Evaluation of Expanded Criteria Donors Using the Kidney Donor Profile Index and the Preimplantation Renal Biopsy.

The increasing comorbidity of kidney transplant (KT) donors make it necessary to develop scores to correctly assess the quality of kidney grafts. This study analyzes the usefulness of the preimplantation biopsy and the Kidney Donor Profile Index (KDPI) as indicators of KT survival from expanded criteria donors (ECD). Retrospective study of KT in our center between January 2010 to June 2019 who received a kidney from an ECD and underwent a preimplantation biopsy. 266 KT were included. Graft survival was categorized by KDPI quartiles: Q1 = 86%, Q2 = 95%, Q3 = 99% and Q4 = 100%. KT from KDPI Q1 presented better survival (p = 0.003) and Q4 donors had worse renal function (p = 0.018) and poorer glomerular filtration rate (3rd month; p = 0.017, 1st year; p = 0.010). KT survival was analyzed according to KDPI quartile and preimplantation biopsy score simultaneously: Q1 donors with biopsy score ≤3 had the best survival, especially comparing against Q3 with a biopsy score >3 and Q4 donors (p = 0.014). In multivariable analysis, hyaline arteriopathy, glomerulosclerosis, and KDPI Q4 were predictors for graft survival. High KDPI and a greater histological injury in the preimplantation biopsy, especially glomerular and vascular lesions, were related to a higher rate of KT loss from ECD.

Stability of carboplatin infusion solutions used in desensitization protocol.

Carboplatin hypersensitivity reactions are one of the major clinical challenges in treating patients with relapse/recurrent ovarian malignancies. Desensitization protocols allow the continuation of treatment in patients who have presented hypersensitivity reactions by gradually re-introducing small amounts of the drug up to full therapeutic doses. Carboplatin desensitization protocol is based on three solutions that are usually prepared in the chemotherapy centralized units of hospital pharmacies. First and second solutions are diluted under the established concentration limit to guarantee the stability of the preparation. We developed a specific high-performance liquid chromatography assay to determine the stability of carboplatin infusion solutions that have been diluted to 0.2 mg/mL and 0.02 mg/mL in 250 mL of 5% dextrose in polypropylene infusion bags which were stored 24 h protected from light at room temperature. Samples were withdrawn at t = 0 h, 3 h, 6 h, and 24 h. The analytical column was a Zorbax eclipse XDB-C18 (150 mm × 4.6 mm; 5 µm particle size). The mobile phase had a flow rate of 1 mL/min under isocratic conditions of water-methanol (98:2, v/v). For 0.2 mg/mL solution, the high-performance liquid chromatography assay revealed no significant losses in carboplatin concentration. However, in 0.02 mg/mL solution remaining carboplatin was > 105% the initial dose after 3 h of storage at room temperature. The ultraviolet-visible spectra analysis showed that carboplatin remained intact during the study in 0.2 mg/mL solution, but some changes were detected in 0.02 mg/mL solution. Thus, 0.2 mg/mL carboplatin solution is stable for 24 h at room temperature in 5% dextrose polypropylene infusion bags but stability could not be proved for 0.02 mg/mL solution.

Diagnosis of right bundle branch block: a concordance study.

BACKGROUND: Right bundle branch block is one of the most common electrocardiographic abnormalities. Most cases of right bundle branch block are detected in asymptomatic patients in primary care, so a correct interpretation of electrocardiograms (ECGs) at this level is necessary. The objective of this research is to determine the degree of concordance in the diagnosis of incomplete and complete right bundle branch block between four primary care researchers and a cardiologist. METHODS: The research design is a retrospective cohort study of patients over 18 years of ages of patients over 18 years of ages who underwent an ECG for any reason and were diagnosed with right bundle branch block by their physician. The physicians participating, 4 primary care researchers and a cardiologist were specialized in interpreting electrocardiographic records. The diagnosis of incomplete and complete right bundle branch block was recorded and other secondary variables were analysed. In case of diagnostic discordance between the researchers, the ECGs were reviewed by an expert cardiologist, who interpreted them, established the diagnosis and analysed the possible causes for the discrepancy. RESULTS: We studied 160 patients diagnosed with right bundle branch block by their general practise. The patients had a mean age of 64.8 years and 54% of them were men. The concordance in the diagnosis of incomplete right bundle branch block showed a Fleiss' kappa index (k) of 0.71 among the five researchers and of 0.85 among only the primary care researchers. The k for complete right bundle branch block was 0.93 among the five researchers and 0.96 among only the primary care researchers. CONCLUSION: The interobserver agreement in the diagnosis of right bundle branch block performed by physicians specialized in ECG interpretation (primary care physicians and a cardiologist) was very good. The variability was greater for the diagnosis of incomplete right bundle branch block.

Anthracycline-based triplets do not improve the efficacy of platinum-fluoropyrimidine doublets in first-line treatment of advanced gastric cancer: real-world data from the AGAMEMON National Cancer Registry.

BACKGROUND: Although anthracycline-based triplets are one of the most widely used schedules to treat advanced gastric cancer (AGC), the benefit of including epirubicin in these therapeutic combinations remains unknown. This study aims to evaluate both the efficacy and tolerance of triplets with epirubicin vs. doublets with platinum-fluoropyrimidine in a national AGC registry. METHODS: Patients with AGC treated with polychemotherapy without trastuzumab at 28 hospitals in Spain between 2008 and 2016 were included. The effect of anthracycline-based triplets against doublets was evaluated by propensity score matching (PSM) and Cox proportional hazards (PH) regression. RESULT: A total of 1002 patients were included (doublets, n = 653; anthracycline-based triplets, n = 349). The multivariable Cox PH regression failed to detect significantly increased OS in favor of triplets with anthracyclines: HR 0.90 (95% CI, 0.78-1.05), p = 0.20035. After PSM, the sample contained 325 pairs with similar baseline characteristics. This method was also unable to reveal an increase in OS: 10.5 (95% CI, 9.7-12.3) vs. 9.9 (95% CI, 9.2-11.4) months, HR 0.91 (CI 95%, 0.76-1.083), and (log-rank test, p = 0.226). Response rates (42.1 vs. 33.1%, p = 0.12) and PFS (HR 0.95, CI 95%, 0.80-1.13, log-rank test, p = 0.873) were not significantly higher with epirubicin-based regimens. The triplets were associated with greater grade 3-4 hematological toxicity, and increased hospitalization due to toxicity by 68%. The addition of epirubicin is viable, but 23.7% discontinued treatment because of adverse effects or patient decision. CONCLUSION: Anthracyclines added to platinum-fluoropyrimidine doublets did not improve the response rate or survival outcomes in patients with AGC but entailed greater toxicity.

Nomogram-based prediction of survival in patients with advanced oesophagogastric adenocarcinoma receiving first-line chemotherapy: a multicenter prospective study in the era of trastuzumab.

BACKGROUND: To develop and validate a nomogram and web-based calculator to predict overall survival (OS) in Caucasian-advanced oesophagogastric adenocarcinoma (AOA) patients undergoing first-line combination chemotherapy. METHODS: Nine hundred twenty-four AOA patients treated at 28 Spanish teaching hospitals from January 2008 to September 2014 were used as derivation cohort. The result of an adjusted-Cox proportional hazards regression was represented as a nomogram and web-based calculator. The model was validated in 502 prospectively recruited patients treated between October 2014 and December 2016. Harrell's c-index was used to evaluate discrimination. RESULTS: The nomogram includes seven predictors associated with OS: HER2-positive tumours treated with trastuzumab, Eastern Cooperative Oncology Group performance status, number of metastatic sites, bone metastases, ascites, histological grade, and neutrophil-to-lymphocyte ratio. Median OS was 5.8 (95% confidence interval (CI), 4.5-6.6), 9.4 (95% CI, 8.5-10.6), and 14 months (95% CI, 11.8-16) for high-, intermediate-, and low-risk groups, respectively (P<0.001), in the derivation set and 4.6 (95% CI, 3.3-8.1), 12.7 (95% CI, 11.3-14.3), and 18.3 months (95% CI, 14.6-24.2) for high-, intermediate-, and low-risk groups, respectively (P<0.001), in the validation set. The nomogram is well-calibrated and reveals acceptable discriminatory capacity, with optimism-corrected c-indices of 0.618 (95% CI, 0.591-0.631) and 0.673 (95% CI, 0.636-0.709) in derivation and validation groups, respectively. The AGAMENON nomogram outperformed the Royal Marsden Hospital (c-index=0.583; P=0.00046) and Japan Clinical Oncology Group prognostic indices (c-index=0.611; P=0.03351). CONCLUSIONS: We developed and validated a straightforward model to predict survival in Caucasian AOA patients initiating first-line polychemotherapy. This model can contribute to inform clinical decision-making and optimise clinical trial design.

Increased Expression of Aquaporin 9 in Trophoblast From Gestational Diabetic Patients.

Gestational diabetes mellitus is the most frequent pathophysiological alteration in pregnancy, increasing the incidence of complications in both mother and fetus. The macrosomia that occurs in these fetuses may be related with some changes in nutrient transport mechanism in placenta. The presence of aquaporin 9, an aquaglyceroporin, has previously been demonstrated in placenta. We raised the question whether aquaporin 9 expression may be upregulated in placenta from gestational diabetes, thus providing a faster transport of glycerol and water through placenta. We studied 21 placentas (13 controls and 8 gestational diabetes) from cesarean delivery at term. The expression of aquaporin 9 was analyzed by quantitative PCR, immunoblot, and immunohistochemistry. The median values from quantitative PCR were compared by nonparametric tests for independent samples (Mann-Whitney U-test). We have found that trophoblast from gestational diabetes express higher amount of aquaporin 9, which was found statistically significant (p<0.05). The increase in aquaporin 9 expression was confirmed by immunoblot, and localization in the syncytiotrophoblast was checked by immunohistochemistry. The increase in aquaporin 9 expression in placenta from gestational diabetes may contribute to the higher transport rate in this pathology of pregnancy.

Elevated expression levels of lysyl oxidases protect against aortic aneurysm progression in Marfan syndrome.

Patients with Marfan syndrome (MFS) are at high risk of life-threatening aortic dissections. The condition is caused by mutations in the gene encoding fibrillin-1, an essential component in the formation of elastic fibers. While experimental findings in animal models of the disease have shown the involvement of transforming growth factor-ß (TGF-ß)- and angiotensin II-dependent pathways, alterations in the vascular extracellular matrix (ECM) may also play a role in the onset and progression of the aortic disease. Lysyl oxidases (LOX) are extracellular enzymes, which initiates the formation of covalent cross-linking of collagens and elastin, thereby contributing to the maturation of the ECM. Here we have explored the role of LOX in the formation of aortic aneurysms in MFS. We show that aortic tissue from MFS patients and MFS mouse model (Fbn1(C1039G/+)) displayed enhanced expression of the members of the LOX family, LOX and LOX-like 1 (LOXL1), and this is associated with the formation of mature collagen fibers. Administration of a LOX inhibitor for 8weeks blocked collagen accumulation and aggravated elastic fiber impairment, and these effects correlated with the induction of a strong and rapidly progressing aortic dilatation, and with premature death in the more severe MFS mouse model, Fbn1(mgR/mgR), without any significant effect on wild type animals. This detrimental effect occurred preferentially in the ascending portion of the aorta, with little or no involvement of the aortic root, and was associated to an overactivation of both canonical and non-canonical TGF-ß signaling pathways. The blockade of angiotensin II type I receptor with losartan restored TGF-ß signaling activation, normalized elastic fiber impairment and prevented the aortic dilatation induced by LOX inhibition in Fbn1(C1039G/+) mice. Our data indicate that LOX enzymes and LOX-mediated collagen accumulation play a critical protective role in aneurysm formation in MFS.

PstS-1, the 38-kDa Mycobacterium tuberculosis glycoprotein, is an adhesin, which binds the macrophage mannose receptor and promotes phagocytosis.

Mycobacterium tuberculosis, the primary causative agent of tuberculosis, infects macrophages and transforms the hostile intracellular environment into a permissive niche. M. tuberculosis infects macrophages using a variety of microbial ligand/cell receptor systems. In this study, binding assays with biotin-labelled mycobacterial cell wall proteins revealed five Concanavalin A-reactive proteins that bind macrophages. Among these proteins, we identified PstS-1, a 38-kDa M. tuberculosis mannosylated glycolipoprotein, and characterized it as an adhesin. Inhibition assays with mannan and immunoprecipitation demonstrated that PstS-1 binds the mannose receptor. We purified PstS-1 to 95.9% purity using ion exchange chromatography. The presence of mannose in purified PstS-1 was demonstrated by Concanavalin A interaction, which was abolished in the presence of sodium m-periodate and α-D-mannosidase. Gas chromatography revealed that purified PstS-1 contained 1% of carbohydrates by weight, which was mainly mannose. Finally, we used fluorescent microbeads coated with purified PstS-1 in phagocytosis assays and discovered that microbead uptake was inhibited by the pre-incubation of cells with GlcNAc, mannan and α-methyl mannoside. The interaction of PstS-1 coated beads with the mannose receptor was confirmed by confocal colocalization studies that showed high Pearson and Manders's colocalization coefficients. Our findings contribute to a better understanding of the strategies M. tuberculosis uses to infect host cells, the critical first step in the pathogenesis of tuberculosis.

The tidepool shrimp, Palaemon ritteri Holmes, constitutes a novel host to the white spot syndrome virus.

The white spot syndrome virus (WSSV) is a lethal and contagious pathogen for penaeid shrimp and a growing number of other crustacean species. To date, there are no effective prophylactic or therapeutic treatments commercially available to interfere with the occurrence and spread of the disease. In addition, the significance of alternative vectors on the dispersal of this disease has been largely ignored and therefore the ecological dynamics of the WSSV is still poorly understood and difficult to ascertain. Thus, an important issue that should be considered in sanitary programmes and management strategies is the identification of species susceptible to infection by WSSV. The results obtained provide the first direct evidence of ongoing WSSV replication in experimentally infected specimens of the tidepool shrimp Palaemon ritteri. Viral replication was detected using a validated set of primers for the amplification by RT-PCR of a 141 bp fragment of the transcript encoding the viral protein VP28. It is therefore conceivable that this shrimp may play a significant role in the dispersal of WSSV.

[Intensive care unit profesionals's knowledge about non invasive ventilation comparative analysis]. / Análisis comparativo de los conocimientos en ventilación mecánica no invasiva de profesionales de cuidados intensivos.

AIMS: The literature highlights the lack of noninvasive ventilation (NIV) protocols and the variability of the knowledge of NIV between intensive care units (ICU) and hospitals, so we want to compare NIV nurses's Knowledge from 4 multipurpose ICU and one surgical ICU. METHODS: Multicenter, crosscutting, descriptive study in three university hospitals. The survey instrument was validated in a pilot test, and the calculated Kappa index was 0.9. Returning a completed survey is an indication of informed consent. Analysis by Chi square test. RESULTS: 117 responded (65%) nurses, 11±9.7 years of experience in ICU and 9.2±7.2 in use of NIV. One of the multipurpose ICU, was initiated NIV an average of 6 years later than the others (95% CI [3.3 to 8.6], P<.001). Only 23.1% of nurses would place a non-vented mask (with no exhalation port) by conventional ventilator, the rest any kind of face mask. 12.7% believed that the mask must be adjusted to the "2-finger" fit while 29% would seal the mask to the patient's face and cover the mask opening where air escapes to facilitate patient/ventilator synchronization. In the surgical ICU agitation identifies mostly as a complication of NIV compared with multipurpose UCIs (31.6% vs 1.8%, P<.001). 56.4% of nurses do not consider respiratory physiotherapy as nursing care, with no difference between units. CONCLUSIONS: Knowledge about types of interface is very dependent on the material of the unit. More training for complications of NIV as agitation and handling secretions it is necessary.

Patient satisfaction and quality of life following laparoscopic Nissen fundoplication.

BACKGROUND: Today, antireflux surgery has an established position in the management of gastroesophageal reflux disease. Some case series have shown good short-term results, but there is still little information regarding long-term results. Studies have recently focused on evaluating residual symptomatology and its impact on quality of life. OBJECTIVES: To determine the postoperative quality of life and degree of satisfaction in patients that underwent laparoscopic Nissen fundoplication. PATIENTS AND METHODS: A total of 100 patients (59 women and 41 men) were studied after having undergone laparoscopic Nissen fundoplication. The variables analyzed were level of satisfaction, gastrointestinal quality of life index (GIQLI), residual symptoms, and the Visick scale. RESULTS: No variation was found in relation to sex; 49 men and 51 women participated in the study. The mean age was 49 years. The degree of satisfaction encountered was: satisfactory in 81 patients, moderate in 3, and bad in 2 patients. More than 90% of the patients would undergo the surgery again or recommend it. The Carlsson score showed improvement at the end of the study (p<0.05). In relation to the GIQLI, a median of 100.61 points±21.624 was obtained. Abdominal bloating, regurgitation, and early satiety were the most frequent residual symptoms. The effect on lifestyle measured by the Visick scale was excellent. CONCLUSIONS: The level of satisfaction and quality of life obtained were comparable with reported standards; and the residual symptoms after antireflux surgery were easily controlled.

[Retroperitoneal mucinous cystadenocarcinoma presenting seven years after appendectomy and right hemicolectomy]. / Cistoadenocarcinoma mucinoso retroperitoneal ocurrido a los 7 años de apendicectomía y hemicolectomía derecha.

Retroperitoneal mucinous cystadenocarcinoma (RMC) with no involvement of neighboring structures is very rare, regardless of whether the lesion is primary or secondary. We report the case of a patient who presented RMC seven years after appendectomy and right hemicolectomy. We discuss both diagnostic possibilities.

CuH-ZSM-5 as hydrocarbon trap under cold start conditions.

Cold start tests are carried out to evaluate the performance of copper-exchanged zeolites as hydrocarbon traps under simulated gasoline car exhaust gases, paying special attention to the role of copper in the performance of these zeolites. It is concluded that the partial substitution of the protons in the parent H-ZSM-5 zeolite is highly beneficial for hydrocarbon trapping due to the formation of selective adsorption sites with specific affinity for the different exhaust components. However, it is also observed that uncontrolled exchanging process conditions could lead to the presence of CuO nanoparticles in the zeolite surface, which seem to block the pore structure of the zeolite, decreasing the hydrocarbon trap efficiency. Among all the zeolites studied, the results point out that a CuH-ZSM-5 with a partial substitution of extra-framework protons by copper cations and without any detectable surface CuO nanoparticles is the zeolite that showed the best performance under simulated cold start conditions due to both the high stability and the hydrocarbon retaining capacity of this sample during the consecutive cycles.

Acupuncture improves sleep in postmenopause in a randomized, double-blind, placebo-controlled study.

BACKGROUND: Insomnia increases in frequency as women approach and pass through menopause. Studies have not shown acupuncture efficacy for insomnia in postmenopausal women. OBJECTIVES: The aim of this study was to evaluate the effectiveness of acupuncture therapy on sleep parameters, depression symptoms and quality of life in postmenopausal women with insomnia. METHODS: This study included 18 postmenopausal women aged 50-67 years old. Participants had a body mass index ≤ 30 kg/m(2), presented a diagnosis of insomnia according to the DSM-IV criteria, had experienced at least 1 year of amenorrhea and had a follicle stimulating hormone level ≥ 30 mIU/ml. Participants were not using antidepressants, hypnotics or hormonal therapy. This study was randomized, double-blind and placebo-controlled. The sample was divided into two groups: acupuncture and 'sham' acupuncture. We performed ten sessions of acupuncture and 'sham' acupuncture during a period of 5 weeks. A polysomnography exam (PSG) and questionnaires (WHOQOL-BREF, Beck Depression Inventory and Pittsburgh Sleep Quality Index) were completed by all patients before and after the treatment period. RESULTS: Anthropometric, polysomnographic, and questionnaire data were similar among the groups at baseline. Comparison of baseline and post-treatment data of the acupuncture group showed that treatment resulted in significantly lower scores on the Pittsburgh Questionnaire and an improvement in psychological WHOQOL. The acupuncture group had a higher percentage of the N3 + 4 stage than the sham group in PSG findings. CONCLUSION: Acupuncture was effective in improving reported sleep quality and quality of life in postmenopausal women with insomnia.

[Work setting, satisfaction and burnout of the nurses in critical care units and hospitalization units. RN4CAST-Spain project]. / Entorno laboral, satisfacción y burnout de las enfermeras de unidades de cuidados críticos y unidades de hospitalización. Proyecto RN4CAST-España.

OBJECTIVE: To know if there are differences between the critical care units and the medical-surgical care units regarding the perception of the nurses working in National Health System hospitals about their work environment, burnout level and job satisfaction. METHOD: A cross-sectional study was conducted with 6,417 nurses from the medical-surgical care units and with 1,122 nurses from critical care units of 59 Spanish hospitals with more than 150 beds. Socio-demographic, job satisfaction, perception of work environment (Practice Environment Scale of the Nursing Work Index [PES-NWI questionnaire]) and burnout measured with the Maslach Burnout Inventory (MBI) data were collected. RESULTS: The PES-NWI showed differences in 4 out of its 5 factors. It showed better values in medical-surgical units in all the factors, except for Staffing and resource adequacy (P<.001), where critical care units showed a mean level of agreement of 2.41 versus 2.19 for the medical-surgical units. Regarding burnout, this was higher in the medical-surgical care units (P=.039) where 23% (952) of the nurses had high levels. Job satisfaction was lower in the critical care units (P=.044) with 70% (578) of nurses being very or strongly satisfied. CONCLUSIONS: The opinion of the nurses, working in critical care units about their hospital is unfavorable. They showed lower levels of burnout than those working in medical-surgical units.

Arena surface vertical impact forces vary with surface compaction.

Mechanical properties of arena surfaces are extrinsic factors for musculoskeletal injury. Vertical impact forces of harrowed and compacted cushion were measured at five locations on 12 arena surfaces (five dirt, seven synthetic [dirt and fiber]). Eight variables related to impact force, displacement, and acceleration were calculated. Surface temperature, cushion depth and moisture content were also measured. The effects of surface material type (dirt/synthetic) and cushion compaction (harrowed/compacted) on vertical impact properties were assessed using an analysis of variance. Relationships of manageable surface properties with vertical impact forces were examined through correlations. Compacted cushion exhibited markedly higher vertical impact force and deceleration with lower vertical displacement than harrowed cushion (P < 0.001), and the effect was greater on dirt than synthetic surfaces (P = 0.039). Vertical displacement (P = 0.021) and soil rebound (P = 0.005) were the only variables affected by surface type. Surface compaction (harrowed, compacted) had a significantly greater effect on vertical impact forces than surface type (dirt, synthetic). By reducing surface compaction through harrowing, extrinsic factors related to musculoskeletal injury risk are reduced. These benefits were more pronounced on dirt than synthetic surfaces. These results indicate that arena owners should regularly harrow surfaces, particularly dirt surfaces.

Shear ground reaction force variation among equine arena surfaces.

Shear forces at the surface-hoof interface affect hoof slide, surface grip, forces transferred to the limb, and injury risk. However, the variation in shear forces among surfaces with different compositions have not been quantified. Shear ground reaction forces were measured on five dirt and seven synthetic arena surfaces. Cohesion/adhesion and angle of internal friction/coefficient of friction were calculated. Surface composition, surface temperature, cushion depth, and moisture content were also measured. The effects of surface material (dirt/synthetic) on shear properties were assessed using analysis of variance (ANOVA; P < 0.05). The relationships between surface composition or management properties and shear properties were analyzed using linear correlation. Shear properties were not different between dirt and synthetic surface categories; however, surface fiber content was correlated with adhesion and coefficient of friction. These correlations predict that more fiber will decrease soil adhesion (r = -0.75; P < 0.01) and increase the coefficient of friction (r = 0.81; P < 0.01). Furthermore, maximum shear force was significantly correlated with cushion depth (r = 0.61; P < 0.01) and moisture content (r = 0.57; P < 0.01), where shear force was greater on surfaces with thicker cushion layers or higher moisture content. The findings suggest that shear mechanical behavior is more dependent on surface composition than surface material categories (dirt/synthetic) and also indicate that arena owners can influence shear forces by adjusting either surface composition or management.

Development and validation of a real-time PCR assay protocol for the specific detection and quantification of pelagiphages in seawater samples.

Earth is inhabited by numerous adaptations of cellular forms shaped by the persistent scrutiny of natural selection. Thus, as natural selection has fixed beneficial adaptations of functional traits, cellular life has conquered almost all environmental niches on our planet. However, cellular life succumbs in number and genetic diversity to viruses. Among all viruses, phages are highly prevalent in diverse environments, and due to their vast genetic diversity and abundance, their relevant role as significant players in several ecological processes is now fully recognized. Pelagiphages, bacteriophages infecting bacteria of the SAR11 clade, are the most abundant viruses in the oceans. However, the ecological contribution of pelagiphages on populations of Pelagibacterales remains largely underestimated. An essential aspect of estimating the impact of bacteriophages is their absolute and precise quantification, which provides relevant information about the host-virus interactions and the structure of viral assemblages. Consequently, due to its abundance and claimed influence in the biogeochemical cycling of elements, the accurate quantification of pelagiphages results in an essential task. This study describes the development and validation of a sensitive, specific, accurate and reproducible qPCR platform targeting pelagiphages. Moreover, this method allowed the detection and quantification of pelagiphages in the Gulf of California for the first time.

ß-hydroxybutyrate is a metabolic regulator of proteostasis in the aged and Alzheimer disease brain.

Loss of proteostasis is a hallmark of aging and Alzheimer disease (AD). Here, we identify ß-hydroxybutyrate (ßHB), a ketone body, as a regulator of protein solubility in the aging brain. ßHB is a small molecule metabolite which primarily provides an oxidative substrate for ATP during hypoglycemic conditions, and also regulates other cellular processes through covalent and noncovalent protein interactions. We demonstrate ßHB-induced protein insolubility across in vitro, ex vivo, and in vivo mouse systems. This activity is shared by select structurally similar metabolites, is not dependent on covalent protein modification, pH, or solute load, and is observable in mouse brain in vivo after delivery of a ketone ester. Furthermore, this phenotype is selective for pathological proteins such as amyloid-ß, and exogenous ßHB ameliorates pathology in nematode models of amyloid-ß aggregation toxicity. We have generated a comprehensive atlas of the ßHB-induced protein insolublome ex vivo and in vivo using mass spectrometry proteomics, and have identified common protein domains within ßHB target sequences. Finally, we show enrichment of neurodegeneration-related proteins among ßHB targets and the clearance of these targets from mouse brain, likely via ßHB-induced autophagy. Overall, these data indicate a new metabolically regulated mechanism of proteostasis relevant to aging and AD.