Search

Abstract Background Inappropriate use of antibiotics is a major cause of antimicrobial resistance (AMR). Inadequate knowledge about AMR among healthcare students could affect their practice of antimicrobial stewardship as future healthcare professionals. This study aims to assess the use of antibiotics and knowledge of AMR among future healthcare professionals of a Nigerian University. Methods Respondents’ knowledge of antimicrobial resistance, use of antibiotics, and source of antibiotics in the past 12 months was explored using a self-administered questionnaire. Multivariate analyses were used to evaluate the relationship between specific variables and respondents’ knowledge. Results Of the 939 questionnaires administered to the students, 866 were filled given a response rate of 92.2 %. A total of (765; 88.3 %) of the respondents were aware that antimicrobial resistance makes it harder to eliminate the infection from the body as existing drugs become less effective. In all 824, (95.2 %) of the respondents had use antibiotics in the past 12 months. The use of antibiotics to treat malaria was self-reported by (175; 21.2 %). About half (432; 52.4 %) purchased the antibiotics from community pharmacies, while others obtained their antibiotics from the hospitals (192; 23.3 %), patent medicine stores (150; 18.2 %), and friends and family (50; 6.1 %) in the last 12 months. In all 506, (58.4 %) had good knowledge of antimicrobial resistance. Logistic regression shows that students in 3rd to 6th year 9.29 [AOR = 9.29, 95 % CI: (3.7–22.96)], had greater knowledge of antimicrobial resistance. Conclusions The healthcare students demonstrated a moderate knowledge of AMR. This underscores the need to adopt several educational tactics to introduce the concepts of AMR to the students and ensure there are strict policies to regulate the flow of antibiotics.

The goal of this paper is to build a relationship for the prediction of (α)-factor based on the Taguchi approach. The tests have been carried out in an experimental pilot that comprises a rectangular tank of 84.6 L, fed with synthetic wastewater in the presence of air in order to preserve the biomass proliferation. The observations reveal that an increase of MLSS to 16 mg. L-1 reduces the (α)-factor from 0.79 to 0.1. The interpretation of the biological and the physical factors impacts on (α)-factor and this leads to elaborate a surface response RSM and multiple linear regression MLR models at a confidence level of 95%. The R2 of the MLR models has showed significant values of about =80%. It has been concluded that the limits of the operating conditions and the design geometry play a key role in the model's ability to explain the experimental change of (α)-factor. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the effect of different fertigation levels and intervals on growth, yield and flower quality of red cut rose (hybrid tea rose) under a semi-controlled condition at the Horticulture Department, College of Agricultural Engineering Sciences, University of Sulaimani. The experiment was laid out in RCBD design with six fertigation treatments, each with three replicates. The fertigation treatments comprised of 0, 0.5, 1 and 1.5 g/L of compound fertilizer (NPK+micro-nutrients) with two intervals (1 and 2 weeks). The results indicated that fertigation rate of 1.5 g/L and 1 g/L increased the flower yield and quality of flower significantly. Additionally, the main and interaction effects of fertigation rates and intervals was clearly observed in this study. [ABSTRACT FROM AUTHOR]

Purpose: The study sought to compare the normative amplitude of accommodation (AoA) in school-going children from studies in the 21st century, based on pooled estimates from meta-analyses, to assess their agreement to Hofstetter’s average formula. Methods: A PRISMA checklist was used to conduct the review. PubMed, EBSCOHOST and Medline electronic databases were employed, and hand searching resulting in 259 studies up to July 2021. After title and abstract screening, 12 studies underwent full-text screening, resulting in five studies for data extraction. The pooled effect size was determined using meta-analyses for sub-groups by age. A one-sample t-test was used to compare the pool-effect size estimates (monocular) to the expected AoA from Hofstetter’s average formula. Results: The comparison of pool estimates of AoA with the expected Hofstetter’s average formula for the age sub-groups showed significant mean differences for: six-year olds: mean difference of –3.4 D (95% CI: –5.85; –1.04; p = 0.025); nine-year olds: mean difference of –4.1D (95% CI: –7.95; –0.20; p = 0.043); ten-year olds: mean difference of –4.6D (95% CI: –8.57; –0.54; p = 0.035) and 11-year olds: mean difference of –5.2 D (95% CI: –8.06; –2.40; p = 0.005). According to the quality assessment tool used, overall, the body of evidence was of good quality. Conclusion: Hofstetter’s prediction of normative amplitude of accommodation today may over-estimate for children aged six, nine, 10 and 11. The observed under-accommodation estimates from these comparisons may warrant consideration in assessing for a larger lag of accommodation in these age groups with myopia or pre-myopia, as part of the surveillance for progression.

Angus (A) × Bos indicus (B; Brahman or Nellore) reciprocal backcross, embryo transfer calves belonging to 28full-sib families were evaluated for differences in feedyard initial BW, feedyard final BW, carcass weight, LM area, adjusted fat thickness, intramuscular fat, and Warner-Bratzler shear force. Two methods of analysis were investigated; method I made no distinction between how the F1 parents were produced, whereas method II distinguished the 2 types of F1 parents (AB vs. BA, corresponding to A × B vs. B × A, respectively). No significant reciprocal differences for these weight and carcass traits were detected under method I analyses, although the same trend existed for subsequent BW rankings as for birth weight and weaning weight. For each weight phase, the cross that involved a larger proportion of B in the sire in relation to the amount in the dam (F1 × A and B × F1) ranked heavier than the respective reciprocal cross (A × F1 and F1 × B). As a whole; A backcross calves had larger (P < 0.001) LM area, more (P < 0.001) marbling, and lower (P < 0.001) Warner-Bratzler shear force than B backcross calves, but no consistent trends were detected between reciprocal crosses for any of these traits, in contrast with the trends observed for the weight traits. Furthermore, males were heavier than females entering (P < 0.001) and leaving (P < 0.001) the feedyard, produced a heavier carcass (P < 0.001), and had larger LM area (P < 0.05) with less adjusted fat (P < 0.001). No difference existed between the sexes for Warner-Bratzler shear force or marbling. No interactions involving sex, sire type, and dam type were observed for any of these traits. The results were similar under methods I and II analyses, with the exception that a significant sire type × dam type interaction was observed for initial feedyard BW. Results from this study suggest that for weight-related traits, both the breed constitution of the embryo transfer calf and the cross that produces the calf play an important role in its ultimate performance for B crossbred calves. For body composition and meat-related traits, it appears that the breed makeup of the embryo transfer calf itself is more important to animal performance than the specific cross used to produce the calf. [ABSTRACT FROM AUTHOR]

Not natural reserves in both the goals and objectives; some of which is dedicated strict protection, especially those that have a scientific interest, some of which is dedicated to attracting visitors and tourist activities and the protection of the environment and cultural values, and this is what is called sustainable tourism, which is one of the most important types of tourism; because they are based primarily to protect the ecological balance, and the contemplation of nature, flora and fauna, and provides human comfort; Vtaatgely feature in sustainable tourism in generating opportunities for local parks, and business in the country, while protecting the environmental and cultural values in the protected areas. From this perspective, research the topic of sustainable tourism treated in Talila protected by shedding light on the development of tourism projects, which is a basic need and economic engine of the state and society at the same time, especially in the Syrian desert, as well as the ability of tourism projects to secure a sustainable financial resources, through answer the following questions: - What positive effects of sustainable tourism in nature reserves? - What is the importance of sustainable tourism in the conservation of natural resources? - How sustainable tourism contribute to raising the attention of local and international political and economic circles in reserves?

Background Context: Isolated decompression and decompression with instrumented fusion are accepted surgical treatments for lumbar spondylolisthesis. Although isolated decompression is a less costly solution with similar patient-reported outcomes, it is associated with higher rates of reoperation than primary fusion., Purpose: To determine the costs associated with primary decompression, primary fusion, and decompression and fusion for degenerative spondylolisthesis. We further sought to establish at what revision rate is primary decompression still a less costly surgical treatment for degenerative lumbar spondylolisthesis., Study Design/setting: A retrospective database study of the Medicare Provider Analysis and Review (MEDPAR) limited data set., Patient Sample: Patients who underwent single-level fusion or decompression for degenerative spondylolisthesis., Outcome Measures: Cost of surgical care., Methods: All inpatient stays that underwent surgery for single-level lumbar/lumbosacral degenerative spondylolisthesis in the 2019 calendar year (n=6,653) were queried from the MEDPAR limited data set. Patients were stratified into three cohorts: primary decompression (n=300), primary fusion (n=5,757), and revision fusion (n=566). Univariate analysis was conducted to determine cost differences between these groups and results were confirmed with multivariable regression. An economic analysis was then done to determine at what revision rate would primary decompression still be a less costly treatment choice., Results: on univariate analysis, the cost of primary single-level decompression for spondylolisthesis was $14,690±9,484, the cost of primary single-level fusion was $26,376±11,967, and revision fusion was $26,686±11,309 (p<0.001). on multivariate analysis, primary fusion was associated with an increased cost of $3,751, and revision fusion was associated with increased cost of $7,502 (95%ci: 2,990-4,512, p<0.001). economic analysis found that a revision rate less than or equal to 43.8% would still result in primary decompression being less costly for a practice than primary fusion for all patients., Conclusions: Isolated decompression for degenerative lumbar spondylolisthesis is a less costly treatment choice even with rates of revision fusion as high as 43.8%. This was true even with an assumed revision rate of 0% after primary fusion. This study solely looks at cost data, however, and many patients may still benefit from primary fusion when appropriately indicated., Competing Interests: Declaration of competing interest One or more of the authors declare financial or professional relationships on ICMJE-TSJ disclosure forms., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Peroxisomes are eukaryotic organelles that are essential for multiple metabolic pathways, including fatty acid oxidation, degradation of amino acids, and biosynthesis of ether lipids. Consequently, peroxisome dysfunction leads to pediatric-onset neurodegenerative conditions, including Peroxisome Biogenesis Disorders (PBD). Due to the dynamic, tissue-specific, and context-dependent nature of their biogenesis and function, live cell imaging of peroxisomes is essential for studying peroxisome regulation, as well as for the diagnosis of PBD-linked abnormalities. However, the peroxisomal imaging toolkit is lacking in many respects, with no reporters for substrate import, nor cell-permeable probes that could stain dysfunctional peroxisomes. Here we report that the BODIPY-C12 fluorescent fatty acid probe stains functional and dysfunctional peroxisomes in live mammalian cells. We then go on to improve BODIPY-C12, generating peroxisome-specific reagents, PeroxiSPY650 and PeroxiSPY555. These probes combine high peroxisome specificity, bright fluorescence in the red and far-red spectrum, and fast non-cytotoxic staining, making them ideal tools for live cell, whole organism, or tissue imaging of peroxisomes. Finally, we demonstrate that PeroxiSPY enables diagnosis of peroxisome abnormalities in the PBD CRISPR/Cas9 cell models and patient-derived cell lines., (© 2024. The Author(s).)

Background: Disparities in the utilization of orthopaedic surgery based on race and ethnicity continue to be reported. We examined the impact of sociodemographic factors on treatment recommendation by hand surgeons for carpal tunnel syndrome (CTS) of similar disease severity., Methods: Patients with electrodiagnostic study (EDS)-confirmed CTS were evaluated at a single institution between 2016 and 2020. Data including patient age, sex, race/ethnicity, ZIP Code, and EDS severity were collected. The primary outcome was the recommended treatment by the hand surgeon at the first clinic visit according to patient race/ethnicity and the Social Deprivation Index (SDI). Secondary outcomes included the treatment selected by patients (nonsurgical or surgical) and the time to surgery., Results: The 949 patients had a mean age of 58 years (range, 18 to 80 years); 60.5% (n = 574) were women. The race/ethnicity of the patient cohort was 9.8% (n = 93) Black non-Hispanic, 11.2% (n = 106) Hispanic/Latino, 70.3% (n = 667) White non-Hispanic, and 8.7% (n = 83) "other." Overall, Black non-Hispanic patients (38.7%; odds ratio, [OR] 0.62; 95% confidence interval [CI], 0.40 to 0.96) and Hispanic/Latino patients (35.8%; OR, 0.55; 95% CI, 0.36 to 0.84) were less likely to have surgery recommended at their first visit compared with White non-Hispanic patients (50.5%). This was no longer apparent after adjusting for demographic and clinical variables including EDS severity and SDI (Black non-Hispanic patients: adjusted odds ratio [aOR], 0.67; 95% CI, 0.4 to 1.11; Hispanic/Latino patients: aOR, 0.69: 95% CI, 0.41 to 1.14). Across all categories of EDS severity, surgeons were less likely to recommend surgery to patients with a higher SDI (aOR: 0.66, 0.64, and 0.54 for quintiles 2, 3 and 4, respectively). When surgery was recommended, patients in the highest SDI quintile were less likely to proceed with surgery (p = 0.032). There was no association between patient race/ethnicity and the treatment selected by the patient or time to surgery (p = 0.303 and p = 0.725, respectively)., Conclusions: Patients experiencing higher levels of social deprivation were less likely to be recommended for CTS surgery and were less likely to proceed with surgery, regardless of patient race/ethnicity. Additional investigation into the social factors influencing both surgeon and patient selection of treatment for CTS, including the impact of patient socioeconomic background, is warranted., Level of Evidence: Prognostic Level III . See Instructions for Authors for a complete description of levels of evidence., Competing Interests: Disclosure: The Disclosure of Potential Conflicts of Interest forms are provided with the online version of the article ( http://links.lww.com/JBJS/H607 )., (Copyright © 2023 by The Journal of Bone and Joint Surgery, Incorporated.)

Objective: Despite growing interest in cervical disc replacement (CDR) for conditions such as cervical radiculopathy, limited data exists describing the impact of obesity on early postoperative outcomes and complications. These data are especially important as nearly half of the adult population in the United States is expected to become obese (body mass index [BMI] ≥ 30 kg/m2) by 2030. The goal of this study was to compare the demographics, perioperative variables, and complication rates following CDR., Methods: The 2005-2020 American College of Surgeons National Surgical Quality Improvement Program datasets were queried for patients who underwent primary 1- or 2-level CDR. Patients were divided into 3 cohorts: Nonobese (BMI: 18.5-29.9 kg/m2), Obese class-I (BMI: 30-34.9 kg/m2), Obese class-II/III (BMI ≥ 35 kg/m2). Morbidity was defined as the presence of any complication within 30 days postoperatively. Rates of 30-day readmission, reoperation, morbidity, individual complications, length of stay, frequency of nonhome discharge disposition were collected., Results: A total of 5,397 patients were included for analysis: 3,130 were nonobese, 1,348 were obese class I, and 919 were obese class II/III. There were more 2-level CDRs performed in the class II/III cohort compared to the nonobese group (25.7% vs. 21.5%, respectively; p < 0.05). Class-II/III had more nonhome discharges than class I and nonobese (2.1% vs. 0.5% vs. 0.7%, respectively; p < 0.001). Readmission rates differed as well (nonobese: 0.5%, class I: 1.1%, class II/III: 2.1%; p < 0.001) with pairwise significance between class II/II and nonobese. Class II/III obesity was an independent risk factor for both readmission (odds ratio [OR], 3.32; p = 0.002) and nonhome discharge (OR, 2.51; p = 0.02). Neither 30-day reoperation nor morbidity rates demonstrated significance. No mortalities were reported., Conclusion: Although obese class-II/III were risk factors for 30-day readmission and nonhome discharge, there was no significant difference in reoperation rates or morbidity. CDR procedures can continue to be safely preformed independent of obesity status.

To promote infections, pathogens exploit host cell machineries such as structural elements of the plasma membrane. Studying these interactions and identifying molecular players are ideal for gaining insights into the fundamental biology of the host cell. Here, we used the anthrax toxin to screen a library of 1,500 regulatory, cell-surface, and membrane trafficking genes for their involvement in the intoxication process. We found that endoplasmic reticulum (ER)-Golgi-localized proteins TMED2 and TMED10 are required for toxin oligomerization at the plasma membrane of human cells, an essential step dependent on localization to cholesterol-rich lipid nanodomains. Biochemical, morphological, and mechanistic analyses showed that TMED2 and TMED10 are essential components of a supercomplex that operates the exchange of both cholesterol and ceramides at ER-Golgi membrane contact sites. Overall, this study of anthrax intoxication led to the discovery that lipid compositional remodeling at ER-Golgi interfaces fully controls the formation of functional membrane nanodomains at the cell surface., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Stress granules (SGs) are ribonucleoprotein functional condensates that form under stress conditions in all eukaryotic cells. Although their stress-survival function is far from clear, SGs have been implicated in the regulation of many vital cellular pathways. Consequently, SG dysfunction is thought to be a mechanistic point of origin for many neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). Additionally, SGs are thought to play a role in pathogenic pathways as diverse as viral infection and chemotherapy resistance. There is a growing consensus on the hypothesis that understanding the mechanistic regulation of SG physical properties is essential to understanding their function. Although the internal dynamics and condensation mechanisms of SGs have been broadly investigated, there have been fewer investigations into the timing of SG formation and clearance in live cells. Because the lifetime of SG persistence can be a key factor in their function and tendency toward pathological dysregulation, SG clearance mechanisms deserve particular attention. Here we show that resveratrol and its analogues piceatannol, pterostilbene, and 3,4,5,4'-tetramethoxystilbene induce G3BP-dependent SG formation with atypically rapid clearance kinetics. Resveratrol binds to G3BP, thereby reducing its protein-protein association valency. We suggest that altering G3BP valency is a pathway for the formation of uniquely transient SGs.

The formation of stress granules (SGs) is an essential aspect of the cellular response to many kinds of stress, but its adaptive role is far from clear. SG dysfunction is implicated in aging-onset neurodegenerative diseases, prompting interest in their physiological function. Here, we report that during starvation stress, SGs interact with mitochondria and regulate metabolic remodeling. We show that SG formation leads to a downregulation of fatty acid β-oxidation (FAO) through the modulation of mitochondrial voltage-dependent anion channels (VDACs), which import fatty acids (FAs) into mitochondria. The subsequent decrease in FAO during long-term starvation reduces oxidative damage and rations FAs for longer use. Failure to form SGs, whether caused by the genetic deletion of SG components or an amyotrophic lateral sclerosis (ALS)-associated mutation, translates into an inability to downregulate FAO. Because metabolic dysfunction is a common pathological element of neurodegenerative diseases, including ALS, our findings provide a direction for studying the clinical relevance of SGs., Competing Interests: Declaration of interests D.K. is a co-founder and CSO of 1Base Pharmaceuticals., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Stress granules (SGs) are dynamic condensates associated with protein misfolding diseases. They sequester stalled mRNAs and signaling factors, such as the mTORC1 subunit raptor, suggesting that SGs coordinate cell growth during and after stress. However, the molecular mechanisms linking SG dynamics and signaling remain undefined. We report that the chaperone Hsp90 is required for SG dissolution. Hsp90 binds and stabilizes the dual-specificity tyrosine-phosphorylation-regulated kinase 3 (DYRK3) in the cytosol. Upon Hsp90 inhibition, DYRK3 dissociates from Hsp90 and becomes inactive. Inactive DYRK3 is subjected to two different fates: it either partitions into SGs, where it is protected from irreversible aggregation, or it is degraded. In the presence of Hsp90, DYRK3 is active and promotes SG disassembly, restoring mTORC1 signaling and translation. Thus, Hsp90 links stress adaptation and cell growth by regulating the activity of a key kinase involved in condensate disassembly and translation restoration., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)

Metabolic regulation is a necessary component of all stress response pathways, because all different mechanisms of stress-adaptation place high-energy demands on the cell. Mechanisms that integrate diverse stress response pathways with their metabolic components are therefore of great interest, but few are known. We show that stress granule (SG) formation, a common adaptive response to a variety of stresses, is reciprocally regulated by the pathways inducing lipid droplet accumulation. Inability to upregulate lipid droplets reduces stress granule formation. Stress granule formation in turn drives lipid droplet clustering and fatty acid accumulation. Our findings reveal a novel connection between stress response pathways and new modifiers of stress granule formation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Amen and Kaganovich.)

Photoconversion enables real-time labeling of protein sub-populations inside living cells, which can then be tracked with submicrometer resolution. Here, we detail the protocol of comparing protein dynamics inside membraneless organelles in live HEK293T cells using a CRISPR-Cas9 PABPC1-Dendra2 marker of stress granules. Measuring internal dynamics of membraneless organelles provides insight into their functional state, physical properties, and composition. Photoconversion has the advantage over other imaging techniques in that it is less phototoxic and allows for dual color tracking of proteins. For complete details on the use and execution of this protocol, please refer to Amen and Kaganovich (2020)., Competing Interests: The authors declare no competing interests., (© 2020 The Author(s).)

Vimentin is one of the first cytoplasmic intermediate filaments to be expressed in mammalian cells during embryogenesis, but its role in cellular fitness has long been a mystery. Vimentin is acknowledged to play a role in cell stiffness, cell motility, and cytoplasmic organization, yet it is widely considered to be dispensable for cellular function and organismal development. Here, we show that Vimentin plays a role in cellular stress response in differentiating cells, by recruiting aggregates, stress granules, and RNA-binding proteins, directing their elimination and asymmetric partitioning. In the absence of Vimentin, pluripotent embryonic stem cells fail to differentiate properly, with a pronounced deficiency in neuronal differentiation. Our results uncover a novel function for Vimentin, with important implications for development, tissue homeostasis, and in particular, stress response.

Stress Granule formation has been linked to the resistance of some cancer cells to chemotherapeutic intervention. A number of studies have proposed that certain anti-tumor compounds promote cancer cell survival by inducing Stress Granule formation, leading to increased cellular fitness and apoptosis avoidance. Here we show that a potent fatty acid synthase inhibitor, fasnall, known for its anti-tumor capabilities, triggers the formation of atypical Stress Granules, independently of fatty acid synthase inhibition, characterized by high internal mobility and rapid turnover., Competing Interests: The authors declare no competing interests., (© 2020 The Author(s).)

As the physical barrier between the cell and the outside environment, the plasma membrane is well-positioned to be the first responder to stress. The membrane is also highly vulnerable to many types of perturbation, including heat, force, osmotic pressure, lipid shortage, and starvation. To determine whether the structural changes in the plasma membrane of Saccharomyces cerevisiae brought about by nutrient stress can be communicated to regulatory networks within the cell, we identified proteins that interact with stress granules (SGs), subcellular structures composed of proteins, and nontranslated RNAs that form when cells are stressed. We found that SG proteins interacted with components of eisosomes, which are subcortical membrane structures with a distinct lipid and protein composition. In response to starvation-triggered phosphorylation of eisosome proteins, eisosomes clustered and recruited SG components, including active Pkc1. The absence of eisosomes impaired SG formation, resulting in delayed recovery from nutrient deprivation. Thus, eisosome clustering is an example of interdomain communication in response to stress and identifies a previously unknown mechanism of SG regulation., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Background: Ifosfamide has recently used as the intravenous component of bidirectional intraoperative chemotherapy (BDIC) with hyperthermic intraperitoneal chemotherapy (HIPEC) using cisplatin plus doxorubicin. Little is known about the systemic toxicities of this BDIC regimen. Therefore, this study aimed to assess the toxicities of this treatment., Methods: A prospective, cohort study, of patients who underwent the BDIC using intravenous ifosfamide 1,300 mg/m 2 and a HIPEC regimen of cisplatin 50 mg/m 2 plus doxorubicin 15 mg/m 2 , at King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia. Incidences and severity of leukopenia, neutropenia, thrombocytopenia, and erythrocytopenia were assessed over 45 days after BDIC. Nephrotoxicity was assessed according to the RIFLE (Risk, Injury, Failure, Loss of kidney function, and End-stage kidney disease) classification system. Haemorrhagic cystitis was assessed by cystoscopy., Results: A total of 18 patients were enrolled in the study. Grade 1 leukopenia developed in 11.1% of the patients, with 5.5% developed neutropenia. Thrombocytopenia developed in 61.1% of patients; it was grade 1 or 2 in most patients, but grade 3 in 1 (5.5%) patient. All patients developed erythrocytopenia after BDIC. Leukopenia, neutropenia, and thrombocytopenia resolved without treatment in all patients. Nephrotoxicity developed in 33.3% of the patients. One patient progressed to the End-stage kidney disease classification. No patient developed haemorrhagic cystitis., Conclusions: Intravenous ifosfamide combined with HIPEC using cisplatin plus doxorubicin yielded low rates of mild leukopenia. Mild thrombocytopenia was frequent, but severe suppression of platelets was uncommon. Nephrotoxicity developed in one-third of the patients, and haemorrhagic cystitis was absent., Competing Interests: Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication., (© 2019 Hakeam et al., published by De Gruyter.)

We present a set of vectors containing integrative modules for efficient genome integration into the commonly used selection marker loci of the yeast Saccharomyces cerevisiae . A fragment for genome integration is generated via PCR with a unique set of short primers and integrated into HIS3 , URA3 , ADE2 , and TRP1 loci. The desired level of expression can be achieved by using constitutive ( TEF1p , GPD1p ), inducible ( CUP1p , GAL1/10p ), and daughter-specific ( DSE4p ) promoters available in the modules. The reduced size of the integrative module compared to conventional integrative plasmids allows efficient integration of multiple fragments. We demonstrate the efficiency of this tool by simultaneously tagging markers of the nucleus, vacuole, actin, and peroxisomes with genomically integrated fluorophores. Improved integration of our new pDK plasmid series allows stable introduction of several genes and can be used for multi-color imaging. New bidirectional promoters ( TEF1p-GPD1p , TEF1p-CUP1p , and TEF1p-DSE4p ) allow tractable metabolic engineering., Competing Interests: Conflict of interest: The authors declare no conflict of interest.

Age can be reset during mitosis in both yeast and stem cells to generate a young daughter cell from an aged and deteriorated one. This phenomenon requires asymmetry-generating genes (AGGs) that govern the asymmetrical inheritance of aggregated proteins. Using a genome-wide imaging screen to identify AGGs in Saccharomyces cerevisiae, we discovered a previously unknown role for endocytosis, vacuole fusion, and the myosin-dependent adaptor protein Vac17 in asymmetrical inheritance of misfolded proteins. Overproduction of Vac17 increases deposition of aggregates into cytoprotective vacuole-associated sites, counteracts age-related breakdown of endocytosis and vacuole integrity, and extends replicative lifespan. The link between damage asymmetry and vesicle trafficking can be explained by a direct interaction between aggregates and vesicles. We also show that the protein disaggregase Hsp104 interacts physically with endocytic vesicle-associated proteins, such as the dynamin-like protein, Vps1, which was also shown to be required for Vac17-dependent sequestration of protein aggregates. These data demonstrate that two physiognomies of aging-reduced endocytosis and protein aggregation-are interconnected and regulated by Vac17., (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)

Competing Interests: Conflict of interest: The authors declare no conflict of interest.

Bacteria display an array of contact-dependent interaction systems that have evolved to facilitate direct cell-to-cell communication. We have previously identified a mode of bacterial communication mediated by nanotubes bridging neighboring cells. Here, we elucidate nanotube architecture, dynamics, and molecular components. Utilizing Bacillus subtilis as a model organism, we found that at low cell density, nanotubes exhibit remarkable complexity, existing as both intercellular tubes and extending tubes, with the latter frequently surrounding the cells in a "root-like" fashion. Observing nanotube formation in real time showed that these structures are formed in the course of minutes, displaying rapid movements. Utilizing a combination of super-resolution, light, and electron microscopy, we revealed that nanotubes are composed of chains of membranous segments harboring a continuous lumen. Furthermore, we discovered that a conserved calcineurin-like protein, YmdB, presents in nanotubes and is required for both nanotube production and intercellular molecular trade., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Cells are often exposed to physical or chemical stresses that can damage the structures of essential biomolecules. Stress-induced cellular damage can become deleterious if not managed appropriately. Rapid and adaptive responses to stresses are therefore crucial for cell survival. In eukaryotic cells, different stresses trigger post-translational modification of proteins with the small ubiquitin-like modifier SUMO. However, the specific regulatory roles of sumoylation in each stress response are not well understood. Here, we examined the sumoylation events that occur in budding yeast after exposure to hyperosmotic stress. We discovered by proteomic and biochemical analyses that hyperosmotic stress incurs the rapid and transient sumoylation of Cyc8 and Tup1, which together form a conserved transcription corepressor complex that regulates hundreds of genes. Gene expression and cell biological analyses revealed that sumoylation of each protein directs distinct outcomes. In particular, we discovered that Cyc8 sumoylation prevents the persistence of hyperosmotic stress-induced Cyc8-Tup1 inclusions, which involves a glutamine-rich prion domain in Cyc8. We propose that sumoylation protects against persistent inclusion formation during hyperosmotic stress, allowing optimal transcriptional function of the Cyc8-Tup1 complex.

The human von Hippel-Lindau (VHL) tumor suppressor is a marginally stable protein previously used as a model substrate of eukaryotic refolding and degradation pathways. When expressed in the absence of its cofactors, VHL cannot fold and is quickly degraded by the quality control machinery of the cell. We combined computational methods with in vivo experiments to examine the basis of the misfolding propensity of VHL. By expressing a set of randomly mutated VHL sequences in yeast, we discovered a more stable mutant form. Subsequent modeling suggested the mutation had caused a conformational change affecting cofactor and chaperone interaction, and this hypothesis was then confirmed by additional knockout and overexpression experiments targeting a yeast cofactor homolog. These findings offer a detailed structural basis for the modulation of quality control fate in a model misfolded protein and highlight burial mode modeling as a rapid means to detect functionally important conformational changes in marginally stable globular domains., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Exposing cells to folding stress causes a subset of their proteins to misfold and accumulate in inclusion bodies (IBs). IB formation and clearance are both active processes, but little is known about their mechanism. To shed light on this issue, we performed a screen with over 4,000 fluorescently tagged yeast proteins for co-localization with a model misfolded protein that marks IBs during folding stress. We identified 13 proteins that co-localize to IBs. Remarkably, one of these IB proteins, the uncharacterized and conserved protein Iml2, exhibited strong physical interactions with lipid droplet (LD) proteins. Indeed, we here show that IBs and LDs are spatially and functionally linked. We further demonstrate a mechanism for IB clearance via a sterol-based metabolite emanating from LDs. Our findings therefore uncover a function for Iml2 and LDs in regulating a critical stage of cellular proteostasis., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Neurodegenerative diseases and other proteinopathies constitute a class of several dozen illnesses etiologically linked to pathological protein misfolding and aggregation. Because of this strong association with disease pathology, cell death, and aging, accumulation of proteins in aggregates or aggregation-associated structures (inclusions) has come to be regarded by many as a deleterious process, to be avoided if possible. Recent work has led us to see inclusion structures and disordered aggregate-like protein mixtures (which we call dynamic droplets) in a new light: not necessarily as a result of a pathological breakdown of cellular order, but as an elaborate cellular architecture regulating function and stress response. In this review, we discuss what is currently known about the role of inclusion structures in cellular homeostasis, stress response, toxicity, and disease. We will focus on possible mechanisms of aggregate toxicity, in contrast to the homeostatic function of several inclusion structures.

Aging is associated with the accumulation of several types of damage: in particular, damage to the proteome. Recent work points to a conserved replicative rejuvenation mechanism that works by preventing the inheritance of damaged and misfolded proteins by specific cells during division. Asymmetric inheritance of misfolded and aggregated proteins has been shown in bacteria and yeast, but relatively little evidence exists for a similar mechanism in mammalian cells. Here, we demonstrate, using long-term 4D imaging, that the vimentin intermediate filament establishes mitotic polarity in mammalian cell lines and mediates the asymmetric partitioning of damaged proteins. We show that mammalian JUNQ inclusion bodies containing soluble misfolded proteins are inherited asymmetrically, similarly to JUNQ quality-control inclusions observed in yeast. Mammalian IPOD-like inclusion bodies, meanwhile, are not always inherited by the same cell as the JUNQ. Our study suggests that the mammalian cytoskeleton and intermediate filaments provide the physical scaffold for asymmetric inheritance of dynamic quality-control JUNQ inclusions. Mammalian IPOD inclusions containing amyloidogenic proteins are not partitioned as effectively during mitosis as their counterparts in yeast. These findings provide a valuable mechanistic basis for studying the process of asymmetric inheritance in mammalian cells, including cells potentially undergoing polar divisions, such as differentiating stem cells and cancer cells.