Early Stages Of Progression Research Articles

The earliest events in BRAF-mutant colorectal cancer: exome sequencing of sessile serrated lesions with a tiny focus dysplasia or cancer reveals recurring mutations in two distinct progression pathways.

Around 15–30% of colorectal cancers (CRC) develop from sessile serrated lesions (SSLs). After many years of indolent growth, SSLs can develop dysplasia and rapidly progress to CRC through events that are only partially understood. We studied molecular events at the very early stages of progression of SSLs via the MLH1‐proficient and deficient pathways to CRC. We collected a cohort of rare SSLs with a small focus (<10 mm) of dysplasia or cancer from the pathology archives of three hospitals. Whole‐exome sequencing was performed on DNA from nonprogressed and progressed components of each SSL. Putative somatic driver mutations were identified in known cancer genes that were differentially mutated in the progressed component. All analyses were stratified by MLH1 proficiency. Forty‐five lesions with a focus dysplasia or cancer were included, of which 22 (49%) were MLH1‐deficient. Lesions had a median diameter of 10 mm (interquartile range [IQR] 8–15), while the progressed component had a median diameter of 3.5 mm (IQR 1.75–4.75). Tumor mutational burden (TMB) was high in MLH1‐deficient lesions (23.9 mutations per MB) as compared to MLH1‐proficient lesions (6.3 mutations per MB). We identified 34 recurrently mutated genes in MLH1‐deficient lesions. Most prominently, ACVR2A and RNF43 were affected in 18/22 lesions, with mutations clustered in three hotspots. Most lesions with RNF43 mutations had concurrent mutations in ZNRF3. In MLH1‐proficient lesions APC (10/23 lesions) and TP53 (6/23 lesions) were recurrently mutated. Our results show that the mutational burden is exceptionally high even in the earliest MLH1‐deficient lesions. We demonstrate that hotspot mutations in ACVR2A and in the RNF43/ZNRF3 complex are extremely common in the early progression of SSLs along the MLH1‐deficient serrated pathway, while APC and TP53 mutations are early events in the the MLH1‐proficient pathway. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Roles of TGF-β in cancer hallmarks and emerging onco-therapeutic design.

Transforming growth factor-beta (TGF-β) is a double-edged sword in cancer treatment because of its pivotal yet complex and roles played during cancer initiation/development. Current anti-cancer strategies involving TGF-β largely view TGF-β as an onco-therapeutic target that not only substantially hinders its full utilisation for cancer control, but also considerably restricts innovations in this field. Thereby, how to take advantages of therapeutically favourable properties of TGF-β for cancer management represents an interesting and less investigated problem. Here, by categorising cancer hallmarks into four critical transition events and one enabling characteristic controlling cancer initiation and progression, and delineating TGF-β complexities according to these cancer traits, we identify the suppressive role of TGF-β in tumour initiation and early-stage progression and its promotive functionalities in cancer metastasis as well as other cancer hallmarks. We also propose the feasibility and possible scenarios of combining cold atmospheric plasma (CAP) with onco-therapeutics utilising TGF-β for cancer control given the intrinsic properties of CAP against cancer hallmarks.

Confocal Laser Microscopy for VM Analysis with DAPI and Phalloidin Staining.

Confocal laser scanning microscopy (CLSM) is one of the most prevalent fluorescence microscopy techniques for assessing the progression of cancer cells in three-dimensional structures, such as vasculogenic mimicry (VM). We show a basic approach for using DAPI and phalloidin dyes to detect the early stages of progression and VM of melanoma tumor cells grown in a 3D environment, as well as demonstrating how to acquire images and improve them by changing the software acquisition parameters.

Hepatic echinococcosis: difficulties in diagnosis at the early stages of progression and with complications (literature review)

To demonstrate the possible difficulties in the diagnosis of hepatic echinococcosis, that a specialist in ultrasound diagnostics and a radiologist may encounter in everyday practice, russian and foreign literature as well as our own experience have been reviewed. With high accuracy and specificity of ultrasonography in the differential diagnosis of echinococcal cysts, incorrect interpretation is possible in the presence of mural hemorrhages and (or) hemorrhagic clots in a nonparasitic cyst, which aren’t also visible in ultrasonography and can imitate elements of the chitinous membrane. The degree of prevalence of the process, the exclusion of damage to the lungs and other organs of the abdominal cavity are important. In such cases CT scan is the most convenient and accurate diagnostic method due to its high spatial and temporal resolution, standardization and availability. In the most difficult cases the capabilities of MRI are used in the comparison of diffusion-weighted images and apparent diffusion coefficient, which also makes it possible to diagnose cystic neoplasms. Ultrasonography is a diagnostic technology that is still the most commonly used to differentiate cavity structures; however, all dubious clinical observations should be further examined by using CT or MRI.

DNA polymer films used as drug delivery systems to early-stage diagnose and treatment of breast cancer using 3D tumor spheroids as a model

The present study examines the design of DNA polymeric films (DNA-PFs) associated with aluminum chloride phthalocyanine (AlClPc) (DNA-PFs-AlClPc), as a promising drug delivery system (DDS), applicable for breast cancer treatment and early-stage diagnosis using photodynamic therapy (PDT). This study starts evaluating (MCF7) as a model for breast cancer cell behavior associated with DNA-PFs. Analyses of the morphological behaviors, biochemical reaction, and MCF7 cell adhesion profile on DNA-PFs were evaluated. SEM and AFM analysis allowed the morphological characterization of the DNA-PFs. Cell viability and cell cycle kinetics studies indicate highly biocompatible material capable of anchoring MCF7 cells, allowing the attachment and support of cell in the same structure where the insertion of AlClPc (DNA-PFs-AlClPc). The application of visible light photoactivation based on classical PDT protocol over the DNA-PFs-AlClPc showed a reduction in cell viability with increased cell death proportional to the fluency energy range from 600, 900, and 1800 mJ cm−2. The 3D organoid system mimics the tumor microenvironment which was precisely observed in human breast cancer in early-stage progression in the body. The results observed indicate that the viability was reduced by more than 80% in monolayer culture and around 50% in the 3D organoid cell culture at the highest energy fluency (1800 mJ cm−2). With low energy fluency (100 mJ cm−2,), the DNA-PFs-AlClPc did not show a cytotoxic effect on MCF7 cells, enabling the potential for photodiagnosis of early-stage human breast cancer detection in the initial stage of progression.

Impact of solitons on the progression of initial lesion in aortic dissection

Aortic dissection (AD) is the most common catastrophic disease reported at cardiovascular emergency in hospitals. Herein, a tear in the tunica intima results into separation of layers of aortic wall leading to rupture and torrential bleed. Hypoxia and oxidative stress are associated with AD. The release of hypoxia inducible factor (HIF)-[Formula: see text] from the initial flap lesion in the tunica intima is the basis for aneurysmal prone factors. We framed a boundary value problem (BVP) to evaluate homeostatic saturation for oxygen dynamics using steady-state analysis. We prove uniqueness and existence of the solution of the BVP for gas exchange at capillary–tissue interface as a normal physiological function. Failure of homeostatic mechanism establishes hypoxia, a new quasi-steady-state in AD. We model permeation of two-layer fluid comprised of blood and HIF-[Formula: see text] through tunica media as a generalized [Formula: see text]-dimensional nonlinear evolution equation and solve it using Lie group of transformations method. We note that the two-layer fluid permeates the tunica media as solitary wave including solitons such as bright soliton, dark soliton, peregrine soliton, topological soliton, kink soliton, breather soliton and multi-soliton complex. Also, we introduce the main result and discuss the implications of soliton solution, using graphic interpretation, to describe the early stage of progression of AD.

Identification of hub genes and their novel diagnostic and prognostic significance in pancreatic adenocarcinoma.

Objective:The main reasons for the poor prognoses of pancreatic adenocarcinoma (PA) patients are rapid early-stage progression, advanced stage metastasis, and chemotherapy resistance. Identification of novel diagnostic and prognostic biomarkers of PA is therefore urgently needed.Methods:Three mRNA microarray datasets were obtained from the Gene Expression Omnibus database to select differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses for hub genes were performed using DAVID. Correlations between expression levels of hub genes and cancer-infiltrating immune cells were investigated by TIMER. Cox proportional hazard regression analyses were also performed. Serum hub genes were screened using the HPA platform and verified for diagnostic value using ELISAs.Results:We identified 59 hub genes among 752 DEGs. GO analysis indicated that these 59 hub genes were mainly involved in the defense response to viruses and the type I interferon signaling pathway. We also discovered that RSAD2 and SMC4 were associated with immune cell infiltration in the PA microenvironment. Additionally, DLGAP5 mRNA might be used as an independent risk factor for the prognoses of PA patients. Furthermore, the protein encoded by ISG15, which exists in peripheral blood, was validated as a potential diagnostic biomarker that distinguished PA patients from healthy controls (area under the curve: 0.902, 95% confidence interval: 0.819–0.961).Conclusions:Our study suggested that RSAD2 and SMC4 were associated with immune cell infiltration in the PA microenvironment, while DLGAP5 mRNA expression might be an independent risk factor for the survival prognoses of PA patients. Moreover, ELISAs indicated that serum ISG15 could be a potential novel diagnostic biomarker for PA.

Future Role of the Clinical Lab in Population Health.

Future Role of the Clinical Lab in Population Health.

Modeling nonalcoholic fatty liver disease on a liver lobule chip with dual blood supply

Nonalcoholic fatty liver disease (NAFLD) has emerged as a public health concern. To date, the mechanism of NAFLD progression remains unclear, and pharmacological treatment options are scarce. Traditional animal NAFLD models are limited in helping address these problems due to interspecies differences. Liver chips are promising for modeling NAFLD. However, pre-existing liver chips cannot reproduce complex physicochemical microenvironments of the liver effectively; thus, NAFLD modeling based on these chips is incomplete. Herein, we develop a biomimetic liver lobule chip (LC) and then establish a more accurate on-chip NAFLD model. The self-developed LC achieves dual blood supply through the designed hepatic portal vein and hepatic artery and the microtissue cultured on the LC forms multiple structures similar to in vivo liver. Based on the LC, NAFLD is modeled. Steatosis is successfully induced and more importantly, changing lipid zonation in a liver lobule with the progression of NAFLD is demonstrated for the first time on a microfluidic chip. In addition, the application of the induced NAFLD model has been preliminarily demonstrated in the prevention and reversibility of promising drugs. This study provides a promising platform to understand NAFLD progression and identify drugs for treating NAFLD. Statement of significanceLiver chips are promising for modeling nonalcoholic fatty liver disease. However, on-chip replicating liver physicochemical microenvironments is still a challenge. Herein, we developed a liver lobule chip with dual blood supply, achieving self-organized liver microtissue that is similar to in vivo tissue. Based on the chip, we successfully modeled NAFLD under physiologically differentiated nutrient supplies. For the first time, the changing lipid zonation in a single liver lobule with the early-stage progression of NAFLD was demonstrated on a liver chip. This study provides a promising platform for modeling liver-related diseases.

Frequency of repositioning for preventing pressure ulcers in patients hospitalized in ICU: protocol of a cluster randomized controlled trial

BackgroundDespite being considered preventable, ulcers due to pressure affect between 30 and 50% of patients at high and very high risk and susceptibility, especially those hospitalized under critical care. Despite a lack of evidence over the efficacy in prevention against ulcers due to pressure, hourly repositioning in critical care as an intervention is used with more or less frequency to alleviate pressure on patients’ tissues. This brings up the objective of our study, which is to evaluate the efficacy in prevention of ulcers due to pressure acquired during hospitalization, specifically regarding two frequency levels of repositioning or manual posture switching in adults hospitalized in different intensive care units in different Colombian hospitals.MethodsA nurse-applied cluster randomized controlled trial of parallel groups (two branches), in which 22 eligible ICUs (each consisting of 150 patients), will be randomized to a high-frequency level repositioning intervention or to a conventional care (control group). Patients will be followed until their exit from each cluster. The primary result of this study is originated by regarding pressure ulcers using clusters (number of first ulcers per patient, at the early stage of progression, first one acquired after admission for 1000 days). The secondary results include evaluating the risk index on the patients’ level (Hazard ratio, 95% IC) and a description of repositioning complications. Two interim analyses will be performed through the course of this study. A statistical difference between the groups < 0.05 in the main outcome, the progression of ulcers due to pressure (best or worst outcome in the experimental group), will determine whether the study should be put to a halt/determine the termination of the study.ConclusionThis study is innovative in its use of clusters to advance knowledge of the impact of repositioning as a prevention strategy against the appearance of ulcers caused by pressure in critical care patients. The resulting recommendations of this study can be used for future clinical practice guidelines in prevention and safety for patients at risk.Trial registrationPENFUP phase-2 was Registered in Clinicaltrials.gov (NCT04604665) in October 2020.

Prodromal neuroinvasion of pathological α-synuclein in brainstem reticular nuclei and white matter lesions in a model of α-synucleinopathy.

Neuropathological observations in neurodegenerative synucleinopathies, including Parkinson disease, implicate a pathological role of α-synuclein accumulation in extranigral sites during the prodromal phase of the disease. In a transgenic mouse model of peripheral-to-central neuroinvasion and propagation of α-synuclein pathology (via hindlimb intramuscular inoculation with exogenous fibrillar α-synuclein: the M83 line, expressing the mutant human Ala53Thr α-synuclein), we studied the development and early-stage progression of α-synuclein pathology in the CNS of non-symptomatic (i.e. freely mobile) mice. By immunohistochemical analyses of phosphroylated α-synuclein on serine residue 129 (p-S129), our data indicate that the incipient stage of pathological α-synuclein propagation could be categorized in distinct phases: (i) initiation phase, whereby α-synuclein fibrillar inoculum induced pathological lesions in pools of premotor and motor neurons of the lumbar spinal cord, as early as 14 days post-inoculation; (ii) early central phase, whereby incipient α-synuclein pathology was predominantly detected in the reticular nuclei of the brainstem; and (iii) late central phase, characterized by additional sites of lesions in the brain including vestibular nuclei, deep cerebellar nuclei and primary motor cortex, with coincidental emergence of a sensorimotor deficit (mild degree of hindlimb clasping). Intriguingly, we also detected progressive α-synuclein pathology in premotor and motor neurons in the thoracic spinal cord, which does not directly innervate the hindlimb, as well as in the oligodendroglia within the white matter tracts of the CNS during this prodromal phase. Collectively, our data provide crucial insights into the spatiotemporal propagation of α-synuclein pathology in the nervous system of this rodent model of α-synucleinopathy following origin in periphery, and present a neuropathological context for the progression from pre-symptomatic stage to an early deficit in sensorimotor coordination. These findings also hint towards a therapeutic window for targeting the early stages of α-synuclein pathology progression in this model, and potentially facilitate the discovery of mechanisms relevant to α-synuclein proteinopathies. In a rodent model of synucleinopathy, Ferreira et al., delineate the spatiotemporal progression of incipient α-synuclein pathology (of peripheral origin) in the CNS. The authors show early affection of brainstem reticular nuclei in non-paralyzed mice, and pathological white matter lesions in relation to the neuronal pathology.

APPROPRIATE LABORATORY AND IMAGING APPROACH FOR EARLY DIAGNOSIS IN HOSPITAL SETTING

The main objective of global health is to reduce preventable death equally throughout the world. One important strategy is primary prevention through health promotion and early detection. Early diagnosis is defined as several methods to determine in patients the nature of a disease or disorder at its early stage of progression. In a referral health system, such as Indonesia, early diagnosis could be implemented both in primary care and secondary or tertiary referral hospital. Primary care centres screens fatal diseases, such as cancer and cardiovascular risk factors, mostly in asymptomatic patients with identified risk of having the diseases. Accordingly, the early detection programme has been designed for large population in a community setting. It is uncommon for hospitals in Indonesia applying primary prevention where most of the patients are referred with at least a couple of differential diagnoses. Subsequently, early diagnosis in hospital setting would be defined either as accelerating the diagnosis confirmation or detecting any uncommon findings on patients with atypical presentation. In some situation, early detection of life-threatening condition at hospital would prolong or even prevent death. Aside from accurate history taking and comprehensive physical examination, advance laboratory procedure and imaging might assist in early detection at hospital setting. This time, our issue presents case reports on the benefit of early detection in the management of rare disease, anaemia, and COVID-19 pandemic among patients without symptoms or with atypical presentation.&#x0D; In developed countries, most of known congenital diseases were screened prenatally or immediately after delivery. The most common practice in Indonesia is a regular prenatal ultrasound and through physical examination of the baby in their first week. However, this approach would still miss many rare diseases with delayed presentation such as Legg-Calve-Perthes Diseases (LCPD). Pradyana and Astuti1 discussed a paediatric case of LCPD highlighting the difficulty in diagnosis confirmation on a limp gait symptom. Serial imaging not only important in case follow-up, but also to detect newly existing abnormalities which verify the presence of LCPD.&#x0D; Iron deficiency anaemia (IDA) primarily affects women and children in Indonesia. Currently, school-based screening of haemoglobin in teenage girls could prevent the sequalae of IDA through iron supplementation. Early detection in anaemia would determine the presence of IDA before the declined haemoglobin. Recently, international investigators discovered novel biomarker to detect the presence of IDA in asymptomatic patients. Fenty described the potential of using reticulocyte haemoglobin equivalent (Ret-He) to characterize IDA amongst asymptomatic patients in hospital settings.2 This approach would be beneficial for screening purpose in high-risk population, both in community and hospital setting.&#x0D; During COVID-19 pandemic, early diagnosis is a necessity to reduce the infection rate with early quarantine. However, challenging situation might come in patients with atypical presentation, such as gastritis. Widiastana et al describe the later COVID positive result on patients with gastritis which nonresponsive to general treatment.2 The patients did not have any classical signs and symptoms of COVID-19, hence in the community, misdiagnosis of similar condition might exacerbate the rising infection cases.

A Transcriptomics-Based Meta-Analysis Combined With Machine Learning Identifies a Secretory Biomarker Panel for Diagnosis of Pancreatic Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is generally incurable due to the late diagnosis and absence of markers that are concordant with expression in several sample sources (i.e., tissue, blood, plasma) and platforms (i.e., Microarray, sequencing). We optimized meta-analysis of 19 PDAC (tissue and blood) transcriptome studies from multiple platforms. The key biomarkers for PDAC diagnosis with secretory potential were identified and validated in different cohorts. Machine learning approach i.e., support vector machine supported by leave-one-out cross-validation was used to build and test the classifier. We identified a 9-gene panel (IFI27, ITGB5, CTSD, EFNA4, GGH, PLBD1, HTATIP2, IL1R2, CTSA) that achieved ∼0.92 average sensitivity and ∼0.90 average specificity in distinguishing PDAC from healthy samples in five training sets using cross-validation. These markers were also validated in proteomics and single-cell transcriptomics studies suggesting their prognostic role in the diagnosis of PDAC. Our 9-gene classifier can not only clearly discriminate between better and poor survivors but can also precisely discriminate PDAC from chronic pancreatitis (AUC = 0.95), early stages of progression [Stage I and II (AUC = 0.82), IPMA and IPMN (AUC = 1), and IPMC (AUC = 0.81)]. The 9-gene marker outperformed the previously known markers in blood studies particularly (AUC = 0.84). The discrimination of PDAC from early precursor lesions in non-malignant tissue (AUC > 0.81) and peripheral blood (AUC > 0.80) may assist in an early diagnosis of PDAC in blood samples and thus will also facilitate risk stratification upon validation in clinical trials.

Evaluation of Salivary Lactate Dehydrogenase as a Prognostic Biomarker in Tobacco Users with and without Potentially Malignant Disorders of the Oral Cavity.

Background Oral squamous cell carcinoma (OSCC) is a deadly disease that develops in a multistage process and is often preceded by oral potentially malignant disorders (PMDs), of which many are caused by tobacco usage. It is associated with a shift from an aerobic to anaerobic glycolytic pathway, and hence an increase in lactate dehydrogenase (LDH) levels is seen. The objective of this study was to estimate and correlate the level of salivary LDH in healthy individuals with tobacco users (with or without PMDs). Methods A total of 78 patients were selected from the Department of Oral Medicine and Radiology, Dayananda Sagar College of Dental Sciences, Bengaluru, Karnataka, India, and were divided into three groups of 26 patients each, namely controls, tobacco users without PMD, and tobacco users with PMD. A total of 1 mL of unstimulated saliva was collected from each patient using the spit method. Sample was subjected to centrifugation at 2,500 rpm, and salivary LDH was quantified by a standard kit (LDH-P kit, DIALAB, Neudorf, Austria) using an autoanalyzer. Results There was a statistically significant ( p < 0.001) gradual increase in the level of LDH in controls (267 ± 27.64 U/L), tobacco users without PMD (391 ± 80.53 U/L), and tobacco users with PMD (706.1 ± 199 U/L). Increase in the LDH level was also noted with increased duration and frequency of the habit. Conclusion LDH can potentially be used as a promising biomarker in the very early stages of progression toward oral cancer caused by tobacco use.

Crosstalk Between Prostate Cancer Cells and Tumor-Associated Fibroblasts Enhances the Malignancy by Inhibiting the Tumor Suppressor PLZF.

Although prostate cancer is clinically manageable during the early stages of progression, metastatic progression severely compromises the prognosis and leads to mortality. Constitutive activation of STAT3 has been connected to prostate cancer malignancy, and abolishing the STAT3 activity may diminish tumor growth and metastasis. However, its suppressor genes and pathways have not been well established. In this study, we show that promyelocytic leukemia zinc finger (PLZF) has a putative tumor-suppressor function in prostate cancer by inhibiting phosphorylation of STAT3. Compared with a benign prostate, high-grade prostate cancer patient tissue was negatively correlated with PLZF expression. PLZF depletion accelerated proliferation and survival, migration, and invasion in human prostate cancer cells. Mechanistically, we demonstrated a novel role of PLZF as the transcriptional regulator of the tyrosine phosphatase SHP-1 that inhibits the oncogenic JAKs–STAT3 pathway. These results suggest that the collapse of PLZF expression by the CCL3 derived from fibroblasts accelerates the cell migration and invasion properties of prostate cancer cells. Our results suggest that increasing PLZF could be an attractive strategy for suppressing prostate cancer metastasis as well as for tumor growth.

Help, I Need Somebody! Business and Technology Advice in Emerging Science-Based Ventures

Early-stage science-based ventures (SBVs) require a wide range of intellectual resources and know-how to successfully commercialize their technologies. Often entrepreneurs gain this knowledge through advisory relationships providing commercial and technical guidance. We explore the effects of those dual advice domains – business and technology – on the early-stage progress of SBVs. Using longitudinal data from 112 emerging SBVs, we find that business advice has a positive impact on SBV progress, while technology advice does not, and in fact, delays the process of finding a market application for the SBV’s technology. We further find that advice complementary to the entrepreneur’s experience, thus creating greater knowledge variety in the SBV, will lead to a more positive outcome. These insights help unpack the underexplored mechanisms of advice absorption in SBVs.

Breast cancer mortality trends in Europe among women in perimenopausal and postmenopausal age (45+)

IntroductionThe aim of the study was to analyze breast cancer (BC) mortality trends among women at the age of 45 years old and older (45+) in the 28 European Union (EU) countries, as well as in 3 non-EU countries – Norway, Switzerland and the Russian Federation (control group) within the period 1959–2017.Material and methodsMortality and population data were sourced from the World Health Organization (WHO) database, and age-standardized mortality rates were calculated using the standard world population. Changes in mortality trends were analyzed using Joinpoint Trend Analysis Software.ResultsThe majority of analyzed countries showed a meaningful decrease in BC mortality among women aged 45+. However, the results of our study suggest that there are 4 EU countries – Croatia, Poland, Romania and Slovakia – where increasing BC mortality trends started to be visible in the analyzed age group. Currently, the observed increase is still not significant, but the obtained data suggest the possibility of further continuation of the observed trend in the future. Moreover, in Bulgaria we also noted continuation of the increase in BC mortality (statistically significant).ConclusionsDue to the availability of better treatment options, as well as presence of effective tools for detecting BC at the early stages of progression, BC mortality is falling in most analyzed European countries. To maintain this situation and to stop BC mortality increase in the analyzed age group in Bulgaria, Croatia, Poland, Romania and Slovakia, immediate actions for improvement of BC management in the European health care systems should be considered.

Validation of a Fast, Robust, Inexpensive, Two-Tiered Neonatal Screening Test algorithm on Dried Blood Spots for Spinal Muscular Atrophy.

Spinal muscular atrophy (SMA) is one of the leading genetic causes of infant mortality with an incidence of 1:10,000. The recently-introduced antisense oligonucleotide treatment improves the outcome of this disease, in particular when applied at an early stage of progression. The genetic cause of SMA is, in >95% of cases, a homozygous deletion of the survival motor neuron 1 (SMN1) gene, which makes the low-cost detection of SMA cases as part of newborn screening programs feasible. We developed and validated a new SALSA MC002 melting curve assay that detects the absence of the SMN1 exon 7 DNA sequence without detecting asymptomatic carriers and reliably discriminates SMN1 from its genetic homolog SMN2 using crude extracts from newborn screening cards. Melting curve analysis shows peaks specific for both the SMN1 gene and the disease modifying SMN2 homolog. The detection of the SMN2 homolog, of which the only clinically relevant difference from the SMN1 gene is a single nucleotide in exon 7, was only used to confirm a correct reaction in samples that lacked the SMN1 gene, and not for SMN2 quantification. We retrieved 47 DBS samples from children with genetically-confirmed SMA, after informed consent from parents, and 375 controls from the national archive of the Dutch National Institute for Public Health and the Environment (RIVM). The assay correctly identified all anonymized and randomized SMA and control samples (i.e., sensitivity and specificity of 100%), without the detection of carriers, on the three most commonly-used PCR platforms with melting curve analysis. This test’s concordance with the second-tier ‘golden standard’ P021 SMA MLPA test was 100%. Using the new P021–B1 version, crude extracts from DBS cards could also be used to determine the SMN2 copy number of SMA patients with a high level of accuracy. The MC002 test showed the feasibility and accuracy of SMA screening in a neonatal screening program.

Epigenetic regulation of mammalian Hedgehog signaling to the stroma determines the molecular subtype of bladder cancer.

In bladder, loss of mammalian Sonic Hedgehog (Shh) accompanies progression to invasive urothelial carcinoma, but the molecular mechanisms underlying this cancer-initiating event are poorly defined. Here, we show that loss of Shh results from hypermethylation of the CpG shore of the Shh gene, and that inhibition of DNA methylation increases Shh expression to halt the initiation of murine urothelial carcinoma at the early stage of progression. In full-fledged tumors, pharmacologic augmentation of Hedgehog (Hh) pathway activity impedes tumor growth, and this cancer-restraining effect of Hh signaling is mediated by the stromal response to Shh signals, which stimulates subtype conversion of basal to luminal-like urothelial carcinoma. Our findings thus provide a basis to develop subtype-specific strategies for the management of human bladder cancer.

Memory Performance of People with Different Dementia Severity for Different Semantic Hierarchies

ABSTRACTBackground: Memory disorder is a significant symptom during early-stage Alzheimer’s disease (AD). Changes in semantic memory are frequently seen in terms of forgetting names, loss of word meanings, and difficulties in linguistic expression. Significant semantic degeneration is not a normal phenomenon in elderly persons, and it may be an important sign in the early stages of progression of AD.Methods: Thirty-four participants aged between 60 and 86 years were recruited for an experiment with a 3 × 4 × 2 factorial design that was conducted to explore the differences in semantic memory performance among controls with normal cognitive performance (NC), individuals classified as mildly cognitively impaired (MCI), and individuals with AD.Results: The performance of participants diagnosed with mild AD was poorest for the attribute category, and there was no difference in response to different word frequencies. Although those diagnosed with MCI performed similarly to healthy elderly participants in terms of semantic memory, their performance profiles for different semantic hierarchies were similar to those of participants with AD.Conclusion: Semantic memory had degraded among participants with AD and MCI, and the rate of semantic degeneration was different in different semantic hierarchies.