Clinical characteristics and treatment of IMP-type carbapenemase-producing Enterobacteriaceae bacteremia: Case series and literature review.

BACKGROUND: Several carbapenemases have been identified globally in Enterobacteriaceae. In Japan, IMP-type carbapenemase is the most prevalent, although cases of carbapenemase-producing Enterobacteriaceae (CPE) bacteremia are still scarce. The present case series and literature review aimed to elucidate the clinical characteristics and treatment strategies for IMP-type CPE bacteremia. METHODS: Clinical data on pediatric cases of IMP-type CPE bacteremia at the Tokyo Metropolitan Children's Medical Center between 2010 and 2020 were collected, and a review of past studies of IMP-type CPE bacteremia has been provided. RESULTS: Five pediatric episodes of IMP-type CPE bacteremia were identified. Our review of previous literature on IMP-type CPE bacteremia revealed 24 adult patients, but no pediatric patients. All 29 cases had underlying diseases, and 23 (79%) received combination therapy. The median duration of antibiotic therapy was 14 days (interquartile range: 9-14 days). The overall mortality rate was 38% (11/29). The mortality rates associated with monotherapy and combination therapy were 67% (4/6) and 30% (7/23), respectively. CONCLUSIONS: We report the first case series of IMP-type CPE bacteremia in children. Our review of past studies suggests that combination therapy might lead to better survival outcomes in patients with IMP-type CPE bacteremia. Further research is needed to establish an optimal treatment strategy for IMP-type CPE bacteremia.

Effects of temporary rearing with organic selenium on the muscle flavor and texture properties of largemouth bass (Micropterus salmonides).

In this study, the influences of organic selenium (Se, 0.002 mg/L) on the muscle flavor and texture properties of Micropterus salmonides under fasting temporary rearing (8 weeks) was investigated. Electronic nose and headspace solid-phase microextraction-gas chromatography-mass spectrometry analysis suggested that organic Se regulated the types and contents of volatile compounds, especially aldehydes and ketones, which were increased in the early temporary rearing but decreased in the late stage. Organic Se significantly increased the content of 5'-inosine monophosphate by approximately 15 % (p < 0.05), and decreased the content of hypoxanthine and hypoxanthine ribonucleoside by more than 20 % (p < 0.05). After the 8th temporary rearing week, muscle hardness and springiness increased by at least 10 % (p < 0.01), resilience and gumminess improved by at least 18 % (p < 0.05) and 5.9 % (p < 0.05), respectively. In conclusion, organic Se ameliorates the flesh quality of M. salmonides during long-term temporary rearing.

Psychometric Properties of the Spanish-language Version of the Interpersonal Mindfulness in Parenting (IM-P) Scale among Mothers of Preschool Children in Chile / Propiedades psicométricas de la versión en español de la Escala de Atención Plena (Mindfulness) Interpersonal en la Parentalidad en madres de preescolares en Chile

Abstract The Interpersonal Mindfulness in Parenting (IM-P) scale is one of the first measures that specifically assesses mindful parenting, a specific application of mindfulness, that has been defined as paying attention to your child and parenting in a particular way, intentionally, in the present moment, and non-judgmentally. Psychometric properties of a Spanish-language version of the IM-P scale were examined in a sample of 111 mothers of preschool-age children living in Santiago, Chile. The original IM-P model with five factors and 31 items showed indicators of goodness of fit within acceptable ranges, however two items presented extremely low factor loadings that suggest a lack of fit to the model. Also, there was a high correlation between two factors which were theoretical and conceptually very related: Compassion for the self and child and Non-judgmental Acceptance of the self and child. Therefore, it was considered appropriate to test a new four-factor model in which these two factors were merged into one, and items loading low in the previous model were eliminated. This new model showed a slightly better fit than the five-factor model. The resultant four-factor version and its subscales showed good internal consistencies. Construct validity of the IM-P scale was investigated by calculating correlations with general mindfulness (Five Facet Mindfulness Questionnaire, FFMQ). As expected, a significant positive correlation was found between the two measures (r=0.73, p<0.01), and among almost all subscales. In general, the results present sound psychometric properties of the Spanish translation of the IM-P in Chilean mothers of preschool children.

Resumen Las propiedades psicométricas de la versión en español de la escala de Atención Plena (Mindfulness) Interpersonal en la Parentalidad (IM-P) fueron examinadas en una muestra de 111 madres de niños y niñas de edad preescolar en Santiago de Chile. La estructura de cinco factores de la escala IM-P original no fue completamente respaldada mediante análisis factorial confirmatorio. Por lo tanto, se testeó una estructura de cuatro factores. Dos de los factores encontrados fueron coherentes con aquellos originalmente hipotetizados. Los otros consistieron en una re-agrupación de items de las subescalas Consciencia Emocional de si misma y del hijo(a), Aceptación sin juicio de si misma y del hijo(a) y Compasión hacia si misma y hacia su hijo(a), en dos factores: uno que da cuenta de compasión y no-juicio de la madre hacia si misma y, el otro que da cuenta de la compasión, no-juicio y consciencia emocional de la madre hacia su hijio. La versión resultante de cuatro factores y sus sub-escalas presentaron buena consistencia interna. Se analizó la validez de constructo mediante cálculo de correlaciones con mindfulness general (Cuestionario de cinco dimensiones de Mindfulness, FFMQ). Como se esperaba, se encontró una correlación positiva significativa entre las dos medidas (r=0.71, p<0.01), y entre la mayor parte de las subescalas. En general, los resultados dan cuenta de buenas propiedades psicométricas de la versión en español de la escala IM-P en madres de niños y niñas preescolares en Chile.

Idiopathic mesenteric phlebosclerosis: one case report and systematic literature review of 240 cases.

AIM: This research is both a case study and a systematic literature review. Our goal was to learn more about the pathophysiology, clinical features, and therapy options for idiopathic mesenteric phlebosclerosis (IMP). METHODS: A case of IMP was treated by surgery in our department. Combined with the data of 240 cases of IMP retrieved from PubMed and Wanfang Data, we conducted a systematic review of this less well-known disease. RESULTS: These 240 cases of IMP mainly occurred in East Asia. Among the patients, 78.7% of them had a history of herbal medicine consumption; 15.2% had no obvious symptoms and their diagnosis was confirmed through characteristic CT findings and colonoscopic manifestations; 58.4% were cured by conservative treatment; 41.6% had severe symptoms and underwent surgical treatment. Additionally, more patients treated with surgical treatment had lesions involving the cecum (59/82 vs. 53/115, P=0.0003) and sigmoid colon (20/82 vs. 10/115, P=0.0025) compared with those treated conservatively. CONCLUSION: The occurrence of IMP is associated with the long-term consumption of herbal medicines. Early diagnosis of IMP could be determined by CT and colonoscopy and conservative or surgical treatment was chosen based on of the severity of the condition (e.g., the involved bowel segment).

Idiopathic mesenteric phlebosclerosis: clinical and CT imaging characteristics.

BACKGROUND: The aim of our study was to evaluate the clinical characteristics and computed tomography (CT) imaging findings of idiopathic mesenteric phlebosclerosis (IMP). METHODS: From January 2013 to May 2019, the clinical data of 10 patients diagnosed with IMP were analyzed retrospectively. Computed tomography angiography (CTA) and colonoscopy were performed in all 10 patients. All CT imaging findings were evaluated by three radiologists, including the form and distribution of calcification, the bowel's thickness, and the surrounding fat gap. The calcification score was calculated according to the extent of the involved mesenteric veins. The colonic wall thickness was defined as the average value of the thickest and thinnest regions of the intestinal wall. The correlation between the calcification scores and the colonic wall thickness was analyzed using Spearman's correlation analysis. RESULTS: All 10 patients were male with an average age of 59.6 years (range, 51-83 years). The average smoking index was 712 (range, 0-1,800). Among them, 7 patients had a history of long-term excessive daily intake of medicinal liquor or Chinese herbal medicine. Clinical symptoms of abdominal pain, diarrhea, bloating, and nausea were found. Colonoscopy showed dark purple discolorations of the edematous mucosa, engorged blood vessels, extensive erosion, ulceration, and multi-focal nodular surface in all patients. CT demonstrated colonic wall thickening, calcification along the mesenteric vein, and blurry surrounding fat gap in all 10 patients. Mesenteric venous calcification involved the terminal ileum, the ascending and transverse colon in all patients, and the descending colon and sigmoid colon's involvement in two patients. A total of 33 segments of the intestinal wall were involved. The median calcification score was 6 points, the mean thickness of the colonic wall was 10.73±3.22 mm, and there was no significant correlation (P=0.782) between calcification score and thickness of the colonic wall. CONCLUSIONS: The main features of IMP are mesenteric venous calcification, colonic wall thickness, and pericolic fat stranding, and there is no correlation between calcification score and colonic wall thickness. Therefore, CT imaging combined with colonoscopy can improve the diagnostic accuracy of IMP.

Effects of Yuanhu- Zhitong tablets on alcohol-induced conditioned place preference in mice.

OBJECTIVES: This study aimed at determining the synergistic effects of Yuanhu Zhitong tablets (YHZTP) on alcohol-induced conditioned place preference (CPP) in mice, in addition, the intervention mechanism was preliminarily explored based on traditional Chinese Medicine (TCM) network pharmacology on alcohol addiction. METHODS: Alcohol-induced CPP mice were used to evaluate the effects of either YHZTP or levo-tetrahydropalmatine (l-THP) plus imperatorin (IMP) administration on animal behavior. The network pharmacological strategy was used to establish the "compound-target" and "disease-drug-target" network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed on the shared targets between the compound and the disease. Twelve algorithms on CytoHubba were used to find the hub genes that were verified by qPCR. RESULTS: Systemic administration (2 g/kg, i.p.) of ethanol (EtOH) to mice was used to induce CPP. YHZTP On its own did not induce CPP or conditioned place aversion (CPA) at the doses of 0.3 g/kg or 0.6 g/kg (i.g.), but attenuated the acquisition and expression of EtOH-induce CPP in mice. In addition, YHZTP (0.3 or 0.6 g/kg) did not exhibit any effect on the motor activity of mice. Acquisition of alcohol-induced CPP was blocked by a combination of l-THP (5 mg/kg, i.g.) + IMP (2.5 mg/kg, i.g.) or l-THP (10 mg/kg, i.g.) + IMP (5 mg/kg, i.g.). However, the combination of l-THP (2.5 mg/kg, i.g.) + IMP (1.25 mg/kg, i.g.) or mono-administration of l-THP and IMP did not exhibit any effect on alcohol-induced CPP. YHZTP was also shown to reverse the up-regulation of Gabra1, Ptgs2, Mapk1, Mapk8, Mapk14, Nr3c, Prkca and Sirt1 genes and the down-regulation of Hhtr2a and Drd2 genes in the prefrontal cortex of EtOH induced CPP mice. These genes were associated with neuroactive ligand-receptor interactions, activation of the sphingolipid, calcium, cAMP, ErbB, NF-kappa B and MAPK signaling pathways. CONCLUSION: YHZTP inhibits EtOH-induced CPP behavior in mice while a combination of l-THP and IMP exerts a synergistic effect on the reduction of EtOH-induced CPP. Possible pharmacological mechanisms include inhibition of the expression of inflammatory factors and regulation of neurotransmitter receptor levels. Therefore, YHZTP is a novel candidate for the treatment of alcohol addiction.

Activity of aztreonam in combination with ceftazidime-avibactam against serine- and metallo-ß-lactamase-producing Pseudomonas aeruginosa.

Existing data support the combination of aztreonam and ceftazidime-avibactam against serine-ß-lactamase (SBL)- and metallo-ß-lactamase (MBL)-producing Enterobacterales, although there is a paucity of data against SBL- and MBL-producing Pseudomonas aeruginosa. In this study, 5 SBL- and MBL-producing P. aeruginosa (1 IMP, 4 VIM) were evaluated against aztreonam and ceftazidime-avibactam alone and in combination via broth microdilution and time-kill analyses. All 5 isolates were nonsusceptible to aztreonam, aztreonam-avibactam, and ceftazidime-avibactam. Combining aztreonam with ceftazidime-avibactam at subinhibitory concentrations produced synergy and restored bactericidal activity in 4/5 (80%) isolates tested. These results suggest that the combination of aztreonam and ceftazidime-avibactam may be a viable treatment option against SBL- and MBL-producing P. aeruginosa.

The CRISPR-Cas9 crATIC HeLa transcriptome: Characterization of a novel cellular model of ATIC deficiency and ZMP accumulation.

In de novo purine biosynthesis (DNPS), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase (EC 2.1.2.3)/inosine monophosphate cyclohydrolase (EC 3.5.4.10) (ATIC) catalyzes the last two reactions of the pathway: conversion of 5-aminoimidazole-4-carboxamide ribonucleotide [aka Z-nucleotide monophosphate (ZMP)] to 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR) then to inosine monophosphate (IMP). Mutations in ATIC cause an untreatable and devastating inborn error of metabolism in humans. ZMP is an adenosine monophosphate (AMP) mimetic and a known activator of AMP-activated protein kinase (AMPK). Recently, a HeLa cell line null mutant for ATIC was constructed via CRISPR-Cas9 mutagenesis. This mutant, crATIC, accumulates ZMP during purine starvation. Given that the mutant can accumulate ZMP in the absence of treatment with exogenous compounds, crATIC is likely an important cellular model of DNPS inactivation and ZMP accumulation. In the current study, we characterize the crATIC transcriptome versus the HeLa transcriptome in purine-supplemented and purine-depleted growth conditions. We report and discuss transcriptome changes with particular relevance to Alzheimer's disease and in genes relevant to lipid and fatty acid synthesis, neurodevelopment, embryogenesis, cell cycle maintenance and progression, extracellular matrix, immune function, TGFß and other cellular processes.

Effects of family integrated care on refractory cancer-related pain receiving intrathecal morphine pump therapy: a cohort study.

BACKGROUND: Family integrated care (FIC), originating from the neonatal intensive care unit, has proved to be an effective therapy for the care of several diseases in adults. However, it is still unclear whether it is successful in the treatment of refractory cancer-related pain using an intrathecal morphine pump (IMP). METHODS: A prospective study was performed to enroll patients with refractory cancer-related pain treated by IMP. Medical records, including demographic data, types of cancer, doses of opioids before the implantation, were gathered at baseline. Besides, numerical pain rating scales (NRS) were measured to assess the severity of pain. Karnofsky performance scores (KPS), self-rating anxiety scale (SAS) scores and selfrating depression scale (SDS) scores were used to assess the physical and psychological status of patients. Follow-up lasted for at least 12 months. The changes in NRS, KPS, SAS, and SDS scores were recorded, as well as adverse events and survival rates. RESULTS: Totally, 33 patients were enrolled and divided into traditional care groups and FIC groups. The results showed that the implantation of IMP relieved the pain in both groups. Moreover, patients in the FIC group had significantly more relief of pain than those in the traditional group (P=0.04). Similarly, KPS raised more, and scores of SAS and SDS decreased more in the FIC group. Besides, fewer adverse events occurred in the FIC group, but the survival rate showed no significant difference between the two groups. CONCLUSIONS: FIC enhances the effects of IMP on the relief of refractory cancer-related pain and improves the physical and psychological status of patients.

Furbellow (Brown Algae) Extract Increases Lifespan in Drosophila by Interfering with TOR-Signaling.

Algal products are well known for their health promoting effects. Nonetheless, an in depth understanding of the underlying molecular mechanisms is still only fragmentary. Here, we show that aqueous furbelow extracts (brown algae, Saccorhiza polyschides) lengthen the life of both sexes of the fruit fly Drosophila melanogaster substantially, if used as nutritional additives to conventional food. This life prolonging effect became even more pronounced in the presence of stressors, such as high-fat dieting of living under drought conditions. Application of the extracts did not change food intake, excretion, or other major physiological parameters. Nevertheless, effects on the intestinal microbiota were observed, leading to an increased species richness, which is usually associated with healthy conditions. Lifespan extension was not observed in target of rapamycin (TOR)-deficient animals, implying that functional TOR signaling is necessary to unfold the positive effects of brown algae extract (BAE) on this important trait. The lack of life lengthening in animals with deregulated TOR signaling exclusively targeted to body fat showed that this major energy storage organ is instrumental for transmitting these effects. In addition, expression of Imaginal morphogenesis protein-Late 2 (Imp-L2), an effective inhibitor of insulin signaling implies that BAE exerts their positive effects through interaction with the tightly interwoven TOR- and insulin-signaling systems, although insulin levels were not directly affected by this intervention.

Efficacy of purified nucleotide supplements on the growth performance and immunity of hybrid striped bass Morone chrysops x Morone saxatilis.

Fishmeal is being increasingly replaced in aquatic animal diets with alternative plant protein feedstuffs such as soybean meal which have lower concentrations of nucleotides; therefore, supplemental sources of exogenous nucleotides in diets could become increasingly important. A 9-week feeding trial was conducted with triplicate groups of juvenile hybrid striped bass (average initial body weight ± standard deviation, 5.6 ± 0.1 g) to determine the effects of supplementing single purified nucleotides on the growth performance and immune parameters. The basal diet, which utilized menhaden fishmeal (25%) and soybean meal (75%) as protein sources, contained 44% protein, 10% lipid and an estimated digestible energy level of 3.5 kcal g-1. Single additions of 5'- adenosine monophosphate (AMP), 5'- uridine monophosphate (UMP), 5'- cytidine monophosphate (CMP), 5'- guanosine monophosphate (GMP), and 5'- inosine monophosphate (IMP) disodium salts (Chem-Impex International, Wood Dale, Illinois, USA) were evaluated with each nucleotide added to the basal diet at 0.5% of dry weight at the expense of cellulose. A positive control diet in this trial was a diet containing 5'- AMP from Sigma-Aldrich also supplemented at 0.5% by weight. Results showed significantly (P < 0.05) improved weight gain between fish fed AMP-supplemented diets and the basal diet. No statistical significance (P > 0.05) was detected in whole-body proximate composition and protein retention of fish fed any of the dietary treatments. The respiratory burst of whole blood phagocytes also was significantly (P < 0.05) higher in fish fed the AMP Sigma diet compared to the other dietary treatments. Dietary IMP and AMP both significantly (P < 0.05) enhanced the capacity of isolated phagocytes to generate extracellular superoxide anion compared to all other dietary treatments. No significant differences were seen in other innate immune parameters such as plasma lysozyme, total plasma protein, and total immunoglobulin. The ability of isolated B lymphocytes to proliferate prompted by the presence of lipopolysaccharides was significantly (P < 0.05) different among dietary treatments with the highest simulation index observed in fish fed the diets containing AMP Sigma and UMP; however, it was not significantly different from that of fish fed the basal diet. Based on all the measured responses, it is concluded that AMP at 0.5% of diet had the most positive influence on growth performance and innate immunostimulation of hybrid striped bass.

The Interpersonal Mindfulness in Parenting Scale in Mothers of Children and Infants: Factor Structure and Associations With Child Internalizing Problems.

Objectives: Mindful parenting, measured by the Interpersonal Mindfulness in Parenting scale (IMP), is beneficial for parents and children. However, the IMP has not been validated in English-speaking parents. Further, little is known about whether mindful parenting is similar in parents of children vs. infants, or how it reduces child internalizing problems. We sought to validate the IMP in English-speaking mothers of children and infants, and to examine relationships between the facets of mindful parenting, child internalizing problems and parent variables related to internalizing. Methods: Using confirmatory factor analyses, we examined the fit of various models of mindful parenting in English-speaking community-recruited mothers of children aged 3-18 years (n = 396) and infants aged 0-2 years (n = 320). We used regression analyses to investigate relationships between the facets of mindful parenting, child internalizing problems, and parent variables including parental experiential avoidance, unhelpful beliefs about child anxiety and accommodation of child anxiety. Results: Mindful parenting can be measured in English-speaking mothers, using either a 5- or 6-factor, 29-item version of the IMP. These versions of the IMP operate similarly for mothers of children and infants. Child internalizing problems and related parent variables were best predicted by non-judgmental acceptance of parenting in mothers of children, and emotional self-awareness and non-reactivity in mothers of infants. Conclusions: The IMP is a valid measure of mindful parenting in English-speaking mothers of children and infants. Mindful parenting predicts child internalizing problems and related parent variables, suggesting that mindful parenting programs could benefit families of children with internalizing problems, potentially by reducing parental experiential avoidance, unhelpful beliefs about or accommodation of child anxiety.

Measurement of cerebral vascular reserves with I-123 IMP SPECT without an arterial input function using the microsphere model and radiopharmaceutical dose calibration.

OBJECTIVES: Cerebral vascular reserve (CVR) is an important indicator for the management of and therapy for cerebral arterial occlusive disease (CAOD). Vasodilatory function is measured using the standard IMP-ARG method. The IMP autoradiography (IMP-ARG) method employed here uses a standardized input function, which was derived from 12 patients between 31 and 71 years of age. Because the population of elderly patients continues to increase in Japan, additional therapies are required to assess CVR in elderly patients with chronic cardiopulmonary disease or a history of smoking, in particular. Despite its popularity, alternatives to the IMP-ARG method are necessary. Here, we proposed the microsphere (MS) method without an input function. METHOD: Using this method and the IMP-ARG method, we measured the CVRs of 18 CAOD patients. RESULTS: The CVRs derived with these two methods were significantly and linearly correlated (r = 0.89, p < 0.01). CVRs categorized by severity were also found to correspond between the two methods (κ = 0.87). CONCLUSIONS: Thus, the method proposed here may serve as a supplemental to and be compatible with the IMP-ARG method for the assessment of CVR. Furthermore, the two methods, when used in conjunction, may result in less error than either would alone.

Comparison of five methods for detection of carbapenemases in Enterobacterales with proposal of a new algorithm.

OBJECTIVES: The aim of this study was to evaluate the performance of five different carbapenemase tests and to develop an algorithm which will permit the detection of most common and rare carbapenemases in routine microbiology laboratories. METHODS: The immunochromatographic tests CARBA-5 (NG), RESIST-4 O.K.N.V. (Coris), the colorimetric ß-CARBA (BioRad), a newly developed carbapenem-inactivation method (CIM) supplemented with zinc (zCIM), and the Xpert Carba-R (Cepheid) were challenged with a collection of 189 molecularly characterized Enterobacterales isolates, including 146 carbapenemase producers (CPE): VIM (n = 48), OXA-48-like (n = 40), NDM (n = 29), KPC (n = 13), IMI (n = 9), IMP (n = 9), OXA-58 (n = 2), and GES (n = 2). RESULTS: The overall sensitivity/specificity values for the five carbapenemase detection tests were 84.2% (CI 77.6-89.2%)/100% (CI 91.8-100%) for RESIST-4, 88.2% (CI 82.1-92.4%)/100% (CI 91.8-100%) for CARBA-5, 88.2% (CI 82.1-92.4%)/100% (CI 91.8-100%) for Xpert Carba-R, 73.7% (CI 66.2-80.0%)/100% (CI 93.4-99.0%) for ß-CARBA, and 97.4% (CI 87.9-99.6%)/97.7% (CI 87.9-99.6%) for zCIM. The four common carbapenemases (KPC, OXA-48-like, NDM, and VIM) were detected with ≥97.6% sensitivity by all tests except for ß-CARBA (76.6% (CI 68.4-83.2%)). IMI and GES were only detected by zCIM (sensitivity 90.9% (CI 62.3-98.4%)). Based on these results a new algorithm was developed, consisting of an immunochromatographic assay as the first test followed by zCIM, which allows detection of 99.3% of all carbapenemases assessed. CONCLUSIONS: Except for ß-CARBA, all methods showed excellent sensitivity/specificity for the detection of the four most frequent carbapenemases. With the new algorithm, rare variants can also be detected. It is rapid, simple, and inexpensive and can be performed in any microbiology laboratory, as no PCR equipment is required.

Aztreonam plus Clavulanate, Tazobactam, or Avibactam for Treatment of Infections Caused by Metallo-ß-Lactamase-Producing Gram-Negative Bacteria.

Metallo-ß-lactamase (MBL)-producing Gram-negative bacteria are often extremely resistant, leading to a real therapeutic dead end. Here, we evaluated the in vitro and in vivo efficacy of aztreonam in combination with ceftazidime-avibactam, ceftolozane-tazobactam, or amoxicillin-clavulanate for the treatment of infections caused by MBL-producing Enterobacteriaceae, MBL-producing Pseudomonas aeruginosa, and extremely drug-resistant Stenotrophomonas maltophilia First, we report two clinical cases, namely, a urinary tract infection caused by an NDM-5-producing Escherichia coli isolate and a pulmonary infection caused by a S. maltophilia isolate efficiently treated with the association of aztreonam-ceftazidime-avibactam and aztreonam-amoxicillin-clavulanate, respectively. Then, a total of 50 MBL-producing Enterobacteriaceae isolates, 3 MBL-producing P. aeruginosa isolates, and 5 extremely drug-resistant S. maltophilia isolates were used to test aztreonam susceptibility in combination with ceftolozane-tazobactam, ceftazidime-avibactam, or amoxicillin-clavulanate. The Etest strip superposition method was used to determine the MICs of the aztreonam/inhibitor combinations. According to CLSI breakpoints, aztreonam susceptibility was fully restored for 86%, 20%, and 50% of the MBL-producing Enterobacteriaceae isolates when combined with ceftazidime-avibactam, ceftolozane-tazobactam, and amoxicillin-clavulanate, respectively. In P. aeruginosa, the aztreonam-ceftazidime-avibactam combination was the most potent, even though the reduction in MICs was at most 2-fold. With the 5 S. maltophilia isolates, aztreonam-ceftazidime-avibactam and aztreonam-amoxicillin-clavulanate were found to be equal (100% susceptibility). Overall, aztreonam-ceftazidime-avibactam was the most potent combination to treat infections caused by MBL producers compared with aztreonam-amoxicillin-clavulanate and aztreonam-ceftolozane-tazobactam. However, in many cases aztreonam-amoxicillin-clavulanate was found to be as efficient as aztreonam-ceftazidime-avibactam, offering the main advantage to be markedly cheaper. We also confirmed the validity of Etest superpositions as a very simple method to determine MICs of aztreonam combinations.

Effects of inosine 5'-monophosphate supplementation in high fishmeal and high soybean diets on growth, immune-related gene expression in gibel carp (Carassius auratus gibelio var. CAS â ¢), and its challenge against Aeromonas hydrophila infection.

The present study was conducted to evaluate dietary inosine 5'-monophosphate (5'-IMP) on growth, immune genes expression and disease resistance against Aeromonas hydrophila in juvenile gibel carp (Carassius auratus gibelio var. CAS Ⅲ) (initial body weight: 7.48 g). Six diets were formulated containing exogenous 5'-IMP at three gradient levels (0, 0.1% and 0.2%) in the high dietary fishmeal group (15% fishmeal: D1, D2, D3) and in the high dietary soybean meal group (33% soybean meal: D4, D5, D6). Each diet was randomly allotted to triplicate tanks in a recirculating system. After the feeding trial, fish were exposed to Aeromonas hydrophila challenge. Hematological and immunological responses were analyzed before and after challenge. The results indicated that feeding rate in all 5'-IMP supplemented treatments (D2, D3, D5 and D6) and daily growth coefficient in D5 and D6 were reduced compared with those of respective control treatments (D1 and D4) without 5'-IMP addition (P < 0.05). The cumulative survival rates were numerically improved by dietary 5'-IMP supplementation (P > 0.05). Compared with the respective control treatment, in the high fishmeal group, plasma SOD and MPO were significantly elevated in D3 at the end of feeding trial (P < 0.05), plasma SOD and lysozyme were significantly increased in D3 after bacterial challenge (P < 0.05); in high soybean meal group, plasma lysozyme activity was significantly elevated in D5 post bacterial challenge (P < 0.05). Most of the expression of immune related genes (intelectin, major histocompatibility complex class II ß (MHC II ß), Complement 3 (C3), Complement component C7-1 (ccC7), lysozyme C, Interleukin 1ß (IL-1ß), Tumor necrosis factor α1 (TNF-α1), Transforming growth factor-beta (TGF-ß) and Interleukin 8 (IL-8)) in spleen, kidney and liver of the fish were significantly affected by supplementation of 5'-IMP at the end of feeding trial and post bacterial challenge. Additionally, adding 5'-IMP in high soybean meal diets exerted further effects of promoting immunity than counterparts in high fishmeal diets. Considering enhanced disease resistance, the immunopotentiation of 5'-IMP was manifested when the addition level was 0.1% in high soybean meal diets and 0.2% in high fishmeal diets.

Whole Proteome Profiling of N-Myristoyltransferase Activity and Inhibition Using Sortase A.

N-myristoylation is the covalent addition of a 14-carbon saturated fatty acid (myristate) to the N-terminal glycine of specific protein substrates by N-myristoyltransferase (NMT) and plays an important role in protein regulation by controlling localization, stability, and interactions. We developed a novel method for whole-proteome profiling of free N-terminal glycines through labeling with S. Aureus sortase A (SrtA) and used it for assessment of target engagement by an NMT inhibitor. Analysis of the SrtA-labeling pattern with an engineered biotinylated depsipeptide SrtA substrate (Biotin-ALPET-Haa, Haa = 2-hydroxyacetamide) enabled whole proteome identification and quantification of de novo generated N-terminal Gly proteins in response to NMT inhibition by nanoLC-MS/MS proteomics, and was confirmed for specific substrates across multiple cell lines by gel-based analyses and ELISA. To achieve optimal signal over background noise we introduce a novel and generally applicable improvement to the biotin/avidin affinity enrichment step by chemically dimethylating commercial NeutrAvidin resin and combining this with two-step LysC on-bead/trypsin off-bead digestion, effectively eliminating avidin-derived tryptic peptides and enhancing identification of enriched peptides. We also report SrtA substrate specificity in whole-cell lysates for the first time, confirming SrtA promiscuity beyond its recognized preference for N-terminal glycine, and its usefulness as a tool for unbiased labeling of N-terminal glycine-containing proteins. Our new methodology is complementary to metabolic tagging strategies, providing the first approach for whole proteome gain-of signal readout for NMT inhibition in complex samples which are not amenable to metabolic tagging.

Prevalence of metallo-ß-lactamase acquired genes among carbapenems susceptible and resistant Gram-negative clinical isolates using multiplex PCR, Khartoum hospitals, Khartoum Sudan.

BACKGROUND: The increased prevalence of carbapenem-resistant Gram-negative isolates caused by Metallo-ß-lactamase (MBL) is worrisome in clinical settings worldwide. The mortality rate associated with infections caused by MBLs producing organisms ranging from 18 to 67%. This study aimed to determine the prevalence of Metallo-ß-lactamase genes among some Gram-negative clinical isolates (Carbapenems susceptible and resistant). METHODS: This paper describes a descriptive cross-sectional study carried out to detect MBL genes such as (blaVIM, blaIMP and blaNDM) by multiplex PCR mixture reaction among 200 Gram-negative clinical isolates (Citrobacter spp, Escherichia coli, Enterobacter spp, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Proteus valgaris). Khartoum hospitals during 2015 to 2016. LIMITATION: The study organisms were not evaluated for non-MBL carbapenemases, such as KPC and OXA-48. RESULTS: The prevalence of MBL genes by multiplex PCR assays among 200 Gram-negative clinical isolates was 72(36.1%). MBL positive genes among 100 carbapenems sensitive and 100 resistant isolates were 27(27%) and 45(45%) respectively. There was a statistically, significant association between the antimicrobial susceptibility and the presences of MBL genes (P.value = 0.008). E.coli was the predominant species possessing MBL genes 26(36.1%), with 22(30.7%) species having a combination of MBL genes. Verona integron Metallo beta-lactamase (VIM) was the most frequent genes 28(38.9%) out of 72 MBL detected genes, followed by imipenemase (IMP) was 19(26.4%), and consequently, New Delhi Metallo beta lactamase was 3(4.2%). CONCLUSION: This study revealed a high prevalence of MBL genes in some Gram-negative isolates from Khartoum State Hospitals which were not previously established in these hospitals.

Hydrophilic interaction liquid chromatography coupled with quadrupole-orbitrap ultra high resolution mass spectrometry to quantitate nucleobases, nucleosides, and nucleotides during white tea withering process.

Nucleotides, nucleosides, and nucleobases are important bioactive compounds. Recent studies suggested that they possess taste activity. However, it remains unknown about their presence in white tea and how they change during white tea manufacture. Here, we first established method based on hydrophilic interaction liquid chromatography coupled with quadrupole-orbitrap ultra high resolution mass spectrometry (HILIC-Quadrupole-Orbitrap-UHRMS) platform, then applied it to study the dynamic changes of nucleotides, nucleosides, and nucleobases during white tea withering process. Five compounds, including adenosine 5'-monophosphate monohydrate (AMP), guanosine 5'-monophosphate disodium salt hydrate (GMP), adenosine, cytidine, thymine and uracil, were detected from withering samples. They showed a general decline trend during white tea withering process, however, considerable amount of them was retained after withering for 48 h except adenosine which was below detection limit after withering for 21 h. This study provided a complete picture about nucleotides, nucleosides and nucleobases changes during white tea withering process.

Structural analysis and taste evaluation of γ-glutamyl peptides comprising sulfur-containing amino acids.

The structures, flavor-modifying effects, and CaSR activities of γ-glutamyl peptides comprising sulfur-containing amino acids were investigated. The chemical structures, including the linkage mode of the N-terminal glutamic acid, of γ-L-glutamyl-S-(2-propenyl)-L-cysteine (γ-L-glutamyl-S-allyl-L-cysteine) and its sulfoxide isolated from garlic were established by comparing their NMR spectra with those of authentic peptides prepared using chemical methods. Mass spectrometric analysis also enabled determination of the linkage modes in the glutamyl dipeptides by their characteristic fragmentation. In sensory evaluation, these peptides exhibited flavor-modifying effects (continuity) in umami solutions less pronounced but similar to that of glutathione. Furthermore, the peptides exhibited intrinsic flavor due to the sulfur-containing structure, which may be partially responsible for their flavor-modifying effects. In CaSR assays, γ-L-glutamyl-S-methyl-L-cysteinylglycine was most active, which indicates that the presence of a medium-sized aliphatic substituent at the second amino acid residue in γ-glutamyl peptides enhances CaSR activity.