukat Khanum Memorial Cancer Hospital, Children Hospital and Jinnah Hospital Lahore and twenty-five healthy demographically matched children (control group) participated in the study. Participants performed task switching experiment as a measure of cognitive flexibility and emotion regulation questionnaire for children and adolescents. In contrast to healthy control group, ALL survivors showed impaired cognitive flexibility and emotion regulation. Results have implications for cognitive rehabilitation to improve functions of frontal-cerebellar system in childhood survivors of ALL.

Zinc oxide nanoparticles and Klebsiella sp. SBP-8 alleviates chromium toxicity in Brassica juncea by regulation of antioxidant capacity, osmolyte production, nutritional content and reduction in chromium adsorption.

Heavy metals are one of the most damaging environmental toxins that hamper growth of plants. These noxious chemicals include lead (Pb), arsenic (As), nickel (Ni), cadmium (Cd) and chromium (Cr). Chromium is one of the toxic metal which induces various oxidative processes in plants. The emerging role of nanoparticles as pesticides, fertilizers and growth regulators have attracted the attention of various scientists. Current study was conducted to explore the potential of zinc oxide nanoparticles (ZnONPs) alone and in combination with plant growth promoting rhizobacteria (PGPR) Klebsiella sp. SBP-8 in Cr stress alleviation in Brassica juncea (L.). Chromium stress reduced shoot fresh weight (40%), root fresh weight (28%), shoot dry weight (28%) and root dry weight (34%) in B. juncea seedlings. Chromium stressed B. juncea plants showed enhanced levels of malondialdehyde (MDA), electrolyte leakage (EL), hydrogen peroxide (H2O2) and superoxide ion (O2• -). However, co-supplementation of ZnONPs and Klebsiella sp. SBP-8 escalated the activity of antioxidant enzymes i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) in B. juncea grown in normal and Cr-toxic soil. It is further proposed that combined treatment of ZnONPs and Klebsiella sp. SBP-8 may be useful for alleviation of other abiotic stresses in plants.

The Programmed Cell Death Ligand 1 and Lipocalin 2 Expressions in Primary Breast Cancer and Their Associations with Molecular Subtypes and Prognostic Factors.

Purpose: Breast cancers exhibit molecular heterogeneity, leading to diverse clinical outcomes and therapeutic responses. Immune checkpoint inhibitors targeting PD-L1 have shown promise in various malignancies, including breast cancer. Lipocalin 2 (LCN2) has also been associated with tumor aggressiveness and prognostic potential in breast cancers. However, the expression of PD-L1 and LCN2 in breast cancer subtypes and their prognostic implications remains poorly investigated. Methods: A retrospective analysis of 89 primary breast cancer cases was conducted to assess PD-L1 and LCN2 expressions using immunohistochemistry. Cases were classified into four different molecular subtypes based on ER, PR, HER2, and Ki-67 status. Associations between PD-L1 and LCN2 expressions and various prognostic factors were examined. Results: Although low expression of LCN2 (Allred score of <3) was observed even in normal breast tissue, LCN2 expression with increasing Allred score (≥3) positively correlated with the histological grade, high Ki-67 proliferation index, and ER/PR negativity. Significant elevations of LCN2 and PD-L1 expressions were observed in triple-negative and HER2-positive breast cancers. Conclusion: The results of the study highlight the association of LCN2 with known prognostic factors and molecular subtypes. To identify potential immunotherapy recipients, it would be useful to evaluate LCN2 as well as PD-L1 immune targets in different subgroups of breast cancer patients. Further studies with larger patient numbers are warranted to validate these observations and establish standardized scoring criteria for LCN2 expression assessment.

Calcium oxide nanoparticles mitigate lead stress in Abelmoschus esculentus though improving the key antioxidative enzymes, nutritional content and modulation of stress markers.

Lead (Pb) is thought to be one of most injurious metals on the earth. Lead stress in plants enhances synthesis of highly toxic reactive oxygen species (ROS). During present research, impact of calcium-oxide nanoparticles (CaO-NPs) was observed on antioxidative defense mechanism in Abelmoschus esculentus plants prone to Pb stress. A CRD experiment was employed with 5 replicates having four treatments (T0 = Control, T1 = Pb stress (200 ppm), T2 = CaO-NPs and T3 = Pb + CaO-NPs). Pb-stressed seedlings exhibited decreased root growth, shoot growth, chlorophyll concentration and biomass accumulation. Moreover, higher synthesis of hydrogen-peroxide (H2O2), malondialdehyde (MDA) and electrolyte leakage (EL) resulting in cellular injuries were noted in plants growing in Pb spiked conditions. Similarly, stressed plants showed higher accumulation of total soluble sugar and proline content besides elevated activity of antioxidative enzymes counting catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX). On the contrary side, CaO-NPs alleviated the Pb induced phytotoxicity through improving activity of antioxidative enzymes. The elevated activity of antioxidant enzymes reduced biosynthesis of H2O2 and MDA which was revealed through the increased growth parameters. In addition, CaO-NPs persuaded enhancement in plant defence machinery by decreased chlorophyll deprivation and augmented the uptake of plant nutrients including K and Ca content. Hence, CaO-NPs can be potent regulators of the antioxidative enzymes and stress markers to ameliorate abiotic stresses.

Disruption of KLHL6 Fuels Oncogenic Antigen Receptor Signaling in B-cell Lymphoma.

Pathomechanisms that activate oncogenic B-cell receptor (BCR) signaling in diffuse large B-cell lymphoma (DLBCL), are largely unknown. Kelch-like family member 6 (KLHL6) encoding a substrate-adapter for Cullin-3-RING E3 ubiquitin-ligase (CRL) with poorly established targets is recurrently mutated in DLBCL. By applying high-throughput protein interactome screens and functional characterization, we discovered that KLHL6 regulates BCR by targeting its signaling subunits CD79A and CD79B. Loss of physiological KLHL6 expression pattern was frequent among the MCD/C5-like activated B-cell DLBCLs and was associated with higher CD79B levels and dismal outcome. Mutations in the BTB domain of KLHL6 disrupted its localization and heterodimerization, and increased surface BCR levels and signaling, whereas Kelch domain mutants had the opposite effect. Malfunctions of KLHL6 mutants extended beyond proximal BCR signaling with distinct phenotypes from KLHL6 silencing. Collectively, our findings uncover how recurrent mutations in KLHL6 alter BCR signaling and induce actionable phenotypic characteristics in DLBCL.

Transgenerational Seed Exposure to Elevated CO2 Involves Stress Memory Regulation at Metabolic Levels to Confer Drought Resistance in Wheat.

Drought is the worst environmental stress constraint that inflicts heavy losses to global food production, such as wheat. The metabolic responses of seeds produced overtransgenerational exposure to e[CO2] to recover drought's effects on wheat are still unexplored. Seeds were produced constantly for four generations (F1 to F4) under ambient CO2 (a[CO2], 400 µmol L-1) and elevated CO2 (e[CO2], 800 µmol L-1) concentrations, and then further regrown under natural CO2 conditions to investigate their effects on the stress memory metabolic processes liable for increasing drought resistance in the next generation (F5). At the anthesis stage, plants were subjected to normal (100% FC, field capacity) and drought stress (60% FC) conditions. Under drought stress, plants of transgenerational e[CO2] exposed seeds showed markedly increased superoxide dismutase (16%), catalase (24%), peroxidase (9%), total antioxidants (14%), and proline (35%) levels that helped the plants to sustain normal growth through scavenging of hydrogen peroxide (11%) and malondialdehyde (26%). The carbohydrate metabolic enzymes such as aldolase (36%), phosphoglucomutase (12%), UDP-glucose pyrophosphorylase (25%), vacuolar invertase (33%), glucose-6-phosphate-dehydrogenase (68%), and cell wall invertase (17%) were decreased significantly; however, transgenerational seeds produced under e[CO2] showed a considerable increase in their activities in drought-stressed wheat plants. Moreover, transgenerational e[CO2] exposed seeds under drought stress caused a marked increase in leaf Ψw (15%), chlorophyll a (19%), chlorophyll b (8%), carotenoids (12%), grain spike (16%), hundred grain weight (19%), and grain yield (10%). Hence, transgenerational seeds exposed to e[CO2] upregulate the drought recovery metabolic processes to improve the grain yield of wheat under drought stress conditions.

Perceptions of childhood sexual abuse survivors: development and initial validation of a new scale to measure stereotypes of adult survivors of childhood sexual abuse.

The Childhood Sexual Abuse Stereotypes Scale was developed to assess stereotypes of adult survivors of childhood sexual abuse. Scale items were derived from two studies that elicited cultural and personal beliefs about, and emotions experienced towards adult childhood sexual abuse survivors among university undergraduates. Two scales, Emotions and Characteristics, were developed and administered online to 182 participants. Exploratory factor analysis produced 7 factors, 4 for the Emotions Scale and 3 for the Characteristics Scale. Study 2 replicated this factor structure using confirmatory factor analysis (N = 457). Reliability and validity analyses suggest that the Childhood Sexual Abuse Stereotypes Scale has satisfactory psychometric properties. The Childhood Sexual Abuse Stereotypes Scale can be used to examine stereotypes of legal and health care professionals likely to work with survivors as well as in stereotyping research.

Proliferation and migration of ML1 follicular thyroid cancer cells are inhibited by IU1 targeting USP14: role of proteasome and autophagy flux.

USP14 is a deubiquitinating enzyme involved in protein degradation by interacting with the proteasome and removal of poly-ubiquitin chains on target proteins. USP14 can influence cellular processes such as cell survival, DNA repair, ER stress, endocytosis, and the inflammatory response. USP14 further plays a role in tumor growth, and the inhibition of USP14 by compounds such as IU1 may affect cancer cell migration and invasion. Here we have studied the mechanisms for the action of IU1 in ML1 follicular thyroid cancer cells, comparing them with control, primary thyroid cells. Treatment with IU1 reduced proliferation of ML1 cells in a concentration-dependent manner, and more prominently than in control cells. IU1 decreased basal migration of ML1 cells, and after stimulation of cells with the bioactive compound, sphingosine-1-phosphate. The sphingosine-1-phosphate receptor 3 was increased in ML1 cells as compared with control thyroid cells, but this was not influenced by IU1. Further studies on the mechanism, revealed that IU1 enhanced the proteasome activity as well as LC3B-dependent autophagy flux in ML1 cells with an opposite effect on control thyroid cells. This indicates that IU1 elicits a cell-type dependent autophagy response, increasing it in ML1 cancer cells. The IU1-mediated stimulation of autophagy and proteasomes can likely contribute to the reduced cell proliferation and migration observed in ML1 cells. The precise set of proteins affected by IU1 in ML1 thyroid and other cancer cells warrant further investigations.

N-Glycomic Profiling of Microsatellite Unstable Colorectal Cancer.

Aberrant glycosylation affects cancer progression and immune evasion. Approximately 15% of colorectal cancers (CRCs) demonstrate microsatellite instability (MSI) and display major differences in outcomes and therapeutic responses, as compared to corresponding microsatellite stable (MSS) tumors. We compared the N-glycan profiles of stage II and IV MSI CRC tumors, further subdivided into BRAFV600E wild-type and mutated subgroups (n = 10 in each subgroup), with each other and with those of paired non-neoplastic mucosal samples using mass spectrometry. Further, the N-glycans of BRAFV600E wild-type stage II MSI tumors were compared to corresponding MSS tumors (n = 9). Multiple differences in N-glycan profiles were identified between the MSI CRCs and control tissues, as well as between the stage II MSI and MSS samples. The MSI CRC tumors showed a lower relative abundance of high-mannose N-glycans than did the control tissues or the MSS CRCs. Among MSI CRC subgroups, acidic N-glycans showed tumor stage and BRAF mutation status-dependent variation. Specifically, the large, sulfated/phosphorylated, and putative terminal N-acetylhexosamine-containing acidic N-glycans differed between the MSI CRC subgroups, showing opposite changes in stages II and IV, when comparing BRAF mutated and wild-type tumors. Our results show that molecular subgroups of CRC exhibit characteristic glycan profiles that may explain certain carcinogenic properties of MSI tumors.

BapC autotransporter protein of Bordetella pertussis is an adhesion factor.

Bordetella pertussis, the causative agent of whooping cough, attaches to mucosal surfaces in upper respiratory tract, where it produces, a variety of surface-associated and secreted molecules. Among various secreted products, some of the proteins belonging to autotransporter family; pertactin (Prn), bordetella resistance to killing (BrkA) and a newly identified member, bordetella autotransporter protein-C (BapC), are investigated in this study for their adherence potential to various respiratory and non-respiratory tract specific cell lines. Our results reveal that BapC and Prn mutants adhere significantly less (p < 0.0001 and p < 0.05) respectively to human non-respiratory (HeLa-229) and murine macrophages (P-388 D-1) cells compared to their wild-type strains. Prn, BrkA and BapC share no homology in their passenger domains except existence of common motifs arginine-glycine-asparctic (RGD) and glycosaminoglycan binding site (SGXG). We have shown that RGD and SGXG motifs are present in the coiled region in Prn and BrkA proteins with the exception in BapC where R (463) of RGD and S (597) of SGXG motif were observed in beta sheet of the modeled protein structures. Therefore, there is possibility that such arrangement of motifs can confer greater probability of BapC in better selective adherence to binding sites on the HeLa-229 and P-388 D-1 cell lines.