Evaluation of microwave steam bags for the decontamination of filtering facepiece respirators

Reusing filtering facepiece respirators (FFRs) has been suggested as a strategy to conserve available supplies for home and healthcare environments during an influenza pandemic. For reuse to be possible, used FFRs must be decontaminated before redonning to reduce the risk of virus transmission; however, there are no approved methods for FFR decontamination. An effective method must reduce the microbial threat, maintain the function of the FFR, and present no residual chemical hazard. The method should be readily available, inexpensive and easily implemented by healthcare workers and the general public. Many of the general decontamination protocols used in healthcare and home settings are unable to address all of the desired qualities of an efficient FFR decontamination protocol. The goal of this study is to evaluate the use of two commercially available steam bags, marketed to the public for disinfecting infant feeding equipment, for FFR decontamination. The FFRs were decontaminated with microwave generated steam following the manufacturers' instructions then evaluated for water absorption and filtration efficiency for up to three steam exposures. Water absorption of the FFR was found to be model specific as FFRs constructed with hydrophilic materials absorbed more water. The steam had little effect on FFR performance as filtration efficiency of the treated FFRs remained above 95%. The decontamination efficacy of the steam bag was assessed using bacteriophage MS2 as a surrogate for a pathogenic virus. The tested steam bags were found to be 99.9% effective for inactivating MS2 on FFRs; however, more research is required to determine the effectiveness against respiratory pathogens.     

Modeling the efficiency of UV at 254 nm for disinfecting the different layers within N95 respirators

The study presented a Monte Carlo simulation of light transport in eight commonly used filtered facepiece respirators (FFRs) to assess the efficacy of UV at 254 nm for the inactivation of SARS-CoV-2. The results showed different fluence rates across the thickness of the eight different FFRs, implying that some FFR models may be more treatable than others, with the following order being (from most to least treatable): models 1512, 9105s, 1805, 9210, 1870+, 8210, 8110s and 1860, for single side illumination. The model predictions did not coincide well with some previously reported experimental data on virus inactivation when applied to FFR surfaces. The simulations predicted that FFRs should experience higher log reductions (>>6-log) than those observed experimentally (often limited to ~5-log). Possible explanations are virus shielding by aggregation or soiling, and a lack of the Monte Carlo simulations considering near-field scattering effects that can create small, localized regions of low UV photon probability on the surface of the fiber material. If the latter is the main cause in limiting practical UV viral decontamination, improvement might be achieved by exposing the FFR to UV isotropically from all directions, such as by varying the UV source to the FFR surface angle during treatment.     

A method to determine the available UV-C dose for the decontamination of filtering facepiece respirators

Aims: To develop a method to assess model‐specific parameters for ultraviolet‐C (UV‐C, 254 nm) decontamination of filtering facepiece respirators (FFRs). Methods and Results: UV‐C transmittance was quantified for the distinct composite layers of six N95 FFR models and used to calculate model‐specific α‐values, the percentage of the surface UV‐C irradiance available for the internal filtering medium (IFM). Circular coupons, excised from the FFRs, were exposed to aerosolized particles containing MS2 coliphage and treated with IFM‐specific UV‐C doses ranging from 38 to 4707 J m^?2. Models exposed to a minimum IFM dose of 1000 J m^?2 demonstrated at least a 3 log reduction (LR) in viable MS2. Model‐specific exposure times to achieve this IFM dose ranged from 2 to 266 min. Conclusions: UV‐C transmits into and through FFR materials. LR of MS2 was a function of model‐specific IFM UV‐C doses. Significance and Impact of the Study: Filtering facepiece respirators are in high demand during infectious disease outbreaks, potentially leading to supply shortages. Reuse of disposable FFRs after decontamination has been discussed as a possible remediation strategy, but to date lacks supporting scientific evidence. The methods described here can be used to assess the likelihood that UV‐C decontamination will be successful for specific FFR models.     

Two-channel brain-stem frequency-following responses to pure tone and missing fundamental stimuli

In 2 separate experiments the brain-stem frequency-following response (FFR)was recorded to a pure tone (200 Hz) and complex “missing fundamental” (MF) stimuli differing in temporal fine structure and envelope modulation depth. FFRs were simultaneously recorded in 2 channels with horizontal and vertical dipole orientations. Horizontal electrodes were identical in both experiments (right-left ear), but the vertical configuration was varied (vertex-left ear; vertex-linked mastoids). The horizontal channel yielded a well defined FFR to tone stimulation at a latency consistent with an origin along the auditory nerve. However, there was no horizontal response to MF stimulation. This latter finding provides electrophysiological support for the conclusion that MFs are not directly coded in the peripheral neural response. Vertical recordings, however, showed equally well defined FFRs to tone and MF stimuli. Thus, a representation of the missing fundamental frequency is registered in the brain-stem. Vertical latencies were consistent with a source at the level of the lateral lemniscus. The FFR is well suited to elucidate certain brain-stem mechanisms of auditory information processing. Important additional information results when responses are compared in horizontal and vertical dipole orientations. Thus, the present results provide the first evoked response demonstration of a peripheral-brain-stem dichotomy of MF coding.     

COVID-19 global pandemic planning: Dry heat incubation and ambient temperature fail to consistently inactivate SARS-CoV-2 on N95 respirators

The ongoing pandemic of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has placed a substantial strain on the supply of personal protective equipment, particularly the availability of N95 respirators for frontline healthcare personnel. These shortages have led to the creation of protocols to disinfect and reuse potentially contaminated personal protective equipment. A simple and inexpensive decontamination procedure that does not rely on the use of consumable supplies is dry heat incubation. Although reprocessing with this method has been shown to maintain the integrity of N95 respirators after multiple decontamination procedures, information on the ability of dry heat incubation to inactivate SARS-CoV-2 is largely unreported. Here, we show that dry heat incubation does not consistently inactivate SARS-CoV-2-contaminated N95 respirators, and that variation in experimental conditions can dramatically affect viability of the virus. Furthermore, we show that SARS-CoV-2 can survive on N95 respirators that remain at room temperature for at least five days. Collectively, our findings demonstrate that dry heat incubation procedures and ambient temperature for five days are not viable methods for inactivating SARS-CoV-2 on N95 respirators for potential reuse. We recommend that decontamination procedures being considered for the reuse of N95 respirators be validated at each individual site and that validation of the process must be thoroughly conducted using a defined protocol.     

Development of a Test System To Evaluate Procedures for Decontamination of Respirators Containing Viral Droplets

The aim of this study was to develop a test system to evaluate the effectiveness of procedures for decontamination of respirators contaminated with viral droplets. MS2 coliphage was used as a surrogate for pathogenic viruses. A viral droplet test system was constructed, and the size distribution of viral droplets loaded directly onto respirators was characterized using an aerodynamic particle sizer. The sizes ranged from 0.5 to 15 μm, and the sizes of the majority of the droplets were the range from 0.74 to 3.5 μm. The results also showed that the droplet test system generated similar droplet concentrations (particle counts) at different respirator locations. The test system was validated by studying the relative efficiencies of decontamination of sodium hypochlorite (bleach) and UV irradiation with droplets containing MS2 virus on filtering facepiece respirators. It was hypothesized that more potent decontamination treatments would result in corresponding larger decreases in the number of viable viruses recovered from the respirators. Sodium hypochlorite doses of 2.75 to 5.50 mg/liter with a 10-min decontamination period resulted in approximately 3- to 4-log reductions in the level of MS2 coliphage. When higher sodium hypochlorite doses (≥8.25 mg/liter) were used with the same contact time that was used for the dilute solutions containing 2.75 to 5.50 mg/liter, all MS2 was inactivated. For UV decontamination at a wavelength of 254 nm, an approximately 3-log reduction in the level of MS2 virus was achieved with dose of 4.32 J/cm² (3 h of contact time with a UV intensity of 0.4 mW/cm²), while with higher doses of UV irradiation (≥7.20 J/cm²; UV intensity, 0.4 mW/cm²; contact times, ≥5 h), all MS2 was inactivated. These findings may lead to development of a standard method to test decontamination of respirators challenged by viral droplets.During an infectious disease outbreak widespread panic can result from a limited understanding of the transmission route. Although some research points to a larger role for droplet nuclei (21, 28), other research suggests that droplets are the principal means of transmitting respiratory infections (10, 26). Droplets containing an infectious microorganism are believed to be transmitted to individuals who directly inhale the droplets resulting from coughing by carriers in close proximity or who ingest droplets spread to the mouth or the nose via the hands (7, 15). Large droplets were first defined as droplets more than 100 μm in diameter by Wells (30). Elsewhere, however, droplets more than 5 μm (23) or 10 μm (8) in diameter are often treated as large droplets. In this paper, the term “viral droplet” refers to all virus-containing liquid particles that retain their original size without significant evaporation, regardless of their specific size.N95 filtering facepiece respirators (FFRs) are routinely employed to prevent exposure of workers to biological hazards such as severe acute respiratory syndrome, tuberculosis, and novel H1N1 influenza A (6). In addition to CDC interim guidance (6), a recent report from the National Academies'' Institute of Medicine suggests that healthcare workers who are in close contact with individuals with novel H1N1 influenza illnesses should use fit-tested N95 respirators to reduce the risk of infection (18). This report also recommends increased research on influenza transmission and respiratory protection, which would enable policy makers to update these types of recommendations as additional disease prevention data become available.Current best practices suggest that once an FFR is worn in the presence of an infected patient, it should be considered potentially contaminated and discarded (5). However, during a pandemic outbreak a shortage of FFRs could occur (4). According to another report from the Institute of Medicine, during a 42-day influenza pandemic outbreak over 90 million N95 FFRs will be needed to protect workers in the healthcare sector (4). Furthermore, this report suggested that FFR reuse following decontamination should be considered a possible solution to deal with anticipated FFR shortages. Low-temperature biological decontamination methods have been suggested as a possible solution, but additional research needs to be done to determine whether infectious organisms can survive the decontamination process and if the decontamination method changes respirator fit (29).While it is well known that droplets play a role in the transmission of some respiratory infections, there is a lack of knowledge and data on the effectiveness of decontamination methods applied to respirators and porous personal protective equipment. There are several test methods for evaluation of the effectiveness of decontamination procedures for liquids and for hard porous or nonporous surfaces when they are challenged with viruses (2, 3, 27), while other methods are used to assess the effectiveness of decontamination procedures for FFRs when they are challenged with viral droplet nuclei (12). However, there is no test method to evaluate the effectiveness of biological decontamination procedures for disposable FFRs after they are challenged with viral droplets (liquid droplets containing a virus) whose sizes are similar to the sizes of droplets expelled by humans.Therefore, the aim of the present study was to develop a test system to evaluate the effectiveness of procedures for decontamination of respirators contaminated with viral droplets. The system was validated using two possible decontamination strategies: sodium hypochlorite and UV irradiation. It was hypothesized that the more potent decontamination treatments would result in corresponding larger decreases in the number of viable viruses recovered from the respirators than the less aggressive treatments.     

Cross-correlation and latency compensation analysis of click-evoked and frequency-following brain-stem responses in man

Cross-correlation (CC) and latency compensation (LC) analyses were applied to the human click-evoked brain-stem auditory evoked response (BAER) and the brain-stem frequency-following response (FFR). FFRs were elicited by pure tone stimuli (230 Hz and 460 Hz) or by complex tones derived from the sum of 3rd (920 Hz), 4th (1150 Hz), and 5th (1380 Hz) harmonics of the missing 230 Hz fundamental. The lower and upper harmonics always began in sine phase, while the middle harmonic varied in starting phase, resulting in harmonically complex stimuli with differing amplitude and phase patterns.Cross-correlations were computed between individual trials and a wave form t emplate (smoothed wave V for BAER, pure tone stimulus sinusoids for FFR). Trials were included in the analysis only if values of r² exceeded 0.5 (negative values of r were thus included, which controlled for the chance occurrence of positive correlations). Although brain-stem recordings are noisy, requiring as many as 1000 stimuli/average, correlation analysis consistently identified more positive than negative trials (approximately 2:1 ratio). Trials were also deleted if the lag associated with the selected r² was at the maximum shift position (‘extreme lag’).Averaging trials that satisfied the correlation and lag criteria led to sizeable enhancement of BAER (mean = 114%) and FFR (mean = 68% for 230 Hz stimulus) amplitudes. LC analysis resulted in additional, albeit smaller, increases in amplitude (approximately 10%). FFRs to harmonically complex stimuli were characterized by a clear periodicity at the missing fundamental frequency (230 Hz). However, amplitudes varied according to the modulation depth of the stimulus and, in certain cases, actually exceeded that of the FFR response to a 230 Hz pure tone.The results demonstrate the effectiveness of cross-correlation and, to a lesser degree, latency compensation analysis, applied to two classes of brain-stem potentials. It is anticipated that such techniques will prove useful in the study of auditory signal processing at the level of the brain-stem.     

Factors affecting aerosol SARS-CoV-2 transmission via HVAC systems; a modeling study

The role of heating, ventilation, and air-conditioning (HVAC) systems in the transmission of SARS-CoV-2 is unclear. To address this gap, we simulated the release of SARS-CoV-2 in a multistory office building and three social gathering settings (bar/restaurant, nightclub, wedding venue) using a well-mixed, multi-zone building model similar to those used by Wells, Riley, and others. We varied key factors of HVAC systems, such as the Air Changes Per Hour rate (ACH), Fraction of Outside Air (FOA), and Minimum Efficiency Reporting Values (MERV) to examine their effect on viral transmission, and additionally simulated the protective effects of in-unit ultraviolet light decontamination (UVC) and separate in-room air filtration. In all building types, increasing the ACH reduced simulated infections, and the effects were seen even with low aerosol emission rates. However, the benefits of increasing the fraction of outside air and filter efficiency rating were greatest when the aerosol emission rate was high. UVC filtration improved the performance of typical HVAC systems. In-room filtration in an office setting similarly reduced overall infections but worked better when placed in every room. Overall, we found little evidence that HVAC systems facilitate SARS-CoV-2 transmission; most infections in the simulated office occurred near the emission source, with some infections in individuals temporarily visiting the release zone. HVAC systems only increased infections in one scenario involving a marginal increase in airflow in a poorly ventilated space, which slightly increased the likelihood of transmission outside the release zone. We found that improving air circulation rates, increasing filter MERV rating, increasing the fraction of outside air, and applying UVC radiation and in-room filtration may reduce SARS-CoV-2 transmission indoors. However, these mitigation measures are unlikely to provide a protective benefit unless SARS-CoV-2 aerosol emission rates are high (>1,000 Plaque-forming units (PFU) / min).     

Coronary pressure and FFR predict long-term outcome after PTCA

Although coronary stents have been the most important improvement in percutaneous coronary interventions in the last 10 years, it is well known to interventionalists that many patients after percutaneous transluminal coronary angioplasty (PTCA) have a favourable outcome without stenting. Coronary angiography, however, is not sensitive enough to identify those particular patients and it has been suggested that a combination of angiographic and functional criteria would be more suitable to distinguish patients with a low restenosis chance after plain balloon angioplasty. In the present study, the authors investigated the value of coronary pressure measurement for conditional stenting in 85 patients. It was demonstrated that in patients in whom a high fractional flow reserve (FFR) was present (> 0.90), the incidence of coronary events at two-year follow-up was almost three times lower than in those patients with an FFR below 0.90. Such high FFRs could be obtained in approximately 45% of all patients. In an additional group of patients, it was demonstrated by intravascular ultrasound (IVUS) studies that the mechanism of a high FFR after plain balloon angioplasty is most likely the result of a larger lumen compared with patients with a suboptimal FFR. This means that, in patients in whom both the angiographic and the functional result after PTCA is optimal, a restenosis rate is achieved similar to that achieved by stenting. Obviously, in such patients, additional stenting and a number of problems in the long-term possibly related to stenting can be avoided. Therefore, coronary angiography and coronary pressure measurement have a complementary value in the evaluation of PTCA results and such information can be easily obtained by using a pressure wire instead of a regular guidewire.     

Sensory Dysfunction of Bladder Mucosa and Bladder Oversensitivity in a Rat Model of Metabolic Syndrome

Purpose

To study the role of sensory dysfunction of bladder mucosa in bladder oversensitivity of rats with metabolic syndrome.

Materials and Methods

Female Wistar rats were fed a fructose-rich diet (60%) or a normal diet for 3 months. Based on cystometry, the fructose-fed rats (FFRs) were divided into a group with normal detrusor function or detrusor overactivity (DO). Acidic adenosine triphosphate (ATP) solution (5mM, pH 3.3) was used to elicit reflex micturition. Cystometric parameters were evaluated before and after drug administration. Functional proteins of the bladder mucosa were assessed by western blotting.

Results

Compared to the controls, intravesical acidic ATP solution instillation induced a significant increase in provoked phasic contractions in both FFR groups and a significant decrease in the mean functional bladder capacity of group DO. Pretreatment with capsaicin for C-fiber desentization, intravesical liposome for mucosal protection, or intravenous pyridoxal 5-phosphate 6-azophenyl-2′,4′-disulfonic acid for antagonized purinergic receptors can interfere with the urodynamic effects of intravesical ATP in FFRs and controls. Over-expression of TRPV1, P2X₃, and iNOS proteins, and down-regulation of eNOS proteins were observed in the bladder mucosa of both fructose-fed groups.

Conclusions

Alterations of sensory receptors and enzymes in the bladder mucosa, including over-expression of TRPV1, P2X₃, and iNOS proteins, can precipitate the emergence of bladder phasic contractions and oversensitivity through the activation of C-afferents during acidic ATP solution stimulation in FFRs. The down-regulation of eNOS protein in the bladder mucosa of FFRs may lead to a failure to suppress bladder oversensitivity and phasic contractions. Sensory dysfunction of bladder mucosa and DO causing by metabolic syndrome are easier to elicit bladder oversensitivity to certain urothelium stimuli.     

Individual Differences in the Frequency-Following Response: Relation to Pitch Perception

The scalp-recorded frequency-following response (FFR) is a measure of the auditory nervous system’s representation of periodic sound, and may serve as a marker of training-related enhancements, behavioural deficits, and clinical conditions. However, FFRs of healthy normal subjects show considerable variability that remains unexplained. We investigated whether the FFR representation of the frequency content of a complex tone is related to the perception of the pitch of the fundamental frequency. The strength of the fundamental frequency in the FFR of 39 people with normal hearing was assessed when they listened to complex tones that either included or lacked energy at the fundamental frequency. We found that the strength of the fundamental representation of the missing fundamental tone complex correlated significantly with people''s general tendency to perceive the pitch of the tone as either matching the frequency of the spectral components that were present, or that of the missing fundamental. Although at a group level the fundamental representation in the FFR did not appear to be affected by the presence or absence of energy at the same frequency in the stimulus, the two conditions were statistically distinguishable for some subjects individually, indicating that the neural representation is not linearly dependent on the stimulus content. In a second experiment using a within-subjects paradigm, we showed that subjects can learn to reversibly select between either fundamental or spectral perception, and that this is accompanied both by changes to the fundamental representation in the FFR and to cortical-based gamma activity. These results suggest that both fundamental and spectral representations coexist, and are available for later auditory processing stages, the requirements of which may also influence their relative strength and thus modulate FFR variability. The data also highlight voluntary mode perception as a new paradigm with which to study top-down vs bottom-up mechanisms that support the emerging view of the FFR as the outcome of integrated processing in the entire auditory system.     

Rapid generation of mouse model for emerging infectious disease with the case of severe COVID-19

Since the pandemic of COVID-19 has intensely struck human society, small animal model for this infectious disease is in urgent need for basic and pharmaceutical research. Although several COVID-19 animal models have been identified, many of them show either minimal or inadequate pathophysiology after SARS-CoV-2 challenge. Here, we describe a new and versatile strategy to rapidly establish a mouse model for emerging infectious diseases in one month by multi-route, multi-serotype transduction with recombinant adeno-associated virus (AAV) vectors expressing viral receptor. In this study, the proposed approach enables profound and enduring systemic expression of SARS-CoV-2-receptor hACE2 in wild-type mice and renders them vulnerable to SARS-CoV-2 infection. Upon virus challenge, generated AAV/hACE2 mice showed pathophysiology closely mimicking the patients with severe COVID-19. The efficacy of a novel therapeutic antibody cocktail RBD-chAbs for COVID-19 was tested and confirmed by using this AAV/hACE2 mouse model, further demonstrating its successful application in drug development.     

Analysis of damage due to moist heat treatment of spores of Bacillus subtilis

Aims:  To determine conditions for generation and recovery of Bacillus subtilis spore populations heavily damaged by moist heat treatment.
Methods and Results:  Bacillus subtilis spores were treated with moist heat and spore viability was assessed on different media. A rich medium and several minimal media gave similar spore recoveries after moist heat treatment, but lack of glucose in minimal media greatly decreased spore recovery. High NaCl levels also greatly decreased the recovery of moist heat-treated spores on minimal media, and addition of good osmoprotectants reversed this effect. Moist heat treatment did not decrease spore recovery on minimal media with high salt through DNA damage or by eliminating spore germination, but by affecting spore outgrowth.
Conclusions:  Conditions for generating B. subtilis spore populations with high levels of conditional moist heat damage have been determined. The major conditional damage appears to be in spore outgrowth, perhaps because of damage to one or more important metabolic enzymes.
Significance and Impact of the Study:  This work has provided new insight into the mechanism of B. subtilis spore killing by moist heat.     

Clinical Characteristics of Hospitalized Patients with SARS-CoV-2 and Hepatitis B Virus Co-infection

In addition to the recent emerged SARS-CoV-2, hepatitis B virus (HBV) is one of the viruses which cause a global infection and threat public health. In worldwide, the prevalence of HBsAg is about 3.9% (Polaris Observatory 2018). According to a nationwide epidemiological survey of population whose ages range from 1 to 59 years in China, 2006, the prevalence of HBsAg was 7.2% (Liang et al. 2009). As SARS-CoV-2 and HBV both can cause liver damage (Fan et al. 2020), further understanding of the risk of SARS-CoV-2 on patients with HBV infection is urgently required in order to design an optimized treatment strategy. However, the impacts of SARS-CoV-2 infection on HBV patients are still not clear. For example, we do not yet know whether the SARS-CoV-2 infection is more severe in HBV patients and we also do not have much knowledge about the impact of SARS-CoV-2 on the course of HBV infection. In this retrospective study, we investigated the clinical characterizes of the patients coinfected with SARS-CoV-2 and HBV by analyzing the clinical records and laboratory tests of 123 COVID-19 patients admitted to Zhongnan Hospital of Wuhan University, Wuhan, China, from January 5 to February 20, 2020.     

Seasonal plasticity in auditory processing of the envelope and temporal fine structure of sounds in three songbirds

Songs mediate mate attraction and territorial defence in songbirds during the breeding season. Outside of the breeding season, the avian vocal repertoire often includes calls that function in foraging, antipredator and social behaviours. Songs and calls can differ substantially in their spectral and temporal content. Given seasonal variation in the vocal signals, the sender–receiver matching hypothesis predicts seasonal changes in auditory processing that match the physical properties of songs during the breeding season and calls outside of it. We tested this hypothesis in white-breasted nuthatches, Sitta carolinensis, tufted titmice, Baeolophus bicolor, and Carolina chickadees, Poecile carolinensis. We measured the envelope-following response (EFR), which quantifies phase locking to the amplitude envelope, and the frequency-following response (FFR), which quantifies phase locking to the temporal fine structure of sounds. Because songs and calls of nuthatches are amplitude modulated at different rates, we predicted seasonal changes in EFRs that match the rates of amplitude fluctuation in songs and calls. In chickadees and titmice, we predicted stronger FFRs during the spring and stronger EFRs during the winter because songs are tonal and calls include amplitude-modulated elements. In all three species, we found seasonal changes in EFRs and FFRs. EFRs varied across seasons and matched the amplitude modulations of songs and calls in nuthatches. In addition, female chickadees had stronger EFRs in the winter than in the spring. In all three species, FFRs during the spring tended to be stronger in females than in males. We also found species differences in EFRs and FFRs in both seasons; EFRs and FFRs tended to be higher in nuthatches than in chickadees and titmice. We discuss the potential mechanisms underlying seasonality in EFRs and FFRs and the implications of our results for communication during the breeding season and outside of it, when these three species form mixed-species flocks.     

Ecophysiology of seed germination of wild Dahlia coccinea (Asteraceae) in a spatially heterogeneous fire-prone habitat

Dahlia coccinea grows on fire-prone xerophilous shrubland, on a lava field located in Mexico City. Two kinds of experiments were performed to test the role of fire and environmental heterogeneity on germination. The first experiment tested the effect of environmental conditions (constant and alternating temperatures, cold stratification and light). The second one tested the effects of fire and high temperatures (dry and moist heat) on germination. Seeds of Dahlia were indifferent to light. The seeds showed physiological dormancy, which was lost by after-ripening or by gibberellins. During simulated fires, dry seeds tolerated high temperatures of short duration and also withstood prolonged exposure to 60 °C. Dry heat treatment reduced the mechanical restriction for embryo growth in dormant seeds. Ash and prolonged exposure to moist heat inhibited germination. Exogenous gibberellins reversed the deleterious effects of prolonged exposure to moist heat. The effect of cold stratification was related to the seeds' physiological stage and to light conditions; stratification in the dark reduced germination. Seeds of D. coccinea could tolerate, evade, or be slightly favored by the effects of low intensity fires occurring in their habitat. Seed responses to treatments suggest that the spatially heterogeneous lava field could provide a wide variety of micro-sites where physiological dormancy could be broken and during fires seeds could maintain their viability and subsequently germinate and/or develop a seed bank.     

Regional differences in the force-frequency relation of human left ventricular myocardium in mitral regurgitation: implications for ventricular shape

Sphericalization of the left ventricular (LV) chamber shape in patients with mitral regurgitation (MR) contributes to increased LV wall stress and energy consumption. On the basis of previous observations, we hypothesized the existence of regional differences in the force-frequency relation (FFR) within the LV that may contribute to its shape. Accordingly, in the present study, we assessed regional variation in the FFR in patients undergoing surgery for chronic, nonischemic MR with class II-III heart failure symptoms and related our findings to the in vivo LV shape. FFRs (steady-state isometric twitches, 0.2-3.4 Hz, 37 degrees C) were evaluated in MR myocardium from the LV subepicardial free wall (MR-FW) and papillary muscle (MR-PM) and from the subepicardial free wall in coronary artery bypass graft patients with normal LV contraction patterns [nonfailing (NF)]. Ascending slope, optimal stimulation frequency, and maximal twitch tension of the FFR were depressed in MR-FW and MR-PM compared with NF (P < 0.05). FFR depression was greater in MR-PM than in MR-FW. Between 107 and 134 beats/min, twitch tension became weaker in MR-PM, whereas it increased in MR-FW. Elevation of intracellular cAMP with forskolin eliminated FFR depression in MR-FW but not in MR-PM. MR-PM also had a 35% lower myosin heavy chain content and slowed twitch kinetics. In MR patients, the echocardiographic end-diastolic LV shape (end-diastolic eccentricity index = long axis/short axis) correlated with the ratio of ascending FFR slopes such that the end-diastolic eccentricity index increased 10% per 15% increase in slope ratio (r = 0.88, P = 0.01). These regional differences in the frequency dependence of contractility between the free wall and papillary myocardium may contribute to changes in LV shape in MR as well as during exercise.     

Evaluation of Viral Inactivation on Dry Surface by High Peak Power Microwave (HPPM) Exposure

Previous research has shown that virus infectivity can be dramatically reduced by radio frequency exposure in the gigahertz (GHz) frequency range. Given the worldwide SARS-CoV-2 pandemic, which has caused over 1 million deaths and has had a profound global economic impact, there is a need for a noninvasive technology that can reduce the transmission of virus among humans. RF is a potential wide area-of-effect viral decontamination technology that could be used in hospital rooms where patients are expelling virus, in grocery and convenience stores where local populations mix, and in first responder settings where rapid medical response spans many potentially infected locations within hours. In this study, we used bovine coronavirus (BCoV) as a surrogate of SARS-CoV-2 and exposed it to high peak power microwave (HPPM) pulses at four narrowband frequencies: 2.8, 5.6, 8.5, and 9.3 GHz. Exposures consisted of 2 µs pulses delivered at 500 Hz, with pulse counts varied by decades between 1 and 10,000. The peak field intensities (i.e. the instantaneous power density of each pulse) ranged between 0.6 and 6.5 MW/m², depending on the microwave frequency. The HPPM exposures were delivered to plastic coverslips containing BCoV dried on the surface. Hemagglutination (HA) and cytopathic effect analyses were performed 6 days after inoculation of host cells to assess viral infectivity. No change in viral infectivity was seen with increasing dose (pulse number) across the tested frequencies. Under all conditions tested, exposure did not reduce infectivity more than 1.0 log_10. For the conditions studied, high peak power pulsed RF exposures in the 2–10 GHz range appear ineffective as a virucidal approach for hard surface decontamination. © 2023 Bioelectromagnetics Society.     

Impact of inoculum preparation and storage conditions on the response of Escherichia coli O157:H7 populations to undercooking and simulated exposure to gastric fluid

This study evaluated the impact of inoculum preparation and storage conditions on the response of Escherichia coli O157:H7 exposed to consumer-induced stresses simulating undercooking and digestion. Lean beef tissue samples were inoculated with E. coli O157:H7 cultures prepared in tryptic soy broth or meat decontamination runoff fluids (WASH) or detached from moist biofilms or dried biofilms formed on stainless steel coupons immersed in inoculated WASH. After inoculation, the samples were left untreated or dipped for 30 s each in hot (75 degrees C) water followed by lactic acid (2%, 55 degrees C), vacuum packaged, stored at 4 (28 days) or 12 degrees C (16 days), and periodically transferred to aerobic storage (7 degrees C for 5 days). During storage, samples were exposed to sequential heat (55 degrees C; 20 min) and simulated gastric fluid (adjusted to pH 1.0 with HCl; 90 min) stresses simulating consumption of undercooked beef. Under the conditions of this study, cells originating from inocula of planktonic cells were, in general, more resistant to heat and acid than cells from cultures grown as biofilms and detached prior to meat inoculation. Heat and acid tolerance of cells on meat stored at 4 degrees C was lower than that of cells on nondecontaminated meat stored at 12 degrees C, where growth occurred during storage. Decontamination of fresh beef resulted in injury that inhibited subsequent growth of surviving cells at 12 degrees C, as well as in decreases in resistance to subsequent heat and acid stresses. The shift of pathogen cells on beef stored under vacuum at 4 degrees C to aerobic storage did not affect cell populations or subsequent survival after sequential exposure to heat and simulated gastric fluid. However, the transfer of meat stored under vacuum at 12 degrees C to aerobic storage resulted in reduction in pathogen counts during aerobic storage and sensitization of survivors to the effects of sequential heat and acid exposure.