Targeting the OB-Folds of Replication Protein A with Small Molecules

Replication protein A (RPA) is the main eukaryotic single-strand (ss) DNA-binding protein involved in DNA replication and repair. We have identified and developed two classes of small molecule inhibitors (SMIs) that show in vitro inhibition of the RPA-DNA interaction. We present further characterization of these SMIs with respect to their target binding, mechanism of action, and specificity. Both reversible and irreversible modes of inhibition are observed for the different classes of SMIs with one class found to specifically interact with DNA-binding domains A and B (DBD-A/B) of RPA. In comparison with other oligonucleotide/oligosaccharide binding-fold (OB-fold) containing ssDNA-binding proteins, one class of SMIs displayed specificity for the RPA protein. Together these data demonstrate that the specific targeting of a protein-DNA interaction can be exploited towards interrogating the cellular activity of RPA as well as increasing the efficacy of DNA-damaging chemotherapeutics used in cancer treatment

Continuous Risk Management for Industrial IoT: a Methodological View

Emergent cyber-attacks and exploits targeting Operational Technologies (OT) call for a proactive risk management approach. The convergence between OT and the Internet-of-Things in industries introduces new opportunities for cyber-attacks that have the potential to disrupt time-critical and hazardous processes. This paper proposes a methodology to adapt traditional risk management standards to work in a continuous fashion. Monitoring of risk factors is based on incident and event management tools, and misbehaviour detection to address cyber-physical systems’ security gaps. Another source of information that can enhance this approach is threat intelligence. Risks are calculated using Bayesian Networks

Safety and Immunogenicity of a Human Papillomavirus Peptide Vaccine (CIGB-228) in Women with High-Grade Cervical Intraepithelial Neoplasia: First-in-Human, Proof-of-Concept Trial

Objective. CIGB-228 is a novel therapeutic vaccine consisting of HLA-restricted HPV16 E7 epitope adjuvated with VSSP. This trial was designed to evaluate the toxicity, safety, immunogenicity, HPV clearance, and lesion regression. Methods. Seven women were entered. All were HLA-A2 positive, had biopsy-proven high-grade CIN, histologically positive for HPV16, and beared persistent postbiopsy lesions visible by digital colposcopy. HLA-A2 women with biopsy-proven high-grade CIN, HPV16-positive, and beared persistent postbiopsy lesions visible by digital colposcopy were vaccinated. One weekly injections of CIGB-228 vaccine was given for four weeks. Then, loop electrosurgical excision procedure (LEEP) of the transformation zone was performed. Study subjects were followed for 1 year after LEEP. Results. No toxicity beyond grade 1 was observed during and after the four vaccinations. Five of seven women had complete and partial regression. Cellular immune response was seen in all patients. HPV was cleared in three of the patients with complete response. Conclusion. CIGB-228 vaccination was well tolerated and capable to induce IFNγ-associated T-cell response in women with high-grade CIN. In several patients, lesion regression and HPV clearance were observed

Correction: Clinical epigenomics: genome-wide DNA methylation analysis for the diagnosis of Mendelian disorders (Genetics in Medicine, (2021), 23, 6, (1065-1074), 10.1038/s41436-020-01096-4)

A Correction to this paper has been published: https://doi.org/10.1038/s41436-021-01130-z

De novo variants of NR4A2 are associated with neurodevelopmental disorder and epilepsy

Purpose This study characterizes the clinical and genetic features of nine unrelated patients with de novo variants in the NR4A2 gene. Methods Variants were identified and de novo origins were confirmed through trio exome sequencing in all but one patient. Targeted RNA sequencing was performed for one variant to confirm its splicing effect. Independent discoveries were shared through GeneMatcher. Results Missense and loss-of-function variants in NR4A2 were identified in patients from eight unrelated families. One patient carried a larger deletion including adjacent genes. The cases presented with developmental delay, hypotonia (six cases), and epilepsy (six cases). De novo status was confirmed for eight patients. One variant was demonstrated to affect splicing and result in expression of abnormal transcripts likely subject to nonsense-mediated decay. Conclusion Our study underscores the importance of NR4A2 as a disease gene for neurodevelopmental disorders and epilepsy. The identified variants are likely causative of the seizures and additional developmental phenotypes in these patients

Clinical epigenomics: genome-wide DNA methylation analysis for the diagnosis of Mendelian disorders

Purpose: We describe the clinical implementation of genome-wide DNA methylation analysis in rare disorders across the EpiSign diagnostic laboratory network and the assessment of results and clinical impact in the first subjects tested. Methods: We outline the logistics and data flow between an integrated network of clinical diagnostics laboratories in Europe, the United States, and Canada. We describe the clinical validation of EpiSign using 211 specimens and assess the test performance and diagnostic yield in the first 207 subjects tested involving two patient subgroups: the targeted cohort (subjects with previous ambiguous/inconclusive genetic findings including genetic variants of unknown clinical significance) and the screening cohort (subjects with clinical findings consistent with hereditary neurodevelopmental syndromes and no previous conclusive genetic findings). Results: Among the 207 subjects tested, 57 (27.6%) were positive for a diagnostic episignature including 48/136 (35.3%) in the targeted cohort and 8/71 (11.3%) in the screening cohort, with 4/207 (1.9%) remaining inconclusive after EpiSign analysis. Conclusion: This study describes the implementation of diagnostic clinical genomic DNA methylation testing in patients with rare disorders. It provides strong evidence of clinical utility of EpiSign analysis, including the ability to provide conclusive findings in the majority of subjects tested

Trace elements in hemodialysis patients: a systematic review and meta-analysis

<p>Abstract</p> <p>Background</p> <p>Hemodialysis patients are at risk for deficiency of essential trace elements and excess of toxic trace elements, both of which can affect health. We conducted a systematic review to summarize existing literature on trace element status in hemodialysis patients.</p> <p>Methods</p> <p>All studies which reported relevant data for chronic hemodialysis patients and a healthy control population were eligible, regardless of language or publication status. We included studies which measured at least one of the following elements in whole blood, serum, or plasma: antimony, arsenic, boron, cadmium, chromium, cobalt, copper, fluorine, iodine, lead, manganese, mercury, molybdenum, nickel, selenium, tellurium, thallium, vanadium, and zinc. We calculated differences between hemodialysis patients and controls using the differences in mean trace element level, divided by the pooled standard deviation.</p> <p>Results</p> <p>We identified 128 eligible studies. Available data suggested that levels of cadmium, chromium, copper, lead, and vanadium were higher and that levels of selenium, zinc and manganese were lower in hemodialysis patients, compared with controls. Pooled standard mean differences exceeded 0.8 standard deviation units (a large difference) higher than controls for cadmium, chromium, vanadium, and lower than controls for selenium, zinc, and manganese. No studies reported data on antimony, iodine, tellurium, and thallium concentrations.</p> <p>Conclusion</p> <p>Average blood levels of biologically important trace elements were substantially different in hemodialysis patients, compared with healthy controls. Since both deficiency and excess of trace elements are potentially harmful yet amenable to therapy, the hypothesis that trace element status influences the risk of adverse clinical outcomes is worthy of investigation.</p

Premedicación con Ketorolac para Analgesia Postoperatoria en Cirugía Ginecológica

El objetivo de la investigaci&oacute;n fue verificar la eficacia del uso de Ketorolac en la premedicaci&oacute;n para analgesia postoperatoria en cirug&iacute;a ginecol&oacute;gica. Se estudiaron 50 pacientes programadas para Histerectom&iacute;a Abdominal, divididas en dos grupos en forma aleatoria: Grupo A o control (n=25) y Grupo B o experimental (n=25) que recibi&oacute; Ketorolac 30 mg. EV 20 minutos antes de la inducci&oacute;n de la anestesia, los dos grupos fueron similares respecto a la edad, A.S.A. y tiempo de cirug&iacute;a, en ambos grupos se midieron par&aacute;metros hemodin&aacute;micos durante el pre, trans y postoperatorio, duraci&oacute;n de la analgesia e incidencia de efectos adversos. Las variables num&eacute;ricas se analizaron con la prueba de derivaci&oacute;n est&aacute;ndar y test de student. Se obtuvieron como resultados de param&eacute;tros hemodin&aacute;micos transoperatorios: Grupo A TAS (111.4 +/-11.98), TAD (72.4 +/- 7.03) y FC (86.0 +/- 7.41) y Grupo B: TAS (100.3 +/-5.10), TAD (61.6 +/- 6.26) y FC (72.4 +/- 7.03) P(&lt;0.0001), que indic&oacute; una diferencia muy significativa, de igual manera los par&aacute;metros hemodin&aacute;micos postoperatorios mostraron que exist&iacute;an diferencias extremadamente significativas para el Grupo A:TAS (127.9 +/- 14.3), TAD (84.2 +/- 7.01) y FC (93.5 +/- 9.3) en relaci&oacute;n con el Grupo B: TAS (111.4 +/- 10.5), TAD (73.4 +/- 6.9) y FC (73.4 +/- 6.95). La variable analgesia postoperatoria fue de 0.25+/-0.20 en el Grupo A y de 4.8+/-0.76 para el B, indic&oacute; una diferencia muy significativa (P&lt;0.0001). No se observaron efectos adversos importantes. Se concluye que la administraci&oacute;n de Ketorolac EV en la premedicaci&oacute;n representa una alternativa efectiva para reducir el dolor en el postoperatorio inmediato en Cirug&iacute;a Ginec&oacute;logica con una baja incidencia de efectos adversos.ABSTRACTThe objetive of the investigation was to verify the effectiveness of the use of the Ketorolac in the premedication for post-operative analgesia in Gynecological Surgery. 50 patients were studied programmed for Abdominal Hysterectomy, divided in two groups in aleatory form:Group A or Control (n=25) and group B or Experimental (n=25) that Ketorolac 30 mg received EV 20 minutes before the induction of the anesthesia, both groups were similar regarding the age, A.S.A and time of Surgery, in the two groups parameters hamodynamic pre were measured, trans and postoperative, duration of the analgesia and incidence of adverse effects. The numeric variables will be analyzed with the test of standard desviation and student T. They are obtain as results of parameters hemodynamic transoperatives: Group A TAS (114,6 +/- 11.98), TAD (72.4+/-7.03) and FC (86.0 +/-7.41) and Group B TAS (100.3 +/- 5.10), TAD (61.6 +/- 6.26) and FC (72.4 +/- 7.03) (P&lt;0.0001) that indicated a very significant difference, in a same way the postoperative hemodynamic parameter showed that extremely significant differences exited for the Group A:TAS (127.9 +/- 14.3), TAD (84.2 +/- 7.01) and FC (93.5 +/-9.3) in connection with the Group B:TAS (111.4 +/- 10.5), TAD (73.4 +/- 6.9) and FC (73.4 +/- 6.95). The variable postoperative analgesia was of 0.25 +/- 0.20 in the Group A and of 4.8 +/- 0.76 for the B, it indicated a very significant difference (P&lt;0.0001). Important adverse effects are not observed.You concludes that Ketorolac EV&acute;S administration in the premedication represente an effective alternative to reduce the pain in the postoperative one immediate in Gynecological Surgery with a low incidence of adverse effects