TGFβ1-induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer

Cten (C-terminal tensin-like) is a member of the tensin protein family found in complex with integrins at focal adhesions. It promotes epithelial‐mesenchymal transition (EMT) and cell motility. The precise mechanisms regulating Cten are unknown, although we and others have shown that Cten could be under the regulation of several cytokines and growth factors. Since Transforming growth factor beta 1 (TGF-β1) regulates integrin function and promotes EMT / cell motility, we were prompted to investigate whether TGF-β1 induces EMT and cell motility through Cten signalling in colorectal cancer. TGF-β1 signalling was modulated by either stimulation with TGF-β1 or knockdown of TGF-β1 in the CRC cell lines SW620 and HCT116. The effect of this modulation on expression of Cten, EMT markers and on cellular function was tested. The role of Cten as a direct mediator of TGF-β1 signalling was investigated in a CRC cell line in which the Cten gene had been deleted (SW620ΔCten). When TGF-β1 was stimulated or inhibited, this resulted in, respectively, upregulation and downregulation of Cten expression and EMT markers (Snail, Rock, N-Cadherin, Src). Cell migration and cell invasion were significantly increased following TGF-β1 stimulation and lost by TGF-β1 knockdown. TGF-β1 stimulation of the SW620ΔCten cell line resulted in selective loss of the effect of TGF-β1 signalling pathway on EMT and cell motility whilst the stimulatory effect on cell proliferation was retained. These data suggested Cten may play an essential role in mediating TGF-β1 induced EMT and cell motility and may therefore play a role in metastasis in CRC

Investigations into the role of Cten signalling in colorectal cancer

C-terminal tensin-like (Cten, also known as Tensin4) is the member of the tensin gene family. Cten functions as an oncogene in a variety of cancer types and its expression is commonly associated with poor prognosis and metastasis in colorectal cancer (CRC). Although several studies have shown that Cten has a critical role in the regulation of cell motility and invasion in different tumour tissues, the underlying signalling mechanisms have not been fully elucidated. This thesis investigated the biological activity of Cten in four different ways in order to further elucidate the mechanisms of Cten signalling in CRC cells. Potential downstream targets of Cten signalling involved in the regulation of epithelial-to-mesenchymal transition (EMT) induced cell motility i.e. Rho-associated protein kinase1 (ROCK1), Src and Snail were investigated. Cten expression was manipulated in different cell lines using multiple approaches including forced expression, gene knockdown and constitutive depletion (through Crispr/Cas9 gene deletion) to eliminate artefacts of methodology and cell line specific effects. Snail, Src and ROCK1 were identified as novel downstream targets of Cten signalling and additionally, Cten was shown to increase the stabilisation of both Src and Snail proteins. The functional relevance of Cten-Snail, Cten-Src and Cten-ROCK1 signalling was assessed, and the overall findings demonstrated that Cten could promote cell motility and colony formation directly through the positive regulation of the Src/ROCK1/Snail dependent axis. To gain a deeper insight into the mechanisms of Cten’s biological function, mutations, at two important residues (i.e. arginine 474 and tyrosine 479) in the Src homology 2 (SH2) domain of Cten were introduced into one construct (GFP-CtenR474A+Y479F) using site directed mutagenesis. These two residues in the SH2 domain of Cten were found to not only be important for interacting with Src, ROCK1, or Snail signalling, but also for regulating cell motility and colony formation efficiency. Numerous Cten regulatory factors have been identified, however, little is known about how Cten is activated and regulated in cancer cells. The relationship between transforming growth factor beta 1 (TGFβ1) and Cten was investigated and stimulation of cells with TGFβ1 or knockdown of TGFβ1 resulted in changes in Cten expression as well as its downstream targets of ROCK1, Src, Snail, and N-cadherin. Furthermore, this positive interaction between TGFβ1 and Cten was functionally relevant and caused changes in cell motility. and the nuclear translocation of ROCK1, Src, and Snail protein increased by TGFβ1 is probably mediated via upregulation of the Cten signalling pathway The biological function of Cten in the nucleus was further investigated and shown to increase nuclear localisation of Src, ROCK1, and Snail, further promoting the migratory capability and colony formation efficiency in CRC cells. Finally, Cten expression was shown to positively correlate with both ROCK1 and Src expression in a series of primary CRCs. This correlation was consistent with that observed following manipulation of Cten expression in CRC cell lines. In conclusion, this study has revealed a number of novel findings regarding the biological function of Cten signalling in CRC. However, further validation of the findings may enhance the understanding of the role of Cten in the invasion-metastasis cascade in the future

Cten promotes Epithelial–Mesenchymal Transition (EMT) in colorectal cancer through stabilisation of Src

Cten is an oncogene promoting EMT in many signaling pathways, namely through Snail. We investigated whether Cten function could be mediated through Src. Cten levels were modulated by forced expression in HCT116 and gene knockdown in SW620 CRC (colorectal cancer) cell lines. In all cell lines, Cten was a positive regulator of Src expression. The functional importance of Src was tested by simultaneous Cten overexpression and Src knockdown. This resulted in abrogation of Cten motility-inducing activity and reduction of colony formation ability together with failure to induce Cten targets. In SW620ΔCten reduced Src expression increased following restoration of Cten, also leading to increased cell motility and colony formation, which were lost if Src was concomitantly knocked down. By qRT-PCR we showed modulation of Cten had no effect on Src mRNA. However, a CHX pulse chase assay demonstrated stabilization of Src protein by Cten. Finally, expression of Cten and Src was tested in a series of 84 primary CRCs and there was a significant correlation between them (P = 0.001). We conclude that Src is a novel and functionally important target of the Cten signaling pathway and that Cten protein causes post-transcriptional stabilization of Src in promoting EMT and possibly metastasis in CRC

Attitudes of Elementary Schools Students in Najran District towards Science

This study aims to explore the attitudes of elementary school students, in Najran district in Saudi Arabia, towards science. The modified Attitudes towards Science Inventory questionnaire was used to collect the raw data, with appropriate validity and reliability. The questionnaire of eight items measures only the construct of enjoyment in science using a 3-point Likert scale, which is appropriate for elementary students. This questionnaire was administrated to a sample of 95 students (49 males & 46 females) from two urban and two rural elementary schools. The findings indicated that while there was no significant difference in attitudes towards science between boys and girls. At the same time, grade 5 and 6 students were having more positive attitudes than grade 4 students. Additionally, the urban students showed higher level of attitudes towards science than the rural students. These findings are discussed in the context of science teaching and research. Keywords: Elementary Students, Attitudes towards Scienc

Cten promotes epithelial-mesenchymal transition through post-transcriptional stabilization of Snail

Cten promotes cell migration however the knowledge of underlying signalling pathways is sparse. We have shown that Cten downregulates E-cadherin, a feature of epithelial to mesenchymal transition (EMT). This prompted us to investigate whether Cten further contributed to EMT processes to regulate cell motility.The regulation of Snail by Cten was investigated following overexpression, knockdown (by RNA-interference) or knockout of Cten in HCT116, Caco-2 and SW620 colorectal cancer (CRC) cell lines. Subsequently, the cycloheximide (CHX) pulse chase assay was used to investigate changes in Snail protein stability and the functional relevance of Cten-Snail signalling was investigated.Snail was identified as a downstream target of Cten signalling using multiple approaches of Cten expression manipulation. Furthermore, this activity was mediated through the SH2 domain of Cten. The CHX assay confirmed that Cten was regulating Snail at a post transcriptional level and this was through the prevention of Snail degradation. Cell migration, invasion and colony formation efficiency were increased following forced expression of GFP-Cten but subsequently lost when Snail was knocked down, demonstrating a functional Cten-Snail signalling axis.In conclusion, we have described a novel Cten-Snail signaling pathway that contributes to cell motility in CRC, mediated by the stabilization of Snail protein. This finding potentially furthers the understanding of EMT regulatory networks in cancer metastasis

Dermatological Emergencies in Family Medicine: Recognition, Management, and Referral Considerations

Numerous people with skin disorders who have real dermatologic crises show up at the emergency room. Family doctors need to be able to identify potentially fatal dermatological disorders quickly since they could be the first to encounter patients with these illnesses. The purpose of this review is to provide guidance for early recognition, help identify distinct symptoms, and enable early diagnosis of emerging dermatological conditions. Necrotizing fasciitis, Stevens-Johnson syndrome, toxic epidermal necrolysis, Rocky Mountain spotted fever, and other possible emergencies that might manifest as dermatological symptoms are examples of these conditions. In this article we will be discussing the dermatological emergencies present at primary care settings and encountered by family physician, recognition and management of those emergencies, referral considerations, role of family medicine in dermatological emergencies and other topics

Prevalence, Recognition, And Management Of Mental Disorders In Primary Care

According to latest figures, 50% of individuals will have a minimum of one mental health issue in their lifetime, with at least 25% experiencing a mental condition in the previous year. primary care doctors are overwhelmingly responsible for recognition, evaluation, therapy, and referral, with at least one-third of their consultations containing a direct and obvious mental aspect Primary care physicians are the foundation of the majority of medical care programs that involve recognizing, diagnosing, treating, and referring patients to specialists for every kind of diseases, whether physical, mental, or both. Over the last two decades, there has been a greater emphasis on this position, notably in the treatment of mental problems in primary care

Assessment and Management of Scabies in Primary Care Settings

Scabies is an overlooked tropical illness that yet has significant worldwide effects and lasting health repercussions. The condition is caused by the mite Sarcoptes scabei var. hominis, which is a parasitic organism that dwells on the outer layer of the human skin. Scabies is prevalent in impoverished neighborhoods as a result of the high population density in locations such as nursing homes, correctional facilities, and among homeless and displaced children. Nevertheless, modern nations are also prone to scabies infestations, particularly in cases of institutional outbreaks or mini epidemics occurring after conflict or natural calamities. Scabies diagnosis can be aided by both invasive and noninvasive techniques. This paper reviews assessment diagnosis, and management of scabies in primary health care settings

Impact Of Bariatric Surgery on Gastroesophageal Reflux Disease: A Cross-Sectional Study

Objective: To assess the occurrence of GERD after bariatric surgery and surgery impact on GERD. Methods: This research employs a cross-sectional study design to investigate the impact of bariatric surgery on Gastroesophageal Reflux Disease (GERD) among individuals who have undergone various types of bariatric surgeries. Results: The study included 302 participants. The most frequent weight among them was more than 96 kg (n= 130, 43%) followed by 76-85 kg (n= 51, 16.9%). The most frequent height among study participants was 1.61-1.70 m (n= 100, 33.1%) followed by 1.51-1.60 m (n= 99, 32.8%). The most frequent body mass index (BMI) value among study participants was more than 35 kg/m2 (n= 126, 41.7%) followed by 25-29.9 kg/m2 (n= 67, 22.2%). The most frequent age among study participants was 26-36 years (n= 104, 34.4%) followed by 15-25 years (n= 83, 27.5%). The most frequent gender among study participants was Female (n= 162, 53.6%) followed by Male (n= 140, 46.4%). Participants were asked about the type of obesity surgery. The most frequent was Gastric sleeve (n=222, 73.5%), followed by Gastric bypass (n=33, 10.9%).   Conclusion: Study results showed that most of the study participants are extremely obese according to their BMI. The most common obesity surgery type was a Gastric sleeve followed by a Gastric bypass. The most of participants were a non-smoker. Most of them had weight loss. In addition, most of the study participants had good social connection

Prevalence of Comorbidities Associated With Type 2 Diabetes Mellitus In Ksa: A Cross-Sectional Study

Objective: To determine the prevalence of comorbidities associated with type 2 diabetes mellitus among the Saudi adult population. Methods: A cross-sectional study design will be employed to investigate the prevalence of comorbidities associated with Type 2 Diabetes Mellitus (T2DM) within the Kingdom of Saudi Arabia (KSA) population. This design allows for the collection of data at a single point in time, providing insights into the existing relationship between T2DM and comorbidities.Results: The study included 601 participants. The most frequent body mass index BMI value among study participants was overweight 25-29.9 kg/m2 (n= 196, 33%), followed by Normal 18.5-24.9 kg/m2 (n= 177, 29%). The frequent gender among study participants was male years (n= 356, 59%) and female (n= 245, 41%). The most frequent age among study participants was 40-50 years (n= 145, 24%), followed by 18-28 years (n= 140, 23%). The most frequent marital status among study participants was married (n= 360, 60%), followed by single (n= 160, 27%). The perceived physical activity per week among study participants, most of whom did not do any activity (n=231,38%) followed one-time activity (n=98,16%). Conclusion: The results showed that most of the study participants were overweight according to their BMI. The majority of patients had normal blood pressure. Most of the participants suffer from diabetes. In addition, most of the participants do not do any activity, and the majority of participants are non-smokers