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Nevin Manimala Statistics

Construction of Pseudomonas aeruginosa SDK-6 with synthetic lipase gene cassette and optimization of different parameters using response surface methodology for over-expression of recombinant lipase

Folia Microbiol (Praha). 2024 May 3. doi: 10.1007/s12223-024-01167-y. Online ahead of print.

ABSTRACT

Lipases are industrially important enzymes having vast applications in various fields. Cloning and expression of lipase enzyme-encoding genes in suitable host lead to their widespread use in different fields. The present study represents the first attempt towards the expression of the synthetic lipase gene in Pseudomonas aeruginosa. An alkalophilic lipase gene (GenBank accession number: NP_388152) from Bacillus subtilis was synthetically designed and introduced in the pJN105 vector and subsequently cloned in Pseudomonas aeruginosa SDK-6. Agarose gel electrophoresis confirmed the transformation of SDK-6, exhibiting a band difference of ~ 700 bp between native and recombinant pJN105. Further amplification of cloned lipase gene was confirmed using PCR amplification with Lip 1 and Lip 2 primers respectively, followed by restriction analysis. Approximately 15-fold increase in lipase production was observed in recombinant Pseudomonas as compared to the native strain. One factor at a time (OFAT) analysis revealed L-arabinose, inoculum size (0.5%; v/v), and agitation (120 rpm) as significant factors affecting the over-expression of lipase enzyme. Optimization of enzyme induction conditions by central composite design (CCD) led to 1.60-fold increase in the production of lipase at 0.65% (w/v) inducer concentration, OD600-1.075 before induction and 35 °C post induction temperature with overall lipase production of 50.50 IU/mL. Statistical validation of observed value via ANOVA showed an F-value of 138.70 at p < 0.01 with R2 of 0.9921.

PMID:38700831 | DOI:10.1007/s12223-024-01167-y

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Nevin Manimala Statistics

Evaluation of a prehospital endovascular therapy stroke bypass program

CJEM. 2024 May 3. doi: 10.1007/s43678-024-00685-5. Online ahead of print.

ABSTRACT

INTRODUCTION: Prehospital stroke endovascular therapy bypass transports patients with suspected large vessel occlusion directly to an endovascular therapy capable center. Our objective was to determine if an endovascular therapy bypass protocol improved access to stroke treatments. Secondary objectives were to determine safety, effectiveness, and rate of subsequent interfacility transfers.

METHODS: Endovascular therapy bypass in 2018 was implemented in Eastern Ontario, for patients with a Los-Angeles-Motor-Scale ≥ 4 (positive large vessel occlusion screen) with a 90-min transport time if < 6 h from last seen well. A before-after health record review was conducted from Dec 1, 2017 to Nov 30, 2019. A piloted data form was used to extract demographics, times, primary outcomes (endovascular therapy and intravenous (IV) tissue plasminogen activator (tPA) rate), and secondary outcomes (redirect to closer hospital, airway intervention, and subsequent interfacility transfer). We present descriptive statistics and odds ratios (OR) with 95% confidence intervals (CI) from multivariable logistic regression.

RESULTS: We included 379 stroke patients (165 pre and 214 post-implementation). The endovascular therapy rate between groups was similar (14.1% vs 15.1%). The bypass had an OR of 0.98 (95% CI 0.54-1.78) for receiving endovascular therapy. IV tPA was given to 25.4% of patients pre vs 27.4% post-implementation (OR 1.06, 95% CI 0.65-1.74). No patients became unstable during transport, only one patient had an intubation attempt. The inappropriate bypass (false positive) rate was 12.7% pre vs 12.8% post-implementation (positive predictive value 87%). The bypass protocol had an OR of 1.06 (95% CI 0.58-1.95) for subsequent interfacility transfer with a mean of 2.7 h at the community site before transfer.

CONCLUSIONS: Endovascular therapy stroke bypass with 90-min transport radius and Los-Angeles-Motor-Scale ≥ 4 was safe and well executed by paramedics. Our study did not show any difference in endovascular therapy rate from its implementation. The IV tPA rate was similar between groups despite potentially bypassing thrombolysis capable centers.

PMID:38700785 | DOI:10.1007/s43678-024-00685-5

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Nevin Manimala Statistics

Association between serum levels of GDF-15, suPAR, PIVKA-II, sdLDL and clinical outcomes in hospitalized COVID-19 patients

Intern Emerg Med. 2024 May 3. doi: 10.1007/s11739-024-03630-7. Online ahead of print.

ABSTRACT

To quantify the circulating levels of novel serum biomarkers including GDF-15, PIVKA-II, sdLDL, suPAR, and of CRP in hospitalized COVID-19 patients compared with healthy subjects, and to evaluate their association(s) with outcomes in COVID-19. We considered patients with confirmed COVID-19, hospitalized in an Internal Medicine ward. The clinical characteristics were collected, including the number and type of comorbidities. Serum levels of GDF-15, PIVKA-II, suPAR, sdLDL, as well as CRP were measured. As outcomes, we considered Intensive Care Unit (ICU) transfer or death, as well as the length of stay (days) and in-hospital complications. Data were statistically analyzed, as appropriate, and a p value < 0.05 was considered significant. Ninety-three patients and 20 healthy controls were enrolled. COVID-19 patients vs. controls showed higher median levels of GDF-15 (p < 0.0001), PIVKA-II (p < 0.0001) and sdLDL (p = 0.0002), whereas no difference was observed for suPAR. In COVID-19 patients, the most frequent comorbidities were arterial hypertension (62.4%) and cardiovascular disease (30.1%). GDF-15 levels positively correlated with age (r = 0.433, p < 0.0001), and this correlation was confirmed for suPAR (r = 0.308, p = 0.003) and CRP (Rho = 0.40 p < 0.0001), but not for PIVKA-II and sdLDL. Higher GDF-15 levels were associated with a higher number of comorbidities (p = 0.021). The median length of stay was 22 (15; 30) days. During hospitalization, 15 patients (16%) were ICU transferred, and 6 (6.45%) died. GDF-15 serum levels correlated with the length of stay (rho = 0.27 p = 0.010), and were associated with ICU transfer or death (p = 0.003), as well as PIVKA-II (p = 0.038) and CRP (p < 0.001). Moreover, higher GDF-15 and PIVKA-II serum levels were associated with infectious complications (p = 0.008 and p = 0.017, respectively). In this cohort of hospitalized COVID-19 patients, novel inflammatory biomarkers, including GDF-15, suPAR and PIVKA II were associated with some patient’s clinical characteristics, complications, and poor outcomes.

PMID:38700782 | DOI:10.1007/s11739-024-03630-7

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Nevin Manimala Statistics

Comparison of perioperative and functional outcomes of single-incision versus standard multi-incision robot-assisted laparoscopic radical prostatectomy: a prospective, controlled, nonrandomized trial

J Robot Surg. 2024 May 3;18(1):195. doi: 10.1007/s11701-024-01962-2.

ABSTRACT

To compare perioperative and functional outcomes between improved (port-free) single-site robot-assisted laparoscopic radical prostatectomy (pf-ssRARP) and standard multi-port robot-assisted radical prostatectomy (MPRARP). A total of 372 consecutive patients underwent RARAP using the da Vinci Si® robotic surgical system. Group I (n = 210) included patients undergoing pf-ssRARP and Group II (n = 162) included patients undergoing MPRARP. Demographics and perioperative data including postoperative recovery outcomes were recorded and compared between the two groups. Overall mean operative time was significantly shorter with the pf-ssRARP compared to the MPRARP (p < 0.05). The length of hospitalization after the pf-ssRARP was shorter (p < 0.05). In Group I, the positive surgical margin rate was 15.2%; while in Group II, the positive margin rate was 33.3% (p < 0.05). The rate of instant urinary continence was significantly higher in Group I than in Group II (p < 0.05). The percentage of urinary continence was higher in the pf-ssRARP than in the MPRARP, at 6 months post-surgery (p < 0.05) and 9 months post-surgery (p < 0.05). There was no significant difference in the proportion of erectile function in the pf-ssRARP and MPRARP groups at the time of reaching the endpoint of this study (p > 0.05). The two groups were comparable in terms of total hospitalization costs (p < 0.05). The improved (port-free) single-site robot-assisted laparoscopic radical prostatectomy is a practical and easy technique to implement in clinical practice. Extraperitoneal implementation of the modified technique requires only a small incision, no special PORT, no additional auxiliary foramen creation, increased postoperative aesthetics and reduced hospitalization costs, and a high percentage of early postoperative urinary control recovery.

PMID:38700764 | DOI:10.1007/s11701-024-01962-2

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Nevin Manimala Statistics

Evolution of Cooperation in Spatio-Temporal Evolutionary Games with Public Goods Feedback

Bull Math Biol. 2024 May 3;86(6):67. doi: 10.1007/s11538-024-01296-y.

ABSTRACT

In biology, evolutionary game-theoretical models often arise in which players’ strategies impact the state of the environment, driving feedback between strategy and the surroundings. In this case, cooperative interactions can be applied to studying ecological systems, animal or microorganism populations, and cells producing or actively extracting a growth resource from their environment. We consider the framework of eco-evolutionary game theory with replicator dynamics and growth-limiting public goods extracted by population members from some external source. It is known that the two sub-populations of cooperators and defectors can develop spatio-temporal patterns that enable long-term coexistence in the shared environment. To investigate this phenomenon and unveil the mechanisms that sustain cooperation, we analyze two eco-evolutionary models: a well-mixed environment and a heterogeneous model with spatial diffusion. In the latter, we integrate spatial diffusion into replicator dynamics. Our findings reveal rich strategy dynamics, including bistability and bifurcations, in the temporal system and spatial stability, as well as Turing instability, Turing-Hopf bifurcations, and chaos in the diffusion system. The results indicate that effective mechanisms to promote cooperation include increasing the player density, decreasing the relative timescale, controlling the density of initial cooperators, improving the diffusion rate of the public goods, lowering the diffusion rate of the cooperators, and enhancing the payoffs to the cooperators. We provide the conditions for the existence, stability, and occurrence of bifurcations in both systems. Our analysis can be applied to dynamic phenomena in fields as diverse as human decision-making, microorganism growth factors secretion, and group hunting.

PMID:38700758 | DOI:10.1007/s11538-024-01296-y

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Nevin Manimala Statistics

The Reliability and Quality of YouTube Videos About Isotretinoin Treatment and Suicide

Clin Exp Dermatol. 2024 May 3:llae166. doi: 10.1093/ced/llae166. Online ahead of print.

ABSTRACT

BACKGROUND: Seeking health information online has drastically increased. Isotretinoin is one of the agents used to treat acne.

OBJECTIVES: In this study, we aimed to investigate the reliability and quality of YouTube videos related to acne treatment, which have been misconception to increase suicide.

METHODS: We used the terms “isotretinoin suicide” and “acne treatment suicide” to search YouTube videos. Videos that were not in English, irrelevant, or devoid of audio were not included. The information in the videos was primarily categorized as reliable or unreliable based on its scientific validation. DISCERN and the Global Quality Score were used to evaluate the videos’ overall quality.

RESULTS: 200 videos in total were examined. 112 videos were included in the study. 39 videos (34.8 %) were found to be reliable, and 73 videos (65.2%) were found to be unreliable. DISCERN values of videos uploaded by physicians or professional organizations and health information websites were found to be significantly higher. There was a statistically significant negative correlation between DISCERN score and video length and the length of time the video was on YOUTUBE, while a positive correlation was observed between DISCERN score and subscriber.

CONCLUSION: Although videos created by dermatologists have become widespread in recent years, it is still insufficient. Patient experience videos mostly contain information which does not reflect the reality and emphasize that isotretinoin increases the risk of suicide without evidence. As the number of dermatologists posting videos on YouTube increases, the chances of people accessing correct information will increase.

PMID:38699954 | DOI:10.1093/ced/llae166

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Nevin Manimala Statistics

Simultaneous automated ascertainment of prevalent vertebral fracture and abdominal aortic calcification in clinical practice: role in fracture risk assessment

J Bone Miner Res. 2024 May 3:zjae066. doi: 10.1093/jbmr/zjae066. Online ahead of print.

ABSTRACT

Whether simultaneous automated ascertainments of prevalent vertebral fracture (auto-PVFx) and abdominal aortic calcification (auto-AAC) on vertebral fracture assessment (VFA) lateral spine bone density (BMD) images jointly predict incident fractures in routine clinical practice is unclear. We estimated the independent associations of auto-PVFx and auto-AAC primarily with incident major osteoporotic and secondarily with incident hip and any clinical fractures in 11 013 individuals (mean [SD] age 75.8 [6.8] years, 93.3% female) who had a BMD test combined with VFA between March 2010 and December 2017. Auto-PVFx and auto-AAC were ascertained using convolutional neural networks (CNNs). Proportional hazards models were used to estimate the associations of auto-PVFx and auto-AAC with incident fractures over a mean (SD) follow-up of 3.7 (2.2) years, adjusted for each other and other risk factors. At baseline, 17% (n = 1881) had auto-PVFx and 27% (n = 2974) had a high level of auto-AAC (≥ 6 on scale of 0 to 24). Multivariable-adjusted hazard ratios (HR) for incident major osteoporotic fracture (95% C.I.) were 1.85 (1.59, 2.15) for those with compared to those without auto-PVFx, and 1.36 (1.14, 1.62) for those with high compared to low auto-AAC. The multivariable-adjusted HRs for incident hip fracture were 1.62 (95% C.I. 1.26 to 2.07) for those with compared to those without auto-PVFx, and 1.55 (95% C.I. 1.15 to 2.09) for those high auto-AAC compared to low auto-AAC. The 5-year cumulative incidence of major osteoporotic fracture was 7.1% in those with no auto-PVFx and low auto-AAC, 10.1% in those with no auto-PVFx and high auto-AAC, 13.4% in those with auto-PVFx and low auto-AAC, and 18.0% in those with auto-PVFx and high auto-AAC. While physician manual review of images in clinical practice will still be needed to confirm image quality and provide clinical context for interpretation, simultaneous automated ascertainment of auto-PVFx and auto-AAC can aid fracture risk assessment.

PMID:38699950 | DOI:10.1093/jbmr/zjae066

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Nevin Manimala Statistics

Resistance to torsion of cement vs screw-retained abutments under a tangential load: A pilot study

J Oral Implantol. 2024 May 3. doi: 10.1563/aaid-joi-D-23-00051. Online ahead of print.

ABSTRACT

AIM: To compare the resistance to torsion between two implant systems with internal hexagon connection, one using screw-retained abutments (Titanium Fix) and the other using cementable abutments (ITEC) under a tangential load.

MATERIALS AND METHODS: An in vitro experimental study was carried out. Fourteen implants, seven implants from each system, were included in this study. The implants were placed at a 45-degree angle into metal blocks to simulate their position in the maxilla. Then, implants were subjected to a resistance test on a CMT5L universal testing machine, and the maximum load was applied to each sample. The maximum force with which the torsion was achieved in each sample was analyzed. The data were tested using the Shapiro-Wilk test and showed normal distribution. Student t-test was used to examine statistical significance between the two groups, and the p-value was set at P < 0.05.

RESULTS: There was a statistically significant difference between the two groups (P = 0.001). ITEC implants with a cementable abutment showed greater flexural strength compared to the Titanium fix with a screwed abutment implant system.

CONCLUSIONS: The cemented abutment showed more resistance to torsion against a tangential load in comparison with the screwed abutment.

PMID:38699942 | DOI:10.1563/aaid-joi-D-23-00051

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Health complaints before and at one and five years after removal of dental amalgam restorations – data from a prospective cohort study in Norway

Acta Odontol Scand. 2024 May 3;83:219-229. doi: 10.2340/aos.v83.40260.

ABSTRACT

OBJECTIVE: Health complaints attributed to dental amalgam fillings comprise both intraoral and general health complaints. There are data suggesting that patients with medically unexplained physical symptoms (MUPS) attributed to amalgam fillings show improvement in symptoms after removal of all amalgam fillings. However, data indicating changes of specific health complaints are limited. This study evaluated the changes of health complaints after removal of amalgam restorations in patients with health complaints attributed to dental amalgam fillings.

METHOD: Patients with MUPS attributed to dental amalgam (Amalgam cohort) had all their amalgam fillings removed. The participants indicated an intensity of 11 local and 12 general health complaints on numeric rating scales before the treatment and at follow-up after 1 and 5 years. The comparison groups comprising a group of healthy individuals and a group of patients with MUPS without symptom attribution to dental amalgam did not have their amalgam restorations removed.

RESULTS: In the Amalgam cohort, mean symptom intensity was lower for all 23 health complaints at follow-up at 1 year compared to baseline. Statistically significant changes were observed for specific health complaints with effect sizes between 0.36 and 0.68. At the 5-year follow-up, the intensity of symptoms remained consistently lower compared to before the amalgam removal. In the comparison groups, no significant changes of intensity of symptoms of health complaints were observed.

CONCLUSION: After removal of all amalgam restorations, both local and general health complaints were reduced. Since blinding of the treatment was not possible, specific and non-specific treatment effects cannot be separated.

PMID:38699922 | DOI:10.2340/aos.v83.40260

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Use of spectroscopic process analytical technology for rapid quality evaluation during preparation of CHO cell culture media

Biotechnol Prog. 2024 May 3:e3477. doi: 10.1002/btpr.3477. Online ahead of print.

ABSTRACT

Media preparation parameters contribute significantly to media quality, cell culture performance, productivity, and product quality. Establishing proper media preparation procedures is critical for ensuring a robust CHO cell culture process. Process analytical technology (PAT) enables unique ways to quantify assessments and improve media quality. Here, cell culture media were prepared under a wide range of temperatures (40-80°C) and pH (7.6-10.0). Media quality profiles were compared using three real-time PATs: Fourier-transform infrared (FTIR) spectroscopy, Raman spectroscopy, and excitation-emission matrix (EEM) spectroscopy. FTIR and Raman spectroscopies identified shifts in media quality under high preparation temperature (80°C) and at differing preparation pH which negatively impacted monoclonal antibody (mAb) production. In fed-batch processes for production of three different mAbs, viable cell density (VCD) and cell viability were mostly unaffected under all media preparation temperatures, while titer and cell specific productivity of mAb decreased when cultured in basal and feed media prepared at 80°C. High feed preparation pH alone was tolerated but cell growth and productivity profiles deviated from the control condition. Further, charge variants (main, acidic, basic species) and glycosylation (G0F, afucosylation, and high mannose) were examined. Statistically significant differences were observed for one or more of these quality attributes with any shifts in media preparation. In this study, we demonstrated strong associations between media preparation conditions and cell growth, productivity, and product quality. The rapid evaluation of media by PAT implementation enabled more comprehensive understanding of different parameters on media quality and consequential effects on CHO cell culture.

PMID:38699906 | DOI:10.1002/btpr.3477