bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of epidemiology and novel mutation of Helicobacter pylori antibiotic resistance in South Korea
Authors: SoonYoung Park 
Published: May 2022

BackgroundHelicobacter pylori is a Gram‐negative bacterium associated with upper gastrointestinal disease, peptic ulcer, mucosa‐associated lymphoid tissue lymphoma and eradication treatment is recommended because it is one of the causes of gastric cancer. However, resistance to some antibiotics used for treatment has developed, the eradication rate is declining worldwide. This study focused on antibiotic susceptibility test (AST) of H. pylori isolated from patient clinical specimens. The aim of this study is to identify the resistance epidemiology of H. pylori and to elucidate the resistance mechanism through gene analysis.methodsH. pylori isolated from patient clinical specimens is identified and AST for amoxicillin (AMX) and clarithromycin (CLR) are performed, and the results are analyzed through Whole Genome Sequencing (WGS) and MultiLocus Sequence Typing (MLST), respectively. The strain type of the MLST result is compared with the CLR resistance minimum inhibitory concentration (MIC) value to analyze the significant points.H. pylori is cultured on EYE Agar plate and incubated in Anaerobic Jar for 2~5 days with microaerophilic air condition (5% O2, 10% CO2, 85% N2). Cultured isolates are identified by MALDI‐TOF system and performed AST using Epsilometer test to determining MIC.Results366 H. pylori were isolated from 1952 clinical patients, and 36 AMX‐resistant H. pylori and 101 CLR‐resistant H. pylori were identified. Among the identified H. pylori, 9 AMX high MIC strains were selected and WGS was performed, and 12 CLR high MIC strains were selected and MLST was performed.As the WGS result of 9 clinical strains resistant to AMX, single nucleotide polymorphism (SNP) of the potential target gene was analyzed with reference to ATCC 26695. Six genes, Penicillin binding protein1A (PBP1), PBP2, NhaC, hofH, hofC, and hefC, were selected as potential target genes. As a result of the analysis, 8, 7, 4, 9, 16, and 2 SNPs were found, respectively, and it was specifically confirmed that there were many SNPs in the outer membrane component Gene. However, No correlation was found between AMX MIC value and target SNP.12 For the CLR resistance strain, MLST was performed using 7 housekeeping genes (atpA, efp, trpC, ppa, mutY, yphC, Ure1). In 12 strain, 10 CLR‐resistance H. pylori are not identified strain type(ST) to The National Center for Biotechnology Information(NCBI) database because they have a lots of mutation in 7 housekeeping gene. 2 CLR‐resistance H. pylori are identified the new type strains.ConclusionIn this study, we conclude that hofC, hofH related to outer membrane protein component may be associated to the AMX resistance. And additional research is continued to need about other resistance potential gene, especially hopC.
Epigenetic Age Acceleration as a Risk Predictor of Hepatocellular Carcinoma
Authors: Luna Aesis , Thrush Kyra 
Published: May 2022

IntroductionMortality rates from hepatocellular carcinoma (HCC) have been steadily increasing in the US over the past 15 years. Most patients are diagnosed at advanced stages when the mortality rate is as high as 89%. Early diagnostic indicators are not currently available, and unfortunately, invasive liver core biopsies or resections are the standard of care to confirm HCC diagnosis. While no mutations have have proven to be tumor drivers, novel genome wide methylation studies show clear, cancer‐specific patterns of methylation and identified deregulation of epigenetic signatures. Epigenetic clocks have been shown to be a versatile tool for the prediction of a variety of patient phenotypes. Most commonly used for the prediction of age and mortality risk, these have also been used as indicators of alcohol consumption, smoking, and proxies for various blood‐based biomarkers. Acceleration in a number of clocks has been correlated to increased all‐cause mortality, as well as cancer risk. While much focus in HCC has been on clocks trained to precisely predict chronological age, confidence in the characterization of the notable epigenetic landscape and target region identification will be improved through the use of clocks trained to instead predict biological age with low signal to noise ratio. The goal of this study is to use epigenetic age acceleration to develop early cancer risk detection, improve treatment options, and to understand the relationship between accelerated liver tissue aging and tumorigenesis. Epigenetic aging events are potentially reversible adjuvant. Methylated DNA is also far more stable in serum and tissue than un‐methylated nucleic acids. These factors make epigenetic events attractive candidates for serum and biopsy biomarkers.MethodsGenomic DNA was extracted from IRB‐ approved, post‐operative or snap frozen, clinical patient liver resections or explants. DNA methylation (DNAm) was collected using Illumina BeadChip arrays. DNAm was used to calculate epigenetic age according to PCPhenoAge in over 600 liver samples, 8 tumor infiltrated NK‐cell isolates and 50 cell culture samples. PCPhenoAge is an improved version of the original biological aging clock with increased reliability and reduced susceptibility to technical noise. Multiple regression was used to residualize PCPhenoAge with respect to age and multiple phenotypes, such as alcohol consumption, known to affect epigenetic age. These residuals capture the epigenetic age acceleration independent of known behavioral risk factors for HCC.ResultsPreliminary analysis of 27 of our matched HCC patient tissues‐‐14 Alcohol and 13 Hepatitis C virus (HCV) associated HCC patients‐‐show higher PCPhenoAge in tumor versus non‐tumor tissues. Further, alcohol‐related HCC tumor tissue has significantly higher PCPhenoAge than HCV‐related HCC tumor tissue.
Engineered Nanoparticles as Potential Therapeutics for Acute Myeloid Leukemia
Authors: Syrovets Tatiana , Gaiser Ann‐Kathrin , Schmiech Michael , Simmet Thomas 
Published: May 2022

Acute myeloid leukemia (AML) is a malignancy associated with poor prognosis. Particularly, older patients suffer greatly from the standard chemotherapy and have a 5‐year survival of only 4%. Hence, new therapeutic agents with higher specificity and lower general cytotoxicity are urgently required.We have discovered that positively charged amino‐functionalized polystyrene nanoparticles (NP‐PS+) induce accumulation of acidic vesicular organelles with elevated pH and impaired processing of procathepsin B leading to mTOR inhibition, activation of autophagy, and induction of caspase‐dependent apoptosis in leukemia cells, but not in normal human macrophages. The antileukemic effect of NP‐PS+ was also preserved in vivo, where NP‐PS+ inhibited proliferation and induced apoptosis in leukemia xenografts grown on chick chorioallantoic membranes.Similar to polystyrene particles, amino‐functionalized gold nanoparticles (NP‐Au+) exhibited selective cytotoxicity towards AML cell lines as well as primary patient‐derived AML cells. Thus, NP‐Au+ particles induced cell death in primary human leukemia cells and reduced their colony‐forming potential, whereas normal hematopoietic cells remained unaffected by the treatment with NP‐Au+. NP‐Au+ targeted specifically the oxidative mitochondrial respiration, which is, different to normal hematopoietic cells, the main source of energy production in AML blasts and leukemic stem cells. Different to NP‐Au+, conventional chemotherapeutics such ascytarabine act in a cell cycle‐dependent manner and target only on proliferating AML blasts but not on quiescent leukemic stem cells. In agreement with the in vitro data, NP‐Au+ exhibited antileukemic efficacy against primary human AML xenografted into mice applied either as monotherapy or as a cytarabine combination regimen in the absence of detectable adverse events.Thus, this engineered nanomaterial that targets particularly resistant quiescent leukemic stem cells holds great promise as a novel nanotherapeutic for the treatment of acute myeloid leukemia independent of its cytogenetic profile.
Proteins of the ubiquitin system in the Shoc2 ‐ ERK1/2 signaling axis and Noonan‐like syndrome with loose anagen hair (NSLAH) RASopathy
Authors: Galperin Emilia , Wilson Patricia , Abdelmoti Lina , Norcross Rebecca , Palayam Malathy 
Published: May 2022

The ERK1/2 signaling pathway is critical in organismal development and tissue morphogenesis. Deregulation of this pathway leads to congenital abnormalities with severe developmental dysmorphisms, i.e. RASopathies. The core ERK1/2 cascade relies on scaffold proteins such as Shoc2 to guide and fine‐tune its signals. Mutations in shoc2 lead to the development of the pathology termed Noonan‐like Syndrome with Loose Anagen Hair (NSLAH) RASopathy. However, the mechanisms underlying the functions of Shoc2 and its contributions to disease progression remain unclear.We found that Shoc2 assembles an elegant multi‐component complex that incorporates several proteins of the ubiquitin system. To fine‐tune the amplitude of ERK1/2 signal transmitted via the complex, Shoc2 tethers the E3 ligase HUWE1, the (AAA+) ATPases, PSMC5 and VCP/p97, and the deubiquitinating enzyme, USP7. All of these enzymes are integral to the intricate feedback mechanism.Our recent studies demonstrated that ERK1/2 pathway activation triggers the interaction of Shoc2 with the ubiquitin‐specific protease USP7. We identified that in the Shoc2 module, USP7 functions as a molecular “switch” that controls the E3 ligase HUWE1 and the HUWE1‐induced regulatory feedback loop. We also found that disruption of Shoc2‐USP7 binding leads to aberrant activation of the Shoc2‐ERK1/2 axis. The zebrafish vertebrate model was then used to show that Shoc2 congenital mutations affecting Shoc2 interaction with USP7 lead to aberrant Shoc2 ubiquitination and signal transmission. Thus, our studies reveal a role for USP7 in the pathogenic mechanisms underlying NSLAH extending our understanding of how ubiquitin‐specific proteases regulate intracellular signaling.In summary, our studies are the first to demonstrate that the Shoc2 scaffold employs multi‐protein enzymatic machinery to govern the amplitude of Shoc2‐ERK1/2 signals. We also uncover novel molecular mechanisms underlying the pathogenesis of Noonan‐like syndrome with loose anagen hair. Overall, these studies significantly advance our understanding of the mechanisms by which non‐enzymatic scaffolds regulate the specificity and dynamics of the ERK1/2 signaling networks.
Discerning a Potential Link between MEMO1’s Role in Vascular, Bone, and Craniofacial Development
Authors: Wuebker Samantha , Van Otterloo Eric 
Published: May 2022

Embryonic development of craniofacial structures requires the tight integration of multiple tissues and cell‐types. For example, the intersection of neural crest derived mesenchyme with mesoderm derived endothelium (i.e., vasculature) is necessary for craniofacial bone development and mineralization. Despite craniofacial anomalies being the 2nd leading congenital anomaly worldwide, our understanding of how these tissues interact is limited.Through a forward genetic screen, we previously identified a novel role for the protein encoded by the gene mediator of ERBB2‐driven cell motility 1 (Memo1) in craniofacial development. Global loss of Memo1 resulted in a range of defects, including hypo‐mineralization of the craniofacial skeleton, fully penetrant cleft secondary palate, coupled with widespread vascular abnormalities. Interestingly, while neural crest cell specific deletion of Memo1 recapitulated some aspects of global loss of Memo1, several defects were less apparent—suggesting MEMO1 functions in tissues outside the neural crest during craniofacial development. Further, molecular profiling of craniofacial elements in global Memo1mutants identified significantly altered Vegfa expression, coupled with disrupted vascular development, relative to controls. Based on these observations, along with Memo1’s semi‐ubiquitous expression during development, we hypothesized that MEMO1 functions within the endothelial derived vasculature and this function is necessary for proper craniofacial development.To test this hypothesis, we have generated a Memo1 endothelial specific mouse knock‐out model. Similar to Memo1 global knock‐outs, and in contrast to Memo1 neural crest knock‐outs and controls, Memo1 endothelial knock‐outs develop vascular hemorrhaging and edema with variable penetrance. Vascular defects are accompanied by gross alterations to craniofacial shape. We are currently utilizing µCT, cell sorting coupled with gene expression profiling, and ex vivo ‘sprouting’ assays to quantify the impact endothelial loss of Memo1 has on craniofacial bone development and mineralization, MEMO1’s molecular role in craniofacial endothelial cells, and MEMO1’s functional role in endothelial cells, respectively.Overall, in addition to MEMO1’s neural crest cell specific role, our findings extend MEMO1’s function into an additional cell‐type, endothelial cells, during craniofacial development. These data illuminate the molecular pathology for how disrupted neural crest or endothelial cells may lead to human craniofacial disorders.
Where is the primate transverse tarsal joint? Chopart vs. Lisfranc
Authors: Greiner Thomas M. 
Published: May 2022

The concept of the transverse tarsal joint is typically associated with accommodative twists of the foot, and with a midfoot flexion known as the midtarsal break. The specific joints associated for these motions are often assumed, but are rarely validated. The two most common candidates are “Chopart’s Joint,” which separates the cuboid and navicular from the calcaneus and talus, and “Lisfranc’s Joint,” formed by a combination of all the tarsometatarsal joints. Both of these joints were initially identified as sites for surgical amputations and were not based on any presumed function. This study investigates the utility of these composite joints as sites of foot motion for humans and for some non‐human primates.Data were drawn from 3D kinematic evaluations of the cadaveric feet of 12 humans, 11 chimpanzees, and 11 baboons. Data were collected in response to an anterior‐posterior and medial‐lateral driving motion of the leg over the foot. Data were derived from rigid cluster tracking of individual foot bones during an anterior‐posterior motion of the leg, typically associated with dorsiflexion‐plantarflexion, and a medial‐lateral leg motion, associated with inversion‐eversion and/or supination‐pronation. Data were analyzed using the Functional Alignment procedure to derive the orientation of axes of rotation for the involved joints.Results show that the axis orientation of Chopart’s Joint is not a good compromise between the very different motions of the talonavicular and calcaneocuboid joints in any primate. This observation applies to results drawn from both anterior‐posterior and medial‐lateral driving motions. The rotational axis of Lisfranc’s Joint in the different primates is also quite divergent from the individual tarsometatarsal joint axes when responding to the anterior‐posterior driving action, but the axis is more parallel to those of the tarsometatarsal joints during the medial‐lateral driving action. However, it should be noted that the joint axes of the individual tarsometatarsal joints are almost identical, while the Lisfranc’s Joint axis stands apart.These results show that the individual joint actions are more complicated than their composite forms expressed as Chopart’s or Lisfranc’s joints. The concept of single “transverse tarsal” joint may not be functionally justified, within the human animal or as a basis for comparison of humans and non‐human primates. Researchers should reassess how well these traditional joint concepts apply to their specific questions of foot biomechanics.The author conveys his respect and gratitude to the persons whose remains are associated with this project.
Chemogenetic activation of glutamatergic preBötzinger Complex neurons increases breathing in mice
Authors: Parzei Natalka , Scarpellini Carolina , Montandon Gaspard 
Published: May 2022

ObjectiveRespiratory rhythm is generated by complex neural circuits in the medulla. At the core of these circuits is the preBötzinger Complex (preBötC), a collection of cells generating respiratory rhythm in the ventral part of the medulla. In addition to its role in generating respiratory rhythm, the preBötC also plays a significant role in opioid‐induced respiratory depression. The preBötC is made up of a heterogenous group of neurons, with a large portion being glutamatergic and expressing µ‐opioid receptors. A marker for these glutamatergic neurons is vesicular glutamate transporter 2 (vGLUT2). Here we propose that targeting vGLUT2‐expressing preBötC neurons using chemogenetics can stimulate respiratory rhythm and potentially alleviate opioid‐induced respiratory depression.HypothesisActivation of vGLUT2 preBötC neurons stimulates breathing and alleviates respiratory depression by fentanyl in mice.MethodsUsing a cre‐lox recombination approach, we expressed the excitatory hM3Dq (a modified version of the excitatory M3 muscarinic receptor) into vGLUT2 preBötC neurons. To this aim, we injected into the preBötC recombinant adeno‐associated virus transcriptionally activated by Cre‐recombinase containing hM3Dq and mCherry DNA flanked by lox sites in vglut2‐IRES‐Cre male mice (Jackson Labs). Four weeks after adeno‐associated virus, mice were anesthetized with isoflurane and respiratory activity was measured through electrodes implanted into the diaphragm and the genioglossus muscle. Mice were given consecutive intramuscular injections of saline and the hM3Dq ligand, clozapine‐N‐oxide (CNO; 0.5 and 1 mg/kg), while respiratory responses were monitored. Brains were collected for histological analysis with expression of hM3Dq determined by expression of mCherry.ResultsInjection of 0.5mg/kg CNO elicited a 24.55% increase in respiratory rate 8 minutes following CNO injection. Injection of 1mg/kg CNO destabilized breathing within 6‐25 minutes following injection. Histology confirmed expression of mCherry in the region of the preBötC. Further studies will determine whether stimulation of vGLUT2 preBötC neurons can prevent or reverse respiratory rate depression elicited by fentanyl.ConclusionsOur preliminary data showed that stimulation of glutamatergic preBötC neurons can increase respiratory rate in anesthetized conditions. Further studies will determine whether stimulation of glutamatergic preBötC cells also increases respiratory rate in freely‐behaving mice and whether the glutamatergic preBötC neurons are viable targets for the development of preventive drug therapies for respiratory depression during opioid overdose.
Sex‐Dependent Changes in Vascular Function in Adult Rats Following Prenatal Exposure to Methamphetamine
Authors: Chavva Hasitha , Belcher Adam M. , Brazeau Daniel A. , Rorabaugh Boyd R. 
Published: May 2022

BackgroundPrenatal exposure to central nervous system stimulants such as cocaine, nicotine, and caffeine alters cardiovascular function in adult offspring. Methamphetamine use during pregnancy is associated with negative consequences in the offspring. However, most studies of methamphetamine use during pregnancy have focused on behavioral and neurological outcomes. Relatively little is known regarding the impact of prenatal methamphetamine exposure on the adult cardiovascular system. This study examined the hypothesis that methamphetamine use during pregnancy alters contractile function of the vasculature in the adult offspring. We also investigated the role of perivascular adipose tissue (PVAT) in methamphetamine‐induced changes in vascular function.MethodsPregnant female rats received daily injections of saline or methamphetamine (5 mg/kg) throughout gestation. Agonist‐induced contraction and relaxation responses were measured in aortas of 5 month old offspring in the presence and absence of perivascular adipose tissue.ResultsPrenatal methamphetamine significantly attenuated acetylcholine‐induced relaxation in male (but not female) aortas when PVAT remained intact. However, prenatal methamphetamine had no impact on acetylcholine‐induced relaxation when PVAT was removed. Nitroprusside‐induced relaxation was unaffected by prenatal methamphetamine. Prenatal exposure to methamphetamine had no impact on acetylcholine‐induced relaxation in third order mesenteric arteries of male or female offspring regardless of the presence of PVAT. Angiotensin II‐induced contractile responses were significantly potentiated in male (but not female) aortas regardless of the presence of PVAT. This effect was abolished by L‐Nitro arginine methyl ester (L‐NAME). Contractile responses to phenylephrine and serotonin were unaffected in both male and female aortas. Basal blood pressure in adult male and female offspring was also unaffected by prenatal exposure to methamphetamine.ConclusionsPrenatal exposure to methamphetamine sex‐dependently alters PVAT function in the aorta. Methamphetamine also enhances angiotensin II‐induced contraction through a mechanism that involves suppression of nitric oxide signaling. These data provide evidence that methamphetamine use during pregnancy induces sex‐dependent changes in the vasculature that may increase the risk of cardiovascular disease in adult offspring.
Sex Differences in Estimates of Cardiac Autonomic Function Using Time Domain based Method of Heart Rate Variability: Effects of Oral Capsaicin
Authors: Zaleski Kendall S. , Gyampo Abena , Lora Brian , Tomasi Tawn , Lynch Meaghan , Giuriato Gaia , Basso Emma , Finegan Emma , Schickler Jack , Venturelli Massimo , Ives Stephen J. 
Published: May 2022

Heart rate variability (HRV) estimates autonomic nervous system (ANS) influence on the heart and is sex‐specific. Sensory afferents exhibit sex‐specificity; although, there is a paucity of data on the potential effects of Capsaicin, an agonist for transient receptor potential vanilloid channel‐1 (TRPV1), on cardiac ANS activity and if the effect is sex‐dependent. Given the predictive nature of HRV on risk of developing hypertension, understanding the sex differences in factors governing HRV is paramount.PURPOSETherefore, this study sought to determine the sex‐specificity in the effect of capsaicin on cardiac autonomic function estimated through HRV. It was hypothesized that females would have lower HRV than the age‐matched males and that capsaicin could attenuate these sex differences.METHODSHRV was measured in 38 young males (M: n=25) and females (F: n=13), in a blinded crossover design, after acute ingestion of placebo or capsaicin capsules. Resting measurements of HR, RR interval, root mean square of successive differences (RMSSD), natural log‐transformed RMSSD (LnRMSSD), standard deviation of n‐n intervals (SDNN), number of pairs of successive n‐n intervals that differ by more than 50 msec (NN50), and percent NN50 to total n‐n intervals (PNN50) were obtained using standard techniques.RESULTSUnder placebo, males had significantly lower minimum HR (M: 49±9.7 vs. F: 58±16 beats/min, p=0.038, d=‐0.738) and significantly higher NN50 (M: 141±118 vs. F: 33±23, p=0.003, d=‐0.129) than females. There was a main effect of sex on HR (M: 59±9.3 vs. F: 65±12 beats/min, p=0.036, η2=0.098), minimum HR (M: 47±8.3 vs. F: 56±12 beats/min, p=0.014, η2=0.124), and NN50 (M: 177±143 vs. F: 29±17, p<0.001, η2=0.249). There was a significant interaction of sex and treatment (p=0.02, η2=0.027) for RMSSD, where males increased (Placebo: 78±55 vs. Capsaicin: 91±64 ms), and females decreased (Placebo: 105±83 vs. Capsaicin 76±43 ms).CONCLUSIONThis study recapitulates previously documented sex differences in HR and HRV. Acute ingestion of capsaicin increased RMSSD in men, but decreased RMSSD in women, suggesting a sexual dimorphism in parasympathetic response, perhaps due to differences in TRPV1‐sensitive afferents or sensitivity. The physiological ramifications of these findings, specifically in the regulation of blood pressure and ultimately cardiovascular disease risk remain to be explored, especially in the transition to menopause in women.