Median Lifespan Research Articles

Optimising the mass-rearing of Aphytis sp. (Hymenoptera: Aphelinidae) to biologically control San Jose scales

ABSTRACT Augmentative biological control promotes sustainable food production via reducing reliance on chemical insecticides. The San Jose scale, Comstockaspis perniciosa (Hemiptera: Diaspididae), is a major pest in deciduous fruit agriculture worldwide. Surveys in Uruguay have identified a species of Aphytis to be the main natural enemy of this pest; this genus of parasitoids is mostly used for their effectiveness in regulating scale populations. This study aimed to enhance the biological control of San Jose scale via evaluating the biological characteristics of Aphytis sp. to optimise its mass-rearing. The host development stage with the highest oviposition and offspring emergence was determined. Longevity, daily egg production throughout life, and daily offspring emergence in the first 10 days of life were studied. Offspring production at different parasitoid densities was evaluated, incorporating parasitoid interference and using scales for host-feeding. Results showed greater oviposition and emergence in adult female scales compared to the second nymphal instar, regardless of scales being young or pre-reproductive females. The parasitoids had a median adult life span of 10 days, with the highest oviposition on the first day. However, offspring emergence was low during the first five days. Mass production levels approached those of individual parasitism at a density of 13 scales per parasitoid per day. These findings suggest an optimised production scheme for Aphytis sp., enhancing profitability for producers and contributing to sustainable agroecosystems via reducing the need for chemical insecticides.

Genotype and sex affect the combined impact of temperature and low-dose insecticide exposure on insect survival.

Insecticide contamination and climate change are key factors driving the global decline in insect populations. However, how these factors interact to impact insect survival remains uncertain. In this study, we examined the effects of sex and genotype on the response to long-term low insecticide exposure at two temperatures, 18 °C and 28 °C, using the Drosophila melanogaster model. We focused on a polymorphic gene, Cyp6g1, known for conferring broad insecticide resistance. We found that while temperature and insecticide have a synergistic effect on mortality of susceptible flies (Cyp6g1-M allele), they act additively on resistant flies (Cyp6g1-BA allele). And whereas the mortality of BA flies exposed to insecticides is strongly dependent on sex at 18 °C, no sex bias is found at 28 °C. Under no insecticide exposure, BA females showed shorter median lifespan than males regardless of temperature, possibly reflecting a cost associated with the resistant allele. Surprisingly, across all genotypes, females showed lower Cyp6g1 gene expression levels than males, which contrasts with their higher insecticide tolerance. Temperature and insecticide exposure had small effects on Cyp6g1 expression levels, suggesting the presence of additional mechanisms of resistance. Our results indicate that the effect of high insecticide doses on insect mortality cannot be used to predict how insects will respond to low contaminating doses, especially when considering the strong interactions between sex, temperature, and genotype. The combined effects of temperature and long-term low insecticide exposure are complex and can have major impacts on insect population dynamics and survival.

Comparative lifespan and healthspan of nonhuman primate species common to biomedical research.

There is a critical need to generate age- and sex-specific survival curves to characterize chronological aging consistently across nonhuman primates (NHP) used in biomedical research. Sex-specific Kaplan-Meier survival curves were computed in 12 translational aging models: baboon, bonnet macaque, chimpanzee, common marmoset, coppery titi monkey, cotton-top tamarin, cynomolgus macaque, Japanese macaque, pigtail macaque, rhesus macaque, squirrel monkey, and vervet/African green. After employing strict inclusion criteria, primary results are based on 12,269 NHPs that survived to adulthood and died of natural/health-related causes. A secondary analysis was completed for 32,616 NHPs that died of any cause. Results show a pattern of reduced male survival among catarrhines (African and Asian primates), especially macaques, but not platyrrhines (Central and South American primates). For many species, median lifespans were lower than previously reported. An important consideration is that these analyses may offer a better reflection of healthspan than lifespan since research NHPs are typically euthanized for humane welfare reasons before their natural end of life. This resource represents the most comprehensive characterization of sex-specific lifespan and age-at-death distributions for 12 biomedically relevant species, to date. These results clarify relationships among NHP ages and provide a valuable resource for the aging research community, improving human-NHP age equivalencies, informing investigators of expected survival rates, providing a metric for comparisons in future studies, and contributing to understanding of factors driving lifespan differences within and among species.

Comparative lifespan and healthspan of nonhuman primate species common to biomedical research.

There is a critical need to generate age- and sex-specific survival curves to characterize chronological aging consistently across nonhuman primates (NHP) used in biomedical research. Sex-specific Kaplan-Meier survival curves were computed in 12 translational aging models: baboon, bonnet macaque, chimpanzee, common marmoset, coppery titi monkey, cotton-top tamarin, cynomolgus macaque, Japanese macaque, pigtail macaque, rhesus macaque, squirrel monkey, and vervet/African green. After employing strict inclusion criteria, primary results are based on 12,269 NHP that survived to adulthood and died of natural/health-related causes. A secondary analysis was completed for 32,616 NHP that died of any cause. Results show a pattern of reduced male survival among catarrhines (African and Asian primates), especially macaques, but not platyrrhines (Central and South American primates). For many species, median lifespans were lower than previously reported. An important consideration is that these analyses may offer a better reflection of healthspan than lifespan since research NHP are typically euthanized for humane welfare reasons before their natural end of life. This resource represents the most comprehensive characterization of sex-specific lifespan and age-at-death distributions for 12 biomedically relevant species, to date. These results clarify relationships among NHP ages and provide a valuable resource for the aging research community, improving human-NHP age equivalencies, informing investigators of expected survival rates, providing a metric for comparisons in future studies, and contributing to understanding of factors driving lifespan differences within and among species.

Development and validation of a nomogram for circuit lifespan of regional citrate anticoagulation-continuous renal replacement therapy in intensive care patients with acute kidney injury.

Regional citrate anticoagulation-continuous renal replacement therapy (RCA-CRRT) has a wide range of applications in clinical practice, but unplanned downtime due to RCA-CRRT circuit coagulation is as high as 15.75%-66.70%. To build a nomogram model for predicting the lifespan circuits of RCA-CRRT. A prospective observational cohort study was conducted in Sichuan, China. The collected clinical data from 404 RCA-CRRT sessions involving 135 patients were utilized. The patients' basic information, laboratory indicators and RCA-CRRT parameters were used as independent variables, and the survival status and survival time of RCA-CRRT circuits were used as dependent variables. A Cox multivariate analysis was performed to build the nomogram model for predicting the lifespan of RCA-CRRT circuits. The model was validated internally and externally. The median lifespan of RCA-CRRT circuits was 28.0 (12.0-46.5) h, and the unplanned downtime rate was 23.76%. In the Cox multivariate analysis, venous pressure, haemoglobin, Sequential Organ Failure Assessment (SOFA), lactate, and blood transfusion were identified as statistically significant predictive factors for the lifespan of RCA-CRRT circuits (p < .05). Subsequently, a nomogram model for predicting the lifespan of RCA-CRRT circuits was developed. The AUC values for internal and external validation within the 12-72-h timeframe ranged from 0.648 to 0.816 and 0.613 to 0.956, respectively. Both the calibration curve and clinical decision curve demonstrated the model's good performance. The nomogram model developed in this study demonstrates its efficacy in accurately predicting the lifespan circuits of RCA-CRRT. Clinical nurses can use the prediction model to assess the lifespan of RCA-CRRT circuits, so as to formulate a personalized RCA-CRRT treatment plan for patients, thus reducing the unplanned downtime of RCA-CRRT.

Lifespan of male and female APP/PS1 and APPNL-F/NL-F mouse models of Alzheimer's disease.

Alzheimer's disease (AD) disproportionately affects women, yet most preclinical research studies are male-centric. We performed lifespan analyses of male and female AD mouse models (APP/PS1 and APPNL-F/NL-F) and their shared genetic background control (C57BL/6). Survival curves support significant sex differences between within genotypes. Minimal longevity revealed increased age in male APP/PS1, and decreased age in APPNL-F/NL-F mice. Maximal longevity revealed an increased average age in males. Furthermore, median lifespan differed between sex and genotype. This study supports sexual dimorphic survival in two mouse models of AD, emphasizing the need to examine mechanisms and treatments in both sexes.

Knockout of &lt;i&gt;Hsp70&lt;/i&gt; genes modulates age-related transcriptomic changes in leg muscles, reduces locomotion speed and lifespan of &lt;i&gt;Drosophila melanogaster&lt;/i&gt;

The study investigated the effect of knockout of six Hsp70 genes (orthologues of the mammalian genes Hspa1a, Hspa1b, Hspa2 and Hspa8) on age-related changes in gene expression in the legs of Drosophila melanogaster, which contain predominantly skeletal muscle bundles. For this, the leg transcriptomic profile was examined in males of the w1118 control line and the Hsp70– line on the 7th, 23rd and 47th days of life. In w1118 flies, an age-related decrease in the locomotion (climbing) speed (a marker of functional state and endurance) was accompanied by a pronounced change in the transcriptomic profile of the leg skeletal muscles, which is conservative in nature. In Hsp70– flies, the median lifespan was shorter and the locomotion speed was significantly lower compare to the control; at the same time, complex changes in the age-related dynamics of the skeletal muscle transcriptome were observed. Mass spectrometry-based quantitative proteomic showed that 47-day-old Hsp70– flies, compared with w1118, demonstrated multidirectional changes in the content of key enzymes of glucose metabolism and fat oxidation (glycolysis, pentose phosphate pathway, Krebs cycle, beta-oxidation and oxidative phosphorylation). Such dysregulation may be associated with a compensatory increase in the expression of other genes encoding chaperones (small Hsp, Hsp40, 60, and 70), which regulate specific sets of target proteins. Taken together, our data show that knockout of six Hsp70 genes slightly reduces the median lifespan of flies, but significantly reduces the locomotion speed, which may be associated with complex changes in the transcriptome of the leg skeletal muscles and with multidirectional changes in the content of key enzymes of energy metabolism.

Heating alters the nutritional and antioxidant characteristics of lotus root

To investigate the impact of heating on the nutritional and antioxidant properties of lotus root, a comparative analysis was conducted between its heated and unheated whole powders. The whole powder of heated lotus root (HLRWP) exhibited a higher level of total soluble sugars but a lower level of free phenolic compounds compared to the whole powder of fresh lotus root (FLRWP). Meanwhile, they showed significant differences in metabolite composition encompassing phenolic compounds, flavonoids, amino acids, vitamins and lipids. The replacement of sugar and corn in the medium with 25% HLRWP effectively enhanced the activities of antioxidant enzymes (total superoxide dismutase and catalase), while reducing malonaldehyde content in Drosophila melanogaster males. The average, median and maximum lifespans of the fruit flies were extended by 35.90%, 23.49% and 24.00% respectively, accompanied by an improvement in climbing ability. Additionally, their resistance to oxidative stress induced by hydrogen peroxide or paraquat was enhanced. The antioxidant capacity of HLRWP in vivo was found to be obviously stronger compared to FLRWP, which may have a crucial association with the regulation of L-arginine and adenosine monophosphate levels. The results indicate that heating can enhance the nutritional quality of lotus root products by strengthening their antioxidant capacity.

Roles of Growth Hormone-Dependent JAK-STAT5 and Lyn Kinase Signaling in Determining Lifespan and Cancer Incidence.

In rodents, loss of growth hormone (GH) or its receptor is associated with extended lifespan. We aimed to determine the signaling process resulting in this longevity using GH receptor (GHR)-mutant mice with key signaling pathways deleted and correlate this with cancer incidence and expression of genes associated with longevity. GHR uses both canonical janus kinase (JAK)2-signal transducer and activator of transcription (STAT) signaling as well as signaling via the LYN-ERK1/2 pathway. We used C57BL/6 mice with loss of key receptor tyrosines and truncation resulting in 1) loss of most STAT5 response to GH; 2) total inability to generate STAT5 to GH; 3) loss of Box1 to prevent activation of JAK2 but not LYN kinase; or 4) total knockout of the receptor. For each mutant we analyzed lifespan, histopathology to determine likely cause of death, and hepatic gene and protein expression. The extended lifespan is evident in the Box1-mutant males (retains Lyn activation), which have a median lifespan of 1016 days compared to 890 days for the Ghr-/- males. In the females, GhrBox1-/- mice have a median lifespan of 970 days compared to 911 days for the knockout females. Sexually dimorphic GHR-STAT5 is repressive for longevity, since its removal results in a median lifespan of 1003 days in females compared to 734 days for wild-type females. Numerous transcripts related to insulin sensitivity, oxidative stress response, and mitochondrial function are regulated by GHR-STAT5; however, LYN-responsive genes involve DNA repair, cell cycle control, and anti-inflammatory response. There appears to be a yin-yang relationship between JAK2 and LYN that determines lifespan.

Levels of Amyloid Beta (Aβ) Expression in the Caenorhabditis elegans Neurons Influence the Onset and Severity of Neuronally Mediated Phenotypes.

A characteristic feature of Alzheimer's disease (AD) is the formation of neuronal extracellular senile plaques composed of aggregates of fibrillar amyloid β (Aβ) peptides, with the Aβ1-42 peptide being the most abundant species. These Aβ peptides have been proposed to contribute to the pathophysiology of the disease; however, there are few tools available to test this hypothesis directly. In particular, there are no data that establish a dose-response relationship between Aβ peptide expression level and disease. We have generated a panel of transgenic Caenorhabditis elegans strains expressing the human Aβ1-42 peptide under the control of promoter regions of two pan-neuronal expressed genes, snb-1 and rgef-1. Phenotypic data show strong age-related defects in motility, subtle changes in chemotaxis, reduced median and maximum lifespan, changes in health span indicators, and impaired learning. The Aβ1-42 expression level of these strains differed as a function of promoter identity and transgene copy number, and the timing and severity of phenotypes mediated by Aβ1-42 were strongly positively correlated with expression level. The pan-neuronal expression of varying levels of human Aβ1-42 in a nematode model provides a new tool to investigate the in vivo toxicity of neuronal Aβ expression and the molecular and cellular mechanisms underlying AD progression in the absence of endogenous Aβ peptides. More importantly, it allows direct quantitative testing of the dose-response relationship between neuronal Aβ peptide expression and disease for the first time. These strains may also be used to develop screens for novel therapeutics to treat Alzheimer's disease.

Stochasticity in Dietary Restriction-Mediated Lifespan Outcomes in Drosophila.

Dietary restriction (DR) is widely considered to be one of the most potent approaches to extend healthy lifespan across various species, yet it has become increasingly apparent that DR-mediated longevity is influenced by biological and non-biological factors. We propose that current priorities in the field should include understanding the relative contributions of these factors to elucidate the mechanisms underlying the beneficial effects of DR. Our work conducted in two laboratories, represents an attempt to unify DR protocols in Drosophila and to investigate the stochastic effects of DR. Across 64 pairs of survival data (DR/ad libitum, or AL), we find that DR does not universally extend lifespan. Specifically, we observed that DR conferred a significant lifespan extension in only 26.7% (17/64) of pairs. Our pooled data show that the overall lifespan difference between DR and AL groups is statistically significant, but the median lifespan increase under DR (7.1%) is small. The effects of DR were overshadowed by stochastic factors and genotype. Future research efforts directed toward gaining a comprehensive understanding of DR-dependent mechanisms should focus on unraveling the interactions between genetic and environmental factors. This is essential for developing personalized healthspan-extending interventions and optimizing dietary recommendations for individual genetic profiles.

Stochasticity in Dietary Restriction-Mediated Lifespan Outcomes in Drosophila.

Dietary restriction (DR) is widely considered to be one of the most potent approaches to extend healthy lifespan across various species, yet it has become increasingly apparent that DR-mediated longevity is influenced by biological and non-biological factors. We propose that current priorities in the field should include understanding the relative contributions of these factors to elucidate the mechanisms underlying the beneficial effects of DR. Our work conducted in two laboratories, represents an attempt to unify DR protocols in Drosophila and to investigate the stochastic effects of DR. Across 64 pairs of survival data (DR/ad libitum, or AL), we find that DR does not universally extend lifespan. Specifically, we observed that DR conferred a significant lifespan extension in only 26.7% (17/64) of pairs. Our pooled data show that the overall lifespan difference between DR and AL groups is statistically significant, but the median lifespan increase under DR (7.1%) is small. The effects of DR were overshadowed by stochastic factors and genotype. Future research efforts directed toward gaining a comprehensive understanding of DR-dependent mechanisms should focus on unraveling the interactions between genetic and environmental factors. This is essential for developing personalized healthspan-extending interventions and optimizing dietary recommendations for individual genetic profiles.

Crocetin Delays Brain and Body Aging by Increasing Cellular Energy Levels in Aged C57BL/6J Mice.

Aging is usually accompanied by mitochondrial dysfunction, reduced energy levels, and cell death in the brain and other tissues. Mitochondria play a crucial role in maintaining cellular energy through oxidative phosphorylation (OXPHOS). However, OXPHOS is impaired as the mitochondrial oxygen supply decreases with age. We explored whether pharmacologically increased oxygen diffusion by crocetin can restore OXPHOS and help delay the aging of the brain and other vital organs. We found that aged mice treated with crocetin for four months displayed significantly improved memory behavior, neuromuscular coordination, and ATP and NAD+ levels in the brain and other vital organs, leading to an increased median life span. The transcriptomic analysis of hippocampi from crocetin-treated mice revealed that enhanced brain energy level was caused by the upregulation of genes linked to OXPHOS, and their expression was close to that in young mice. The chronic treatment of aged astrocytes also showed improved mitochondrial membrane potential and energy state of the cells. Moreover, chronic treatment with crocetin did not cause any oxidative stress. Our data suggest that restoring OXPHOS and the normal energy state of the cell can delay aging and enhance longevity. Therefore, molecules such as crocetin should be further explored to treat age-related diseases.

WP5.5 - Investigating the Optimal Timing for Definitive Treatment in Sigmoid Volvulus: A Retrospective Cohort Study

Abstract Aims The evidence base is replete of guidance on the optimal timing for definitive surgical management of acute sigmoid volvulus (ASV), with limited understanding or long-term outcomes for those managed conservatively. This study aimed to assess the management and outcomes of this understudied patient population. Methods A retrospective cohort study was conducted in a UK tertiary centre between January 2018 and December 2022. Results 69 patients presented with ASV (54M:15F, age 73.7 SD +/- 15.0 years). The median number of acute admissions per patient was 2 (IQR 1-4.5), with 29 patients (42.0%) presenting only once. 44 patients (63.8%) were managed conservatively. Of those patients, 22 (50.0%) were readmitted within the study period. 25 patients (26.2%) underwent operative management, with 18 (72.0%) of these patients undergoing emergency surgery (6 during their index admission, 33.3%). Elective surgical management was performed in 7 patients after an average of 3 previous admissions. All-cause mortality amongst the cohort was 47.8% (n=33), with a median life span of 1.1 years from index admission (IQR 0.3-2.6); 24.0% in the operative group (5 acute, 1 elective) versus 61.4% in the conservatively managed group. Conservatively managed patients were at greater odds of mortality compared to acutely operated patients (OR 4.129, 95% CI 1.25-13.67, p=0.02) and electively operated patients (OR 9.529, 95% CI 1.05-86.20, p=0.045). Acute operations had no significant greater odds of mortality compared to elective operations (p=0.49). Conclusions Conservative management of ASV was associated with greater odds of all-cause mortality compared to patients who received an operation. Operations performed in the acute setting had no greater mortality odds than in the elective setting.

How can we achieve more accurate reporting of average dog lifespan?

Despite major advances in our understanding of dogs as a biological system (including genetics/epigenetics, physiology, cognition, and behavior), the veterinary field lacks consensus around a critical piece of information: namely, the average lifespan/life expectancy of a domestic dog. This deficiency is due in part to unavailable and/or inconsistent collection of dog mortality data. In an effort to review historical and current reports of dog lifespan to determine whether the domestic dog's lifespan has changed over time, we found that incongruous data were prohibitive to conducting a formal meta-analysis of dog lifespan reports. However, in examining several different kinds of dog aging and mortality studies covering a span of about 40 years (1981 to 2023), it seems apparent that the median lifespan of domesticated dogs has not recently decreased, as has been reported in the popular press, but rather has increased steadily over that time frame. Still, assessing the validity of these numbers is a challenge, as methodology, cohort, and covariates (such as weight, breed, etc) vary from study to study. We therefore recommend the adoption of a comprehensive, standardized method for reporting and recording dog mortality so that a more accurate understanding of dogs' average lifespan can be obtained in the future.

Bcl-2 Orthologues, Buffy and Debcl, Can Suppress Drp1-Dependent Age-Related Phenotypes in Drosophila.

The relationship of Amyotrophic Lateral Sclerosis, Parkinson's disease, and other age-related neurodegenerative diseases with mitochondrial dysfunction has led to our study of the mitochondrial fission gene Drp1 in Drosophila melanogaster and aspects of aging. Previously, the Drp1 protein has been demonstrated to interact with the Drosophila Bcl-2 mitochondrial proteins, and Drp1 mutations can lead to mitochondrial dysfunction and neuronal loss. In this study, the Dopa decarboxylase-Gal4 (Ddc-Gal4) transgene was exploited to direct the expression of Drp1 and Drp1-RNAi transgenes in select neurons. Here, the knockdown of Drp1 seems to compromise locomotor function throughout life but does not alter longevity. The co-expression of Buffy suppresses the poor climbing induced by the knockdown of the Drp1 function. The consequences of Drp1 overexpression, which specifically reduced median lifespan and diminished climbing abilities over time, can be suppressed through the directed co-overexpression of pro-survival Bcl-2 gene Buffy or by the co-knockdown of the pro-cell death Bcl-2 homologue Debcl. Alteration of the expression of Drp1 acts to phenocopy neurodegenerative disease phenotypes in Drosophila, while overexpression of Buffy can counteract or rescue these phenotypes to improve overall health. The diminished healthy aging due to either the overexpression of Drp1 or the RNA interference of Drp1 has produced novel Drosophila models for investigating mechanisms underlying neurodegenerative disease.

Polygenic prediction of human longevity on the supposition of pervasive pleiotropy

The highly polygenic nature of human longevity renders pleiotropy an indispensable feature of its genetic architecture. Leveraging the genetic correlation between aging-related traits (ARTs), we aimed to model the additive variance in lifespan as a function of the cumulative liability from pleiotropic segregating variants. We tracked allele frequency changes as a function of viability across different age bins and prioritized 34 variants with an immediate implication on lipid metabolism, body mass index (BMI), and cognitive performance, among other traits, revealed by PheWAS analysis in the UK Biobank. Given the highly complex and non-linear interactions between the genetic determinants of longevity, we reasoned that a composite polygenic score would approximate a substantial portion of the variance in lifespan and developed the integrated longevity genetic scores (iLGSs) for distinguishing exceptional survival. We showed that coefficients derived from our ensemble model could potentially reveal an interesting pattern of genomic pleiotropy specific to lifespan. We assessed the predictive performance of our model for distinguishing the enrichment of exceptional longevity among long-lived individuals in two replication cohorts (the Scripps Wellderly cohort and the Medical Genome Reference Bank (MRGB)) and showed that the median lifespan in the highest decile of our composite prognostic index is up to 4.8 years longer. Finally, using the proteomic correlates of iLGS, we identified protein markers associated with exceptional longevity irrespective of chronological age and prioritized drugs with repurposing potentials for gerotherapeutics. Together, our approach demonstrates a promising framework for polygenic modeling of additive liability conferred by ARTs in defining exceptional longevity and assisting the identification of individuals at a higher risk of mortality for targeted lifestyle modifications earlier in life. Furthermore, the proteomic signature associated with iLGS highlights the functional pathway upstream of the PI3K-Akt that can be effectively targeted to slow down aging and extend lifespan.

Regional citrate anticoagulation versus LMWH anticoagulation for CRRT in liver failure patients without increased bleeding risk.

The optimal anticoagulation regimen for continuous renal replacement therapy (CRRT) in liver failure (LF) patients without increased bleeding risk remains controversial. Therefore, we conducted a monocentric retrospective study to evaluate the efficacy and safety of the regional citrate anticoagulation (RCA) versus low molecular weight heparin (LMWH) anticoagulation for CRRT in LF without increased bleeding risk. According to the anticoagulation strategy for CRRT, patients were divided into the RCA and LMWH-anticoagulation groups. The evaluated endpoints were patient survival, filter lifespan, bleeding, citrate accumulation, and totCa/ionCa ratio. Totally 167 and 164 filters were used in the RCA and LMWH group, respectively. The median filter lifespan was significantly longer in the RCA group (34 h (IQR = 24-54) versus 24 h (IQR = 18-45.5) [95%CI, 24.5-33]; p < 0.001). The 4-week mortality rate was significantly higher in the LMWH-anticoagulation group (71 (57.72%) vs 53 (40.46%); p = 0.006). After adjusted the important parameters in the multivariate COX regression model, the mortality risk was significantly reduced in the RCA group (HR = 0.668 [95%CI, 0.468-0.955]; p = 0.027). In the LMWH group, 30 bleeding episodes (24,19%) were observed, whereas only 7 (5.34%) occurred in the RCA group (p < 0.001). Two patients (1.5%) in the RCA group occurred citrate accumulation. In LF patients without increased bleeding risk who underwent CRRT, RCA significantly extended the filter lifespan and improved patient survival rate. There was no significant difference in the rate of adverse events between the two groups.

Assessment of microbial safety of fresh vegetables through Caenorhabditis elegans model

Caenorhabditis elegans is widely used as a model to predict the virulence of animal-origin human pathogens, as C. elegans and humans share similar intestinal epithelial cells. A study was undertaken to assess the feasibility of C. elegans for surveillance of foodborne pathogens in fresh vegetables. First, the virulent strains of four foodborne pathogens viz., Pseudomonas aeruginosa (PAO1), Escherichia coli (O157), Salmonella enterica (CI1), and Staphylococcus aureus (ATCC25923) were fed to the worm as sole diet and compared the survival, phenome traits, and gut integrity with standard E. coli OP50. Secondly, the vegetable surface microbiota (tomato) was enriched with gradient concentrations of PAO1, and the worm's survival and gut distension and leakage were assessed. Thirdly, the total microbiota of fresh vegetables (tomato, brinjal, and lablab) were evaluated for the worm's survival. The gut epithelial cell damage and leakage were monitored by blue dye-based smurf assay. Foodborne pathogens reduced the median lifespan (LT50) of the worm drastically, with PAO1 had 2.5 days, O157 (6 days), CI1 (8 days), ATCC25923 (6 days), while the standard diet OP50 had 16 days. The phenome traits of worms, viz., pharyngeal pumping, defecation cycle, body bends, head thrashes, touch stimulus, paralysis, gut leakage, and reproduction, were significantly negatively affected due to pathogen feeding. The gut epithelial integrity was severely affected, and blue dye leakage was the highest in O157 (73 %), followed by PAO1 (55 %), CI1 (40 %) and ATCC25923 (38 %), while the gut epithelial cells were intact in OP50 fed worms. The pathogen enrichment in tomato surface microbiota diet showed that LT50 of the worms was reduced from 8 days to 2.5 days and 10 to 8 days with increasing gradient of PAO1 in fresh tomato microbiota and 24-hours pre-enriched tomato microbiota, respectively. Smurf test also confirmed that an increase in PAO1 proportion in total microbiota distended the gut with high dye leakage. The total microbiota extracted from tomato, brinjal, and lablab did not affect the worm's health and had an extended median life span (11 – 13 days) when they were devoid of any foodborne pathogens. These findings provide a novel and straightforward approach for assessing foodborne pathogens using animal models.

The ALS-associated KIF5A P986L variant is not pathogenic for Drosophila motoneurons

Amyotrophic lateral sclerosis (ALS) is a devastating paralytic disorder caused by the death of motoneurons. Several mutations in the KIF5A gene have been identified in patients with ALS. Some mutations affect the splicing sites of exon 27 leading to its deletion (Δ27 mutation). KIF5A Δ27 is aggregation-prone and pathogenic for motoneurons due to a toxic gain of function. Another mutation found to be enriched in ALS patients is a proline/leucine substitution at position 986 (P986L mutation). Bioinformatic analyses strongly suggest that this variant is benign. Our study aims to conduct functional studies in Drosophila to classify the KIF5A P986L variant. When expressed in motoneurons, KIF5A P986L does not modify the morphology of larval NMJ or the synaptic transmission. In addition, KIF5A P986L is uniformly distributed in axons and does not disturb mitochondria distribution. Locomotion at larval and adult stages is not affected by KIF5A P986L. Finally, both KIF5A WT and P986L expression in adult motoneurons extend median lifespan compared to control flies. Altogether, our data show that the KIF5A P986L variant is not pathogenic for motoneurons and may represent a hypomorphic allele, although it is not causative for ALS.