Follow-Up Study Confirms the Presence of Gastric Cancer DNA Methylation Hallmarks in High-Risk Precursor Lesions.

To adopt prevention strategies in gastric cancer, it is imperative to develop robust biomarkers with acceptable costs and feasibility in clinical practice to stratified populations according to risk scores. With this aim, we applied an unbiased genome-wide CpG methylation approach to a discovery cohort composed of gastric cancer (n = 24), and non-malignant precursor lesions (n = 64). Then, candidate-methylation approaches were performed in a validation cohort of precursor lesions obtained from an observational longitudinal study (n = 264), with a 12-year follow-up to identify repression or progression cases. H. pylori stratification and histology were considered to determine their influence on the methylation dynamics. As a result, we ascertained that intestinal metaplasia partially recapitulates patterns of aberrant methylation of intestinal type of gastric cancer, independently of the H. pylori status. Two epigenetically regulated genes in cancer, RPRM and ZNF793, consistently showed increased methylation in intestinal metaplasia with respect to earlier precursor lesions. In summary, our result supports the need to investigate the practical utilities of the quantification of DNA methylation in candidate genes as a marker for disease progression. In addition, the H. pylori-dependent methylation in intestinal metaplasia suggests that pharmacological treatments aimed at H. pylori eradication in the late stages of precursor lesions do not prevent epigenome reprogramming toward a cancer signature.

Subcutaneous panniculitis-like T-cell lymphoma, lupus erythematosus profundus, and overlapping cases: molecular characterization through the study of 208 genes.

Subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is a rare cytotoxic cutaneous lymphoma. Differential diagnosis with lupus erythematosus panniculitis (LEP) can be challenging and overlapping cases have been described. In this study, we investigate whether gene expression profiling may or not identify markers that can be used to improve our understanding of the disease and to make a precise differential diagnosis. SPTCL, LEP, and overlapping cases were analyzed using a customized NanoString platform including 208 genes related to T-cell differentiation, stromal signatures, oncogenes, and tumor suppressor genes. Gene expression unsupervised analysis of the samples differentiated SPTCL from LEP samples. Most overlapping cases were clustered with LEP cases. Differentially expressed genes were observed when comparing SPTCL with LEP cases; and overlapping with LEP cases. Gene set enrichment analysis recognized gene sets defining each group. In conclusion, SPTCL and LEP have distinctive molecular profiles and the molecular background of overlapping cases more closely resembles LEP.

Role of endoscopic ultrasound-guided fine-needle aspiration in the diagnosis and classification of lymphomas.

BACKGROUND: endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is a safe and effective technique in the diagnosis of mediastinal and abdominal masses. However, the usefulness of EUS-FNA in the diagnosis and classification of lymphomas is controversial. The aim of this study was to determine the yield of EUS-FNA in the diagnosis and classification of lymphomas. METHODS: a retrospective case series was performed in a tertiary referral center. All consecutive patients referred for EUS-FNA with a suspected diagnosis of lymphoma from March 2013 to June 2019 were included. RESULTS: thirty-five patients (54.3 % women, median age 72 years) were included. The most frequent location of the node was the abdomen (67.9 %). Nodes were punctured using a 22-gauge (85.7 %) and 19-gauge needle (14.3 %) with a slow-pull technique. The number of passes performed were three or more in 82.9 % of patients. The samples were processed by the cellblock method. Adequate samples for immunohistochemical and molecular biological study were obtained in 33 (94.3 %) patients. EUS-FNA correctly diagnosed lymphoma in 30 out of 35 patients (85.7 %), and subclassification was determined in 23 patients (65.7 %). The most frequent diagnosis was non-Hodgkin lymphoma (85.7 %). There was one moderate adverse event (bleeding), which was resolved during the procedure. CONCLUSIONS: EUS-FNA may be a valuable technique in the evaluation of suspected lymphomas with an adequate diagnostic yield and a very low rate of adverse events.

Development of Action Potential Waveform in Hippocampal CA1 Pyramidal Neurons.

CA1 pyramidal neurons undergo intense morphological and electrophysiological changes from the second to third postnatal weeks in rats throughout a critical period associated with the emergence of exploratory behavior. Using whole cell current-clamp recordings in vitro and neurochemical methods, we studied the development of the somatic action potential (AP) waveform and some of the underlying channels in this critical period. At the third postnatal week, APs showed a more hyperpolarized threshold, higher duration and amplitude. Subthreshold depolarization broadened APs and depolarized their peak overshoots more pronouncedly in immature neurons (2 weeks old). These features were mimicked by pharmacologically blocking the fast-inactivating A-type potassium current (IA) and matched well with the higher concentrations of Kv4.2 and Kv4.3 and the lower concentrations of BK and Kv1.2 channels detected by Western blotting. Repetitive stimulation with high frequency trains (50 Hz) reproduced AP broadening associated to inactivation of the A-type current in immature cells. Moreover, repetitive firing showed changes in AP amplitude consistent with the inactivation of both sodium and potassium subthreshold currents, which resulted in higher AP amplitudes in the more immature neurons. We propose that maturation of AP waveform and excitability in this critical developmental period could be related to the onset of exploratory behaviors.

Cholangiocarcinoma, 10-Year Survival After Liver Transplantation: A Case Report.

Cholangiocarcinoma is the second most common neoplasm in the liver, with a very poor, short-term prognosis. Today, surgery associated with or without an adjuvant is the only curative treatment. Liver transplantation (LT) is the best treatment for hepatocellular carcinoma tumor. In recent years, treatment of hilar cholangiocarcinoma by LT associated with neoadjuvant therapy has been studied under a criterion. But could it be possible to apply LT like the curative treatment of intrahepatic cholangiocarcinoma (iCC)? Initially the answer is no, but there are different studies about incidental LT in patients with iCC that demonstrate survival over 40% to 50%. In our center, we conducted a review of 468 transplants completed between 2002 and 2018, and we identified 1 case of incidental LT in a patient with iCC with an overall survival of 10 years. Because there is currently an increase in donors owing to the expansion of the criteria, a study to consider extending the criteria of LT to include iCC would be beneficial.

Eight Years of Extracorporeal Membrane Oxygenation in Liver Transplantation: Our Experience.

Liver grafts from donors after cardiac death (DCD) involve a risk of failure owing to warm ischemia, among other factors. To minimize this important issue, new systems like normothermic regional perfusion have arisen. We report an observational and unicentric study focused on the results of liver transplantation after DCD, performing normothermic regional perfusion using extracorporeal membrane oxygenation. In the period between 2011 and 2018, 33 recipients underwent the procedure, 9 from Maastricht II DCD donor liver transplantation (LT) and 24 from Maastricht III DCD donors. The median recipient survival rose to 67 ± 9 months and 41 ± 7 months, respectively. Only 1 patient suffered from ischemic cholangiopathy needing retransplantation. Therefore, according to our experience, liver grafts from DCD using extracorporeal membrane oxygenation are suitable for LT.

Novel presenilin 1 mutation (p.Thr-Pro116-117Ser-Thr) in a Spanish family with early-onset Alzheimer's disease.

Presenilin 1 (PSEN1) is a γ-secretase component, which is in charge of the amyloid precursor protein (APP) cleavage. APP is believed to play a central role in the pathogenesis of Alzheimer's disease (AD). PSEN1 mutations are the most important causes of familial AD, being related to the earlier onset and rapid progression of the disease. Presenilins and APP mutations represent an extraordinary opportunity to study the pathophysiology of AD. We describe the clinical and genetic study of a 37-year-old male patient with a novel mutation in PSEN1 (p.Thr-Pro116-117Ser-Thr). We have studied the pedigree of his family with a further 9 members affected, all of them with onset in their 30s. We have also described the clinical data and results of brain biopsies in 2 of them. DNA sequencing of a tissue sample from an uncle of the patient, who died of AD in the 80s, showed the same mutation as in the patient. These data and predictive analysis indicate the pathogenicity of the mutation.

Role of low-voltage-activated calcium current and extracellular calcium in controlling the firing pattern of developing CA1 pyramidal neurons.

The firing pattern of individual neurons is an important element for information processing and storing. During the first weeks of development, there is a transitional period during which CA1 pyramidal neurons display burst-spiking behavior in contrast to the adult regular-firing pattern. Spike after-depolarizations (ADPs) constitute a major factor underlying burst-spiking behavior. Using current-clamp recordings, we studied ADP waveforms and firing patterns in CA1 pyramidal neurons of Wistar rats from 9 to 19 postnatal days (P9-19). The percentage of burst-spiking neurons increased up to P16, in correlation with the emergence of an active component in the ADP. The application of low-voltage-activated (LVA) calcium channel blockers such as nickel or mibefradil suppressed the generation of the active ADP component and burst-spiking behavior. In agreement with the development of the ADP waveform and burst-spiking behavior, voltage-clamp experiments in dissociated pyramidal neurons showed an increase in the LVA calcium current in P16-19 vs P9-12. Finally, we found that a reduction of extracellular calcium levels decreases the percentage of burst-spiking cells due to a reduction in the active component of the ADP. We conclude that a major contribution of LVA calcium channels to ADP determines the bursting capability of CA1 pyramidal neurons during a transitional postnatal period in contrast to adulthood.

Lysine Restriction and Pyridoxal Phosphate Administration in a NADK2 Patient.

We report the case of a 10-year-old Spanish girl with mutations in NADK2 Prenatal central nervous system abnormalities showed ventriculomegaly, colpocephaly, and hypoplasia of the corpus callosum. At birth, axial hypotonia, uncoordinated movements, microcephaly, and generalized cerebellar atrophy were detected. Metabolic investigations revealed high lysine, lactate, and pipecolic acid levels in blood and cerebrospinal fluid. Pyruvate carboxylase and pyruvate dehydrogenase activity in fibroblasts were normal. Beginning at birth she received biotin, thiamine, and carnitine supplementation. A lysine-restricted diet was started when she was 1 month old. Because pipecolic acid was high, pyridoxine was added to treatment. At 3 years old, astatic myoclonic epilepsy appeared, with no response to levetiracetam. We switched pyridoxine to pyridoxal phosphate, with electroclinical improvement. Because the activity of mitochondrial respiratory chain complexes III and IV was slightly low in muscle, other cofactors such as ubidecarenone, idebenone, vitamin E, and creatine were added to the treatment. At 8 years old, plasma acylcarnitine testing was performed, and high levels of 2-trans, 4-cis-decadienoylcarnitine were found. Whole exome sequencing identified a homozygous splice site mutation in NADK2 (c.956+6T>C; p.Trp319Cysfs*21). This substitution generates exon skipping, leading to a truncated protein. In fact, NADK2 messenger RNA and the corresponding protein were almost absent. Now, at 10 years of age she presents with ataxia and incoordination. She has oromotor dysphasia but is able to understand fluid language and is a very friendly girl. We hypothesize that the patient's clinical improvement could be due to her lysine-restricted diet together with cofactors and pyridoxal phosphate administration.

Relationships between bone density values from cone beam computed tomography, maximum insertion torque, and resonance frequency analysis at implant placement: a pilot study.

PURPOSE: The aim of the present study was to determine bone density in designated implant sites using cone beam computed tomography (CBCT) and to evaluate possible correlations between age, gender, insertion torque measurements, and resonance frequency analysis (RFA) values. MATERIALS AND METHODS: Completely and partially edentulous patients were treated with implants between 2007 and 2008 and evaluated retrospectively. The preoperative examination included a panoramic radiograph, CBCT, diagnostic casts, and a clinical examination of the jaws. With the CBCT scans, bone densities were recorded in Hounsfield units (HU). Insertion torque values and implant stability measurements (via RFA) were also noted. RESULTS: Mean bone density and insertion torque values were 623 ± 209 HU and 42.4 ± 4 Ncm, respectively, for the 82 implants placed. Mean primary stability (implant stability quotient) was 62.4 ± 8. The differences in mean bone density at implant sites in the mandible (717 ± 204 HU) and the maxilla (490 ± 128 HU) were statistically significant for all patients (P < .05). There was a statistically significant relationship between bone density values and insertion torque measurements for implant sites in the anterior mandible (r = 0.562, P < .05), as well as between bone density and RFA values for men (r = 0.412, P < .05). CONCLUSIONS: Bone density measurements using preoperative CBCT may be helpful as an objective diagnostic tool. These values, in conjunction with RFA values and insertion torque measurements, can provide the implant surgeon with an objective assessment of bone quality and may be especially useful where poor-quality bone is suspected.

Salivary gland application of botulinum toxin for the treatment of sialorrhea.

Sialorrhea or excessive salivation, and drooling, are common and disabling manifestations in different neurological disorders. A review is made of the literature, based on a PubMed search, selecting those articles describing clinical trials involving the injection of botulinum toxin A in the salivary glands of patients with different diseases characterized by sialorrhea. The most frequently treated diseases were infant cerebral palsy (30%), Parkinson's disease (20%) and amyotrophic lateral sclerosis (15%). Over half of the authors injected the product into the parotid glands, 9.5% into the submaxillary glands, and 38% into both. The total doses of toxin injected varied from 10-100 units of Botox or 30-450 units of Dysport according to the different authors. A reduction was observed in the production of saliva following these injections, and the duration of the therapeutic effect was 1.5-6 months. Six articles (30%) described the presence of adverse effects such as dysphagia, xerostomia and chewing difficulties. Most of the clinical studies involved small patient samples, with no blinding or randomization, and no control group. Moreover, no data are available on the efficacy and adverse effects of treatment in the context of long-term prospective studies. The effective therapeutic dose and ideal form of application remain to be established, and require the conduction of further controlled clinical trials involving large sample sizes.

Salivary gland application of botulinum toxin for the treatment of sialorrhea

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Sialorrhea or excessive salivation, and drooling, are common and disabling manifestations in different neurological disorders. A review is made of the literature, based on a PubMed search, selecting those articles describing clinical trialsinvolving the injection of botulinum toxin A in the salivary glands of patients with different diseases characterized by sialorrhea.The most frequently treated diseases were infant cerebral palsy (30%), Parkinson’s disease (20%) and amyotrophic lateral sclerosis (15%). Over half of the authors injected the product into the parotid glands, 9.5% into the submaxillary glands, and 38% into both. The total doses of toxin injected varied from 10-100 units of Botox® or 30-450 units of Dysport®according to the different authors. A reduction was observed in the production of saliva following these injections, and the duration of the therapeutic effect was 1.5-6 months. Six articles (30%) described the presence of adverse effects such as dysphagia, xerostomia and chewing difficulties.Most of the clinical studies involved small patient samples, with no blinding or randomization, and no control group.Moreover, no data are available on the efficacy and adverse effects of treatment in the context of long-term prospective studies. The effective therapeutic dose and ideal form of application remain to be established, and require the conduction of further controlled clinical trials involving large sample sizes (AU)

Simultaneous HPLC quantification of monoamines and metabolites in the blood-free rat cochlea.

Monoamine quantification in peripheral sensory receptors, such as the cochlea, is of major interest since monoamines could play a role in neurotransmission. A three-step biochemical protocol was developed to analyze monoamine content within the cochlea. Removal of the blood by aortic perfusion was carried out with an anticoagulant solution prior to the dissection of the cochlea from the temporal bone. The cochlear monoamines and some of their metabolites were then quantified, from homogenated cochlear tissue, by a new application of high performance liquid chromatography coupled to electrochemical detection. This method demonstrated enough sensitivity to detect norepinephrine (NE), dopamine (DA), serotonin (5-HT) and some of their metabolites (3,4-dihydroxyphenylacetic acid, DOPAC; homovanillic acid, HVA; and 5-hydroxyindole-3-acetic acid, 5-HIAA). Furthermore, it enabled the demonstration of noise-induced changes in the cochlear concentrations of NE, DA, DOPAC and HVA. In addition, the aortic perfusion allowed removal of the blood-borne 5-HT from the cochlea without inducing systemic alterations or monoamine degradation, as shown by the absence of effects on NE, DA, DOPAC, HVA or 5-HIAA concentrations. The present methodology may constitute a useful strategy to analyze monoamine turnover in the cochlea and other peripheral sensory receptors.