Defects in the central metabolism prevent thymineless death in Escherichia coli, while still allowing significant protein synthesis.

Starvation of E. coli thyA auxotrophs for the required thymine or thymidine leads to cessation of DNA synthesis and, unexpectedly, to thymineless death (TLD). Previously, TLD-alleviating defects were identified by the candidate gene approach, for their contribution to replication initiation, fork repair, or SOS induction. However, no TLD-blocking mutations were ever found, suggesting a multifactorial nature of TLD. Since (until recently) no unbiased isolation of TLD suppressors was reported, we used enrichment after insertional mutagenesis to systematically isolate TLD suppressors. Our approach was validated by isolation of known TLD-alleviating mutants in recombinational repair. At the same time, and unexpectedly for the current TLD models, most of the isolated suppressors affected general metabolism, while the strongest suppressors impacted the central metabolism. Several temperature-sensitive (Ts)-mutants in important/essential functions, like nadA, ribB or coaA, almost completely suppressed TLD at 42°C. Since blocking protein synthesis completely by chloramphenicol prevents TLD, while reducing protein synthesis to 10% alleviates TLD only slightly, we measured the level of protein synthesis in these mutants at 42°C and found it to be 20-70% of the WT, not enough reduction to explain TLD prevention. We conclude that the isolated central metabolism mutants prevent TLD by affecting specific TLD-promoting functions.

Neisserial adhesin A (NadA) binds human Siglec-5 and Siglec-14 with high affinity and promotes bacterial adhesion/invasion.

Neisserial adhesin A (NadA) is a meningococcal surface protein included as recombinant antigen in 4CMenB, a protein-based vaccine able to induce protective immune responses against Neisseria meningitidis serogroup B (MenB). Although NadA is involved in the adhesion/invasion of epithelial cells and human myeloid cells, its function in meningococcal physiology is still poorly understood. To clarify the role played by NadA in the host-pathogen interaction, we sought to identify its cellular receptors. We screened a protein microarray encompassing 2,846 human and 297 mouse surface/secreted recombinant proteins using recombinant NadA as probe. Efficient NadA binding was revealed on the paired sialic acid-binding immunoglobulin-type lectins receptors 5 and 14 (Siglec-5 and Siglec-14), but not on Siglec-9 therein used as control. The interaction was confirmed by biochemical tools with the determination of the KD value in the order of nanomolar and the identification of the NadA binding site by hydrogen-deuterium exchange coupled to mass spectrometry. The N-terminal domain of the Siglec-5 that recognizes the sialic acid was identified as the NadA binding domain. Intriguingly, exogenously added recombinant soluble Siglecs, including Siglec-9, were found to decorate N. meningitidis surface in a NadA-dependent manner. However, Siglec-5 and Siglec-14 transiently expressed in CHO-K1 cells endorsed NadA binding and increased N. meningitidis adhesion/invasion while Siglec-9 did not. Taken together, Siglec-5 and Siglec-14 satisfy all features of NadA receptors suggesting a possible role of NadA in the acute meningococcal infection.IMPORTANCEBacteria have developed several strategies for cell colonization and immune evasion. Knowledge of the host and pathogen factors involved in these mechanisms is crucial to build efficacious countermoves. Neisserial adhesin A (NadA) is a meningococcal surface protein included in the anti-meningococcus B vaccine 4CMenB, which mediates adhesion to and invasion of epithelial cells. Although NadA has been shown to bind to other cell types, like myeloid and endothelial cells, it still remains orphan of a defined host receptor. We have identified two strong NadA interactors, Siglec-5 and Siglec-14, which are mainly expressed on myeloid cells. This showcases that NadA is an additional and key player among the Neisseria meningitidis factors targeting immune cells. We thus provide novel insights on the strategies exploited by N. meningitidis during the infection process, which can progress to a severe illness and death.

Auricular Therapy for Migraine.

PURPOSE OF REVIEW: Migraine brings hours or even days of disability, affecting 15% of the US population and one billion people worldwide. Migraine treatments have improved over the years and there is now a range of non-pharmacologic therapies that can be administered as monotherapy, combined with pharmacologic therapy or combined with other non-pharmacologic therapies to give greater options for those who do not tolerate, do not respond to, or who wish to reduce or avoid pharmacologic treatments. RECENT FINDINGS: We conducted a review of the literature on auricular therapy as acute or preventive treatment for migraine, searching the databases of MEDLINE and ClinicalTrials.gov from 2013 to 2023. A total of 43 articles contained at least one search term, with three studies specific to acute or prevention of migraine (one for acute only, one for prevention only and one for both acute and prevention). The population was limited to, adults with migraine ages 18 or older, with the administration of auricular therapy as the intervention. While there have been studies on the use of auricular therapy for pain on two specific standardized auricular therapies, Battlefield Acupuncture (BFA) and National Acupuncture Detoxification Association (NADA), neither of these protocols were utilized in any of the studies specific to migraine management. Each of the three studies used different techniques, with one using acupuncture needles and five specific points and two using semi-permanent needles (remained in for a few days) that were placed in areas that showed high activity. Each of these studies showed auricular therapy to have benefit for the management of migraine. However, the authors of each of the studies recommended further studies. Auricular therapy may be a helpful adjunctive treatment to abort a current migraine attack or aid in reducing the frequency or severity of migraine attacks.

NADA Acupuncture in Specialized Palliative Care: Patients' and Family Caregivers' Experiences.

CONTEXT: Over the last 30 years, non-pharmacological treatment with ear acupuncture, including National Acupuncture Detoxification Association (NADA) acupuncture, has become popular in Western society to relieve cancer-related symptoms. OBJECTIVES: This study aim is to explore whether NADA acupuncture plays a role in relieving suffering experienced by patients hospitalized in a specialized palliative care ward and their family caregivers and whether it contributes to their coping skills. METHODS: A qualitative study with a hermeneutic approach utilizing inductive thematic content analysis. Purposeful sampling took place in a specialized palliative care ward in Denmark and all data were collected through semi-structured interviews with patients and family caregivers as either individual or family interviews. RESULTS: A total of 10 patients and 15 family caregivers participated in interviews. Four themes emerged: (1) communication about treatment with NADA acupuncture, (2) relief of suffering generates extra energy and inner strength, (3) sharing the experience with the family is beneficial, and (4) physical and psychological effects after receiving NADA acupuncture. CONCLUSION: Patients in palliative care and their family caregivers experienced relief of suffering after receiving NADA acupuncture, as well as improved well-being, and extra energy and inner strength to cope with their life situation. The ability to share NADA acupuncture gave family caregivers the feeling of being cared for and contributed to feelings of fellowship and togetherness within the family, strengthening their ability to communicate and cope.

NADA Ear Acupuncture and Medical Acupuncture for Pain- and Health-Related Quality of Life among Older Patients with Chronic Nonspecific Low Back Pain: A Prospective Clinical Trial.

BACKGROUND: The purpose of this study was to investigate the efficacy and safety of the NADA (National Acupuncture Detoxification Association)-standardized ear acupuncture protocol in comparison to medical acupuncture (MA) in the treatment of chronic nonspecific low back pain (LBP) in older adults. METHODS: This was a prospective, clinical, single center, open label, comparative study. A total of 60 older patients with chronic nonspecific LBP were enrolled in the study. The patients were divided into two groups. The MA group received treatment with medical acupuncture (MA), while the NADA group received NADA ear acupuncture once a day for 20 min, for a total of 10 sessions. The co-primary outcome measures were the reduction in pain intensity evaluated by the Numeric Rating Scale (NRS) compared to baseline and improvement in patients' quality of life (QOL) assessed in the SF-36 questionnaire before and after treatment; this was compared between the two groups. RESULTS: After two weeks of treatment, a significant reduction compared to baseline was observed in the NRS scores following treatment with medical acupuncture as well as after the utilization of NADA ear acupuncture protocol: NRS score for average pain experienced by the patients over the previous week (NRSa) MA: p = 0.002; NADA: p < 0.001, maximum NRS score in the past week (NRSm) MA: p < 0.001; NADA: p < 0.001, and NRS score at the time of examination (NRSe) MA: p = 0.001; NADA: p < 0.001. Reduction of the NRSa score compared to baseline was significantly greater in the NADA group (p = 0.034). Significant improvements in the QOL of patients according to the SF-36 questionnaire compared to baseline were observed in the MA group in the following domains: PF (p = 0.003), RP (p = 0.002), SF (p = 0.041), RE (p = 0.005), MH (p = 0.043), HT (p = 0.013), PCS (p = 0.004), and MCS (p = 0.025); and in the NADA group, in the following domains: PF (p = 0.004), RP (p = 0.048), BP (p = 0.001), VT (p = 0.035), RE (p = 0.006), MH (p < 0.001), HT (p = 0.003), PCS (p < 0.001), and MCS (p < 0.001). There were minor complications observed in 35% of patients (total of 20 participants); 31% (9 patients) in the MA group and 39% (11 patients) in the NADA group. These were minor and quickly resolved, including insertion point pain, minor bleeding after needle removal, and one instance of fainting. No patients in either group reported worsening of LBP. These complications occurred in 4.14% of MA sessions (12 times/290 sessions) and in 6.07% of NADA acupuncture sessions (16 times/280 sessions). CONCLUSION: The outcomes of this study suggest that both MA and NADA ear acupuncture could be a valuable and personalized component of a comprehensive approach to managing chronic nonspecific LBP in older patients. Incorporation of MA and NADA ear acupuncture into the clinical management of chronic nonspecific LBP in elderly patients has the potential to reduce pain intensity and improve the overall quality of life of affected individuals. However, further studies are needed to confirm our findings.

The Role of Acupuncture in Treating a Patient with a Gambling Disorder.

Background: Addiction is a chronic/relapsing disorder characterized by compulsive seeking and continuous involvement despite dangerous consequences. It causes long-term changes in the brain. Gambling disorder is a nonsubstance behavioral addiction. An important feature of gambling disorder is maladaptive gambling behavior that is persistent and repetitive, and interferes with patients' personal lives, families, and/or activities. Acupuncture is a nonpharmacologic therapeutic modality for managing addiction, with good results. Case: A 32-year-old man with a gambling disorder was referred from a psychiatry department. The patient had a history of gambling since high school. He began to gamble in larger amounts in 2019. The patient got antiseizure, antidepressant, and antipsychotic medications, and cognitive behavioral therapy in the psychiatry department. Manual acupuncture therapy was performed at GV 20, Ex-HN 1, Ex-HN 3, PC 6, ST 40, and LR 3. Electroacupuncture was performed at LI 4, LI 11, ST 36, SP 6, and ST 25. Laser acupuncture was performed at NADA protocol bilateral ear points. The patient also received scalp acupuncture for extrapyramidal symptoms After this combination of acupuncture therapy for 8 sessions, with pharmacotherapy and psychotherapy, the patient's condition improved. Conclusions: Acupuncture produces positive results in patient with gambling disorders. It helps regulate the reward system; stimulates release of neurotransmitters in the brain; produces neuroprotective effects; and activates certain brain areas to suppress impulsivity and craving and to balance emotions. Acupuncture is thought to be related to increased blood flow in the frontal lobe, thereby increasing nerve metabolism, as well as regulating dopamine in the basal ganglia. A combination of acupuncture, pharmacologic agents, and psychotherapy has a positive synergistic effect in patients with gambling disorder.

Ameliorating effect of 6-week swimming exercise on mice with experimental autoimmune encephalomyelitis (EAE) by reducing fetuin-A and increasing AMPK & NAD⁺ levels in liver tissue.

Objectives: Multiple sclerosis (MS) is a chronic inflammatory disease affecting sensory and motor function in the central nervous system. Physical activities in the prevention and treatment of such conditions have shown promising results. However, their mechanisms of action have not been fully known yet and need further study. The present study aimed to evaluate the preventive effect of swimming exercise on some liver factors involved in inflammation and MS. Materials and Methods: In this study, experimental autoimmune encephalomyelitis was induced in C57BL/6 mice, and the effect of a 6-week swimming exercise on the levels of fetuin-A, AMP-activated protein kinase (AMPK), and Nicotinamide adenine dinucleotide (NAD+) in their liver tissue was investigated by western blot analysis and NAD+ colorimetric assay. Results: The study showed that EAE induction substantially (3.5 - fold) enhanced the fetuin-A levels and caused a reduction in AMPK and NAD+ amount. This is when doing 6 weeks of swimming exercise reduced fetuin-A to slightly above control. Also, levels of AMPK and NAD+ markedly increased in C57BL/6 mice with EAE. Conclusion: Doing regular exercise may limit the body's inflammatory responses and reduce the severity of MS by regulating the expression of fetuin-A and increasing AMPK and NAD⁺ levels in liver tissue.

Ear acupuncture as an adjunct in a treatment protocol for anorexia nervosa: utilization rate and nurses' experience.

BACKGROUND: Anorexia nervosa (AN) is a life-threatening illness. Sometimes long inpatient treatment is necessary, increasing the anxiety that comes with hospitalization and the necessary weight gain. Safe, non-pharmacological adjunctive therapies that improve subjective health are called for. OBJECTIVE: The aim of this non-randomized, mixed-methods observational study was to describe the utilization rate and nurses' experiences of ear acupuncture in a highly specialized clinic for eating disorders in Sweden, in which acupuncture had been implemented as part of routine clinical care. Twenty-five patients with AN-treated voluntarily or by law were included. The semi-standardized National Acupuncture Detoxification Association (NADA) ear acupuncture protocol, sometimes combined with needling at 2 traditional acupuncture point locations on the body, had been implemented as a voluntary adjunct to usual care, twice weekly. To evaluate the acceptance of acupuncture, the study examined how often patients chose acupuncture when offered on schedule, and how often they asked for extra acupuncture sessions. Patients rated their subjective health using the hospital anxiety and depression scale (HADS) and visual analogue scale (VAS), treatment satisfaction with usual care and acupuncture, and health-related quality of life with the RAND36 instrument. A credibility/expectancy questionnaire (CEQ) was used to measure confidence in treatment. Body mass index (BMI) was calculated to follow the patients' recovery. Nurses' experiences of giving acupuncture as a part of routine care were captured in interviews, and analysed with content analysis. Ethical approval was obtained. RESULTS: Despite an initially moderate level of trust in acupuncture, the utilization rate of the scheduled acupuncture was 89% and patients asked for extra acupuncture sessions on 28 occasions. No serious side effects were reported. Nurses' experiences of providing acupuncture were positive. They were generally enthusiastic, although they reported finding it difficult to organize group treatments and to find time for acupuncture sessions if they were not scheduled. CONCLUSION: Further research into the effectiveness and costs of acupuncture in psychiatric care is needed. This study provides relevant information for clinicians as well as researchers planning future randomized controlled trials.

Desejo, Vontade, nada: de Schopenhauer a Freud, Freud com Schopenhauer / Deseo, Voluntad, nada: desde Schopenhauer hasta Freud, Freud con Schopenhauer / Desire, Will, nothingness: from Schopenhauer to Freud, Freud with Schopenhauer

O artigo visa estabelecer uma articulação entre as categorias de Vontade - referida à filosofia da Schopenhauer - e de desejo - recuperada desde o Wunsch proposto por Freud em seu Projeto para uma psicologia científica -, para, com isso, refazer o caminho percorrido por eles e indicar sua lógica interna diante de outra categoria: o nada, a elas intimamente ligada. Concluímos que há maior solidariedade entre elas do que se supõe, pois, se a Vontade é "sem extensão" e se o desejo é uma ficção cuja mecânica exige alívio, então sua própria satisfação seria uma negação interna da Vontade.

The article aims to establish an articulation between the categories of Will - referred to Arthur Schopenhauer's philosophy - and desire - recovered from the Wunsch proposed by Freud in his Project for a scientific psychology - in order to retrace their path and to indicate their internal logic in the face of another category: nothingness, internally linked with them. We conclude that these concepts seem to be more intimate than it is supposed, because if Will is "without extension" and desire is a fiction whose mechanics demand relief, then its own satisfaction would be an inner denial of the Will.

El artículo intenta establecer una articulación entre los conceptos de Voluntad - referido a la filosofía de Schopenhauer - y de deseo - recuperado desde el Wunsch propuesto por Freud en su Proyecto para una psicología científica. Intentamos rehacer el camino para indicar su lógica interna frente a otra categoría: la nada, íntimamente ligada. Llegamos a la conclusión de que voluntad, deseo y nada parecen ser más íntimos de lo que se supone, pues si la Voluntad es "sin extensión" y el deseo una ficción cuya mecánica exige alivio, entonces la propia satisfacción sería una negación interna de la Voluntad.

Complete mitochondrial genome of the Pacific limpet Cellana nigrolineata (Gastropoda: Patellogastropoda) determined by shotgun sequencing using the Illumina NGS platform.

The Pacific limpet Cellana nigrolineata is one of the most commonly found limpets in the intertidal shores of Japan. Here, we report the full mitogenome sequence of an individual specimen of the species, which was collected from the intertidal rocky beach in the Nada beach of Gobo City, Wakayama, Japan (33.8316 N, 135.1751 E), in 2018. The sequence was determined by the shotgun sequencing method using the NGS Illumina MiSeq platform. The genomic structure of C. nigrolineata is the same as the previously reported congener, C. radiata, which shows a representative Nacellidae and metazoan mitogenomic structures. The mitogenome has all of its 37 genes included in its 16,153 bp, with one control region located between the tRNA-Cys and tRNA-Gly genes. In order to clarify the phylogenetic position of C. nigrolineata in Gastropoda, a data set including the mitogenomes of 10 patellogastropods, 10 non-patellogastropod gastropods, and four outgroups were used in maximum likelihood inferences. Although with some exceptions, the resulting phylogeny supported the monophylies of traditionally accepted gastropod subclasses, and thus confirms the position of C. nigrolineata in Patellogastropoda.

Genetic Diversity of Meningococcal Serogroup B Vaccine Antigens among Carriage Isolates Collected from Students at Three Universities in the United States, 2015-2016.

Carriage evaluations were conducted during 2015 to 2016 at two U.S. universities in conjunction with the response to disease outbreaks caused by Neisseria meningitidis serogroup B and at a university where outbreak and response activities had not occurred. All eligible students at the two universities received the serogroup B meningococcal factor H binding protein vaccine (MenB-FHbp); 5.2% of students (181/3,509) at one university received MenB-4C. A total of 1,514 meningococcal carriage isolates were obtained from 8,905 oropharyngeal swabs from 7,001 unique participants. Whole-genome sequencing data were analyzed to understand MenB-FHbp's impact on carriage and antigen genetic diversity and distribution. Of 1,422 isolates from carriers with known vaccination status (726 [51.0%] from MenB-FHbp-vaccinated, 42 [3.0%] from MenB-4C-vaccinated, and 654 [46.0%] from unvaccinated participants), 1,406 (98.9%) had intact fHbp alleles (716 from MenB-FHbp-vaccinated participants). Of 726 isolates from MenB-FHbp-vaccinated participants, 250 (34.4%) harbored FHbp peptides that may be covered by MenB-FHbp. Genogroup B was detected in 122/1,422 (8.6%) and 112/1,422 (7.9%) isolates from MenB-FHbp-vaccinated and unvaccinated participants, respectively. FHbp subfamily and peptide distributions between MenB-FHbp-vaccinated and unvaccinated participants were not statistically different. Eighteen of 161 MenB-FHbp-vaccinated repeat carriers (11.2%) acquired a new strain containing one or more new vaccine antigen peptides during multiple rounds of sample collection, which was not statistically different (P = 0.3176) from the unvaccinated repeat carriers (1/30; 3.3%). Our findings suggest that lack of MenB vaccine impact on carriage was not due to missing the intact fHbp gene; MenB-FHbp did not affect antigen genetic diversity and distribution during the study period.IMPORTANCE The impact of serogroup B meningococcal (MenB) vaccines on carriage is not completely understood. Using whole-genome sequencing data, we assessed the diversity and distribution of MenB vaccine antigens (particularly FHbp) among 1,514 meningococcal carriage isolates recovered from vaccinated and unvaccinated students at three U.S. universities, two of which underwent MenB-FHbp mass vaccination campaigns following meningococcal disease outbreaks. The majority of carriage isolates recovered from participants harbored intact fHbp genes, about half of which were recovered from MenB-FHbp-vaccinated participants. The distribution of vaccine antigen peptides was similar among carriage isolates recovered from vaccinated and unvaccinated participants, and almost all strains recovered from repeat carriers retained the same vaccine antigen profile, suggesting insignificant vaccine selective pressure on the carriage population in these universities.

Epitope and Paratope Mapping by HDX-MS Combined with SPR Elucidates the Difference in Bactericidal Activity of Two Anti-NadA Monoclonal Antibodies.

Characterization of antigen-antibody interactions is crucial for understanding antibody-mediated protection against pathogens, biopharmaceutical development, as well as evaluation of the immune response post vaccination. Bexsero is a multicomponent vaccine against Neisseria meningitidis serogroup B in which one of the key vaccine antigens is Neisserial adhesin A (NadA), a trimeric coiled-coil protein. Two NadA-specific monoclonal antibodies (mAbs) isolated from Bexsero-vaccinated individuals have been shown to have similar binding affinity and appear to recognize a similar antigen region, yet only one of the mAbs is bactericidal. In this study, we use hydrogen/deuterium exchange mass spectrometry (HDX-MS) to perform an in-depth study of the interaction of the two mAbs with NadA antigen using a combined epitope and paratope mapping strategy. In addition, we use surface plasmon resonance (SPR) to investigate the stoichiometry of the binding of the two mAbs to NadA. While epitope mapping only identifies a clear binding impact of one of the mAbs on NadA, the paratope mapping analyses shows that both mAbs are binding to NadA through several complementarity determining regions spanning both heavy and light chains. Our results highlight the advantage of combined epitope and paratope mapping HDX-MS experiments and supporting biochemical experiments to characterize antigen-antibody interactions. Through this combined approach, we provide a rationale for how the binding stoichiometry of the two mAbs to the trimeric NadA antigen can explain the difference in bactericidal activity of the two mAbs.

Molecular typing of group B Neisseria meningitidis'subcapsular antigens directly on biological samples demonstrates epidemiological congruence between culture-positive and -negative cases: A surveillance study of invasive disease over a 13-year period.

BACKGROUND: Surveillance of serogroup B Neisseria meningitidis (MenB) subcapsular antigen variant distribution in invasive disease (IMD) is fundamental for multicomponent vaccine coverage prediction. IMD incidence in Tuscany in 2018 was 0.37/100,000 inhabitants, with MenB representing 57% of cases. More than 50% of MenB responsible for IMD cannot be grown in culture, and molecular characterization of these cases is often lacking. The aim of the present study was to describe the distribution of MenB subcapsular antigens, comparing their distribution in culture-positive and culture-negative cases. METHODS: Molecular data regarding clonal complexes and subcapsular antigen variants of the 55 MenB-IMD occurring in Tuscany from 2007 to 2019 were made available, and their distribution between culture-positive and culture-negative cases was compared. Genetic-MATS and MenDeVAR prediction systems were used to assess multicomponent vaccine coverage predictions. RESULTS: Culture-positive and culture-negative cases presented a similar percentage representation of fHbp subfamilies. Clonal complex 162 was almost constantly associated with fHbp B231/v1.390, Neisserial-heparin-binding-antigen (NHBA) peptide 20, and PorinA P1.22,14 (BAST-3033): these were the most represented antigenic variants, both in culture-positive and culture-negative groups. Point-estimate 4CMenB coverage prediction was 88.5% (84.6%-92.3%). CONCLUSIONS: Our data demonstrate that non-cultivable meningococci, responsible for IMD, possess genetic variants of subcapsular antigens that are representative of what has been observed in culture. The vaccine-related antigenic epidemiology of MenB is thus similar in both groups. One of the first on-field applications of gMATS and MenDeVAR identifies their major advantage in their accessibility and in the possibility of dynamic data implementation that must be pursued continuously in the future.

Auricular acupuncture for the treatment of nonepileptic seizures: A pilot study.

OBJECTIVE: The aim of this pilot study was to examine the safety and tolerability of an eight-week treatment of twice weekly auricular acupuncture as a therapeutic intervention for the treatment of psychogenic nonepileptic seizures (PNES). The secondary aim was to determine if auricular acupuncture was effective at reducing PNES event frequency. Psychogenic nonepileptic seizures are as common as multiple sclerosis and are highly resistant to intervention. Unfortunately, there is no standard of care therapy, although mental health treatment referrals are commonly made. Acupuncture is felt to be a safe and cost-effective treatment for many diseases and symptoms including mental health disorders such as posttraumatic stress disorder (PTSD), anxiety, and conversion disorders. We propose to examine auricular acupuncture as a treatment for PNES. METHODS: A single-center, 16-week trial of auricular acupuncture was designed: a four-week baseline period, followed by an eight-week active treatment period during which subjects received twice weekly auricular acupuncture, then followed by a four-week washout period. Tolerability was defined as completion of 12 of 16 treatments. Successful compliance with seizure diary was defined as >80%, or 100 of 120 days completed. Safety profile was established and seizure count per week was also calculated. RESULTS: A total of 29 subjects enrolled from a consecutive sample of 73 PNES diagnoses discharging from our epilepsy unit. Twenty-two subjects entered the study with 91% completing more than 12 of 16 treatments, 91% meeting the endpoint of completing >80% of diary entries. Adverse events were mild, and only two subjects were withdrawn due to increased PNES frequency. Although short-term in design, using one-way repeated measures analysis of variance (ANOVA), we observed a statistically significant reduction in seizure count from baseline at treatment month one, treatment month two, and at the washout period (p = 0.0083). Eleven of 12 subjects (52.3%) who received at least one treatment recorded a 50% frequency reduction. Three of 21 subjects (14%) were PNES-free at the end of the study. SIGNIFICANCE: Auricular acupuncture is well-tolerated in a population with PNES and exhibits similar event frequency reductions as cognitive behavioral therapy (CBT). We believe that this is the first literature report examining auricular acupuncture for the treatment of PNES.

Flow Chemistry System for Carbohydrate Analysis by Rapid Labeling of Saccharides after Glycan Hydrolysis.

This study demonstrates the utilization of a flow chemistry system for continuous glycan hydrolysis and saccharide labeling to assist with the existing methods in glycan structural analysis. Acidic hydrolysis of glycans could be accelerated in a flow system. Aldoses and α-ketoacid-type saccharides were effectively labeled with naphthalene-2,3-diamine (NADA) at 60 °C for 10 min to form the fluorescent naphthimidazole (NAIM) and quinoxalinone (QXO) derivatives, respectively. The NADA-labeled derivatives improved the structural determination and composition analysis for their parent saccharides by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), liquid chromatography mass spectrometry (LC-MS), and nuclear magnetic resonance (NMR). Furthermore, this protocol was applied to determine the SA-Gal-Glc sequence of GM3-sugar out of six possible permutations.

Endocannabinoids and endocannabinoid-like compounds modulate hypoxia-induced permeability in CaCo-2 cells via CB1, TRPV1, and PPARα.

BACKGROUND AND PURPOSE: We have previously reported that endocannabinoids modulate permeability in Caco-2 cells under inflammatory conditions and hypothesised in the present study that endocannabinoids could also modulate permeability in ischemia/reperfusion. EXPERIMENTAL APPROACH: Caco-2 cells were grown on cell culture inserts to confluence. Trans-epithelial electrical resistance (TEER) was used to measure permeability. To generate hypoxia (0% O2), a GasPak™ EZ anaerobe pouch system was used. Endocannabinoids were applied to the apical or basolateral membrane in the presence or absence of receptor antagonists. KEY RESULTS: Complete hypoxia decreased TEER (increased permeability) by ~35% after 4 h (recoverable) and ~50% after 6 h (non-recoverable). When applied either pre- or post-hypoxia, apical application of N-arachidonoyl-dopamine (NADA, via TRPV1), oleamide (OA, via TRPV1) and oleoylethanolamine (OEA, via TRPV1) inhibited the increase in permeability. Apical administration of anandamide (AEA) and 2-arachidonoylglycerol (2-AG) worsened the permeability effect of hypoxia (both via CB1). Basolateral application of NADA (via TRPV1), OA (via CB1 and TRPV1), noladin ether (NE, via PPARα), and palmitoylethanolamine (PEA, via PPARα) restored permeability after 4 h hypoxia, whereas OEA increased permeability (via PPARα). After 6 h hypoxia, where permeability does not recover, only basolateral application PEA sustainably decreased permeability, and NE decreased permeability. CONCLUSIONS AND IMPLICATIONS: A variety of endocannabinoids and endocannabinoid-like compounds modulate Caco-2 permeability in hypoxia/reoxygenation, which involves multiple targets, depending on whether the compounds are applied to the basolateral or apical membrane. CB1 antagonism and TRPV1 or PPARα agonism may represent novel therapeutic targets against several intestinal disorders associated with increased permeability.

Nicotinamide riboside, an NAD+ precursor, attenuates the development of liver fibrosis in a diet-induced mouse model of liver fibrosis.

OBJECTIVE: Liver fibrosis is part of the non-alcoholic fatty liver disease (NAFLD) spectrum, which currently has no approved pharmacological treatment. In this study, we investigated whether supplementation of nicotinamide riboside (NR), a nicotinamide adenine dinucleotide (NAD+) precursor, can reduce the development of liver fibrosis in a diet-induced mouse model of liver fibrosis. METHODS: Male C57BL/6 J mice were fed a low-fat control (LF), a high-fat/high-sucrose/high-cholesterol control (HF) or a HF diet supplemented with NR at 400 mg/kg/day (HF-NR) for 20 weeks. Features of liver fibrosis were assessed by histological and biochemical analyses. Whole-body energy metabolism was also assessed using indirect calorimetry. Primary mouse and human hepatic stellate cells were used to determine the anti-fibrogenic effects of NR in vitro. RESULTS: NR supplementation significantly reduced body weight of mice only 7 weeks after mice were on the supplementation, but did not attenuate serum alanine aminotransferase levels, liver steatosis, or liver inflammation. However, NR markedly reduced collagen accumulation in the liver. RNA-Seq analysis suggested that the expression of genes involved in NAD+ metabolism is altered in activated hepatic stellate cells (HSCs) compared to quiescent HSCs. NR inhibited the activation of HSCs in primary mouse and human HSCs. Indirect calorimetry showed that NR increased energy expenditure, likely by upregulation of ß-oxidation in skeletal muscle and brown adipose tissue. CONCLUSION: NR attenuated HSC activation, leading to reduced liver fibrosis in a diet-induced mouse model of liver fibrosis. The data suggest that NR may be developed as a potential preventative for human liver fibrosis.

Potentiation of Phase Variation in Multiple Outer-Membrane Proteins During Spread of the Hyperinvasive Neisseria meningitidis Serogroup W ST-11 Lineage.

BACKGROUND: Since 2009, increases in the incidence of invasive meningococcal disease have occurred in the United Kingdom due to a sublineage of the Neisseria meningitidis serogroup W ST-11 clonal complex (hereafter, the "original UK strain"). In 2013, a descendent substrain (hereafter, the "2013 strain") became the dominant disease-causing variant. Multiple outer-membrane proteins of meningococci are subject to phase-variable switches in expression due to hypermutable simple-sequence repeats. We investigated whether alterations in phase-variable genes may have influenced the relative prevalence of the original UK and 2013 substrains, using multiple disease and carriage isolates. METHODS: Repeat numbers were determined by either bioinformatics analysis of whole-genome sequencing data or polymerase chain reaction amplification and sizing of fragments from genomic DNA extracts. Immunoblotting and sequence-translation analysis was performed to identify expression states. RESULTS: Significant increases in repeat numbers were detected between the original UK and 2013 strains in genes encoding PorA, NadA, and 2 Opa variants. Invasive and carriage isolates exhibited similar repeat numbers, but the absence of pilC gene expression was frequently associated with disease. CONCLUSIONS: Elevated repeat numbers in outer-membrane protein genes of the 2013 strain are indicative of higher phase-variation rates, suggesting that rapid expansion of this strain was due to a heightened ability to evade host immune responses during transmission and asymptomatic carriage.

Ear Acupuncture according to the NADA (National Acupuncture Detoxification Association).

This editorial is a brief report on the National Acupuncture Detoxification Association (NADA) ear acupuncture and is intended to briefly summarize the main scientific work. The complementary addiction-detoxification auricular acupuncture method has not been sufficiently experimentally explored in many areas. There have been clinical studies, some of which contradict the success. A total of 27 referenced publications were found that refer to the method that has existed for many decades and should be briefly listed here.

Carriage meningococcal isolates with capsule null locus dominate among high school students in a non-endemic period, Italy, 2012-2013.

Meningococcal disease incidence in Italy remains quite low in the overall population except for infants. Within a study on carriage isolates among high school students we aimed to define: i) the prevalence of carriage isolates, ii) the phenotypic and iii) the molecular features of meningococci by Whole Genome Sequencing (WGS). A total of 1697 pharyngeal samples from undergraduate students (age range 14-19 years) were collected from 2012 to 2013 from six larger cities in Italy. One hundred and twenty culture positive meningococci (7%) were analyzed. Carriage isolates were sent to the National Reference Laboratory for invasive meningococcal disease (IMD) for PCR-based serogroup identification, Multilocus Sequence Typing, PorA and FetA typing. Moreover, factor H binding protein (fHbp), Neisseria Heparin Binding Antigen (NHBA) and Neisserial adhesin A (NadA) were typed. Core genome MLST (cgMLST) was performed on a subsample of 75 carriage isolates. Capsule null locus (cnl) predominated (47%), followed by serogroup B (27%). The antimicrobial susceptibility profile revealed an high prevalence of reduced susceptibility to penicillin G (54%) and a full susceptibility to ceftriaxone, ciprofloxacin and rifampicin. Carriage isolates presented a high genetic diversity: the clonal complexes (ccs) cc1136, cc198 and cc41/44, were the predominant. An high heterogeneity was also observed for PorA and FetA types. The fhbp and nhba genes were identified in all the carriage isolates; only 5% of the carriage isolates presented the nadA gene. The core genome MLST analysis revealed that the majority of the cnl isolates clustered in a distinct group. The evidence gathered during this study provides the estimate of carriage isolates in high school students in a non-epidemic period in Italy that was lower than expected. Moreover, the highest proportion of carriage isolates were cnl and, overall, they were molecular heterogeneous.