Higher Dosage of the Epidermal Growth Factor Receptor Mutant Allele in Lung Adenocarcinoma Correlates with Younger Age, Stage IV at Presentation, and Poorer Survival

IntroductionThe clinical significance of epidermal growth factor receptor (EGFR) mutant allele specific imbalance (MASI) in lung adenocarcinomas is unknown.MethodsEGFR MASI was characterized by sequencing electropherograms (SEs) and EGFR fluorescence in situ hybridization (FISH) in 96 prospectively tested lung adenocarcinoma patients with a median follow-up of 20 months (all cases were EGFR mutation-positive).ResultsIn 25 cases, the mutant allele (MA) peak was higher than the wild-type allele (WA) peak, indicating the presence of EGFR MASI (25/96, 26%). The adenocarcinomas with EGFR MASI had a 4.4-fold higher average EGFR/Chromosome Enumeration Probe 7 ratio than carcinomas without MASI (7.9 ± 3.8 versus 1.8 ± 0.6, p = 0.01). A high degree of correlation between the MA/WA ratio (SE) and the EGFR/CEP7 ratio (FISH) (ρ = 0.757, p = 0.003) validated the quantitative nature of SE. Amplification was the most common mechanism of EGFR MASI (13/21, 62%). EGFR MASI was more commonly associated with exon 19 mutations than with exon 21 mutations (19/53, 36%, versus 6/43, 14%, p = 0.015, odds ratio [OR] = 3.4) and in patients younger than 65 years (17/46, 37%, versus 8/50, 16%, p = 0.019, OR = 3.1). Patients with EGFR MASI presented with stage IV disease more frequently (p = 0.01, OR = 3.5) and had a poorer disease-specific survival rate (p = 0.021, 54% versus 83% at 31 months).ConclusionsEGFR MASI in lung adenocarcinomas can be assessed based on SE and can be used to identify younger patients with more aggressive disease

Epidermal growth factor receptor variant III mediates head and neck cancer cell invasion via STAT3 activation.

Epidermal growth factor receptor (EGFR) is frequently overexpressed in head and neck squamous cell carcinoma (HNSCC) where aberrant signaling downstream of this receptor contributes to tumor growth. EGFR variant III (EGFRvIII) is the most commonly altered form of EGFR and contains a truncated ligand-binding domain. We previously reported that EGFRvIII is expressed in up to 40% of HNSCC tumors where it is associated with increased proliferation, tumor growth and chemoresistance to antitumor drugs including the EGFR-targeting monoclonal antibody cetuximab. Cetuximab was FDA-approved in 2006 for HNSCC but has not been shown to prevent invasion or metastasis. This study was undertaken to evaluate the mechanisms of EGFRvIII-mediated cell motility and invasion in HNSCC. We found that EGFRvIII induced HNSCC cell migration and invasion in conjunction with increased signal transducer and activator of transcription 3 (STAT3) activation, which was not abrogated by cetuximab treatment. Further investigation showed that EGF-induced expression of the STAT3 target gene HIF1-α, was abolished by cetuximab in HNSCC cells expressing wild-type EGFR under hypoxic conditions, but not in EGFRvIII-expressing HNSCC cells. These results suggest that EGFRvIII mediates HNSCC cell migration and invasion by increased STAT3 activation and induction of HIF1-α, which contribute to cetuximab resistance in EGFRvIII-expressing HNSCC tumors

In vitro FRAP reveals the ATP-dependent nuclear mobilization of the exon junction complex protein SRm160

We present a new in vitro system for characterizing the binding and mobility of enhanced green fluorescent protein (EGFP)-labeled nuclear proteins by fluorescence recovery after photobleaching in digitonin-permeabilized cells. This assay reveals that SRm160, a splicing coactivator and component of the exon junction complex (EJC) involved in RNA export, has an adenosine triphosphate (ATP)-dependent mobility. Endogenous SRm160, lacking the EGFP moiety, could also be released from sites at splicing speckled domains by an ATP-dependent mechanism. A second EJC protein, RNPS1, also has an ATP-dependent mobility, but SRm300, a protein that binds to SRm160 and participates with it in RNA splicing, remains immobile after ATP supplementation. This finding suggests that SRm160-containing RNA export, but not splicing, complexes have an ATP-dependent mobility. We propose that RNA export complexes have an ATP-regulated mechanism for release from binding sites at splicing speckled domains. In vitro fluorescence recovery after photobleaching is a powerful tool for identifying cofactors required for nuclear binding and mobility

Structura afecțiunilor biliare la copii

Background. Gallbladder diseases in children nationwide are rarely diagnosed, they have a share of 9.2-8% in the structure of chronic digestive diseases. Objective of the study. To determine the rate and nosological structure of gallbladder diseases in children with chronic digestive diseases hospitalized in the pediatric-gastroenterology clinic of MCCH "Valentin Ignatenco" in 2019. Material and Method. The study was a retrospective, with analysis of clinical data, laboratory, EGDS and abdominal sonography of 694 medical records of children aged 2-18 years treated in the nominated subdivision with chronic digestive pathology. In 28 cases the morphofunctional test with breakfast was performed. Results. Gallbladder diseases in children with chronic digestive pathology were found in 7.6% (53) of cases, the average age being 9.8 years and the predominance of girls (36). All children showed pain in the right hypochondrium and positive signs of gallbladder damage, hepatomegaly <2 cm with normal ALT and AST (18). The nosological structure of the diagnosed gallbladder pathology included: functional disorders of the gallbladder - 88% (47) cases (motor dyskinesias - 28 and spasm of the Oddi sphincter of the gallbladder type - 19) age between 10-14 years; chronic calculous cholecystitis - 7.5% (4) and calculous 3.8% (2) cases. Bilateral sludge was found in 28 children at the same time. Conclusion. Gallbladder diseases in children with chronic digestive pathology are more frequently dominated by motor functional disorders. Introducere. Afecțiunile veziculei biliare la copii în plan național sunt rar diagnosticate, acestea având o pondere de 9,2-8% în structura bolilor digestive cronice. Scopul lucrării. A determina rata și structura nosologică a afecțiunilor veziculei biliare la copiii cu maladii cronice digestive, spitalizați în Clinica pediatrie-gastroenterologie a SCMC „Valentin Ignatenco" în 2019. Material și Metode. Studiul realizat a fost unul retrospectiv, cu analiza datelor clinice, de laborator, EGDS și sonografiei abdominale a 694 de fișe medicale a copiilor cu vârsta 2-18 ani, tratați în subdiviziunea nominalizată cu patologie cronică digestivă. În 28 de cazuri a fost efectuat testul morfofuncțional cu dejun. Rezultate. Afecțiunile veziculei biliare la copiii cu patologie cronică digestivă a fost constatată în 7,6% (53) dintre cazuri, vârsta medie fiind de 9,8 ani și cu predominarea fetelor (36). Toți copiii au prezentat dureri în hipocondrul drept și semne pozitive de afectare a veziculei biliare, hepatomegalie<2cm cu ALT și AST normale (18). Structura nosologică a patologiei veziculei biliare diagnosticate a inclus: dereglări funcționale ale veziculei biliare- 88% (47) cazuri (dischinezii motorice- 28 și spasm al sfincterului Oddi de tip biliar- 19) vârsta între 10-14 ani; colecistită cronică acalculoasă- 7,5%(4) și calculoasă 3,8%(2) cazuri. La 28 de copii concomitent a fost constatat sludge-ul biliar. Concluzii. Afecțiunile veziculei biliare la copiii cu patologie cronică digestivă sunt mai frecvent dominate de dereglările funcționale motorice

COVID-19 infection and liver damage in children. Clinical case study

Introduction. COVID-19 is currently considered a systemic disease affecting the immune system, primarily, lungs, heart, central nervous system, kidneys, intestines, liver and spleen. Impaired liver function and the presence of biochemical changes in liver can be found in approximately 14-53% of adults infected with SARS-CoV-2. Impaired liver function in patients infected with COVID-19 may occur due to a direct effect of the virus on hepatocytes, as well as being secondary to factors such as a systemic inflammatory response of the infected host, the onset of hypoxia (associated with lung damage), multiple organ failure, or due to abusive treatment using overlapping and hepatotoxic drugs. The purpose of this article is to describe a clinical case study regarding the clinical and paraclinical manifestations of liver damage in a 12-year-old child infected with SARSCoV-2, hospitalized at the Municipal Children’s Clinical Hospital „Valentin Ignatenco”, Republic of Moldova. Material and methods. The epidemiological, clinical and paraclinical data were used to highlight this study, followed by the conclusions of multidisciplinary specialists, retrieved from the inpatient medical records of the 12-yearold child with moderate COVID-19 infection, who was admitted for emergency treatment. Results. A 12-year-old patient F. was admitted to the „Covid-19” subunit, complaining of severe general malaise, fever up to 39°C, cough, rhinorrhea. The objective clinical examination revealed hepatomegaly and lack of splenomegaly. Laboratory findings determined leukocytosis 15.88 x109 /l, neutrophilia 72.2%, lymphopenia 26.1%, increased ESR (Erythrocyte Sedimentation rate) – 20 mm/h, increased CRP (C-reactive protein) >12.0 mg/l, increased ALT (alanine aminotransferase) by 16 (50.9-487-764 U/l) values compared to the normal reference and a 3-fold increase in AST (aspartate aminotransferase) that is higher than the normal range (55.8 - 113 - 181 U/l), an increased fibrinogen - 5.3 g/l, increased ferritin - 2834 pmol/l and D-dimer levels - 762 ng/ml. Hepatomegaly was detected on abdominal ultrasound. Covid-19 infection was confirmed by a rapid test of nasopharyngeal exudate for SARS-CoV-2 antigens. Conclusions. Patients with the novel coronavirus (COVID-19) show varying degrees of liver dysfunction, especially those with increased levels of AST and ALT. A question arises within the clinical practice, as whether the liver damage occurred due to direct viral hepatotoxicity or due to the drugs used in COVID-19 treatment

Inducible changes in cell size and attachment area due to expression of a mutant SWI/SNF chromatin remodeling enzyme

The SWI/SNF enzymes belong to a family of ATP-dependent chromatin remodeling enzymes that have been functionally implicated in gene regulation, development, differentiation and oncogenesis. BRG1, the catalytic core subunit of some of the SWI/SNF enzymes, can interact with known tumor suppressor proteins and can act as a tumor suppressor itself. We report that cells that inducibly express ATPase-deficient versions of BRG1 increase in cell volume, area of attachment and nuclear size upon expression of the mutant BRG1 protein. Examination of focal adhesions reveals qualitative changes in paxillin distribution but no difference in the actin cytoskeletal structure. Increases in cell size and shape correlate with over-expression of two integrins and the urokinase-type plasminogen activator receptor (uPAR), which is also involved in cell adhesion and is often over-expressed in metastatic cancer cells. These findings demonstrate that gene expression pathways affected by chromatin remodeling enzymes can regulate the physical dimensions of mammalian cell morphology

JAK Kinase Inhibition Abrogates STAT3 Activation and Head and Neck Squamous Cell Carcinoma Tumor Growth

AbstractAberrant activation of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) 3 has been implicated in cell proliferation and survival of many cancers including head and neck squamous cell carcinoma (HNSCC). AZD1480, an orally active pharmacologic inhibitor of JAK1/JAK2, has been tested in several cancer models. In the present study, the in vitro and in vivo effects of AZD1480 were evaluated in HNSCC preclinical models to test the potential use of JAK kinase inhibition for HNSCC therapy. AZD1480 treatment decreased HNSCC proliferation in HNSCC cell lines with half maximal effective concentration (EC50) values ranging from 0.9 to 4 μM in conjunction with reduction of pSTAT3Tyr705 expression. In vivo antitumor efficacy of AZD1480 was demonstrated in patient-derived xenograft (PDX) models derived from two independent HNSCC tumors. Oral administration of AZD1480 reduced tumor growth in conjunction with decreased pSTAT3Tyr705 expression that was observed in both PDX models. These findings suggest that the JAK1/2 inhibitors abrogate STAT3 signaling and may be effective in HNSCC treatment approaches

A profile of traumatic brain injury within hospital emergency departments — a retrospective study in the Republic of Moldova

Abstract Background: Traumatic brain injury (TBI) is a critical public health and socio- economic problem throughout the world, making epidemiological monitoring of incidence, prevalence, and outcome of TBI necessary. TBI is a major cause of mortality and morbidity in adolescents, young adults, and the elderly, one of the leading causes being road traffic accidents. Methods: A retrospective study was conducted among patients with TBI within 2 medical institutions in Chisinau municipality: Emergency Medicine Institute (EMI) and Valentin Ignatenco Municipal Children’s Hospital (MCH). A questionnaire was applied, completed on the basis of medical records according to the International Classification of Diseases (ICD) 10 codes. The collection period was August 1-October 31, 2018. Data were uploaded using the existing electronic data collection tool—Red Cap and analyzed through Microsoft Excel. Data collection was performed by a resident neurosurgery and a scientific researcher. The ethics committee’s approval has been obtained. Results: There have been identified 150 patients: 57 cases (38.5%) of TBI among children and 93 cases (61.5%) among adults aged between 18 - 73 years old. A large majority (62%) of head injuries were among patients from the urban area (most in adults—60% and males—74%). The most common mechanisms of head injury were falls (53.3%) and road traffic injuries (24%), followed by assault (14.7%) and struck by/or against (8%). The distributions by place of occurrence highlighted that most injuries occurred at home (33.4%) and in transport area (25.3%). Most head injuries were registered among men 121 (81.2%) with a predominance of minor Glasgow Coma Scale (GCS) (65.1%), followed by moderate GCS (9.4%), while in women all cases with GCS minor (18.8%). Conclusion: The data obtained could be useful for the hospital administration in managing the necessary resources and for conducting information campaigns among the high-risk groups