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Purpose: To describe the frequency of head and/or pancreas uncinate process uptake of 99mTc-HYNIC-TOC, to study its nature, and analyze its diagnostic value. Materials and methods: Retrospective evaluation of 47 consecutive 99mTc-HYNIC-TOC examinations was conducted. Head and/or pancreas uncinate process uptake was considered to be physiological in patients with normal CT at the same episode and in follow-up. It was analyzed if age or diabetes mellitus was justifying the existence or not of uptake. Results: 32.5% patients showed uptake; 73% of them were mild. 84.6% patients with uptake have no pathology and 4% had neuroendocrine pancreatic disease at CT. Neither the age nor the diabetes mellitus established differences in patients without lesion. Conclusions: Near one-third of patients show physiological uptake by head and/or pancreas uncinate process at 99mTc-HYNIC-TOC scintigraphy. It seems that neither the diabetes nor the ages are factors that determine this physiological uptake. [ABSTRACT FROM AUTHOR]

Resistance to cancer treatment remains a major clinical hurdle. Here, we demonstrate that the CoREST complex is a key determinant of endocrine resistance and ER +  breast cancer plasticity. In endocrine-sensitive cells, CoREST is recruited to regulatory regions co-bound to ERα and FOXA1 to regulate the estrogen pathway. In contrast, during temporal reprogramming towards a resistant state, CoREST is recruited to AP-1 sites. In reprogrammed cells, CoREST favors chromatin opening, cJUN binding to chromatin, and gene activation by controlling SWI/SNF recruitment independently of the demethylase activity of the CoREST subunit LSD1. Genetic and pharmacological CoREST inhibition reduces tumorigenesis and metastasis of endocrine-sensitive and endocrine-resistant xenograft models. Consistently, CoREST controls a gene signature involved in invasiveness in clinical breast tumors resistant to endocrine therapies. Our studies reveal CoREST functions that are co-opted to drive cellular plasticity and resistance to endocrine therapies and tumorigenesis, thus establishing CoREST as a potential therapeutic target for the treatment of advanced breast cancer., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Aims: In an effort to gain further insight into the underlying mechanisms tied to disease onset and progression of Gulf War Illness (GWI), our team evaluated GWI patient response to stress utilizing RNA-Seq., Main Methods: The protocol included blood collection before exercise challenge (baseline), at maximal exertion, and after exercise challenge (recovery - four hours post-exercise challenge). Peripheral blood mononuclear cell (PBMC) transcriptomics data were analyzed to understand why GWI patients process stressors differently from their healthy counterparts., Key Findings: Our findings validate previously identified dysregulation of immune and inflammatory pathways among GWI patients as well as highlight novel immune and inflammatory markers of disease activity. These results provide a foundation for future research efforts in understanding GWI pathophysiology and creating targeted treatments., Significance: Gulf War Illness is a complex, chronic, and debilitating multi-system illness impacting 25%-30% of the U.S. troops deployed to the 1990-1991 Gulf War. The condition is characterized by medically unexplained fatigue and affects multiple organ systems. Because the underlying mechanisms are largely unknown, patients receive symptom-based treatment, rather than targeting fundamental biological processes. To the best of our knowledge, this is the first study that applies RNA-Seq to analyze the effect of GWI, and the response to stressors in GWI, on the transcriptomic changes in circulating immune cells., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a multisystem illness characterized by medically unexplained debilitating fatigue with suggested altered immunological state. Our study aimed to explore peripheral blood mononuclear cells (PBMCs) for microRNAs (miRNAs) expression in ME/CFS subjects under an exercise challenge. The findings highlight the immune response and inflammation links to differential miRNA expression in ME/CFS. The present study is particularly important in being the first to uncover the differences that exist in miRNA expression patterns in males and females with ME/CFS in response to exercise. This provides new evidence for the understanding of differential miRNA expression patterns and post-exertional malaise in ME/CFS. We also report miRNA expression pattern differences associating with the nutritional status in individuals with ME/CFS, highlighting the effect of subjects' metabolic state on molecular changes to be considered in clinical research within the NINDS/CDC ME/CFS Common Data Elements. The identification of gender-based miRNAs importantly provides new insights into gender-specific ME/CFS susceptibility and demands exploration of sex-suited ME/CFS therapeutics., (© 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)

Background: Mesenchymal stromal cells (MSCs), adult stromal cells most commonly isolated from bone marrow (BM), are being increasingly utilized in various therapeutic applications including tissue repair via immunomodulation, which is recognized as one of their most relevant mechanism of action. The promise of MSC-based therapies is somewhat hindered by their apparent modest clinical benefits, highlighting the need for approaches that would increase the efficacy of such therapies. Manipulation of cellular stress-response mechanism(s) such as autophagy, a catabolic stress-response mechanism, with small molecules prior to or during MSC injection could improve MSCs' therapeutic efficacy. Unfortunately, limited information exists on how manipulation of autophagy affects MSCs' response to inflammation and subsequent immunoregulatory properties., Methods: In this study, we exposed BM-MSC precursor cells, "marrow-isolated adult multilineage inducible" (MIAMI) cells, to autophagy modulators tamoxifen (TX) or chloroquine (CQ), together with IFN-γ. Exposed cells then underwent RNA sequencing (RNAseq) to determine the effects of TX or CQ co-treatments on cellular response to IFN-γ at a molecular level. Furthermore, we evaluated their immunoregulatory capacity using activated CD4+ T cells by analyzing T cell activation marker CD25 and the percentage of proliferating T cells after co-culturing the cells with MIAMI cells treated or not with TX or CQ., Results: RNAseq data indicate that the co-treatments alter both mRNA and protein levels of key genes responsible for MSCs' immune-regulatory properties. Interestingly, TX and CQ also altered some of the microRNAs targeting such key genes. In addition, while IFN-γ treatment alone increased the surface expression of PD-L1 and secretion of IDO, this increase was further enhanced with TX. An improvement in MIAMI cells' ability to decrease the activation and proliferation of T cells was also observed with TX, and to a lesser extent, CQ co-treatments., Conclusion: Altogether, this work suggests that both TX and CQ have a potential to enhance MIAMI cells' immunoregulatory properties. However, this enhancement is more pronounced with TX co-treatment.

Gulf War Illness (GWI) affects about 25% of Persian Gulf veterans with a cluster of chronic symptoms, including immune dysfunction and neurological issues. Recent studies implicate gene expression changes in immune function to be associated with GWI. Since DNA methylation can regulate such changes in gene expression, and disruption of DNA methylation pattern is implicated in various immune and neurological diseases, we aimed to study the DNA methylation patterns in peripheral blood mononuclear cells from GWI patients. Global DNA methylation levels were similar in GWI patients and controls. However, the genome-wide microarray technology detected 10,767 differentially methylated CpG sites across gene regulatory elements and within coding regions. Approximately 88% of them were hypermethylated in GWI patients. The separate analysis found 776 differentially methylated gene promoters (DMP), which were predominantly hypermethylated. Pyrosequencing validation confirmed microarray results. Functional analysis revealed that majority of the DMPs belonged to genes responsible for metabolism and immune system. This is the first pilot human study characterizing genome-wide epigenetic changes associated with GWI. It suggests a significant contribution of epigenetic dysfunction in GWI. Moreover, it supports the dysregulation of immune function in GWI. Lastly, it suggests studies with the larger cohort to validate our findings.

Background: Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a complex condition involving multiple organ systems and characterized by persistent/relapsing debilitating fatigue, immune dysfunction, neurological problems, and other symptoms not curable for at least 6 months. Disruption of DNA methylation patterns has been tied to various immune and neurological diseases; however, its status in ME/CFS remains uncertain. Our study aimed at identifying changes in the DNA methylation patterns that associate with ME/CFS., Methods: We extracted genomic DNA from peripheral blood mononuclear cells from 13 ME/CFS study subjects and 12 healthy controls and measured global DNA methylation by ELISA-like method and site-specific methylation status using Illumina MethylationEPIC microarrays. Pyrosequencing validation included 33 ME/CFS cases and 31 controls from two geographically distant cohorts., Results: Global DNA methylation levels of ME/CFS cases were similar to those of controls. However, microarray-based approach allowed detection of 17,296 differentially methylated CpG sites in 6,368 genes across regulatory elements and within coding regions of genes. Analysis of DNA methylation in promoter regions revealed 307 differentially methylated promoters. Ingenuity pathway analysis indicated that genes associated with differentially methylated promoters participated in at least 15 different pathways mostly related to cell signaling with a strong immune component., Conclusions: This is the first study that has explored genome-wide epigenetic changes associated with ME/CFS using the advanced Illumina MethylationEPIC microarrays covering about 850,000 CpG sites in two geographically distant cohorts of ME/CFS cases and matched controls. Our results are aligned with previous studies that indicate a dysregulation of the immune system in ME/CFS. They also suggest a potential role of epigenetic de-regulation in the pathobiology of ME/CFS. We propose screening of larger cohorts of ME/CFS cases to determine the external validity of these epigenetic changes in order to implement them as possible diagnostic markers in clinical setting., Competing Interests: The authors have declared that no competing interests exist.

Aveolar soft part sarcoma is a rare tumor responsible for about 1% of all soft tissue sarcomas, affecting mostly adolescents and young adults. ASPS has curious patterns of metastatic spread, with seldom lymph node involvement. Lung, bone and brain are the most common metastatic places. Small bowel metastasis are infrequent, having found reported only one case of duodenal metastasis with polypous appearance. We describe a case of duodenal metastasis presenting as abdominal mass five years after initial diagnosis of alveolar soft part sarcoma.

Background: The aim of this study was to develop molecular biology techniques as hybridization with oligonucleotide probes to characterize TEM-1 and TEM-2 beta-lactamases in strains of Salmonella spp., Methods: Twenty seven strains of Salmonella spp. were selected. Twenty six were resistant to ampicillin due to the production of beta-lactamases enzymes of pl of 5.4 and/or 5.6 corresponding to TEM-type. Initially, they were submitted to colony hybridization with a 420 bp. TEM probe obtained from plasmid pBR322. The strains with positive signal were selected to perform colony hybridization and Southern blot with oligonucleotide probes for TEM-1 (Gln 37) and TEM-2 (Lys 37). Finally, polymerase chain reaction technique (PCR) was developed to obtain DNA., Results: Only 17 out of the 26 beta-lactamase producing strains gave positive signals with the TEM intragenic probe. Experiments with the oligonucleotide probes in colony hybridization did not allow us discriminate positive from negative signals. Southern blot of DNA obtained from alkaline lysates did not work as we could not obtain any signals in the filters. To resolve these problems and to obtain enough DNA to perform Southern blot we developed PCR. This way it was able discriminate the bla-TEM-1 from the bla-TEM-2 genes., Conclusions: PCR technique plus oligonucleotide probes are a good alternative for the specific characterization of TEM-1 and TEM-2 beta-lactamases in Salmonella spp.

Background: The purpose of this study was to develop a satisfactory technique to detect punctual mutations in blaTEM genes., Methods: The strains [E. coli HB 101 pBR322 (TEM-1), E. coli J62 RP4 (TEM-2)] were submitted to PCR with primers PL1 and PL2 which amplify the genetic region susceptible of punctual mutations. Then, we developed Southern blot and hybridization with oligonucleotide probes (GIn 37, Lys 37 y Thr 261), corresponding to first and last mutations., Results: A region of 841 bp was amplified using the primers previously described. Hybridization experiments with the Thr 261 probe gave positive signal with both strains (both carry the mutation); GIn 37 only hybridized with TEM-1 and Lys 37 only with TEM-2., Conclusions: The use of primers which amplify all the region susceptible of mutations in blaTEM and oligonucleotide probes allows the specific detection of point modifications in the original genes by the use of digoxigenin-labeled oligonucleotide probes.

Background: The purpose of this work was to study the molecular basis of beta-lactamase production in ampicillin-resistant strains of Salmonella spp., Methods: It was performed analytical isoelectric focusing of beta-lactamases produced by a group of 33 strains selected in basis of their resistance phenotype. Plasmid profile analysis and assays of transferable drug resistance were developed. The study was completed by hybridization experiments with an intragenic TEM probe which allowed the location of the bla-TEM gene., Results: By analytical isoelectrofocusing we found that 26 out of the 27 ampicillin-resistant strains produced beta-lactamases with pl 5.4 and/or 5.6 corresponding to TEM-1 and/or TEM-2 type. Analysis of plasmid DNA revealed in almost all strains plasmids ranging in size from 1.1 to 125 Mdal. This plasmids were responsible of the resistance and, moreover, were able to transfer the resistance by conjugation mechanisms. Southern blot analysis detected the gene that code the TEM beta-lactamase at the 125, 8 and 5.8 Mdal plasmids., Conclusions: Resistance to ampicillin in the strains of Salmonella studied was due to the presence of TEM type beta-lactamases coded by conjugative plasmids. These plasmids coded also resistance to other antimicrobial agents. Our results showed that the use of a DNA probe to the detect TEM-type beta-lactamases using a non radioactive probe, could be a suitable alternative to isoelectric focusing.

The aim of this work has been the production of specific monoclonal antibodies against HBV-antigens and their utilisation in order to study their distribution on liver tissue. The monoclonal antibodies anti-HBc and anti-HBs were obtained by the modified hybridoma technique. This study was performed on 50 patients affected by several chronic hepatopathies. For the detection of the antigens, avidin-biotin-peroxidase complex immunostaining was used. Both cytoplasmic and membranous HBsAg were detected in 15 out of 16 HBsAg+ patients; 8 of 12 HBsAg-/anti-HBc+ patients and 1 HBsAg-/antiHBc- patient. Cytoplasmic and nuclear HBcAg was observed in 12 of 16 HBsAg+ patients and 4 of 20 HBsAg- patients. Although the presence of serum HBsAg is an index of liver infection, in some HBsAg-/antiHB+ patients (20%) with undetectable levels of HBsAg, hepatic injury may be disclosed by the detection of other markers of active viral replication.

We study retrospectively the viral replication state (HBV) of 50 patients with chronic hepatic alterations. The seric DNA-HBV and/or intrahepatic (molecular hybridization), the intrahepatic distribution of HBV antigens (specific monoclonal antibodies labelled with immunoperoxidase), conventional seric HBV markers (commercial enzymoimmunoessay) and the different histopathologic features. We found a correlation between DNA-HBV "in situ" and HBcAg intrahepatic and the seric DNA-HBV production. 81% of the patients with HBsAg (+) had intrahepatic HBcAg and 85% (11/13) of them showed the antigen in their cytoplasms. Patients with HBcAg also had seric and liver DNA-HBV (+). The lack of seric HBsAg did not mean that non-active replication of HBV did not exist because 20% of the patients with HBsAg (-) showed seric and "in situ" DNA-HBV and cytoplasmic HBcAg. The detection of DNA-HBV in endothelial cells and vascular elements in hepatic tissue show that the rate of the HBV host cells is greater.