Human C1q Induces Apoptosis in an Ovarian Cancer Cell Line via Tumor Necrosis Factor

Copyright: © 2016 Kaur, Sultan, Murugaiah, Pathan, Alhamlan, Karteris and Kishore. Human C1q is the first recognition subcomponent of the complement classical pathway that plays a vital role in the clearance of immune complexes, pathogens and apoptotic cells. C1q also has a homeostatic role involving immune and non-immune cells; these functions not necessarily involve complement activation. Recently, C1q has been shown to be expressed locally in the microenvironment of a range of human malignant tumours, where it can promote cancer cell adhesion, migration and proliferation, without involving complement activation. C1q has been shown to be present in the ascitic fluid formed during ovarian cancers. In this study, we have examined the effects of human C1q and its globular domain on an ovarian cancer cell line, SKOV3. We show that C1q and the recombinant globular modules induce apoptosis in SKOV3 cells in a dose-and time-dependent manner. C1q expression was not detectable in the SKOV3 cells. Exogenous treatment with C1q and globular heads at the concentration of 10μg/ml induced apoptosis in approximately 55% cells, as revealed by immunofluorescence microscopy and FACS. The qPCR and caspase analysis suggested that C1q and globular head modules activated TNF-α and upregulation of Fas. The genes of mTOR, RICTOR and RAPTOR survival pathways, which are often over-expressed in majority of the cancers, were significantly downregulated within few hours of the treatment of C1q and globular head modules. In conclusion, C1q, via its globular domain, induced apoptosis in an ovarian cancer cell line, SKOV3 via TNF-α induced apoptosis pathway involving upregulation of Bax and Fas. This study highlights a potentially protective role of C1q in certain cancers

Kinking nonlinear elastic solids, nanoindentations, and geology

Physical Review Letters, 92(25): pp. 2555081-4.The physical mechanism responsible for nonlinear elastic, hysteretic, and discrete memory response of nonlinear mesoscopic elastic solids has to date not been identified.We show, by nanoindenting mica single crystals, that this response is most likely due to the formation of dissipative and fully reversible, dislocation-based kink bands. We further claim that solids with high c=a ratios, which per force are plastically anisotropic, should deform by kinking, provided they do not twin. These kinking nonlinear elastic solids include layered ternary carbides, nitrides, oxides, and semiconductors, graphite, and the layered phases, such as mica, present in nonlinear mesoscopic elastic solids

Ready-to-eat food consumption practices, food safety knowledge and relation to gender and education level of consumers in Kuala Lumpur, Malaysia

This study was aimed to evaluate consumers consumption practices of ready-to-eat (RTE) foods, food safety knowledge, and the relationship to gender and education level. A total of 258 consumers in Klang Valley, Kuala Lumpur, Malaysia were given questionnaires through convenience sampling. The results showed that the biggest motivation for consumers in purchasing RTE foods was convenience (47.3%). 40.7% of respondents purchase RTE foods more than twice a week, and 36.8% of respondents consumed RTE food at restaurants and cafes. A total of 54.3% of the respondents purchased RTE foods for lunch, and 48.1% of respondents usually spend RM11-RM20 per person. About 81% of respondents knew that the common symptom of foodborne disease is diarrhea, and 42.2% of respondents knew about the correct temperature for refrigeration. Most of the respondents (66.7%) knew that room temperature encourages the multiplication of bacteria, and 36.4% of respondents knew that bacteria will grow quickly at body temperature. About 60.9% of respondents knew that raw and cooked food should be separated to prevent the transfer of bacteria, and 62% of the respondents knew that placing raw meat in the plastic bag before putting it into the grocery basket decreases the chance of foodborne illness. However, only 10.1% of the respondents knew that the best way to clean a cutting board after using it for raw meat is by washing with bleach and water. Female respondents and those with higher education level generally had better food safety knowledge. This study could benefit RTE food marketers in gaining better insights into consumers’ behavior patterns towards RTE food. This study could also contribute to knowing the baseline knowledge of consumers in food safety as it is essential for the development of effective health educational programmes

Full-length human Surfactant Protein A inhibits Influenza A Virus infection of A549 lung epithelial cells: a recombinant form containing neck and lectin domains promotes infectivity

Hydrophilic lung surfactant proteins have emerged as key immunomodulators aimed at recognition and clearance of pulmonary pathogens. Surfactant protein A (SP-A) is a surfactant-associated innate immune pattern recognition molecule, which is known to interact with a variety of pathogens, and display anti-microbial effects. SP-A, being carbohydrate pattern recognition molecule, has been suggested to have a wide range of innate immune functions against pathogens. In addition, SP-A can work against respiratory pathogens, including influenza A virus (IAV). Some pandemic pH1N1 strains resist neutralization by SP-A due to differences in the N-glycosylation of viral hemagglutinin (HA). Here, we provide evidence, for the first time, that a recombinant form of human SP-A (rfhSP-A) composed of α-helical neck and carbohydrate recognition domains can actually promote the IAV replication, as observed by an upregulation of M1 expression in lung epithelial cell line, A549, when challenged with pH1N1 and H3N2 IAV subtypes. rfhSP-A (10 μg/ml) bound neuraminidase (NA) (˜60 kDa), matrix protein 1 (M1) (˜25 kDa) and M2 (˜17 kDa) in a calcium dependent manner, as revealed by far western blotting, and direct binding ELISA. However, human full length native SP-A downregulated mRNA expression levels of M1 in A549 cells challenged with IAV subtypes. Furthermore, qPCR analysis showed that transcriptional levels of TNF-α, IL-12, IL-6, IFN-α and RANTES were enhanced following rfhSP-A treatment by both IAV subtypes at 6 h post-IAV infection of A549 lung epithelial cells. In the case of full length SP-A treatment, mRNA expression levels of TNF-α, and IL-6 were downregulated during the mid-to-late stage of IAV infection of A549 cells. Multiplex cytokine/chemokine array revealed enhanced levels of both IL-6 and TNF-α due to rfhSP-A treatment in the case of both IAV subtypes tested, while no significant effect was seen in the case of IL-12. Enhancement of IAV infection of pH1N1 and H3N2 subtypes by truncated rfhSP-A, concomitant with infection inhibition by full-length SP-A, appears to suggest that a complete SP-A molecule is required for protection against IAV. This is in contrast to a recombinant form of trimeric lectin domains of human SP-D (rfhSP-D) that acts as an entry inhibitor of IAV.KACST (14-MED258-20

Olfactory dysfunction: a diagnostic symptom of COVID-19

Besides the common respiratory symptoms or viral pneumonia, COVID-19 is also presented with different neurological symptoms. Olfactory dysfunction (OD) or impairment of the sense of smell is one of the common neurological symptoms being reported in infection caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The severity varies from anosmia to microsmia or hyposmia. OD could have a potential early screening and diagnostic value besides other neurological and common respiratory symptoms. In our descriptive concise review, we aimed to elicit the manifestation of olfactory dysfunction as an early predictor of SARS-CoV-2 infection. We are also aiming to establish, OD as a quick and reliable assessment tool of COVID-19 risk, among the healthcare workers and contact tracers, which can justify self-quarantine of the person as well as recommendation for testing on a priority basis. We have also focused on, any effect to produce this symptom by the drug used for treatment in COVID-19, and if the old age of the patient showing any extra influence to develop OD in COVID-19

Detection and quantification of Vibrio parahaemolyticus in vegetables and environmental samples at farm level

The purpose of this study was to detect and quantify total and pathogenic Vibrio parahaemolyticus from vegetables and environmental samples at the farm level in Cameron Highlands, Pahang, Malaysia. Most Probable Number (MPN) – Polymerase Chain Reaction (PCR) method was used to detect toxR, tdh and trh genes and to quantify their concentration in samples. Samples obtained were cabbage (20), carrot (10), cucumber (10), lettuce (31), tomato (18), manure (10), soil (12), surface swab (21) and water (14), with a total of 146 samples. Sampling locations involved were three vegetable farms, two packing houses and one loading bay. Based on the results, overall, 13.7% of samples were present with V. parahaemolyticus toxR (maximum concentration 1100 MPN/g), with the highest detection in cabbage (6%). Vibrio parahaemolyticus tdh was detected in 1.4% samples (maximum concentration 7.3 MPN/g), and V. parahaemolyticus trh could not be detected in any samples. No tdh and trh genes could be detected from the recovered isolates. This finding highlighted that vegetables and environmental samples could potentially be contaminated with V. parahaemolyticus which poses risk to consumers. This study could be useful in future food safety risk communication and management programmes

Complement-independent Modulation of Influenza A virus infection by Factor H

Copyright © 2020 Murugaiah, Varghese, Saleh, Tsolaki, Alrokayan, Khan, Collison, Sim, Nal, Al-Mohanna and Kishore. The complement system is an ancient innate immune defence mechanism that can recognise molecular patterns on the invading pathogens. Factor H, as an inhibitor of the alternative pathway, down-regulates complement activation on the host cell surface. Locally synthesised factor H at the site of infection/injury, including lungs, can act as a pattern recognition molecule without involving complement activation. Here, we report that factor H, a sialic acid binder, interacts with influenza A virus (IAV) and modulates IAV replication, as observed by an upregulation of matrix protein 1 (M1) expression in H3N2-infected A549 cells, while downregulating M1 in H1N1 subtype-infected cells. Far-western blot revealed that factor H binds hemagglutinin (HA, ~70kDa), neuraminidase (NA, ~60kDa), and M1 (~25kDa). IAV-induced transcriptional levels of IFN-α, TNF-α, IL-12, IL-6, IFN-α, while RANTES were reduced following factor H treatment for the H1N1 subtype at 6 h post-infection. However, for the H3N2 subtype, mRNA levels of these pro-inflammatory cytokines were enhanced. Recombinant form of vaccinia virus complement control protein (VCP), which like factor H, contains CCP modules and has complement-regulatory activity, mirrored the results obtained with factor H. Both factor H (25%) and VCP (45%) were found to reduce luciferase reporter activity in MDCK cells transduced with H1N1 pseudotyped lentiviral particles. Factor H (50%) and VCP (30%) enhanced the luciferase reporter activity for H3N2, suggesting an entry inhibitory role of factor H and VCP against H1N1, but not H3N2. Thus, factor H can modulate IAV infection and inflammatory response independent of its complement-related functions.King Saud University, Riyadh, International Scientific Partnership Programme (ISPP) ISPP-145

Hyaluronic Acid Present in the Tumor Microenvironment Can Negate the Pro-apototic Effect of a Recombinant Fragment of Human Surfactant Protein D on Breast Cancer Cells

Copyright © 2020 Murugaiah, Agostinis, Varghese, Belmonte, Vieni, Alaql, Alrokayan, Khan, Kaur, Roberts, Madan, Bulla and Kishore. Human surfactant protein D (SP-D) belongs to the family of collectins that is composed of a characteristic amino-terminal collagenous region and a carboxy-terminal C-type lectin domain. Being present at the mucosal surfaces, SP-D acts as is a potent innate immune molecule and offers protection against non-self and altered self-such as pathogens, allergens, and tumour. Here, we examined the effect of a recombinant fragment of human SP-D (rfhSP-D) on a range of breast cancer lines. Breast cancer has four molecular subtypes characterised by varied expression of oestrogen (ER), progesterone (PR) and EGF receptors (HER2). The cell viability of HER2 over-expressing (SKBR3) and triple-positive (BT474) breast cancer cell lines (but not of triple-negative cell line (BT20), was reduced following rfhSP-D treatment at 24h. Upregulation of p21/p27 cell cycle inhibitors and p53 phosphorylation (Ser15) in rfhSP-D-treated BT474 and SKBR3 cell lines signified G2/M cell cycle arrest. Cleaved caspase 9 and 3 were detected in rfhSP-D- treated BT474 and SKBR3 cells, suggesting an involvement of intrinsic apoptosis pathway. However, rfhSP-D-induced apoptosis was nullified in the presence of hyaluronic acid whose increased level in breast tumor microenvironment is associated with malignant tumor progression and invasion. rfhSP-D bound to solid-phase HA and promoted tumor cell proliferation. rfhSP-D-treated SKBR3 cells in the presence of hyaluronic acid showed decreased transcriptional levels of p53 when compared to SKBR3 cells treated with rfhSP-D only. Thus, hyaluronic acid appears to negate the anti-tumorigenic properties of rfhSP-D against HER2-over-expressing and triple-positive breast cancer cells.King Saud University, Riyadh, International Scientific Partnership Program (ISPP) ISPP-145; Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Trieste, Italy, RC20/16 and 5MILLE15D

C4b Binding Protein Acts as an Innate Immune Effector Against Influenza A Virus

C4b Binding Protein (C4BP) is a major fluid phase inhibitor of the classical and lectin pathways of the complement system. Complement inhibition is achieved by binding to and restricting the role of activated complement component C4b. C4BP functions as a co-factor for factor I in proteolytic inactivation of both soluble and cell surface-bound C4b, thus restricting the formation of the C3-convertase, C4b2a. C4BP also accelerates the natural decay/dissociation of the C3 convertase. This makes C4BP a prime target for exploitation by pathogens to escape complement attack, as seen in Streptococcus pyogenes or Flavivirus. Here, we examined whether C4BP can act on its own in a complement independent manner, against pathogens. C4BP bound H1N1 and H3N2 subtypes of Influenza A Virus (IAV) most likely via multiple sites in Complement Control Protein (CCP) 1-2, 4-5, and 7-8 domains of its α-chain. In addition, C4BP CCP1-2 bound H3N2 better than H1N1. C4BP bound three IAV envelope proteins: Haemagglutinin (~70 kDa), Neuraminidase (~55 kDa), and Matrix protein 1 (~25kDa). C4BP suppressed H1N1 subtype infection into the lung epithelial cell line, A549, while it promoted infection by H3N2 subtype. C4BP restricted viral entry for H1N1 but had the opposite effect on H3N2, as evident from experiments using pseudo-typed viral particles. C4BP downregulated mRNA levels of pro-inflammatory IFN-α, IL-12, and NFκB in the case of H1N1, while it promoted a pro-inflammatory immune response by upregulating IFN- α, TNF-α, RANTES, and IL-6 in the case of H3N2. We conclude that C4BP differentially modulates the efficacy of IAV entry, and hence, replication in a target cell in a strain-dependent manner, and acts as an entry inhibitor for H1N1. Thus, CCP containing complement proteins such as factor H and C4BP may have additional defense roles against IAV that do not rely on the regulation of complement activation