Mutations Research Articles

Psychoanalytic Observations Near the End of Life

ABSTRACT Using psychoanalytic free association, introspection, self-analysis, and his more than 50 years of experience as an adult and child psychoanalyst, the author explores the limited psychoanalytic literature on the individual analyst’s personal reactions to death and dying. Then he describes his experiences of loss of function, diminished psychoanalytic identity, and his personal attitudes toward death and an afterlife.

The Heterotaxy Gene CCDC11 Is Important for Cytokinesis via RhoA Regulation.

Mutations in CCDC11 (cfap53) have been identified in multiple patients with heterotaxy (Htx), a disorder of left-right (LR) patterning of the internal organs. In Xenopus, depletion of Ccdc11 causes defects in LR patterning, recapitulating the patient phenotype. Upon Ccdc11 depletion, monociliated cells of the Left-Right Organizer (LRO) exhibit multiple cilia per cell. Unexpectedly, we found that Ccdc11 is necessary for successful cytokinesis, explaining the multiciliation phenotype observed in Ccdc11-depleted cells. The small GTPase RhoA is critical for cytokinesis, and our Ccdc11 depletion phenotypes are reminiscent of RhoA loss of function. Here, we demonstrate that during cytokinesis CCDC11 is localized to the cytokinetic contractile ring overlapping with RhoA, and CCDC11 regulates total RhoA protein levels. Our results connect CCDC11 to cytokinesis and LR patterning via RhoA regulation, providing a potential mechanism for heterotaxy disease pathogenesis.

A Good Diagnosis Saves Lives: The Importance of Teledermatology: Case Report

Chromoblastomycosis (CBM) is one of the most prevalent subcutaneous mycoses, caused by melanized or pigmented fungi. Considered a tropical or subtropical disease, commonly seen in rural areas after traumatic inoculation, mostly associated with agriculture. The lesions tend to present on the limbs and follow a chronic and progressive pattern, which if left untreated or misdiagnosed can lead to loss of functionality and lower quality of life. Teledermatology (TD) has been used mainly to diagnose, monitor and treat patients at distance in neglected areas. Guatemala has a vast rural area and despite the efforts from both private and public sectors, we still cannot reach all the patients and not all the patients can reach us. We present a case report on how with the aid of teledermatology a life was changed for the good.

ZNF143-mediated upregulation of MEX3C promotes hepatocellular carcinoma progression

BackgroundMicrovascular invasion is strongly associated with aggressive tumor behavior and recurrence in hepatocellular carcinoma (HCC) patients. Zinc finger protein 143(ZNF143) is a transcription factor involved in a wide variety of physiological and developmental processes. This study primarily focuses on the exact biological role and mechanism of ZNF143 in HCC migration and invasion. MethodsThe expression and prognosis of ZNF143 in HCC patients were analyzed. The levels of ZNF143, mex-3 RNA binding family member C (MEX3C) were quantified by western blot and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell migration ability was detected by wound- healing assay. Matrigel transwell assay was conducted to evaluate the invasion of HCC cells. The differential expression genes of ZNF143 overexpression and knockdown were screened by mRNA profiling analysis. Dual luciferase assay was performed to determine the promoter activity of MEX3C. The enrichment of ZNF143 at MEX3C promoter was determined by chromatin immunoprecipitation (ChIP). ResultsZNF143 is overexpressed in HCC tissues and that its overexpression is correlated with poorer prognosis, especially in HCC patients with higher tumor grades and microvascular invasion. Gain- and loss-of function experiments showed that ZNF143 promotes migration and invasion in HCC cells. mRNA profiling showed that ZNF143 significantly upregulates MEX3C. ZNF143 was positively correlated with MEX3C expression in HCC tissue. ZNF143 activates MEX3C transcription by directly binding to its promoter. MEX3C knockdown inhibited migration and invasion induced by ZNF143 overexpression in HCC cells. ConclusionZNF143 promotes HCC cell migration and invasion by binding to MEX3C promoter and activating its expression.

Influence of Mitochondrial NAD(P) + Transhydrogenase (NNT) on Hypothalamic Inflammation and Metabolic Dysfunction Induced by a High-Fat Diet in Mice.

The mitochondrial protein NAD(P)+ transhydrogenase (NNT) has been implicated in the metabolic derangements observed in obesity. Mice with the C57BL/6J genetic background bear a spontaneous mutation in the Nnt gene and are known to exhibit increased susceptibility to diet-induced metabolic disorders. Most of the studies on NNT in the context of diet-induced obesity have compared C57BL/6J mice with other mouse strains, where differences in genetic background can serve as confounding factors. Moreover, these studies have predominantly employed a high-fat diet (HFD) consisting of approximately 60% of calories from fat, which may not accurately mimic real-world fat-rich diets. In this study, we sought to examine the role of NNT in diet-induced hypothalamic inflammation and metabolic syndrome by using a congenic mice model lacking NNT, along with a HFD providing approximately 45% of calories from fat. Our findings indicate that mice lacking NNT were more protected from HFD-induced weight gain but presented a worse performance on glucose tolerance test, albeit not in insulin tolerance test. Interestingly, the brown adipose tissue of HFD-fed Nnt +/+ mice presented a greater mass and a higher whole-tissue ex-vivo oxygen consumption rate. Also, HFD increased the expression of the inflammatory markers Il1β, Tlr4 and Iba1 in the hypothalamus of Nnt -/- mice. In conclusion, our study highlights the importance of NNT in the context of diet-induced obesity and metabolic syndrome, indicating its contribution to mitigate hypothalamic inflammation and suggesting its role in the brown adipose tissue increased mass.

Evaluation of Genotoxic and Hemolytic Effects of Aphanizomenon flos-aquae and Microcystis aeruginosa Biomass Extracts on Human Blood Cells In Vitro

This study explores the in vitro effects of cyanotoxins from the methanolic extract of the cyanobacteria Aphanizomenon flos-aquae and Microcystis aeruginosa on human blood cells, with samples drawn from the Gruža reservoir in Serbia. These cyanobacteria, which made up 98.5% of the reservoir’s phytoplankton, reached densities of 4,656,450 cells mL−1, with A. flos aquae (3,105,120 cells mL−1) as the dominant species, followed by M. aeruginosa (1,480,130 cells mL−1). A cyanotoxin analysis of biomass detected anatoxin-a (3.56 µg g−1), cylindrospermopsin (6.86 µg g−1), microcystin LR (0.87 µg g−1), and microcystin RR (2.47 µg g−1). This study assessed the genotoxic potential of the methanolic extract of the cyanobacterial biomass by evaluating the DNA damage and the Genetic Damage Index (GDI) in peripheral blood lymphocytes (PBLs) from healthy donors. The results showed a dose-dependent increase in the DNA damage, from 35.67 ± 4.93% at 10 µg mL−1 to 95.67 ± 1.53% at 100 µg mL−1, with a corresponding rise in the GDI from 0.61 ± 0.02 to 2.39 ± 0.07. The extract also caused the concentration-dependent hemolysis of red blood cells, with 5.63% hemolysis at the highest concentration (200 µg mL−1). These findings underscore the significant genotoxic risks posed by cyanotoxins from biomass extracts of A. flos aquae and M. aeruginosa, particularly in water sources used for human consumption.

Variants that get straight to your heart – Cardiogenetic secondary findings in exome sequencing

BackgroundExome sequencing has been established as a fundamental tool in genetic diagnostics. It may also provide information about variants in genes unrelated to the primary purpose, so-called secondary findings. Especially, diagnoses of unnoticed inborn cardiac diseases are of high clinical relevance due to therapeutic options in context of prevention of sudden cardiac death. MethodsExome data of 9962 individuals was analysed for relevant cardiogenetic findings. Genes were selected according to ACMG recommendations for secondary findings (v.3.1). First, a filter for (likely) pathogenic variants, published in the ClinVar database, was used. Second, exome data was screened for loss of function (LoF) variants in genes in which LoF is a known disease pathomechanism. All variants were evaluated by geneticists regarding their pathogenicity. ResultsPathogenic or likely pathogenic variants were identified in 136 different individuals (136/9962, 1.4%), with the Low-Density Lipoprotein Receptor gene (LDLR, 24/136, 17.6%) and the Titin gene (TTN, 24/136, 17.6%), being the most frequently affected ones. 31.6% (43/136) of the identified variants had been reported beforehand, while 47.1% (64/136) had not been reported. The remaining cases (29/136, 21.3%) were part of research projects with no written reports. In 26.5% (36/136), the finding would have been missed, if only index patients and not their parents had been screened for secondary findings in case of trio ES. ConclusionAs demonstrated in our study, at least one or two out of one hundred people are likely to carry a pathogenic cardiogenetic variant. Counselling geneticist and clinicians need to be aware of these findings in exome and genome sequencing. Informed consent of the patient regarding the report of secondary findings should absolutely be obtained beforehand.

Temporal recording of mammalian development and precancer

Temporal ordering of cellular events offers fundamental insights into biological phenomena. Although this is traditionally achieved through continuous direct observations1,2, an alternative solution leverages irreversible genetic changes, such as naturally occurring mutations, to create indelible marks that enables retrospective temporal ordering3–5. Using a multipurpose, single-cell CRISPR platform, we developed a molecular clock approach to record the timing of cellular events and clonality in vivo, with incorporation of cell state and lineage information. Using this approach, we uncovered precise timing of tissue-specific cell expansion during mouse embryonic development, unconventional developmental relationships between cell types and new epithelial progenitor states by their unique genetic histories. Analysis of mouse adenomas, coupled to multiomic and single-cell profiling of human precancers, with clonal analysis of 418 human polyps, demonstrated the occurrence of polyclonal initiation in 15–30% of colonic precancers, showing their origins from multiple normal founders. Our study presents a multimodal framework that lays the foundation for in vivo recording, integrating synthetic or natural indelible genetic changes with single-cell analyses, to explore the origins and timing of development and tumorigenesis in mammalian systems.

Genetic changes clinically relevant in canine osteosarcoma

Osteosarcoma (OSA) is a malignant tumour of osteoblasts, forming neoplastic bone and/or osteoid. The cancer is most prevalent in children and canines, although occurring 10 times more frequently in the latter (Fenger et al., 2014). Up to 85% of malignant bone cancers are diagnosed as OSA, characterised by aggressive metastasis and rapid haematogenous dissemination, particularly to the lungs (Nelson and Couto, 2019, Thompson and Dittmer, 2020). These most commonly occur in dogs aged between 7-10 years (Selvarajah et al., 2009). The nature of canine osteosarcoma (CnOSA) gravitates to the appendicular skeleton, with 95% prevalence in dogs over 40kg, and 40% prevalence in dogs under 15kg (Nelson and Couto, 2019). Regardless of the high prevalence of CnOSA, there has been little progression in improving survival rates in the last 50 years (Zapata et al., 2019). This may be a consequence of the spontaneity of the cancer in canines (Davis and Ostrander, 2014). However, recent combined utilisation of genomic/transcriptomic technologies has been of benefit to investigating genetic loci as causes and/or protectors against CnOSA (Davis and Ostrander, 2014). As a result, somatic mutations in the TP53, MYC, CDKN2A/B, PTEN, RUNX2, and DLG2 genes in humans and dogs with osteosarcoma– and KIT and MDM2 in OSA dogs alone have been identified (Zapata et al., 2019). These findings are not only of prognostic importance to the several dog breeds with genetic predispositions to CnOSA (Simpson et al., 2017), but for humans too. This is not purely for better diagnosis of at-risk patients, but for earlier management, and a foundation for novel approaches to treatment targets for OS. Therefore, this literature review aims to identify the genetic changes clinically relevant in dogs with osteosarcoma.

Ocular manifestations in neurofibromatosis: A case series

Background: A collection of inherited diseases known as neurofibromatosis (NF) are distinguished by a broad spectrum of clinical manifestations. A genetic mutation on chromosome 17 (17q11.2) is the cause of NF type 1 (NF1), whereas a mutation on chromosome 22 (22q12.2) is the cause of NF type 2 (NF2). In NF1, ocular symptoms are more frequent. Patients with NF1 commonly appear with globe enlargement, glaucoma, and orbital facial involvement. Palpebral neurofibromas have the potential to grow extensively and, in rare instances, develop into malignant tumors. Optic nerve gliomas can result in strabismus and proptosis. Up to 80% of patients with NF2 may experience early-onset cataracts, optic nerve hamartomas, and mixed pigment retinal and epithelial hamartomas. Case presentation: We present four cases of neurofibromatosis, each displaying typical clinical features of the disorder. · Case 1: A 28-year-old male presented with headaches, reduced vision, and multiple tiny discolorations in both eyes for six months. Examination revealed multiple hyperpigmented skin lesions (2-5 mm) across his body, and ocular assessment showed normal pupillary reactions and distant vision of 6/6 in both eyes. Slit-lamp examination identified multiple dome-shaped lesions on the iris, consistent with Lisch nodules. · Case 2: A 42-year-old female experienced reduced vision for six months and skin lesions for five years. Examination revealed multiple discrete skin-colored papules and café-au-lait macules on her hands. Her vision was 6/12 in both eyes, with normal pupillary reactions. Slit-lamp findings were also consistent with Lisch nodules. o Case 3: A 48-year-old male reported vision loss in both eyes over two months, accompanied by swelling of the right upper eyelid, which was increasing in size. Lisch nodules were found on the iris of the left eye, and he was diagnosed with typical NF. o Case 4: A 38-year-old male presented with diminished vision. Slit-lamp examination revealed multiple dome-shaped lesions on the iris, suggesting Lisch nodules, along with skin lesions on the face and other body areas Conclusion: Managing neurofibromatosis requires a multidisciplinary approach, involving close collaboration between healthcare providers. Monitoring visual prognosis, enrolling patients in clinical trials, and regular disease monitoring are essential steps in managing the condition.

A review of recent clinical trials to evaluate disease-modifying therapies in the treatment of cardiac amyloidosis

Cardiac amyloidosis (CA) is a serious condition that results in infiltrative cardiomyopathy and heart failure with preserved ejection fraction (HFpEF) that is caused by the extracellular deposition of amyloid fibrils within heart tissue. While many important features of CA have been known for years, its prevalence in elderly patients with HF is increasingly being recognized. Plasma cells produce monoclonal immunoglobulin light chains which results in the formation and aggregation of amyloid fibrils that are responsible for AL amyloidosis. CA is classified as originating from either transthyretin (ATTR) or light chain (AL) amyloidosis. ATTR CA may result from a genetic mutation in the TTR gene, which is inherited (ATTRv), or from age-related deposition from wild-type ATTR (ATTRwt). Cardiac involvement in AL amyloidosis is attributed to either of two mechanisms: the extracellular deposition of amyloid fibril in the myocardium, or direct cardiotoxicity from the fibril aggregates. Typing of amyloid fibrils, a critical determinant of therapy, has also improved with wider availability of laser capture and mass spectrometry of histologic specimens. Specific and accurate evaluation of CA is now possible using cardiac magnetic resonance imaging and bone scintigraphy tracers. Survival in CA has improved markedly as novel chemotherapy agents have become available, but challenges remain in advanced disease. Broadening the amyloid-specific therapeutic landscape to include RNA inhibitors, fibril formation stabilizers and inhibitors, and immunotherapeutic targeting of amyloid deposits holds promise and may improve outcomes in systemic and cardiac amyloidoses. Treatment strategies for CA has recently undergone transformative changes, leading to some progress in outcomes for certain patients. Here, we discuss the basic features of CA as well as the emergence of novel, disease-modifying strategies that have been recently evaluated in clinical trials for the treatment of CA.

Risk factors influencing chronic inflammation in neoplastic transition to prostate cancer

Chronic inflammation and its role in driving cellular plasticity have recently been documented as a significant risk factor for prostate cancer. The progression of prostate cancer has been linked to stages of inflammation-driven changes, ranging from simple atrophy to prostatic intraepithelial neoplasia and eventually to low- and high-grade neoplastic forms. Long-term oxidative stress and the genetic damage caused by chronic inflammation are among the well-characterized risk factors in the development of prostate cancer. Both uncontrollable and controllable factors contribute to this transition process. Non-modifiable risk factors for prostate cancer include age, race, ethnicity, family history of obesity, and certain genetic predispositions. Modifiable risk factors, such as a sedentary lifestyle, poor diet, obesogenic habits, and microbial dysbiosis, may further elevate the risk of neoplastic transformation. Additionally, environmental pollutants, like chlordecone and nitrates, can interact with biological factors, potentially influencing cellular plasticity. These factors collectively contribute to an increased risk of prostate cancer and may facilitate neoplastic progression. Certain molecular markers have also been implicated in promoting chronic inflammation, enhancing cellular proliferation, and inhibiting apoptosis, thereby aiding in this transition. This review provides a comprehensive summary of the known modifiable and non-modifiable risk factors that contribute to the neoplastic transition in the prostate and elevate the risk of prostate cancer.

IDH Mutations in Glioma: Molecular, Cellular, Diagnostic, and Clinical Implications

In 2021, the World Health Organization classified isocitrate dehydrogenase (IDH) mutant gliomas as a distinct subgroup of tumors with genetic changes sufficient to enable a complete diagnosis. Patients with an IDH mutant glioma have improved survival which has been further enhanced by the advent of targeted therapies. IDH enzymes contribute to cellular metabolism, and mutations to specific catalytic residues result in the neomorphic production of D-2-hydroxyglutarate (D-2-HG). The accumulation of D-2-HG results in epigenetic alterations, oncogenesis and impacts the tumor microenvironment via immunological modulations. Here, we summarize the molecular, cellular, and clinical implications of IDH mutations in gliomas as well as current diagnostic techniques.

Vulnerability Assessment for Naval Ships against Air-Explosive Impulses: Modified Damage-Extent Method Incorporating Structural Capacity

Abstract This study introduces an enhanced damage-extent method that incorporates the structural load-bearing capacity of the hull to assess the vulnerability of naval ships to explosive loads. This vulnerability assessment predicts the area of damage to the hull structure and calculates the probability of onboard equipment experiencing functional losses due to the explosive load, thus allowing various design alternatives to be evaluated. The proposed methodology improves upon traditional damage-volume-based approaches, such as damage-radius and ellipsoid methods, by considering the hull's structural stiffness and intrinsic damage resistance. It integrates the hull's structural resistance to the load, enhancing the damage assessment process for both the hull and equipment. This approach facilitates damage prediction for different hull designs by comparing the allowable impulse with the explosive pressure. In assessing the functionality loss and vulnerability of the equipment within the damaged hull, the network of equipment functions is considered. An anti-ship cruise missile with a sea-skimming trajectory is investigated as the explosive charge, with procedures established to simulate its trajectory and impact location on the hull. Hundreds of potential internal and external explosion points are generated, predicting the explosive pressure at each location. The shock wave, including incident overpressure, reflection pressure, and quasi-static gas pressure, is converted into impulses, taking into account the configuration of hull compartments to accurately predict these pressures and equivalent impulse. The resulting impulse is compared with the intrinsic damage capacity of each compartment's structure to assess potential damage. System network and fault tree analysis evaluate the loss of function and vulnerability of equipment within the damaged hull. Finally, the proposed capacity-based damage extent method demonstrates more accurate damage assessment compared to traditional methods, overcoming the limitations of damage-radius and ellipsoid approaches by considering hull strength

Cost minimization analysis of chronic kidney disease management: Evaluating economic strategies for early intervention and treatment optimization

Chronic kidney disease is a common global health condition characterized by progressive loss of kidney function and significant cost burden on the healthcare system worldwide. The disease often coexists with conditions such as diabetes and hypertension, further complicates its management and increasing associated costs. Effective economic management, particularly in the early stages of CKD, is crucial for reducing long-term healthcare expenses and improving patient outcomes. This review examines the utility of cost minimization analysis (CMA) as a strategic tool for evaluating economic options in CKD management. CMA is an economic evaluation method that compares the costs of different therapeutic interventions that have demonstrated equivalent clinical effectiveness, thereby aiding healthcare providers and policymakers in identifying the most cost-efficient options to optimize resource utilization. This review highlights the economic impact of CKD, focusing on the increasing costs linked to advanced disease management and the essential role of early diagnosis and intervention in reducing these costs and explores various cost-effective strategies for CKD management, including pharmacological treatments, lifestyle modifications, and patient education, to identify approaches that are both clinically and economically advantageous. Additionally, the broader implications of cost minimization for healthcare policy, clinical decision-making, and resource allocation are discussed for their significance in a resource-limited healthcare setting. Evidence indicates that prioritizing cost-effective CKD management through early intervention and preventive care can significantly lower healthcare expenditures, slow the disease progression, and enhance patient’s quality of life. By adopting such strategies, healthcare systems can achieve better clinical outcomes and ensure financial sustainability.

The MON1-CCZ1 complex plays dual roles in autophagic degradation and vacuolar protein transport in rice.

Autophagy is a highly conserved cellular program in eukaryotic cells which mediates the degradation of cytoplasmic components through the lysosome, also named the vacuole in plants. However, the molecular mechanisms underlying the fusion of autophagosomes with the vacuole remain unclear. Here, we report the functional characterization of a rice (Oryza sativa) mutant with defects in storage protein transport in endosperm cells and accumulation of numerous autophagosomes in root cells. Cytological and immunocytochemical experiments showed that this mutant exhibits a defect in the fusion between autophagosomes and vacuoles. The mutant harbors a loss-of-function mutation in the rice homolog of Arabidopsis thaliana MONENSIN SENSITIVITY1 (MON1). Biochemical and genetic evidence revealed a synergistic interaction between rice MON1 and AUTOPHAGY-RELATED 8a in maintaining normal growth and development. In addition, the rice mon1 mutant disrupted storage protein sorting to protein storage vacuoles. Furthermore, quantitative proteomics verified that the loss of MON1 function influenced diverse biological pathways including autophagy and vacuolar transport, thus decreasing the transport of autophagic and vacuolar cargoes to vacuoles. Together, our findings establish a molecular link between autophagy and vacuolar protein transport, and offer insights into the dual functions of the MON1-CCZ1 (CAFFEINE ZINC SENSITIVITY1) complex in plants.

Immunomodulatory activity of omadacycline in vitro and in a murine model of acute lung injury.

Cystic fibrosis (CF) is characterized by chronic airway obstruction, infection, and inflammation leading to progressive loss of lung function and eventual respiratory failure. Omadacycline, a tetracycline antibiotic, demonstrates in vitro activity against key CF pathogens, substantial lung penetration, and increasing clinical evidence for the treatment of lung infections in people with CF (PwCF). Preliminary in vitro data demonstrate that omadacycline exhibits anti-inflammatory activity. This study aims to determine the anti-inflammatory effects of omadacycline in vitro and in a murine model of lipopolysaccharide (LPS)-induced lung neutrophilia. In vitro, THP-1-derived macrophages were treated with omadacycline (20-100 µg/mL) 30 minutes prior to LPS stimulation. Pro-inflammatory cytokine (TNF-α, IL-1β/6), chemokine (CXCL-1/2), and MMP-9 levels were analyzed after 24 hours by ELISA. Omadacycline's effects on IL-8-induced human neutrophil chemotaxis were also investigated. In vivo, omadacycline (2.5-30 mg/kg), comparators dexamethasone (1 mg/kg), and azithromycin (30 mg/kg) were administered 1 hour before and 6 hours after intranasal LPS challenge, respectively. Leukocyte counts and differentials in bronchoalveolar lavage fluid (BALF), inflammatory mediator levels in BALF and lung homogenates, pulmonary edema markers, and the severity of lung injury were evaluated 24 hours or 48 hours post-challenge. Treatment with omadacycline in vitro resulted in significant, dose-dependent reductions in IL-6, CXCL-1, and MMP-9 expression and inhibition of IL-8-induced neutrophil chemotaxis. In vivo, omadacycline yielded protective and therapeutic effects by reducing the production of proinflammatory cytokines and chemokines and neutrophil infiltration into the lungs, along with modestly improving lung injury severity. These preclinical results suggest that omadacycline may provide dual anti-bacterial and anti-inflammatory activities relevant to chronic lung infection treatment in PwCF.IMPORTANCENontuberculous mycobacteria, particularly Mycobacterium abscessus complex (MABSC), are a major concern for people with cystic fibrosis (PwCF) due to their association with deteriorating lung function. A substantial barrier to effective treatment is the limited number of safe and effective antibiotics. Omadacycline offers a potential advancement in managing MABSC infections in cystic fibrosis due to its activity, effective penetration into pulmonary secretions, improved tolerability, and good oral bioavailability as shown in healthy volunteers. Our study is the first to explore omadacycline's effects in a model of sterile lung inflammation and acute lung injury. We found that omadacycline not only has potent anti-bacterial properties but also exhibits anti-inflammatory effects, reducing lung inflammation and injury in our preclinical models. These findings underscore omadacycline's potential as a dual-action therapy for lung infections in PwCF, indicating significant potential to improve patient outcomes and guide more effective antimicrobial therapy decisions.

Study of acute and subacute toxicities and genotoxic and mutagenic potentials of the lyophilized extract of Campomanesia sessiliflora (O.Berg) mattos leaves in wistar rats

Campomanesia sessiliflora (O.Berg) Mattos is a Brazilian native plant species used in a popular medicinal tea for treating gastrointestinal, urinary, and dermatological pathologies. This study evaluated the toxicity of Campomanesia sessiliflora (O.Berg) Mattos via acute and subacute toxicity tests. It also analyzed mutagenic and genotoxic potentials by the micronucleus test, which detects genetic material damage indicating mutagenicity, and the comet assay, which assesses DNA damage levels as a genotoxicity indicator. The plant extract initially originated from the ultrasonic maceration of Campomanesia sessiliflora (O.Berg) Mattos leaves in a hydroethanolic solution. The involved animals were adult Wistar rats. Ten females were available to evaluate acute toxicity and estimate the LD50, receiving a dose of 2000 mg/kg. The subacute toxicity evaluation used 35 females and 35 males divided into seven groups: negative control (saline control – SC), positive control (cyclophosphamide control – CC), 125 mg/kg (125), 250 mg/kg (250), 500 mg/kg (500), 1000 mg/kg (1000), and the satellite group (ST). Genotoxicity and mutagenicity experiments applied bone marrow micronucleus and comet assays. Acute and subacute toxicity tests did not present behavioral, physical, and physiological changes (p≥0.05). Administering the Campomanesia sessiliflora (O.Berg) Mattos extract reduced spleen size in male and female animals, without histopathological changes. However, doses above 500 mg/kg showed significant genotoxic and mutagenic effects in the comet and micronucleus assays compared to the control group. The extract did not exhibit acute or subacute toxicity, but doses higher than 500 mg/kg indicated some level of genotoxicity and mutagenicity.

First-line therapy for high-risk people with chronic lymphocytic leukemia: a network meta-analysis.

This is a protocol for a Cochrane Review (intervention). The objectives are as follows: Primary objective: to assess the benefits and harms of currently recommended regimens as the first-line therapy in high-risk people with chronic lymphocytic leukemia, using network meta-analysis Secondary objectives: to assess whether the benefits and harms of the recommended regimens differ according to sex, Rai stage, or genetic mutation status to estimate the ranking of treatments for overall survival, progression-free survival, objective response rate, complete response rate, minimal residual disease, and serious adverse events to estimate the overall rate of adverse events and serious adverse events.

A Prospective Observational Study on Analyzing Lung Cancer Gene Mutation Variant Allele Frequency (VAF) and Its Correlation with Treatment Efficacy

A few studies have reported on the variability of the variant allele frequency (VAF) of gene mutations and the relationship between VAF and the therapeutic effect of molecular-targeted drugs. This joint study was conducted from May 2020 to January 2022 between St. Marianna University School of Medicine and the DNA Chip Research Institute. Cytology samples were used to verify the usefulness of a lung cancer compact panel test, which is a high-sensitivity next-generation sequencing (NGS) gene panel test. We analyzed the distribution of VAF and the duration of the initial molecular-targeted drug administration in patients with advanced-stage epidermal growth factor receptor (EGFR) mutations. Of the 196 patients diagnosed with non-small cell carcinoma (NSCLC), gene mutations were detected in 114 (58.2%) cases and in 68.7% of patients with adenocarcinomas (an increased detection rate). A VAF of 30% or more for DNA mutations was confirmed in 35 patients (33.7%), 10% to <30% in 38 (36.5%), and <10% in 31 (29.8%), including 18 patients (17.3%) with <6%, which is considered the lower limit of the LOD for other companion diagnostic kits. EGFR mutations were detected in 59 patients (30%), of whom 35 were in an advanced stage and received molecular-targeted drugs as their initial treatment. Groups A and B comprised patients with a VAF of ≥10% and <10%, respectively, with 27 patients (median VAF 30.6%, range 11.4–77%) in group A and 8 (median VAF 5.4%, range 0.1–8.5%) in group B. The duration of the administration of molecular-targeted drugs was 17 months (range 3–42 months) in group A and 14 months (range 1–45 months) in group B, with no statistically significant difference between the two groups (p-value = 0.7). Twenty-seven percent of patients had a VAF < 10%. Even in patients with a low VAF for gene mutations, sufficient therapeutic effects were observed with molecular-targeted drugs, suggesting the importance of detecting mutations using a highly sensitive method at initial diagnosis. Further prospective validation studies of a larger number of cases are warranted.