Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.

Validity evidence for an instrument for cognitive load for virtual didactic sessions

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Background: COVID necessitated the shift to virtual resident instruction. The challenge of learning via virtual modalities has the potential to increase cognitive load. It is important for educators to reduce cognitive load to optimize learning, yet there are few available tools to measure cognitive load. The objective of this study is to identify and provide validity evidence following Messicks’ framework for an instrument to evaluate cognitive load in virtual emergency medicine didactic sessions.
Methods: This study followed Messicks’ framework for validity including content, response process, internal structure, and relationship to other variables. Content validity evidence included: (1) engagement of reference librarian and literature review of existing instruments; (2) engagement of experts in cognitive load, and relevant stakeholders to review the literature and choose an instrument appropriate to measure cognitive load in EM didactic presentations. Response process validity was gathered using the format and anchors of instruments with previous validity evidence and piloting amongst the author group. A lecture was provided by one faculty to four residency programs via ZoomTM. Afterwards, residents completed the cognitive load instrument. Descriptive statistics were collected; Cronbach’s alpha assessed internal consistency of the instrument; and correlation for relationship to other variables (quality of lecture).
Results: The 10-item Leppink Cognitive Load instrument was selected with attention to content and response process validity evidence. Internal structure of the instrument was good (Cronbach’s alpha = 0.80). Subscales performed well-intrinsic load (α = 0.96, excellent), extrinsic load (α = 0.89, good), and germane load (α = 0.97, excellent). Five of the items were correlated with overall quality of lecture (p < 0.05).
Conclusions: The 10-item Cognitive Load instrument demonstrated good validity evidence to measure cognitive load and the subdomains of intrinsic, extraneous, and germane load. This instrument can be https://biodas.org/ used to provide feedback to presenters to improve the cognitive load of their presentations.

Assessment of the influence of gluten quality on highland barley dough sheet quality by different instruments

  • This study was to compare the results of texture analyzer with those of farinograph and extensograph and determine whether texture analyzer could be used to evaluate the processing quality of highland barley flour (HBF) dough sheet. The farinograph and extensograph tests were used to determine the reconstituted flour properties, a texture analyzer was applied to measure the tensile strength of HBF dough sheet, and the content of glutenin macropolymer (GMP), free sulfhydryl (-SH) and secondary structure of protein and microstructure in HBF dough sheet were investigated. Furthermore, correlations between these parameters were determined by regression analysis and Pearson correlation coefficient.
  • It was suggested that the reconstituted flours with a higher gluten index showed a higher farinograph quality number (FQN) and greater maximum resistance to extension (Rm ). HBF dough sheets with higher gluten index possessed higher GMP and lower free -SH contents, a more ordered secondary structure of protein, resulting in a more compact gluten network and a stronger tensile strength (TS).
  • The regression and correlation analysis showed that TS was positively correlated with FQN and Rm . In addition, it was significantly correlated with the content of GMP, -SH, secondary structure of protein and gluten network. It was concluded that texture analyzer could be an alternative approach to evaluate the processing quality of HBF dough sheet. Moreover, the gluten index of flours could be used to predict the processing quality of HBF dough sheet. This article is protected by copyright. All rights reserved.

Development of an Instrument to Assess the Stability of Cementless Femoral Implants Using Vibration Analysis During Total Hip Arthroplasty

Objective: The level of primary implant fixation in cementless total hip arthroplasty is a key factor for the longevity of the implant. Vibration-based methods show promise for providing quantitative information to help surgeons monitor implant fixation intraoperatively. A thorough understanding of what is driving these changes in vibrational behavior is important for further development and improvement of these methods. Additionally, an instrument must be designed to enable surgeons to leverage these methods. This study addresses both of these issues.
Method: An augmented system approach was used to develop an instrument that improves the sensitivity of the vibrational method and enables the implementation of the necessary excitation and measurement equipment. The augmented system approach took into account the dynamics of the existing bone-implant system and its interaction with the added instrument.
Results: Two instrument designs are proposed, accompanied by a convergence-based method to determine the insertion endpoint. The modal strain energy density distribution was shown to affect the vibrational sensitivity to contact changes in certain areas.
Conclusion: The augmented system approach led to an instrument design that improved the sensitivity to changes in the proximal region of the combined bone-implant-instrument system. This fact was confirmed both in silico and in vitro. Clinical Impact: The presented method and instruments address practical intraoperative challenges and provide perspective to objectively support the surgeon’s decision-making process, which will ensure optimal patient treatment.

Validation of the PAM-13 instrument in the Hungarian general population 40 years old and above

Background: Patient activation comprises the skills, knowledge and motivation necessary for patients’ effective contribution to their care. We adapted and validated the 13-item Patient Activation Measure (PAM-13) in the ≥ 40 years old Hungarian general population.
Methods: A cross-sectional web survey was conducted among 900 respondents selected from an online panel via quota sampling. After 10 days, the survey was repeated on 100 respondents. The distribution, internal consistency, test-retest reliability, factor structure, convergent, discriminant and known-groups validity of PAM-13 were assessed according to the COSMIN guidelines.
Results: The sample comprised 779 respondents. Mean (± SD) age was 60.4 ± 10.6 years, 54% were female and 67% had chronic illness. Mean (± SD) PAM-13 score was 60.6 ± 10.0. We found good internal consistency (Cronbach alpha: 0.77), moderate test-retest reliability (ICC: 0.62; n = 75), a single-factor structure and good content validity: PAM-13 showed moderate correlation with the eHealth Literacy Scale (r = 0.40), and no correlation with age (r = 0.02), education (r = 0.04) or income (ρ = 0.04). Higher PAM-13 scores were associated with fewer lifestyle risks (p < 0.001), more frequent health information seeking (p < 0.001), participation in patient education (p = 0.018) and various online health-related behaviours. When controlling for health literacy, sociodemographic factors and health status, the association of higher PAM-13 scores with overall fewer lifestyle risks, normal body mass index, physical activity and adequate diet remained significant. Similar properties were observed in the subgroup of participants with chronic morbidity, but not in the age group 65+.
Conclusion: PAM-13 demonstrated good validity in the general population. Its properties in clinical populations and the elderly as well as responsiveness to interventions warrant further research.
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Differential gene expression and chemical patterns of an intertidal crab inhabiting a polluted port and an adjacent marine protected area

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The acquisition of data to safeguard marine protected areas located close to ports is important in order to develop plans that allow effective protection from pollution as well as sustainable development of the port. The area Secche della Meloria is a Marine Protected Area (MPA-MEL) three miles from Livorno Harbour (LH), which is characterized by a long history of pollution. Here we studied the bioaccumulation and transcriptomic patterns of the marbled crab, Pachygrapsus marmoratus (Fabricius, 1787) (Crustacea; Brachyura, Grapsidae), inhabiting the two selected sites.
Results showed that the two crab populations are significantly different in their chemical composition of trace elements and Polyciclic Aromatic Hydrocarbons (PAHs), and gene expression patterns (1280 DEGs). Enrichment analysis indicated that crabs at LH had the highest stress response genes, and they were associated with higher levels of bioaccumulation detected in body tissues. We are confident that the significant differential gene expression profiles observed between crabs, characterized by https://biodas.org/ significant chemical differences, is associated with responses to contaminant exposure.

Comparison of Perioperative Outcomes Between Single-Port and Multi-Port Robotic Adrenalectomy

Background: Single-port (SP) robotic surgery has been utilized in several surgical procedures. We aim to describe our institution’s approach and perioperative experience with SP robotic adrenalectomy and compare it to the traditional multi-port (MP) approach.
Methods: We retrospectively reviewed all patients who underwent robotic adrenalectomy by a single surgeon between March 2019 and March 2020. Patient demographic, perioperative factors, and pathologic outcomes were recorded and analyzed using t-tests, chi-square, or Fisher’s exact tests.
Results: Thirty-six patients underwent SP (n = 11) and MP (n = 25) robotic adrenalectomy. Age, body mass index, gender, operative time, major Clavien-Dindo complications, and margin status showed no differences. Patients undergoing SP adrenalectomy had a lower estimated blood loss (18.1 ± 13.0 vs 65.6 ± 95.0 cc, P = .02) and smaller lesion size (2.8 ± 1.3 vs 4.1 ± 1.8 cm, P = .04) compared to those undergoing MP.
Conclusions: SP adrenalectomy appears to be a feasible approach in select adrenal masses. Further studies are needed to establish its safety and cost effectiveness.

Analysis of ocular injury 1-year outcome in survivors of Beirut Port ammonium nitrate blast

Purpose: Ascertain the 1-year outcome of patients who sustained open eye injuries from the Beirut Port ammonium nitrate (AN) explosion, one of the most powerful non-nuclear explosions in history.
Methods: Retrospective chart review of the operated eyes in 2 major eye hospitals.
Results: Out of 42 patients with open globe injury that was originally sutured, 29 patients (34 eyes) were followed at the 1-year mark. The initial vision in logMAR (mean ± SD) was 2.93 ± 0.87 (hand motion equivalent) and the final vision was 1.80 ± 1.47 (counting finger 2 m equivalent). No light perception (NLP) vision was noted in 12 eyes on presentation and 10 eyes remained so, while 2 eyes reached light perception (LP) vision. Eight eyes had an intraoperative expulsive choroidal hemorrhage (7 NLP and 1 LP both pre- and postoperatively), and 6 of the 8 developed phthisis. All eyes that developed phthisis had NLP preoperatively and postoperatively. Ocular Trauma Score (OTS) correlated inversely with both initial and final vision (p < 0.001). Zone of injury inversely correlated with initial vision (p = 0.02) and positively with final vision (p < 0.001). Final vision was significantly worse in zone 3 vs. zones 1 and 2 (3.2 ± 0.5) vs. 0.9 ± 1.1) (p < 0.001) injuries, as was the initial vision (3.3 ± 0.5 vs. 2.7 ± 0.8; p = 0.002).
Conclusion: The OTS, which provides prognostic information for serious ocular trauma, also yields valuable prognostic information for AN-associated ocular injuries. Expulsive choroidal hemorrhage and NLP vision at presentation remain very poor prognostic signs.

Da Vinci SP Single-Port Robotic Surgery in Gynecologic Tumors: Single Surgeon’s Initial Experience with 100 Cases

Purpose: To report preliminary experience of single-port robotic surgery using the da Vinci SP surgical system in gynecologic tumors.
Materials and methods: This was a retrospective study on 100 consecutive patients who underwent da Vinci SP single-port robotic surgery between November 2018 and January 2021. All procedures were performed by an experienced gynecologic surgeon using a single 2.5-cm umbilical incision.
Results: Of the 100 cases, the procedures included myomectomy (n=76), hysterectomy (n=2), endometrial cancer surgical staging (n=14), radical hysterectomy (n=3), radical trachelectomy (n=3), and ovarian cystectomy (n=2). None of the cases was converted to robotic multiport or open surgery. The median docking time was 5.0 minutes [interquartile range (IQR), 3.0-7.0], the median console time was 107.5 minutes (IQR, 78.7-155.8), and the median total operation time was 250.0 minutes (IQR, 215.0-310.0). The median estimated blood loss was 50.0 mL (IQR, 30.0-100.0), and the median change in hemoglobin level was 0.8 g/dL (IQR, 0.3-1.3). The median pain scores rated on a numerical rating scale immediately after and at 6, 12, and 24 hours after surgery were 5, 2, 2, and 2, respectively. The mean duration of postoperative hospitalization was 2.8 days.
Conclusion: Da Vinci SP single-port robotic surgery was successfully performed in various gynecologic tumors without significant complications. Therefore, this surgical system could be applied in patients who want precise gynecologic surgery while minimizing surgical incision.

Anxiety, Depression and PTSD in Children and Adolescents following the Beirut Port Explosion

Background: On August 4, 2020, Beirut’s port experienced one of the strongest non-nuclear explosions in history, killing approximately 200 people, displacing 300,000 persons, and injuring more than 1000 children.
Methods: An online anonymous survey assessed the prevalence of probable mental health disorders (MHDs) and impact of blast-related and other factors controlling for sociodemographics in 801 children aged 8 to 17 years old.
Results: About two thirds (64%) were screened positive for probable anxiety using the Screen for Childhood Anxiety Related Disorder, 52% for probable PTSD using CRIES-13, and 33% for probable depression using the Mood and Feelings Questionnaire (MFQ). Children who resided farthest way from the explosion site or were not in Beirut during blast had a significantly lower odds of anxiety and PTSD. Children who sustained any physical injury (vs. none) or witnessed casualties (vs. not) were at higher odds for PTSD. Children of parents who reported that their homes sustained minor damages (vs. no damages at all) were at higher odds for anxiety and PTSD, and temporary displacement (vs. none) increased odds of PTSD only. Poorer perceived economic status, poorer academic performance, having a family member injured in the blast, and prior mental health care seeking were associated with higher odds for all MHDs.
Conclusion: Our study, the only one to document the mental health impact of the Beirut Port explosion on children, highlights the critical need for an emergency mental health response, prioritizing disadvantaged communities and children with prior mental health problems.
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Comparative thermoresistance of two biological indicators for monitoring steam autoclaves. 3. Comparison performed at 121 degrees C in a hospital prevacuum steam sterilizer

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According to Pharmacopoea Nordica, steam autoclaves should be regularly monitored by a specific Swedish preparation of Bacillus stearothermophilus spores. If another biological indicator (BI) is used for such a control, it should first be calibrated against the Swedish BI (SBI) and the two BIs should be equally thermoresistant. Attest No. 1262 BI (ABI) has previously been shown to be more thermoresistant than the SBI at 134 degrees C, saturated steam. The purpose of the present study was to compare the thermoresistance of the SBI and the ABI at 121 degrees C, saturated steam and prevacuum. Seven hundred and twenty units of each BI were heat-exposed in an Emmer 760 litre prevacuum, pressure-pulsing steam autoclave.
After prevacuum with steam injection (manual or automatic preconditioning), the following incremental heat exposure times were used in triplicate (20 simultaneously tested units of each BI in each cycle) according to a randomized scheme: 5, 6 1/2, 8, 9 1/2, 11, 12 1/2, 14 and 15 min. The intra-chamber pressure and temperature were continuously monitored throughout the test and equilibration cycles.
The heat-exposed BI units were cultivated and read as recommended by the manufacturers. SBI and ABI showed a survival-time of 8 min and 11 min respectively, and a kill-time between 14 min and 15 min for both BIs. Thus, the ABI had the narrower survival-kill window. Probit analysis testing of the results showed that the difference in thermoresistance, at 121 degrees C, saturated steam and https://biodas.org/ prevacuum between Attest No. 1262 BI and the Swedish BI mentioned in Pharmacopea Nordica was not statistically significant.

Comparative thermoresistance of two biological indicators for monitoring steam autoclaves. 2. Comparison performed at 134 degrees C in a hospital prevacuum steam sterilizer

The thermoresistance of various lots of two biological indicators (BIs) for steam sterilization control, a Scandinavian BI (SBI) and the Attest BI (ABI), were compared during sterilization cycles in a hospital prevacuum (pressure-pulsing) steam autoclave at 134 degrees C, saturated steam. ABI No. 1242, ABI No. 1262 (its replacement) and incremental heat exposure times between 0 s and 180 s were used. The intrachamber temperature and pressure were continuously measured and monitored throughout the sterilization cycles.
The results showed that both of the ABIs were more thermoresistant than the SBI, giving 33.1% (ABI No. 1242), 18.9% (ABI No. 1262), and 0% (SBI) autoclave survivors. Because the time needed to reach 134 degrees C (preconditioning time) increased as the day progressed, and varied from day to day, correlation between individual incremental heat exposure times and the number of surviving BI units was not possible. Standardized test conditions are necessary for a true comparison of BIs.

Ozone: A Novel Sterilizer for Personal Protective Equipment

Objective: Personal protective equipment (PPE) is urgently sought during public health crises. It is necessary for the safety of both the patient and the healthcare professional. Yet during the recent COVID-19 pandemic, PPE scarcity in many countries, including the United States, has impacted the level of care for patients and the safety of healthcare personnel. Additionally, the implementation of mandatory mask mandates for the general public in many countries forced individuals to either reuse PPE, which can contribute to poor hygiene, or buy PPE in bulk and thereby contribute to the scarcity of PPE. In this study, we investigate the possibility of using a cost-effective ozone sterilization unit on contaminated N95 masks as an alternative to current sterilization methods.
Method: This protocol examined ozone’s ability to decontaminate N95 mask fabric that was exposed to a surrogate virus (Escherichia coli bacteriophage MS2). Once the sterilization unit achieves an ozone concentration of ~30 ppm, a 60-minute or 120-minute sterilization cycle commences. Following the sterilization cycle, we investigated the amount of viable virus on the slide using a viral plaque assay and compared it to a non-sterilized, control slide. Furthermore, we carried out trials to investigate the safety of an ozone sterilization device, by measuring the levels of ozone exposure that individuals may experience when operating the sterilization unit post-cycle.
Results: We showed that a 120-minute sterilization cycle at ~30 ppm achieves a 3-log reduction in viral activity, thereby complying with industry and U.S. Food and Drug Administration (FDA) standards. Further, we demonstrated that when following our protocol, the ozone exposure levels for a simple sterilization unit to be used at home complied with federal and industry standards.
Conclusion: Ozone may have the potential to decontaminate masks and other PPE.

Monitoring the Effective Sterilization of Low-Temperature Hydrogen Peroxide Gas Plasma Sterilizers in 58 Hospitals – 22 PLADs, China, June 2015-December 2019

What is already known on this topic?: Hydrogen peroxide sterilizeation is widely used for luminal devices. However, the low penetrability of the sterilant is of major concern.
What is added by this report?: This report investigated the effective sterilization of low-temperature hydrogen peroxide gas plasma sterilizers and compared the applicability of different biological monitoring methods based on medical luminal devices.
What are the implications for public health practice?: It is recommended to use a biological process challenge device for monitoring the sterilization of luminal devices with low-temperature hydrogen peroxide gas plasma sterilizers.

Portable sterilizer with microbe content detection device

  • Background: Infectious diseases, such as the latest COVID pandemic, caused by microorganisms like bacteria and virus, wreak havoc shaking human civilization with its rapid infection rate, and high number of mortalities. In case of a contagious disease, the virus can survive on any surface over a period of time and can be transferred to the human host through touching those surfaces unknowingly. Cleaning those possible surfaces to which these microorganisms can cling onto is one of the major ways to curb the spread. The aim of this study was to design a sterilizer which can clean such surfaces of daily used items easily within a certain period of time and can assess the cleaning efficacy by estimating the presence of microbes before and after sanitization.
  • Method development: To achieve this goal, we propose a portable sterilization unit that contains a sterilization chamber fitted with a microbe content detector. The sterilization chamber will cleanse the surfaces off the microbes using ultraviolet radiation. The chamber can be portable and at the same time big enough to accommodate items of daily use, like watch, wallet, clothes, utensils to even foods for single-house application. The microbe content detector will detect the success of the sterilization procedure by examining the time-lapse laser speckle images captured by a high-speed camera by mean of image processing algorithm, such that the user can determine whether further sterilization is required.
  • Conclusions: Microbe content detection device associated with the conventional sterilization procedure will give an assessment of the effectiveness of the sterilization. Successful implementation of sterilization for a wide variety of items of everyday use aided with microbe content detection technique is first of its kind and should be an effective tool for use in large communities, offices and public places for effective sterilization to help fight against the spread of infectious diseases.
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Design and development of heater control circuit without temperature sensor for monitoring hydrogen in argon

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A thin film based tin oxide sensor is developed to monitor low levels of hydrogen (concentration ranging from 5 to 75 ppm) in the cover gas plenum of the fast breeder test reactor. The heater and the sensor patterns are integrated on a miniature alumina substrate, and necessary electrical leads are incorporated into it. For proper functioning of the sensor, the heater has to be maintained at a constant temperature of 350 °C. This paper gives an outline of the electronics developed to measure the sensor signal and to control the heater temperature.
The major challenge in this work is that there was no provision for embedding a temperature sensor on the heater surface due to physical constraints. This constrained the maintenance of a constant heater temperature for the proper functioning of the sensor. This led us to develop and demonstrate a heater control circuit without a temperature sensor to maintain a fixed temperature for https://biodas.org/ monitoring hydrogen in argon, and electronics for the above-mentioned circuitry is discussed.

ERC-ESICM guidelines on temperature control after cardiac arrest in adults

  • The aim of these guidelines is to provide evidence‑based guidance for temperature control in adults who are comatose after resuscitation from either in-hospital or out-of-hospital cardiac arrest, regardless of the underlying cardiac rhythm. These guidelines replace the recommendations on temperature management after cardiac arrest included in the 2021 post-resuscitation care guidelines co-issued by the European Resuscitation Council (ERC) and the European Society of Intensive Care Medicine (ESICM).
  • The guideline panel included thirteen international clinical experts who authored the 2021 ERC-ESICM guidelines and two methodologists who participated in the evidence review completed on behalf of the International Liaison Committee on Resuscitation (ILCOR) of whom ERC is a member society. We followed the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach to assess the certainty of evidence and grade recommendations. The panel provided suggestions on guideline implementation and identified priorities for future research. The certainty of evidence ranged from moderate to low.
  • In patients who remain comatose after cardiac arrest, we recommend continuous monitoring of core temperature and actively preventing fever (defined as a temperature > 37.7 °C) for at least 72 h. There was insufficient evidence to recommend for or against temperature control at 32-36 °C or early cooling after cardiac arrest. We recommend not actively rewarming comatose patients with mild hypothermia after return of spontaneous circulation (ROSC) to achieve normothermia. We recommend not using prehospital cooling with rapid infusion of large volumes of cold intravenous fluids immediately after ROSC.

Thermal Model of an Omnimagnet for Performance Assessment and Temperature Control

An Omnimagnet is an electromagnetic device that enables remote magnetic manipulation of devices such as medical implants and microrobots. It is composed of three orthogonal nested solenoids with a ferromagnetic core at the center. Electrical current within the solenoids leads to undesired temperature increase within the Omnimagnet. If the temperature exceeds the melting point of the wire insulation, device failure may occur. Thus, a study of heat transfer within an Omnimagnet is a necessity, particularly to maximize the performance of the device. A transient heat transfer model that incorporates all three heat transfer modes is proposed and experimentally validated with an average normalized root-mean-square error of less than 4% (data normalized by temperature in degree celsius). The transient model is not computationally expensive and is applicable to Omnimagnets with different structures. The code is applied to calculate the maximum safe operational time at a fixed input current or the maximum safe input current for a fixed time interval. The maximum safe operational time and maximum safe input current depend on size and structure of the Omnimagnet and the lowest critical temperature of all the Omnimagnet materials. A parametric study shows that increasing convective heat transfer during cooling, and during heating with low input currents, is an effective method to increase the maximum operational time of the Omnimagnet. The thermal model is also presented in a state-space equation format that can be used in a real-time Kalman filter current controller to avoid device failure due to excessive heating.

Integrated Temperature and Position Sensors in a Shape-Memory Driven Soft Actuator for Closed-Loop Control

Soft actuators are a promising option for the advancing fields of human-machine interaction and dexterous robots in complex environments. Shape memory alloy wire actuators can be integrated into fiber rubber composites for highly deformable structures. For autonomous, closed-loop control of such systems, additional integrated sensors are necessary. In this work, a soft actuator is presented that incorporates fiber-based actuators and sensors to monitor both deformation and temperature.
The soft actuator showed considerable deformation around two solid body joints, which was then compared to the sensor signals, and their correlation was analyzed. Both, the actuator as well as the sensor materials were processed by braiding and tailored fiber placement before molding with silicone rubber. Finally, the novel fiber-rubber composite material was used to implement closed-loop control of the actuator with a maximum error of 0.5°.

Surface temperature controls the pattern of post-earthquake landslide activity

The patterns and controls of the transient enhanced landsliding that follows strong earthquakes remain elusive. Geostatistical models can provide clues on the underlying processes by identifying relationships with a number of physical variables. These models do not typically consider thermal information, even though temperature is known to affect the hydro-mechanical behavior of geomaterials, which, in turn, controls slope stability. Here, we develop a slope unit-based multitemporal susceptibility model for the epicentral region of the 2008 Wenchuan earthquake to explore how land surface temperature (LST) relates to landslide patterns over time. 3
We find that LST can explain post-earthquake landsliding while it has no visible effect on the coseismic scene, which is dominated by the strong shaking. Specifically, as the landscape progressively recovers and landslide rates decay to pre-earthquake levels, a positive relationship between LST and landslide persistence emerges. This seems consistent with the action of healing processes, capable of restoring the thermal sensitivity of the slope material after the seismic disturbance. Although analyses in other contexts (not necessarily seismic) are warranted, we advocate for the inclusion of thermal information in geostatistical modeling as it can help form a more physically consistent picture of slope stability controls.
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Superhydrophobic paper in the development of disposable labware and lab-on-paper devices

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Traditionally in superhydrophobic surfaces history, the focus has frequently settled on the use of complex processing methodologies using nonbiodegradable and costly materials. In light of recent events on lab-on-paper emergence, there are now some efforts for the production of superhydrophobic paper but still with little development and confined to the fabrication of flat devices. This work gives a new look at the range of possible applications of bioinspired superhydrophobic paper-based substrates, obtained using a straightforward surface modification with poly(hydroxybutyrate). As an end-of-proof of the possibility to create lab-on-chip portable devices, the patterning of superhydrophobic paper with different wettable shapes is shown with low-cost approaches.
Furthermore, we suggest the use of superhydrophobic paper as an extremely low-cost material to design essential nonplanar lab apparatus, including reservoirs for liquid storage and manipulation, funnels, tips for pipettes, or accordion-shaped substrates for liquid transport or mixing. Such devices take the advantage of the self-cleaning and extremely water resistance properties of the surfaces https://biodas.org/ as well as the actions that may be done with paper such as cut, glue, write, fold, warp, or burn. The obtained substrates showed lower propensity to adsorb proteins than the original paper, kept superhydrophobic character upon ethylene oxide sterilization and are disposable, suggesting that the developing devices could be especially adequate for use in contact with biological and hazardous materials.

Contaminating levels of zinc found in commonly-used labware and buffers affect glycine receptor currents

Zinc is an allosteric modulator of glycine receptor function, enhancing the effects of glycine at nM to low μM concentrations, and inhibiting its effects at higher concentrations. Because of zinc’s high potency at the glycine receptor, there exists a possibility that effects attributed solely to exogenously-applied glycine in fact contain an undetected contribution of zinc acting as an allosteric modulator. We found that glycine solutions made up in standard buffers and using deionized distilled water produced effects that could be decreased by the zinc chelator tricine.
This phenomenon was observed in three different vials tested and persisted even if vials were extensively washed, suggesting the zinc was probably present in the buffer constituents. In addition, polystyrene, but not glass, pipets bore a contaminant that enhanced glycine receptor function and that could also be antagonized by tricine. Our findings suggest that without checking for this effect using a chelator such as tricine, one cannot assume that responses elicited by glycine applied alone are not necessarily also partially due to some level of allosteric modulation by zinc.

Labware additives identified to be selective monoamine oxidase-B inhibitors

Plastic labware is used in all processes of modern pharmaceutical research, including compound storage and biological assays. The use of these plastics has created vast increases in productivity and cost savings as experiments moved from glass test tubes and capillary pipettes to plastic microplates and multichannel liquid handlers. One consequence of the use of plastic labware, however, is the potential release of contaminants and their resultant effects on biological assays.
We report herein the identification of biologically active substances released from a commonly used plastic microplate. The active contaminants were identified by gas chromatography-mass spectroscopy as dodecan-1-ol, dodecyl 3-(3-dodecoxy-3-oxopropyl)sulfanylpropanoate, and dodecanoic acid, and they were found to be selective monoamine oxidase-B inhibitors.

Open Labware: 3-D printing your own lab equipment

The introduction of affordable, consumer-oriented 3-D printers is a milestone in the current “maker movement,” which has been heralded as the next industrial revolution. Combined with free and open sharing of detailed design blueprints and accessible development tools, rapid prototypes of complex products can now be assembled in one’s own garage–a game-changer reminiscent of the early days of personal computing. At the same time, 3-D printing has also allowed the scientific and engineering community to build the “little things” that help a lab get up and running much faster and easier than ever before.

3D Printing in the Laboratory: Maximize Time and Funds with Customized and Open-Source Labware

3D printing, also known as additive manufacturing, is the computer-guided process of fabricating physical objects by depositing successive layers of material. It has transformed manufacturing across virtually every industry, bringing about incredible advances in research and medicine. The rapidly growing consumer market now includes convenient and affordable “desktop” 3D printers. These are being used in the laboratory to create custom 3D-printed equipment, and a growing community of designers are contributing open-source, cost-effective innovations that can be used by both professionals and enthusiasts.
User stories from investigators at the National Institutes of Health and the biomedical research community demonstrate the power of 3D printing to save valuable time and funding. While adoption of 3D printing has been slow in the biosciences to date, the potential is vast. The market predicts that within several years, 3D printers could be commonplace within the home; with so many practical uses for 3D printing, we anticipate that the technology will also play an increasingly important role in the laboratory.

3D-Printed Labware for High-Throughput Immobilization of Enzymes

  1. In continuous flow biocatalysis, chemical transformations can occur under milder, greener, more scalable, and safer conditions than conventional organic synthesis. However, the method typically involves extensive screening to optimize each enzyme’s immobilization on its solid support material. The task of weighing solids for large numbers of experiments poses a bottleneck for screening enzyme immobilization conditions.
  2. For example, screening conditions often require multiple replicates exploring different support chemistries, buffer compositions, and temperatures. Thus, we report 3D-printed labware designed to measure and handle solids in multichannel format and expedite screening of enzyme immobilization conditions.
  3. To demonstrate the generality of these advances, alkaline phosphatase, glucose dehydrogenase, and laccase were screened for immobilization efficiency on seven resins. The results illustrate the requirements for optimization of each enzyme’s loading and resin choice for optimal catalytic performance. Here, 3D-printed labware can decrease the requirements for an experimentalist’s time by >95%.
  4. The approach to rapid optimization of enzyme immobilization is applicable to any enzyme and many solid support resins. Furthermore, the reported devices deliver precise and accurate aliquots of essentially any granular solid material.

Adsorption of bacteriophages on polypropylene labware affects the reproducibility of phage research

Hydrophobicity is one of the most critical factors governing the adsorption of molecules and objects, such as virions, on surfaces. Even moderate change of wetting angle of plastic surfaces causes a drastic decrease ranging from 2 to 5 logs of the viruses (e.g., T4 phage) in the suspension due to adsorption on polymer vials’ walls. The effect varies immensely in seemingly identical containers but purchased from different vendors. Comparison of glass, polyethylene, polypropylene, and polystyrene containers revealed a threshold in the wetting angle of around 95°: virions adsorb on the surface of more hydrophobic containers, while in more hydrophilic vials, phage suspensions are stable.
The polypropylene surface of the Eppendorf-type and Falcon-type can accommodate from around 108 PFU/ml to around 1010 PFU/ml from the suspension. The adsorption onto the container’s wall might result in complete scavenging of virions from the bulk. We developed two methods to overcome this issue. The addition of surfactant Tween20 and/or plasma treatment provides a remedy by modulating surface wettability and inhibiting virions’ adsorption. Plastic containers are essential consumables in the daily use of many bio-laboratories. Thus, this is important not only for phage-related research (e.g., the use of phage therapies as an alternative for antibiotics) but also for data comparison and reproducibility in the field of biochemistry and virology.
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Contaminating levels of zinc found in commonly-used labware and buffers affect glycine receptor currents

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Zinc is an allosteric modulator of glycine receptor function, enhancing the effects of glycine at nM to low μM concentrations, and inhibiting its effects at higher concentrations. Because of zinc’s high potency at the glycine receptor, there exists a possibility that effects attributed solely to exogenously-applied glycine in fact contain an undetected contribution of zinc acting as an allosteric modulator. We found that glycine solutions made up in standard buffers and using deionized distilled water produced effects that could be decreased by the zinc chelator tricine.
This phenomenon was observed in three different vials tested and persisted even if vials were extensively washed, suggesting the zinc was probably present in the buffer constituents. In addition, polystyrene, but not glass, pipets bore a contaminant that enhanced glycine receptor function and that could also be antagonized by tricine. Our findings suggest that without checking for this effect using a chelator such as tricine, one cannot assume that responses elicited by glycine applied alone are not necessarily also partially due to some level of allosteric modulation by zinc.

Superhydrophobic paper in the development of disposable labware and lab-on-paper devices

Traditionally in superhydrophobic surfaces history, the focus has frequently settled on the use of complex processing methodologies using nonbiodegradable and costly materials. In light of recent events on lab-on-paper emergence, there are now some efforts for the production of superhydrophobic paper https://biodas.org/ but still with little development and confined to the fabrication of flat devices. This work gives a new look at the range of possible applications of bioinspired superhydrophobic paper-based substrates, obtained using a straightforward surface modification with poly(hydroxybutyrate). As an end-of-proof of the possibility to create lab-on-chip portable devices, the patterning of superhydrophobic paper with different wettable shapes is shown with low-cost approaches.
Furthermore, we suggest the use of superhydrophobic paper as an extremely low-cost material to design essential nonplanar lab apparatus, including reservoirs for liquid storage and manipulation, funnels, tips for pipettes, or accordion-shaped substrates for liquid transport or mixing. Such devices take the advantage of the self-cleaning and extremely water resistance properties of the surfaces as well as the actions that may be done with paper such as cut, glue, write, fold, warp, or burn. The obtained substrates showed lower propensity to adsorb proteins than the original paper, kept superhydrophobic character upon ethylene oxide sterilization and are disposable, suggesting that the developing devices could be especially adequate for use in contact with biological and hazardous materials.

3D Printing in the Laboratory: Maximize Time and Funds with Customized and Open-Source Labware

3D printing, also known as additive manufacturing, is the computer-guided process of fabricating physical objects by depositing successive layers of material. It has transformed manufacturing across virtually every industry, bringing about incredible advances in research and medicine. The rapidly growing consumer market now includes convenient and affordable “desktop” 3D printers. These are being used in the laboratory to create custom 3D-printed equipment, and a growing community of designers are contributing open-source, cost-effective innovations that can be used by both professionals and enthusiasts.
User stories from investigators at the National Institutes of Health and the biomedical research community demonstrate the power of 3D printing to save valuable time and funding. While adoption of 3D printing has been slow in the biosciences to date, the potential is vast. The market predicts that within several years, 3D printers could be commonplace within the home; with so many practical uses for 3D printing, we anticipate that the technology will also play an increasingly important role in the laboratory.

3D-Printed Labware for High-Throughput Immobilization of Enzymes

In continuous flow biocatalysis, chemical transformations can occur under milder, greener, more scalable, and safer conditions than conventional organic synthesis. However, the method typically involves extensive screening to optimize each enzyme’s immobilization on its solid support material. The task of weighing solids for large numbers of experiments poses a bottleneck for screening enzyme immobilization conditions. For example, screening conditions often require multiple replicates exploring different support chemistries, buffer compositions, and temperatures.
Thus, we report 3D-printed labware designed to measure and handle solids in multichannel format and expedite screening of enzyme immobilization conditions. To demonstrate the generality of these advances, alkaline phosphatase, glucose dehydrogenase, and laccase were screened for immobilization efficiency on seven resins. The results illustrate the requirements for optimization of each enzyme’s loading and resin choice for optimal catalytic performance. Here, 3D-printed labware can decrease the requirements for an experimentalist’s time by >95%. The approach to rapid optimization of enzyme immobilization is applicable to any enzyme and many solid support resins. Furthermore, the reported devices deliver precise and accurate aliquots of essentially any granular solid material.

Additive manufactured customizable labware for biotechnological purposes

Yet already developed in the 1980s, the rise of 3D printing technology did not start until the beginning of this millennium as important patents expired, which opened the technology to a whole new group of potential users. One of the first who used this manufacturing tool in biotechnology was Lücking et al. in 2012, demonstrating potential uses 1, 2. This study shows applications of custom-built 3D-printed parts for biotechnological experiments. It gives an overview about the objects’ computer-aided design (CAD) followed by its manufacturing process and basic studies on the used printing material in terms of biocompatibility and manageability. Using the stereolithographic (SLA) 3D-printing technology, a customizable shake flask lid was developed, which was successfully used to perform a bacterial fed-batch shake flask cultivation.
The lid provides Luer connectors and tube adapters, allowing both sampling and feeding without interrupting the process. In addition, the digital blueprint the lid is based on, is designed for a modular use and can be modified to fit specific needs. All connectors can be changed and substituted in this CAD software-based file. Hence, the lid can be used for other applications, as well. The used printing material was tested for biocompatibility and showed no toxic effects neither on mammalian, nor on bacteria cells. Furthermore an SDS-PAGE-comb was drawn and printed and its usability evaluated to demonstrate the usefulness of 3D printing for everyday labware. The used manufacturing technique for the comb (multi jet printing, MJP) generates highly smooth surfaces, allowing this application.
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Mutations in the receptor-binding domain of human SARS CoV-2 spike protein increases its affinity to bind human ACE-2 receptor

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The severe acute respiratory syndrome virus-2 (SARS CoV-2) infection has resulted in the current global pandemic. The binding of SARS CoV-2 spike protein receptor-binding domain (RBD) to the human angiotensin converting enzyme-2 (ACE-2) receptor causes the host infection. The spike protein has undergone several mutations with reference to the initial strain isolated during December 2019 from Wuhan, China. A number of these mutant strains have been reported as variants of concern and as variants being monitored. Some of these mutants are known to be responsible for increased transmissibility of the virus.
The reason for the increased transmissibility caused by the point mutations can be understood by studying the structural implications and inter-molecular interactions in the binding of viral spike protein RBD and human ACE-2. Here, we use the crystal structure of the RBD in complex with ACE-2 available in www.joplink.net/coronavirus-proteins/ the public domain and analyse the 250 ns molecular dynamics (MD) simulations of wild-type and mutants; K417N, K417T, N440K, N501Y, L452R, T478K, E484K and S494P.
The ionic, hydrophobic and hydrogen bond interactions, amino acid residue flexibility, binding energies and structural variations are characterized. The MD simulations provide clues to the molecular mechanisms of ACE-2 receptor binding in wild-type and mutant complexes. The mutant spike proteins RBD were associated with greater binding affinity with ACE-2 receptor. Communicated by Ramaswamy H. Sarma.

COVID-19 and Alzheimer’s disease: Meninges-mediated neuropathology

SARS-CoV-2 the causative agent of COVID-19 displays a broad range of pathophysiology. Cytokine storms associated with COVID-19 damage the blood-brain barrier (BBB) and allow pro-inflammatory factors to invade the brain, further promoting neurodegeneration. While SARS-CoV-2 viral RNA and proteins have been detected in brain tissues, the mechanisms of neuroinvasion remain unknown. COVID-19 has had a disproportionate impact on those suffering from neurodegenerative disorders such as Alzheimer’s disease (AD).
Understanding the mechanisms of SARS-CoV-2 neuroinvasion is crucial to study the long-term neurocognitive effects of COVID-19 on AD pathology.
Viruses can infiltrate the brain through the meninges via infected immune cells. The meninges regulate the immune surveillance of the brain and play a key role in the efflux of pathogens from the brain. Meningeal dysfunction has been demonstrated to exacerbate amyloid-beta pathogenesis. In this study, we explore the neuroinvasion pathway of SARS-CoV-2 through the meninges and its effect on AD pathology.
Method: 5x FAD x hACE2 mice were inoculated intranasally with a sublethal dose of SARS-CoV-2. The mice were maintained for 2 weeks. Mouse brains and meninges were harvested. The tissue was stained and immunofluorescence imaging was conducted to study viral proliferation and immune responses. Histo-cytometry was conducted for quantitative imaging analysis. Gene expression studies were done using Nanostring assays. All experiments involving the SARS-Cov-2 virus were carried out in a BSL3 facility.
Result: This ongoing study demonstrates the proliferation of the SARS-CoV-2 virus in the brain via meningeal lymphatics. SARS-CoV-2 infection resulted in increased neuroinflammation. Additionally, inflammatory responses induced meningeal dysfunction resulting in increased amyloid-beta pathology and cerebrospinal fluid drainage.
Conclusion: Given the increasing evidence for a viral hypothesis of Alzheimer’s Disease it is extremely important to study the neurodegenerative effects of COVID-19 which has affected millions worldwide. We demonstrate that SARS-CoV-2 infiltrates the brain via the meninges promoting neuroinflammation. Furthermore, amyloid-beta pathologies are exacerbated by COVID-19 in animal models providing preclinical evidence of the long-term neurodegenerative effects of COVID-19.

Exosomes Recovered From the Plasma of COVID-19 Patients Expose SARS-CoV-2 Spike-Derived Fragments and Contribute to the Adaptive Immune Response

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by beta-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has rapidly spread across the globe starting from February 2020. It is well established that during viral infection, extracellular vesicles become delivery/presenting vectors of viral material. However, studies regarding extracellular vesicle function in COVID-19 pathology are still scanty. Here, we performed a comparative study on exosomes recovered from the plasma of either MILD or SEVERE COVID-19 patients.
We show that although both types of vesicles efficiently display SARS-CoV-2 spike-derived peptides and carry immunomodulatory molecules, only those of MILD patients are capable of efficiently regulating antigen-specific CD4+ T-cell responses. Accordingly, by mass spectrometry, we show that the proteome of exosomes of MILD patients correlates with a proper functioning of the immune system, while that of SEVERE patients is associated with increased and chronic inflammation.
Overall, we show that exosomes recovered from the plasma of COVID-19 patients possess SARS-CoV-2-derived protein material, have an active role in enhancing the immune response, and possess a cargo that reflects the pathological state of patients in the acute phase of the disease.

Role of SARS-CoV-2 in causing blood-brain barrier leakage and microglial activation as a risk factor of cognitive deterioration in subjects at risk of Alzheimer’s disease

The recent pandemic provides evidence of altered central nervous system (CNS) function in response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-COV-2 invades the CNS by binding angiotensin-converting enzyme 2 (ACE2) expressed on neurons and glia. SARS-COV-2 may have effects on increased permeability of endothelial cells within the blood-brain barrier (BBB) as studies have shown that the S1 protein can transverse the BBB.
This is interesting because leakage of the BBB is implicated in Alzheimer’s disease (AD) pathogenesis. We hypothesized that SARS-CoV-2 infection leads to innate stimulated inflammation, ultimately activating microglial cells and an influx of pro-inflammatory cytokines and leukocytes in the meninges, contributing to increased permeability of the BBB. This BBB permeability increases AD susceptibility in subjects at risk by causing irreversible damage to the BBB and microglial cell activation.
Method: We developed a double transgenic mouse model using mice expressing human ACE2 receptor and 5xFAD mice that exhibit increased neuropathology seen in human AD allowing modeling of AD in SARS-CoV-2 pathogenesis. The hACE2/5xFAD double transgenic mice were intranasally inoculated with a sub-lethal dose of SARS-CoV-2 to test the hypothesis that SARS-COV-2 potentiates AD pathology and cognitive deterioration through impairment of the BBB. Leukocyte and cytokine populations were measured by flow cytometry and single-nuclei RNA sequencing of the meninges for characterization of microglial populations.
Result: SARS-CoV-2 creates a cytotoxic environment in the brain immediately following infection in hACE2/5xFAD mice leading to leakage of the BBB in the meninges. Activation of microglial innate cells by SARS-COV-2 invasion of the CNS will cause neural deterioration having long term implications on cognitive function. The hACE2/5xFAD mouse model allows us to uncover implications for SARS-COV-2 on AD cognitive deterioration.
Conclusion: The hACE2/5xFAD mouse allows modeling of SARS-CoV-2 in developing AD cases and allowed us to determine the immune environment generated in the meninges in response to SARS-CoV-2 infections. This mouse model provides a platform to proactively determine the effects of SARS-CoV-2 in developing AD cases, a methodology to be exploited for future mouse models determining the relationship of other viruses on AD pathology, and the opportunity to address phenotypes with therapeutics for preventative initiatives.
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Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.

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Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.

Platinum-based chemotherapy is the mainstay of front-line therapy of sufferers affected by pluri-metastatic intermediate/excessive grade NeuroEndocrine Neoplasms (NENs). Nevertheless, there aren’t any commonplace second-line remedies at illness development.

Earlier scientific experiences have evidenced that temozolomide (TMZ), an oral analog of dacarbazine, is lively in opposition to NENs at commonplace doses of 150 to 200 mg/mq per day on days 1 to five of a 28-day cycle, even when a major treatment-related toxicity is reported.

METHODS

Metastatic NENs sufferers had been handled on the ENETS (European NeuroEndocrine Tumor Society) heart of excellence of Naples (Italy), from 2014 to 2017 with a second-line various metronomic schedule of TMZ, 75 mg/m2per os “one week on/one week off”. Toxicity was graded with NCI-CTC standards v4.0; goal responses with RECIST v1.1 and efficiency standing (PS) in accordance with ECOG.

Twenty-six consecutive sufferers had been handled. Median age was 65.5 years. The predominant main organs had been pancreas and lung. Grading was G2 in 11 sufferers, G3 in 15. Greater than half of sufferers had a PS 2 (15 vs. 11 with PS 1). The median time-on-temozolomide remedy was 12.2 months (95% CI: 11.4-19.6). No G3/G4 toxicities had been registered.

Full response was obtained in 1 affected person, partial response in 4, steady illness in 19 (illness management price: 92.3%), and progressive illness in 2. The median total survival from TMZ begin was 28.Three months. PS improved in 73% of sufferers.Metronomic TMZ is an acceptable therapy for G2 and G3 NENs significantly in PS 2 sufferers. Potential and bigger trials are wanted to verify these outcomes.

Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.
Security and Exercise of Metronomic Temozolomide in Second-Line Remedy of Superior Neuroendocrine Neoplasms.

Would ivermectin for malaria management be useful in onchocerciasis-endemic areas?

There may be accumulating proof supporting using ivermectin as a malaria management software. Latest findings from the repeat ivermectin mass drug administrations for management of malaria trial demonstrated a diminished incidence of malaria in villages which obtained repeated ivermectin mass drug administration (MDA; six doses) in comparison with those that had just one spherical of ivermectin.

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A number of different research investigating the advantages of ivermectin for malaria functions are ongoing/deliberate.Whereas ivermectin MDA provides promising views within the combat in opposition to malaria, we spotlight the added advantages and anticipated challenges of conducting future research in onchocerciasis-endemic areas, that are confronted with a considerable illness burden together with onchocerciasis-associated epilepsy.

Rising the frequency of ivermectin MDA in such locations could scale back the burden of each malaria and onchocerciasis, and permit for extra entomological investigations on each the Anopheles mosquitoes and the blackflies.

Upfront, acceptability and feasibility research are wanted to evaluate the endorsement by the native populations, in addition to the programmatic feasibility of implementing ivermectin MDA a number of occasions a yr.Onchocerciasis-endemic websites would doubly profit from ivermectin MDA interventions, as these will alleviate onchocerciasis-associated morbidity and mortality, whereas doubtlessly curbing malaria transmission. Involving onchocerciasis applications and different related stakeholders within the malaria/ivermectin analysis agenda would foster the implementation of pluri-annual MDA in goal communities.