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Synthesis, characterization and properties investigation of various solid forms of an anti-glaucoma drug – dichlorphenamide.

Focusing on a reliable supramolecular synthon approach, novel binary and ternary molecular adducts of the antihypertensive drug ketanserin (KTS) with diverse aromatic carboxylic acid derivatives are reported.

This study aimed to estimate the prevalence of depression, anxiety, post-traumatic stress disorder, and insomnia symptoms among frontline health workers during the corona virus disease 19 (COVID-19) pandemic in Bangladesh and to compare these between medical and allied health workers. We conducted a cross-sectional survey between June and August 2020. A total of 479 health care professionals participated in the study. Anxiety and depression were measured using Patient Health Questionnaire-4 (PHQ-4), post-traumatic stress disorder (PTSD) was measured by primary care post-traumatic stress disorder score (PC-PTSD score), and insomnia was measured by Insomnia Severity Index (ISI). We performed logistic regression to assess risk factors associated with psychological symptoms. Overall, prevalence of anxiety, depression, PTSD and insomnia symptoms were 17.6%, 15.5%, 7.6% and 5.9%, respectively. Compared to allied health workers, doctors had significantly higher prevalence of symptoms of anxiety: doctor versus allied health care worker; 21.1% vs 06%, (OR= 4.19; 95% CI= 1.88-9.35; p

SUMMARYCell size and the cell cycle are intrinsically coupled and abnormal increases in cell size are associated with senescence. The mechanism by which overgrowth primes cells to exit the cell cycle remains unclear. We investigate this using CDK4/6 inhibitors that arrest cell cycle progression in G0/G1 and are used to treat ER+/HER2-metastatic breast cancer. We demonstrate that long-term CDK4/6 inhibition promotes cellular overgrowth during the G0/G1 arrest, causing widespread proteome remodeling and p38-p53-p21-dependent cell cycle exit. Cell cycle exit is triggered by two waves of p21 induction. First, overgrowth during a G0/G1 arrest induces an osmotic stress response, producing the first wave of p21 induction. Second, when CDK4/6 inhibitors are removed, a fraction of cells escape G0/G1 arrest and enter S-phase where overgrowth-driven replication stress results in a second wave of p21 induction that causes cell cycle withdrawal from G2, or the subsequent G1. This could explain why cellular hypertrophy is associated with senescence and why CDK4/6 inhibitors have long-lasting anti-proliferative effects in patients.

AimsFrontline health care workers exposed to COVID-19 patients could be at increased risk of developing psychological issues. The study aimed to estimate the prevalence of mental health-related problems, specifically depression, anxiety, post-traumatic stress disorder (PTSD), and insomnia among health care professionals during the COVID-19 pandemic in Bangladesh and to compare these between medical and allied health care professionals.MethodThis cross-sectional survey was conducted using Google Form then subsequent telephone interview between June and August 2020. Using random sampling, a total of 479 health care professionals participated in the study. We collected data on demographics. Anxiety and depression were measured using 4 items Patient Health Questionnaire-4 (PHQ-4), PTSD was measured using 4 items Primary Care (PC)-PTSD-Screen, and insomnia was measured by using a 7-item Insomnia Severity Index (ISI). A multivariable logistic regression analysis was performed to assess risk factors associated with mental health symptoms.ResultOverall, 17.6% of frontline health workers had symptoms of anxiety, 15.5% had depression symptoms, 7.6% had PTSD symptoms and 5.9% had symptoms of insomnia. Compared to allied health professionals (n = 113, 24%), doctors (n = 366, 76%) had significantly higher prevalence of anxiety: 21.1% vs 06%, (OR = 4.19; 95% CI = 1.88–9.35; p-value ConclusionOur study demonstrated a high prevalence of symptoms of depression, anxiety, PTSD, and insomnia among Bangladeshi frontline health workers (particularly among doctors) during the COVID-19 pandemic. There is an urgent need to address the mental health needs of frontline health workers.Funding: Medical Research Council, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh.

Melt curves of the final selected primer. (PDF 861 kb)

Primer pair efficiency. (PDF 211 kb)

Gel image of all the RNA samples used in the study. (DOCX 17394 kb)

E487K point mutation of aldehyde dehydrogenase (ALDH) 2 (ALDH2*2) in East Asians intrinsically lowers ALDH2 activity. ALDH2*2 is associated with diabetic cardiomyopathy. Diabetic patients exhibit heart failure of preserved ejection fraction (HFpEF) i.e. while the systolic heart function is preserved in them, they may exhibit diastolic dysfunction, implying a jeopardized myocardial health. Currently, it is challenging to detect cardiac functional deterioration in diabetic mice. Stress echocardiography (echo) in the clinical set-up is a procedure used to measure cardiac reserve and impaired cardiac function in coronary artery diseases. Therefore, we hypothesized that high-fat diet fed type-2 diabetic ALDH2*2 mutant mice exhibit HFpEF which can be measured by cardiac echo stress test methodology. We induced type-2 diabetes in 12-week-old male C57BL/6 and ALDH2*2 mice through a high-fat diet. At the end of 4 months of DM induction, we measured the cardiac function in diabetic and control mice of C57BL/6 and ALDH2*2 genotypes by conscious echo. Subsequently, we imposed exercise stress by allowing the mice to run on the treadmill until exhaustion. Post-stress, we measured their cardiac function again. Only after treadmill running, but not at rest, we found a significant decrease in % fractional shortening and % ejection fraction in ALDH2*2 mice with diabetes compared to C57BL/6 diabetic mice as well as non-diabetic (control) ALDH2*2 mice. The diabetic ALDH2*2 mice also exhibited poor maximal running speed and distance. Our data suggest that high-fat fed diabetic ALDH2*2 mice exhibit HFpEF and treadmill exercise stress echo test is able to determine this HFpEF in the diabetic ALDH2*2 mice.

The migration of chromosomes during mitosis is mediated primarily by kinesins that bind to the chromosomes and move along the microtubules, exerting pulling and pushing forces on the centrosomes. We report that a DNA replication protein, Sld5, localizes to the centrosomes, resisting the microtubular pulling forces experienced during chromosome congression. In the absence of Sld5, centriolar satellites, which normally cluster around the centrosomes, are dissipated throughout the cytoplasm, resulting in the loss of their known function of recruiting the centrosomal protein, pericentrin. We observed that Sld5-deficient centrosomes lacking pericentrin were unable to endure the CENP-E- and Kid-mediated microtubular forces that converge on the centrosomes during chromosome congression, resulting in monocentriolar and acentriolar spindle poles. The minus-end-directed kinesin-14 motor protein, HSET, sustains the traction forces that mediate centrosomal fragmentation in Sld5-depleted cells. Thus, we report that a DNA replication protein has an as yet unknown function of ensuring spindle pole resistance to traction forces exerted during chromosome congression.

[This corrects the article DOI: 10.1371/journal.pone.0195796.].

In the paper we establish some new results depending on the comparative growth properties of composite entire or meromorphic functions using generalised pL*-type with rate pand generalised pL*-weak type with rate p and wronskians generated by one of the factors.

The primary eukaryotic single-stranded DNA-binding protein, Replication protein A (RPA), binds to single-stranded DNA at the sites of DNA damage and recruits the apical checkpoint kinase, ATR via its partner protein, ATRIP. It has been demonstrated that absence of RPA incapacitates the ATR-mediated checkpoint response. We report that in the absence of RPA, human single-stranded DNA-binding protein 1 (hSSB1) and its partner protein INTS3 form sub-nuclear foci, associate with the ATR-ATRIP complex and recruit it to the sites of genomic stress. The ATRIP foci formed after RPA depletion are abrogated in the absence of INTS3, establishing that hSSB-INTS3 complex recruits the ATR-ATRIP checkpoint complex to the sites of genomic stress. Depletion of homologs hSSB1/2 and INTS3 in RPA-deficient cells attenuates Chk1 phosphorylation, indicating that the cells are debilitated in responding to stress. We have identified that TopBP1 and the Rad9-Rad1-Hus1 complex are essential for the alternate mode of ATR activation. In summation, we report that the single-stranded DNA-binding protein complex, hSSB1/2-INTS3 can recruit the checkpoint complex to initiate ATR signaling.

When mammalian cells experience radiation insult, DNA replication is stalled to prevent erroneous DNA synthesis. UV-irradiation triggers proteolysis of Mcm10, an essential human replication factor, inhibiting the ongoing replication. Here, we report that Mcm10 associates with E3 ubiquitin ligase comprising DNA damage-binding protein, DDB1, cullin, Cul4 and ring finger protein, Roc1. Depletion of DDB1, Roc1 or Cul4 abrogates the UV-triggered Mcm10 proteolysis, implying that Cul4-Roc1-DDB1 ubiquitin ligase mediates Mcm10 downregulation. The purified Cul4-Roc1-DDB1 complex ubiquitinates Mcm10 in vitro, proving that Mcm10 is its substrate. By screening the known DDB1 interacting proteins, we discovered that VprBP is the substrate recognition subunit that targets Mcm10 for degradation. Hence, these results establish that Cul4-DDB1-VprBP ubiquitin ligase mediates the stress-induced proteolysis of replication factor, Mcm10.

Objectives of this study were to investigate the characteristics of metabolic acidosis associated with rotavirus gastroenteritis and to explore them as a tool in the management of dehydration. The study was retrospective, case record based and conducted in a secondary level District General Hospital. Included study population were 133 children who were admitted to the Paediatric Unit with rotavirus gastroenteritis. Degree of dehydration, result of blood gas analysis, C-reactive protein level, urinary ketones, renal function tests, fluid management and duration of hospitalization were recorded and results compared between patients with and without severe metabolic acidosis (serum bicarbonate < 17 mmol/L). Out of 133 patients, blood gas analysis was obtained in 78 (59%) and 73 (94%) of those showed metabolic acidosis (bicarbonate < 22 mmol/L). Thirty five patients developed severe metabolic acidosis. Patients with severe metabolic acidosis showed signs of dehydration more commonly (97% vs. 74%, p < 0.05) and required intravenous rehydration more frequently (94% vs. 63%, p < 0.05) than those who were not severely acidotic. With respect to gender ratio, initial temperature, serum levels of C-reactive protein, chloride, anion-gap, lactate and ketonuria, there were no significant differences between the severely acidotic patients and those who were not. Urea and creatinine levels were higher (46.6 (11.7) versus 37.8 (11.9) micromole/L for creatinine and 5.5 (1.9) versus 4.2 (2.1) mmol/L for urea, p < 0.05) in patients with severe metabolic acidosis and correlated significantly negatively (for both: r = − 0.29, p = 0.02) with serum bicarbonate levels. We concluded that the majority of children attending hospital with rotavirus gastroenteritis had a metabolic acidosis. Severe metabolic acidosis was associated with more severe dehydration and higher urea and creatinine levels and increased requirement for intravenous fluid boluses and intravenous rehydration. A bicarbonate level of > 17 mmol/L excluded severe clinical dehydration and below this level a degree of dehydration was very likely.

When mammalian cells experience DNA damaging stress, they block DNA replication to avoid erroneous replication of the damaged template. The cells that are unable to respond to DNA damage continue faulty DNA replication that results in incorporation of genomic lesions. To understand the regulation of replication machinery during stress, systemic studies have been carried out but they have been restricted to the evaluation of the mRNA levels and therefore have not been able to identify post-transcriptional changes, vital for immediate blocking of the progressing DNA replication. We have recently discovered that an essential replication factor is downregulated by radiation stress. In this study, we have carried out a systematic evaluation of protein levels of entire replication apparatus after different types of DNA damage. We report that, independent of the status of p53 and retinoblastoma protein, mammalian cells choose targets that are essential for prereplication, preinitiation, and elongation phases of replication. We imposed different kinds of stress to discern whether similar or unique responses are invoked, and we propose a model for inhibition of replication machinery in which mammalian cells target specific essential replication factors based on the experienced stress. © 2010 IUBMB IUBMB Life, 62(10): 764–775, 2010

Background Mcm10 protein is essential for initiation and elongation phases of replication. Human cells proteolyze Mcm10 during mitosis, presumably to ensure a single round of replication. It has been proposed that anaphase promoting complex ubiquitinates Mcm10 in late M and early G1 phases. Results In contrast to the previous work, we report that the degradation of Mcm10 is initiated at the onset of mitosis. Immunoblotting and immunofluorescence assays display that Mcm10 levels are low in all phases of mitosis. We report that Mcm10 degradation is not dependent on anaphase promoting complex. Further, the proteolysis in M-phase can be independently mediated by non-overlapping regions of Mcm10, apparently employing a redundant mechanism to ensure downregulation. Conclusions It is believed that the proteolysis of Mcm10 during mitosis is a vital mechanism to prevent aberrant initiation of replication and the present study describes the regulation of Mcm10 during this phase of the cell-cycle.

We report that upon UV radiation insult, mammalian cells specifically down-regulate Mcm10, a protein essential for the initiation and elongation phases of DNA replication. The levels of a majority of replication factors remain unaffected under this condition, implying that Mcm10 is a key node in the regulation of the replication machinery. High doses of ionizing gamma radiation and exposure to a combination of DNA-damaging chemicals do not decrease Mcm10 protein levels, demonstrating that Mcm10 down-regulation is triggered only by UV-specific damage. The decrease of Mcm10 protein levels is not caused by transcriptional inhibition or cleavage by apoptotic enzymes, but results from degradation by the 26 S proteasome. UV-triggered degradation of Mcm10 requires its linker or C-terminal domain. In addition, Mcm10 down-regulation is not limited to cells from a particular lineage. Therefore, our study reveals a mechanism by which mammalian cells effectively inhibit the replication machinery during stress to prevent it from drifting toward a catastrophic path of genomic instability.