The value of lung function assessment and Testin expression detection in clinicopathological features and prognosis of NSCLC patients.

OBJECTIVE: The aim of this study is to investigate the clinical value and potential prognostic significance of lung function assessment and Testin expression in non-small cell lung cancer (NSCLC) patients. METHODS: The NSCLC patients were classified into three groups according to lung function: group of normal lung function, group of PRISm (preserved ratio impaired spirometry) (FEV1, forced expiratory volume during the first second < 80% predicted and FEV1/FVC (forced vital capacity) ≥ 70%) and group of COPD (chronic obstructive pulmonary disease) (FEV1/FVC < 70%). The pre-operational clinicopathological characteristics of these patients were recorded and the markers of systemic inflammatory response, including neutrophil to lymphocyte ratio (NLR), lymphocyte to monocyte ratio (LMR), platelet to lymphocyte ratio (PLR) and eosinophils (EOS), were compared between three groups. The expression of Testin in NSCLC samples was detected by IHC and we further explored the correlation between Testin expression and clinicopathological characteristics and prognosis of NSCLC patients. Finally, Cox regression analysis was conducted to study the prognostic factors of NSCLC patients. RESULTS: Of the 158 NSCLC patients, percentages of normal lung function, PRISm and COPD were 41.4%, 22.8% and 36.1%, respectively. Patients with tumor in the left lung were more likely to have pulmonary dysfunction (PRISm and COPD) than the right lung. The markers of systemic inflammatory response showed differences to various degree in the three groups and NSCLC patients with PRISm or COPD presented more unfavorable prognosis than patients with normal function. The expression of Testin correlated with lymph node metastasis, TNM stage and tumor invasion of NSCLC patients. Moreover, patients with low Testin expression exhibited poorer disease-free survival and overall survival than those with high Testin expression. In Cox regression analysis, we found that PRISm, COPD and Testin expression served as prognostic factors in NSCLC patients. CONCLUSIONS: The presence of COPD or PRISm influenced systemic inflammatory response and prognosis of NSCLC patients. Testin expression correlated with clinicopathological features and could be potentially used as a prognostic marker in NSCLC.

Study on the expression of testin in the testes of dogs.

Introduction: Testin is a protein involved in cell mobility, adhesion and colony formation. In rats, testin presence has been reported in the testes, and its possible role in spermatogenesis has been suggested. Studies in humans also suggest a possible role of testin as a cancer suppressor protein. In the dog, which represents both an important pet species and a good animal model for studying biological and pathological testicular processes, the presence of testin has never been reported. Material and Methods: In the present study, the expression of testin in foetal, prepubertal, adult and aged canine testes was investigated. Testes from 5 adult and 3 aged dogs, from 2 one-month-old puppies and from 2 foetuses miscarried at the end of pregnancy were immunohistochemically examined with a commercial antibody against testin. Results: Testin was intensely expressed in Sertoli cells in every testis examined. Spermatids were also positive for testin in mature dogs and in the testicular areas of the aged ones which were not atrophic. Weak expression of testin was also detected in all testes examined. Conclusion: The present study, the first demonstrating the presence of testin in canine testes, provides the basis for further dog-human comparative research and for studies on the role of this protein in canine physiology, reproduction and testicular pathologies.

Quantifying Strain-Sensing Protein Recruitment During Stress Fiber Repair.

A family of proteins have been identified that recognize damaged, strained actin filaments in stress fibers. These proteins are often referred to as strain- or force-sensing and trigger downstream signaling mechanisms that can facilitate repair at these strain sites. Here we describe a method using high-resolution microscopy to screen and quantify the mechanosensitive recruitment of proteins to these stress fiber strain sites. Strain sites are induced using spatially controlled illumination of UV light to locally damage actin stress fibers. Recruitment of potential strain-sensing proteins can then either be compared to (Blanchoin, Physiol Rev 94, 235-263, 2014) a known control (e.g., zyxin-GFP) or (Hoffman, Mol Biol Cell 23, 1846-1859, 2012) the pre-damaged stress fiber protein distribution. With this method, strain-sensing proteins and their dynamic association with stress fiber strain sites can be reproducibly measured and compared.

Lateral flow assay: a promising rapid point-of-care testing tool for infections and non-communicable diseases.

The point-of-care testing (POCT) approach has established itself as having remarkable importance in diagnosing various infectious and non-communicable diseases (NCDs). The POCT approach has succeeded in meeting the current demand for having diagnostic strategies that can provide fast, sensitive, and highly accurate test results without involving complicated procedures. This has been accomplished by introducing rapid bioanalytical tools or biosensors such as lateral flow assays (LFAs). The production cost of these tools is very low, allowing developing countries with limited resources to utilize them or produce them on their own. Thus, their use has grown in various fields in recent years. More importantly, LFAs have created the possibility for a new era of incorporating nanotechnology in disease diagnosis and have already attained significant commercial success worldwide, making POCT an essential approach not just for now but also for the future. In this review, we have provided an overview of POCT and its evolution into the most promising rapid diagnostic approach. We also elaborate on LFAs with a special focus on nucleic acid LFAs.

The relationship between TESTIN expression and the prognosis of colorectal cancer.

BACKGROUND: Colorectal carcinoma (CRC) represents a most grave healthy burden worldwide. TESTIN has been confirmed as a predictive biomarker for several cancers. In the present study, we sought to assess the expression level and prognostic values of TESTIN in CRC. METHODS: The levels of TESTIN mRNA and protein were detected in 132 paired CRC tissues and noncancerous ones via quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) assays, respectively. Chi-square test was adopted to analyze the association of TESTIN expression with clinicopathological profiles of CRC patients. To explore prognostic value of TESTIN, Kaplan-Meier curve and Cox regression analyses were employed. RESULTS: TESTIN expression was down-regulated among CRC tissues in comparison to bordering cancer-free samples at both protein and mRNA levels (P < 0.001). Decreased TESTIN expression was closely related to poor tumor differentiation (P = 0.001) and advanced TNM stages (P = 0.001). CRC cases with low expression of TESTIN were more likely to undergo dismal overall survivals (log-rank P = 0.003). Multivariate Cox analysis unveiled that down-regulated expression of TESTIN was independently correlated with poor prognosis (HR=2.422, 95% CI=1.294-4.535, P = 0.006). CONCLUSION: The down-regulation of TESTIN may predict dismal prognosis for CRC patients.

[Increased use of genetic health care in Iceland 2012-2017].

INTRODUCTION: A genetic counselling unit at Landspitali hospital (LSH) was established in 2006. Meanwhile, genetic testing has become an integral part of general health care. In this article we detail the outcome of genetic testing at the Department of Genetic and Molecular Medicine (DGM) at Landspitali over a five year period (2012-2017). Factors that were analyzed for the time period were: Number of patients, reason for referral, reason for genetic testing without genetic counselling and yield (proportion of positive tests) of genetic testing. METHODS: Data was analysed from two medical record databases, Shire and Saga, used by the DGM in the time period. RESULTS: The number of individuals coming for genetic counselling increased every year over the time period. Reasons for referral were cancer-related in two-thirds of cases. Other reasons for referral included various other familial disorders. Most common were autosomal dominant disorders like myotonic dystrophy, hypertrophic cardiomyopathy and autosomal recessive disorders like spinal muscular atrophy (SMA) and GM1-gangliosidosis. Most common reasons for genetic testing outside of the LSH GC unit was because of managable diseases like hemochromatosis and F5/Prothrombin-related thrombophilia. Yield of genetic testing was assessed for a) known mutation testing / carrier testing, b) single gene testing, c) gene panel testing and d) whole genome and whole exome sequencing. Known mutation testing was positive in 33% of cases and single gene testing in 46% of cases. The yield of gene panel testing for cancer was found to be lower (20%) than gene panel testing for other disorders (40%). The yield of whole exome and whole genome sequencing was 46%.

Rapid and sensitive detection of Salmonella species targeting the hilA gene using a loop-mediated isothermal amplification assay.

Salmonella species are among the major pathogens that cause foodborne illness outbreaks. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid and sensitive detection of Salmonella species. We designed LAMP primers targeting the hilA gene as a universal marker of Salmonella species. A total of seven Salmonella species strains and 11 non-Salmonella pathogen strains from eight different genera were used in this study. All Salmonella strains showed positive amplification signals with the Salmonella LAMP assay; however, there was no non-specific amplification signal for the non-Salmonella strains. The detection limit was 100 femtograms (20 copies per reaction), which was ~1,000 times more sensitive than the detection limits of the conventional polymerase chain reaction (PCR) assay (100 pg). The reaction time for a positive amplification signal was less than 20 minutes, which was less than one-third the time taken while using conventional PCR. In conclusion, our Salmonella LAMP assay accurately detected Salmonella species with a higher degree of sensitivity and greater rapidity than the conventional PCR assay, and it may be suitable for point-of-care testing in the field.

Preliminary Study on the Expression of Testin, p16 and Ki-67 in the Cervical Intraepithelial Neoplasia.

Cervical cancer is one of the most common malignant cancers in women worldwide. The 5-year survival rate is 65%; nevertheless, it depends on race, age, and clinical stage. In the oncogenesis of cervical cancer, persistent HPV infection plays a pivotal role. It disrupts the expression of key proteins as Ki-67, p16, involved in regulating the cell cycle. This study aimed to identify the potential role of testin in the diagnosis of cervical precancerous lesions (CIN). The study was performed on selected archival paraffin-embedded specimens of CIN1 (31), CIN2 (75), and CIN3 (123). Moderate positive correlation was observed between testin and Ki-67 as well as testin and p16 expression in all dysplastic lesions (r = 0.4209, r = 0.5681; p < 0.0001 for both). Statistical analysis showed stronger expression of the testin in dysplastic lesions vs. control group (p < 0.0001); moreover, expression was significantly higher in HSIL than LSIL group (p < 0.0024). In addition, a significantly stronger expression of testin was observed in CIN3 vs. CIN1 and CIN3 vs. CIN2. In our study, expression of Ki-67, p16, and testin increased gradually as the lesion progressed from LSIL to HSIL. The three markers complemented each other effectively, which may improve test sensitivity and specificity when used jointly.

The Role of Patch Testing with Indian Cosmetic Series in Patients with Facial Pigmented Contact Dermatitis in India.

BACKGROUND: Pigmented contact dermatitis (PCD) is a non-eczematoid variant of contact dermatitis, mainly characterised by hyperpigmentation. It occurs due to contact with a low amount of allergen over a long duration of time. PCD is frequently seen in Indians but is often misdiagnosed or underdiagnosed because of the asymptomatic nature of the entity. The aetiology and the allergens implicated in PCD in the Indian subcontinent is still an enigma because of the limited studies done. MATERIALS AND METHODS: This was an institution-based cross-sectional study, done at a tertiary hospital. Patch testing with Indian Cosmetic Series was conducted in a standardised method. Readings were taken at 48 hrs/72 hrs and on the 7th day [Figure 2a and b]. The International Contact Dermatitis Research Group (ICDRG) scoring system was used to grade the readings. RESULTS: Out of the 38 biopsy proven cases of PCD, 18 (47%) showed lichenoid features, 17 (45%) showed spongiotic features, 3 (8%) showed a mixed lichenoid and spongiotic pattern. Among total 1216 (32 patches × 38 patients) patch applied, 42 (3.4%) showed positivity in 30 patients. Among allergen categories, colorant (PPD) was found to be most common (37%) followed by fragrances (18%), preservatives (15%), anti-microbial (11%) and emulsifier and anti-oxidants (each 8%). CONCLUSION: It is important to identify the allergens implicated in PCD to help in better management of the condition. Patch testing proves to be a non invasive, low cost method and its role is indispensable in identifying the correct allergen.

Chitinase 3-Like 1, Nestin, and Testin Proteins as Novel Biomarkers of Potential Clinical Use in Colorectal Cancer: A Review.

Colorectal cancer is the third most commonly diagnosed cancer in males and the second most common in females. Only 10-20% of patients are diagnosed at the early stage of disease. Recently, the role of novel biomarkers of the neoplastic process in the early detection of colorectal cancer has been widely discussed. In this review, we focused on the three novel biomarkers that are of potential clinical importance in diagnosing and monitoring colorectal cancer. Chitinase 3-like 1 protein, also known as YKL-40, and nestin and testin proteins are produced by colorectal cancer cells. YKL-40 protein is a marker of proliferation, differentiation, and tissue morphogenetic changes. The level of YKL-40 is elevated in about 20% of patients with colorectal cancer. An increased expression of nestin indicates immaturity. It is a marker of angiogenesis in neoplastic processes. Testin protein is a component of cell-cell connections and focal adhesions. The protein is produced in normal human tissues, but not in tumor tissues. Downregulation of testin increases cell motility, spread, and proliferation, and decreases apoptosis. The usefulness and role of these biomarkers, both alone and combined, in the diagnostics of colorectal cancer should be further explored as early cancer detection may substantially improve treatment outcome and patient survival.

Testin protects against cardiac hypertrophy by targeting a calcineurin-dependent signalling pathway.

Multiple organs express testin (TES), including the heart. Nevertheless, current understanding of the influence of TES on cardiovascular diseases, especially on cardiac hypertrophy and its etiology, is insufficient. This study investigated the influence of TES on cardiac hypertrophy and its etiology. Murine models with excessive TES expression specific to the heart were constructed with an adeno-associated virus expression system. Cardiac hypertrophy was stimulated through aortic banding (AB). The severity of cardiac hypertrophy was evaluated through molecular, echocardiographic, pathological, and hemodynamic examination. The findings of our study revealed that TES expression was remarkably suppressed not only in failing human hearts but also in mouse hearts with cardiac hypertrophy. It was discovered that excessive TES expression driven by an adeno-associated viral vector noticeably inhibited hypertrophy triggered by angiotensin II (Ang II) in cultivated cardiomyocytes from newborn rats. It was also revealed that TES knockdown via AdshTES caused the reverse phenotype in cardiomyocytes. Furthermore, it was proved that excessive TES expression attenuated the ventricular dilation, cardiac hypertrophy, dysfunction, and fibrosis triggered by AB in mice. It was discovered that TES directly interacted with calcineurin and suppressed its downstream signalling pathway. Moreover, the inactivation of calcineurin with cyclosporin A greatly offset the exacerbated hypertrophic response triggered by AB in TES knockdown mice. Overall, the findings of our study suggest that TES serves as a crucial regulator of the hypertrophic reaction by hindering the calcineurin-dependent pathway in the heart.

The Role of Testin in Human Cancers.

Testin is a protein expressed in almost all normal human tissues. It locates in the cytoplasm along stress fibers being recruited to focal adhesions. Together with zyxin and vasodilator stimulated protein it forms complexes with various cytoskeleton proteins such as actin, talin and paxilin. They jointly play significant role in cell motility and adhesion. In addition, their involvement in the cell cycle has been demonstrated. Expression of testin protein level correlates positively with percentage of cells in G1 phase, while overexpression can induce apoptosis and decreased colony forming ability. Decreased testin expression associate with loss by cells epithelial morphology and gain migratory and invasive properties of mesenchymal cells. Latest reports indicate that TES is a tumor suppressor gene which can contribute to cancerogenesis but the mechanism of loss TES gene expression is still unknown. Some authors point out hypermethylation of the CpG island as a main factor, however loss of heterozygosity may also play an important role [4, 5]. The altered expression of testin was found in malignant neoplasm, i.a. ovarian, lung, head and neck squamous cell cancer, breast, endometrial, colorectal, prostate and gastric cancers [1-9]. Testin participate in the processes of tumor growth, angiogenesis, and metastasis [10]. Many researchers stated involvement of testin in tumor progression, what suggest its potential usage in immunotherapy [7, 11]. Understanding the molecular functions of testin may be crucial in development personalized treatment. In the present manuscript up-to-date review of literature can be found.

[Role of Testin in nasopharyngeal squamous cancer with high ability of metastasis].

Objective:To explore the influence and regulatory mechanism of TES gene on proliferation and migration of nasopharyngeal squamous cancer(NSPC) 5-8F cell.Method:DNA fragment encoding TES was obtained by RT-PCR method from the human highly metastatic nasopharyngeal squamous carcinoma cell line 5-8F. we identified the recombinant plasmid pEGFP-N1-TES by RT-PCR and DAN sequencing. we stablely transfected the pEGFP-N1-TES into the human highly metastatic nasopharyngeal squamous carcinoma cell line 5-8F, and detected the expression of TES by the RT-PCR and Western-blot method. And detected the impact of 5-8F cells transfection by flow cytometry and scratch tests.Result:Flow cytometry analysis showed that the apoptotic in 5-8F/pEGFP- N1-TES was significantly higher than non-transected TES and 5-8F/pEGFP-N1,and the differences were statistically significant(P<0.05).Cell scratch experiments showed that the 5-8F/pEGFP-N1-TES group cell migration rate was obviously lower than nontransected TES and 5-8F/pEGFP-N1 group in the first 12 h, 24 h and 48 h.The difference was significant(P<0.01).Conclusion:The stable transfectant cell model was established successfully. TES in vitro could significantly increase apoptosis and reduce the athletic ability. And thus TES gene might be a novel candidate of tumor-suppressor.

Testin (TES) as a candidate tumour suppressor and prognostic marker in human astrocytoma.

Astrocytomas are one of the most common brain tumours; however, the current methods used to characterize these tumours are inadequate. The establishment of molecular markers may identify variables required to improve tumour characterization and subtyping, and may aid to specify targets for improved treatment with essential prognostic value for patient survival. One such candidate is testin (TES), which was reported to have prognostic value for glioblastoma patients. However, the role of TES protein in gliomagenesis is currently unknown. In the present study, the methylation status of the TES promoter was investigated in post-operative astrocytoma tumours of different malignancy grade, and its association with the survival of astrocytoma patients was evaluated. In addition, the expression of TES protein was investigated in the same set of astrocytoma tumours tissue, and the association of protein expression with glioma patients survival was evaluated. The methylation status of TES was assessed by methylation-specific polymerase chain reaction in 138 different grade astrocytoma samples. Western blot analysis was used to characterize the expression pattern of TES in 86 different grade astrocytoma specimens: 13 of pathological grade I, 31 of pathological grade II, 17 of pathological grade III and 25 of pathological grade IV (glioblastoma). Statistical analyses were conducted to investigate the association between tumour molecular pattern, patient clinical variables and overall survival. The methylation analysis of the TES promoter exhibited a distinct profile between astrocytomas of different malignancy grade (P<0.001). Furthermore, gene promoter methylation was significantly associated with patients' age, survival and pathological grade (P<0.001). The protein expression level of TES was significantly lower in glioblastoma (grade IV astrocytoma) than in lower grade (II-III) astrocytoma tissue (P=0.028 and P=0.04, respectively). Additionally, short overall survival of patients was markedly associated with low TES protein expression (P=0.007). However, no association between TES methylation and TES protein expression was noticed. The present study demonstrated that decreased expression of TES may be important in tumour progression and prognosis in human astrocytomas. TES may be a useful marker for predicting the clinical outcome of astrocytoma patients.

TES inhibits colorectal cancer progression through activation of p38.

The human TESTIN (TES) gene has been identified as a candidate tumor suppressor based on its location at a common fragile site - a region where loss of heterozygosity has been detected in numerous types of tumors. To investigate its role in colorectal cancer (CRC), we examined TES protein levels in CRC tissue samples and cell lines. We observed that TES was markedly reduced in both CRC tissue and cell lines. Additionally, overexpression of TES significantly inhibited cell proliferation, migration, and invasion, while increasing cell apoptosis in colon cancer cells. By contrast, shRNA-mediated TES knockdown elicited the opposite effects. TES inhibited the progression of CRC by up-regulating pro-apoptotic proteins, down-regulating anti-apoptotic proteins, and simultaneously activating p38 mitogen-activated protein kinase (MAPK) signaling pathways. Collectively, these data indicate that TES functions as a necessary suppressor of CRC progression by activating p38-MAPK signaling pathways. This suggests that TES may have a potential application in CRC diagnosis and targeted gene therapy.

[Expression and clinical significance of Testin in nasopharyngeal carcinoma].

Objective:Our purpose was to investigate the expression of Testin gene,and its possible relationship with the clinicopathological features of human nasopharyngeal carcinoma.Method:The expression of Testin in nasopharyngeal carcinoma tissues were detected by immunohistochemistry methods,semi-quantitative reverse transcriptase polymerase chain reaction and Western blot.The correlations of Testin to clinicopathologic features of nasopharyngeal carcinoma were analyzed.Result:The mRNA level of Testin was down-regulated in human nasopharyngeal carcinoma.The positive rate of Testin protein was significantly lower in human nasopharyngeal carcinoma tissues than that in nomal tissues;The protein level of Testin was down-regulated in cancers as compared with corresponding normal tissues.Testin expression was positively correlated with the differentiation of nasopharyngeal carcinoma.Meanwhile,differences in gender and age were not significance(P>0.05 respectively) .There was a significant correlation between invasion,distant metastasis and differentiation degree and Testin expression(P<0.05 respectively).Conclusion:The decreased expression of Testin gene may play an importmant role in the development of esophageal squamous cancer.Thus Testin gene might be a novel candidate of tumor-suppressor.It may be an objective marker for prognostic factor and malignant level for nasopharyngeal carcinoma.

TESTIN was commonly hypermethylated and involved in the epithelial-mesenchymal transition of endometrial cancer.

We previously reported frequent loss of TESTIN in human endometrial carcinoma, which significantly suppressed tumor proliferation and invasion. Herein, we further explored the mechanisms underlying TESTIN loss and its roles in the epithelial-mesenchymal transition (EMT, a key step for tumor spreading). Methylation-specific PCR was performed to investigate the promoter status of TESTIN in a panel of endometrial cancer and normal endometrium tissues. The expression of TESTIN mRNA was determined by real-time PCR. Up- and down-regulation of TESTIN were achieved by transient transfection with pcDNA3.1-TESTIN and shRNA-TESTIN plasmids, respectively. The EMT alterations were observed under the optical microscope and EMT-related markers were detected by real-time PCR and western blot. Compared to the control (3.6%), TESTIN was hypermethylated in 43.7% endometrial cancer tissues (p < 0.001). Moreover, TESTIN hypermethylation was significantly correlated with advanced tumor stage, deep myometrial invasion and lymphatic node metastasis. In vitro, the demethylating agent dramatically restored the expression of TESTIN. In addition, up-regulation of TESTIN significantly suppressed the EMT procedure; whereas down-regulation of TESTIN enhanced EMT. In conclusion, we demonstrated that loss of TESTIN was mainly caused by hypermethylation, which might be a potent prognostic marker. Furthermore, we proved that TESTIN significantly suppressed the EMT procedure, proposing restoration of TESTIN to be a novel therapeutic strategy for endometrial carcinoma.

Testin interacts with vangl2 genetically to regulate inner ear sensory cell orientation and the normal development of the female reproductive tract in mice.

BACKGROUND: Planar cell polarity (PCP) signaling regulates the coordinated polarization of cells and is required for the normal development and function of many tissues. Previous studies have identified conserved PCP genes, such as Van Gogh-like 2 (Vangl2) and Prickle (Pk), in the regulation of coordinated orientation of inner ear hair cells and female reproductive tract development. Testin shares a PET-LIM homology with Pk. It is not clear whether Testin acts in PCP processes in mammals. RESULTS: We identified Testin as a Vangl2-interacting protein through a 2-hybrid screen with a cochlea cDNA library. Testin is enriched to cell-cell boundaries in the presence of Vangl2 in cultured cells. Genetic inactivation of Testin leads to abnormal hair cell orientation in the vestibule and cellular patterning defects in the cochlea. In addition, Testin genetically interacts with Vangl2 to regulate hair cell orientation in the cochlea and the opening of the vaginal tract. CONCLUSIONS: Our findings suggested Testin as a gene involved in coordinated hair cell orientation in the inner ear and in female reproductive tract development. Furthermore, its genetic interaction with Vangl2 implicated it as a potential molecular link, responsible for mediating the role of Vangl2-containing membranous PCP complexes in directing morphologic polarization.