New dose-response model and SARS-CoV-2 quanta emission rates for calculating the long-range airborne infection risk.

Predictive models for airborne infection risk have been extensively used during the pandemic, but there is yet still no consensus on a common approach, which may create misinterpretation of results among public health experts and engineers designing building ventilation. In this study we applied the latest data on viral load, aerosol droplet sizes and removal mechanisms to improve the Wells Riley model by introducing the following novelties i) a new model to calculate the total volume of respiratory fluid exhaled per unit time ii) developing a novel viral dose-based generation rate model for dehydrated droplets after expiration iii) deriving a novel quanta-RNA relationship for various strains of SARS-CoV-2 iv) proposing a method to account for the incomplete mixing conditions. These new approaches considerably changed previous estimates and allowed to determine more accurate average quanta emission rates including omicron variant. These quanta values for the original strain of 0.13 and 3.8 quanta/h for breathing and speaking and the virus variant multipliers may be used for simple hand calculations of probability of infection or with developed model operating with six size ranges of aerosol droplets to calculate the effect of ventilation and other removal mechanisms. The model developed is made available as an open-source tool.

Experimental study on the exposure level of surgical staff to SARS-CoV-2 in operating rooms with mixing ventilation under negative pressure.

The purpose of this study was to reveal the exposure level of surgical staff to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from the patient's nose and wound during operations on COVID-19 patients. The tracer gas N2O is used to simulate SARS-CoV-2 from the patient's nose and wound. In this study, concentration levels of tracer gas were measured in the breathing zones of these surgical staff in the operating room under three pressure difference conditions: -5 pa-15 pa and -25 pa compared to the adjunction room. These influencing factors on exposure level are analyzed in terms of ventilation efficiency and the thermal plume distribution characteristics of the patient. The results show that the assistant surgeon faces 4 to 12 times higher levels of exposure to SARS-CoV-2 than other surgical staff. Increasing the pressure difference between the OR lab and adjunction room can reduce the level of exposure for the main surgeon and assistant surgeon. Turning on the cooling fan of the endoscope imager may result in a higher exposure level for the assistant surgeon. Surgical nurses outside of the surgical microenvironment are exposed to similar contaminant concentration levels in the breathing zone as in the exhaust. However, the ventilation efficiency is not constant near the surgical patient or in the rest of the room and will vary with a change in pressure difference. This may suggest that the air may not be fully mixed in the surgical microenvironment.

Zonal modeling of air distribution impact on the long-range airborne transmission risk of SARS-CoV-2.

A widely used analytical model to quantitatively assess airborne infection risk is the Wells-Riley model which is limited to complete air mixing in a single zone. However, this assumption tends not to be feasible (or reality) for many situations. This study aimed to extend the Wells-Riley model so that the infection risk can be calculated in spaces where complete mixing is not present. Some more advanced ventilation concepts create either two horizontally divided air zones in spaces as displacement ventilation or the space may be divided into two vertical zones by downward plane jet as in protective-zone ventilation systems. This is done by evaluating the time-dependent distribution of infectious quanta in each zone and by solving the coupled system of differential equations based on the zonal quanta concentrations. This model introduces a novel approach by estimating the interzonal mixing factor based on previous experimental data for three types of ventilation systems: incomplete mixing ventilation, displacement ventilation, and protective zone ventilation. The modeling approach is applied to a room with one infected and one susceptible person present. The results show that using the Wells-Riley model based on the assumption of completely air mixing may considerably overestimate or underestimate the long-range airborne infection risk in rooms where air distribution is different than complete mixing, such as displacement ventilation, protected zone ventilation, warm air supplied from the ceiling, etc. Therefore, in spaces with non-uniform air distribution, a zonal modeling approach should be preferred in analytical models compared to the conventional single-zone Wells-Riley models when assessing long-range airborne transmission risk of infectious respiratory diseases.

Estimating the impact of indoor relative humidity on SARS-CoV-2 airborne transmission risk using a new modification of the Wells-Riley model.

A novel modified version of the Wells-Riley model was used to estimate the impact of relative humidity (RH) on the removal of respiratory droplets containing the SARS-CoV-2 virus by deposition through gravitational settling and its inactivation by biological decay; the effect of RH on susceptibility to SARS-CoV-2 was not considered. These effects were compared with the removal achieved by increased ventilation rate with outdoor air. Modeling was performed assuming that the infected person talked continuously for 60 and 120 min. The results of modeling showed that the relative impact of RH on the infection risk depended on the ventilation rate and the size range of virus-laden droplets. A ventilation rate of 0.5 ACH, the change of RH between 20% and 53% was predicted to have a small effect on the infection risk, while at a ventilation rate of 6 ACH this change had nearly no effect. On the contrary, increasing the ventilation rate from 0.5 ACH to 6 ACH was predicted to decrease the infection risk by half which is remarkably larger effect compared with that predicted for RH. It is thus concluded that increasing the ventilation rate is more beneficial for reducing the airborne levels of SARS-CoV-2 than changing indoor RH. PRACTICAL IMPLICATIONS: The present results show that humidification to moderate levels of 40%-60% RH should not be expected to provide a significant reduction in infection risk caused by SARS-CoV-2, hence installing and running humidifiers may not be an efficient solution to reduce the risk of COVID-19 disease in indoor spaces. The results do however confirm that ventilation has a key role in controlling SARS-CoV-2 virus concentration in the air providing considerably higher benefits. The modified model developed in the present work can be used by public health experts, engineers, and epidemiologists when selecting different measures to reduce the infection risk from SARS-CoV-2 indoors allowing informed decisions concerning indoor environmental control.

Dynamic interaction of a downward plane jet and a cough jet with respect to particle transmission: An analytical and experimental study.

A cough jet can travel beyond the breathing zone of the source person, and thus, infectious viral- and bacterial-laden particles can be transported from the source person to others in close proximity. To reduce the interpersonal transmission of coughed particles, the objective of this study was to analytically and experimentally investigate the performance of downward plane jets with various discharge velocities. Chamber measurements were conducted to examine the interaction between a transient cough jet (discharge velocities of 12 m/sec and 16 m/sec) and a steady downward plane jet (discharge velocities from 1.0-8.5 m/sec) with respect to the transport of and human exposure to coughed particles. The results show that a relatively high-speed cough can easily penetrate a downward plane jet with a discharge velocity of less than 6 m/sec. A downward plane jet with a discharge velocity of 8.5 m/sec can bend the cough jet to a certain extent. In this study, momentum comparison of the cough jet and the downward plane jet shows that the value of personal exposure to coughed particles depends on the ratio of jet momentums. The results show that when the two momentums are equivalent or if the downward plane jet has a greater momentum, the cough jet is deflected downward and does not reach the breathing zone of the target thermal dummy. Using the ratio of the two momentums, it may be estimated whether the transmission of a cough jet can be controlled. A trajectory model was developed based on the ratio of the two momentums of a cough jet and a downward jet and was validated using the experimental data. In addition, the predicted trajectory of the cough jet agreed well with the results from smoke visualization experiments. This model can be used to guide the design of downward plane jet systems for protection of occupants from coughed particles.

Reducing the risk of viral contamination during the coronavirus pandemic by using a protective curtain in the operating room.

BACKGROUND: Airborne transmission diseases can transfer long and short distances via sneezing, coughing, and breathing. These airborne repertory particles can convert to aerosol particles and travel with airflow. During the Coronavirus disease 2019 (COVID-19) pandemic, many surgeries have been delayed, increasing the demand for establishing a clean environment for both patient and surgical team in the operating room. METHODS: This study aims to investigate the hypothesis of implementing a protective curtain to reduce the transmission of infectious contamination in the surgical microenvironment of an operating room. In this regard, the spread of an airborne transmission disease from the patient was evaluated, consequently, the exposure level of the surgical team. In the first part of this study, a mock surgical experiment was established in the operating room of an academic medical center in Norway. In the second part, the computational fluid dynamic technique was performed to investigate the spread of airborne infectious diseases. Furthermore, the field measurement was used to validate the numerical model and guarantee the accuracy of the applied numerical models. RESULTS: The results showed that the airborne infectious agents reached the breathing zone of the surgeons. However, using a protective curtain to separate the microenvironment between the head and lower body of the patient resulted in a 75% reduction in the spread of the virus to the breathing zone of the surgeons. The experimental results showed a surface temperature of 40 ˚C, which was about a 20 ˚C increase in temperature, at the wound area using a high intensity of the LED surgical lamps. Consequently, this temperature increase can raise the patient's thermal injury risk. CONCLUSION: The novel method of using a protective curtain can increase the safety of the surgical team during the surgery with a COVID-19 patient in the operating room.

Modeling the impact of indoor relative humidity on the infection risk of five respiratory airborne viruses.

With a modified version of the Wells-Riley model, we simulated the size distribution and dynamics of five airborne viruses (measles, influenza, SARS-CoV-2, human rhinovirus, and adenovirus) emitted from a speaking person in a typical residential setting over a relative humidity (RH) range of 20-80% and air temperature of 20-25 °C. Besides the size transformation of virus-containing droplets due to evaporation, respiratory absorption, and then removal by gravitational settling, the modified model also considered the removal mechanism by ventilation. The trend and magnitude of RH impact depended on the respiratory virus. For rhinovirus and adenovirus humidifying the indoor air from 20/30 to 50% will be increasing the relative infection risk, however, this relative infection risk increase will be negligible for rhinovirus and weak for adenovirus. Humidification will have a potential benefit in decreasing the infection risk only for influenza when there is a large infection risk decrease for humidifying from 20 to 50%. Regardless of the dry solution composition, humidification will overall increase the infection risk via long-range airborne transmission of SARS-CoV-2. Compared to humidification at a constant ventilation rate, increasing the ventilation rate to moderate levels 0.5 → 2.0 h-1 will have a more beneficial infection risk decrease for all viruses except for influenza. Increasing the ventilation rate from low values of 0.5 h-1 to higher levels of 6 h-1 will have a dominating effect on reducing the infection risk regardless of virus type.

Arabidopsis Hormone Database: a comprehensive genetic and phenotypic information database for plant hormone research in Arabidopsis.

Plant hormones are small organic molecules that influence almost every aspect of plant growth and development. Genetic and molecular studies have revealed a large number of genes that are involved in responses to numerous plant hormones, including auxin, gibberellin, cytokinin, abscisic acid, ethylene, jasmonic acid, salicylic acid, and brassinosteroid. Here, we develop an Arabidopsis hormone database, which aims to provide a systematic and comprehensive view of genes participating in plant hormonal regulation, as well as morphological phenotypes controlled by plant hormones. Based on data from mutant studies, transgenic analysis and gene ontology (GO) annotation, we have identified a total of 1026 genes in the Arabidopsis genome that participate in plant hormone functions. Meanwhile, a phenotype ontology is developed to precisely describe myriad hormone-regulated morphological processes with standardized vocabularies. A web interface (http://ahd.cbi.pku.edu.cn) would allow users to quickly get access to information about these hormone-related genes, including sequences, functional category, mutant information, phenotypic description, microarray data and linked publications. Several applications of this database in studying plant hormonal regulation and hormone cross-talk will be presented and discussed.

The investigation of the influence of thermal plume and breathing on sleeping microenvironment.

Most of humans' lifetime was spent indoor, especially in bedroom. Thus, understanding the characteristics of the sleep microenvironment is a prerequisite for better control and improvement of our sleeping environment. This study investigated the temperature and velocity field above the heads of sleeping people with supine postures, and explored the interactions between the thermal plume and the breathing airflow, where both the thermal manikins and real human subjects were used in our experiment settings. Three different breathing modes were considered in this study, where the non-breathing mode was used to investigate the characteristics of the thermal plume, and the synergy of the mouth and nose breathing mode on the thermal plume was also investigated. The results showed that the thermal plume of a supine posture person was not strong compared to that of a standing or sitting person, and the breathing airflow could influence the development of the thermal plume. Over the head of a sleeping person, the velocity of the thermal plume could be increased by both of the breathing modes, but no significant difference in the velocity and temperature field was found for the two breathing modes. It was also found that pollutants near the bed surface could be brought to the breathing zone with low velocity airflow, but could be blocked by the nasal exhalation jet. The findings in this study could provide theoretical support and guidance to improve the air quality in the breathing zone.

Experimental and simulated evaluations of airborne contaminant exposure in a room with a modified localized laminar airflow system.

The traditional mixing ventilation is not an energy effective approach to remove indoor air pollutants, maintain breath zone air quality, and control the airborne transmission. This study investigated the potential of a localized laminar airflow ventilation system to alleviate human exposure to pollutants. Breathing thermal manikins with sitting posture and supine posture were used to simulate the human. N2O was used as the tracer gas to simulate the indoor pollutant emission. The contaminant exposure index (εexp) and intake fraction index (IF) were used to assess the risk of human pollutant exposure for various supply air velocities given different emission source positions. Enhanced pollutant removal efficiency (Eff) (from the result) showed the qualification and desirability of the localized laminar airflow ventilation system in improving the breath zone air quality. The results showed that the CFD results could fit well with the experimental data and found out the interaction between thermal plume and supply air. The results also indicated a low εexp and IF, with over 90%, all of which were highly correlated with the supply velocity. Human's different breathing methods have little influence on the pollutant exposure so as to the location of the pollution source. This study found that localized laminar airflow ventilation system could efficiently provide fresh air to the breathing zone without sacrificing the thermal environment around human. It can be used for small region air quality control such as that in the bedroom and living room where desired air quality is favored.

Type I renal tubular acidosis caused by Sjögren's syndrome with hypokalemia as the first symptom: a case report.

Sjögren's syndrome is a chronic inflammatory autoimmune disease characterized by exocrine gland involvement and marked lymphocytic infiltration. Numerous reports of patients with Sjögren's syndrome have described kidney damage, mainly involving distal tubule dysfunction, severe renal calcification, kidney stones, and rickets. We herein describe a patient with primary Sjögren's syndrome who developed type I renal tubular acidosis with hypokalemia as the first symptom. This case highlights the possibility that an underlying autoimmune disorder should be considered in a patient presenting with distal tubular acidosis or recurrent hypokalemic periodic paralysis because treatment of the primary disease improves the outcome.

Can CO2 sensors in the ventilation system of a pool facility help reduce the variability in the trihalomethane concentration observed in indoor air?

Volatile and hazardous compounds are formed during the chlorination of pool water. Monitoring components in the air, such as the four trihalomethanes; chloroform, dichlorobromomethane, dibromochloromethane and bromoform (tTHM), is challenging. Carbon dioxide (CO2) sensors are used for controlling air quality in different buildings and can be installed in ventilation systems for continuous surveillance and monitoring purposes. However, such sensors are not used in indoor swimming facilities. In this study, samples of tTHM and CO2 were collected and analysed, along with other air and water quality parameters such as combined chlorine, to evaluate whether CO2 sensors could be used to explain the observed variability in the tTHM concentration in an indoor swimming facility and thereby reduce the exposure of individuals utilising the pool to tTHM. Random intercept models were built for the tTHM and CO2 concentrations, respectively, and the results show that the relationships between combined chlorine in the water, CO2 in the air and number of occupants explain 52% of the variability in tTHM. The correlation between occupancy and CO2 concentration (ρ = 0.65, p ≤ 0.01) suggests that CO2 sensors should be used so that the air supply corresponds to the demand of the users.

Involvement of an R2R3-MYB transcription factor gene AtMYB118 in embryogenesis in Arabidopsis.

MYB transcription factors play important roles in various developmental processes in plants. Here we report the characterization of AtMYB118, a gene encoding a putative R2R3-type MYB transcription factor, which expresses predominantly in siliques. Real-time quantitative PCR analysis and in situ hybridization showed that the transcripts of AtMYB118 were mainly detected in developing embryos. Constitutive over-expression of AtMYB118 resulted in pleiotropic phenotypes, including dwarfism, compact rosettes, backward curly-leaves, smaller flowers and siliques, and premature seed dehydration at the tip of siliques. Microarray analysis showed that many genes encoding proteins accumulated during embryogenesis were remarkably up-regulated in AtMYB118-over-expressed transgenic plants, including late embryogenesis abundant proteins (LEA proteins), storage proteins, seed maturation proteins, and proteins related with seed dehydration, desiccation and ABA signaling pathway. These results suggest that AtMYB118 may play an important role during embryogenesis and seed maturation.

Laminar airflow and mixing ventilation: Which is better for operating room airflow distribution near an orthopedic surgical patient?

BACKGROUND: There has been little research on the performance of laminar airflow (LAF) and mixing ventilation (MV) systems regarding clean airflow distribution near a surgical patient in operating rooms (ORs). The objective of this study was to examine the performance of LAF and MV systems in ORs at St Olavs Hospital in Norway. METHODS: Experimental measurements were conducted in 2 ORs equipped with LAF and MV systems. RESULTS: Under real operating conditions, airflow distribution from the LAF system was disrupted, and airflow velocity became significantly lower than that of MV above the lying patient. Airflow pattern was observed as distributed vertically downward and horizontally with LAF and MV, respectively. Turbulence intensity of supply airflow from LAF was much lower than that of MV. CONCLUSIONS: The airflow distribution by LAF system in close proximity to a patient is greatly affected by thermal plumes generated above incisions by both patients and surgical facilities. The effect of surgical facilities on airflow distribution by using MV is not significant compared to LAF ventilation. New guidelines are needed for the design of clean airflow distribution systems in the vicinity of surgical patients in ORs.

Simulation and Experiment on Droplet Formation and Separation for Needle-Type Micro-Liquid Jetting Dispenser.

The needle-type droplet jetting dispenser has wide applications in the field of microelectronic packaging, and for which the good quality of droplet formation and separation is the key to successful dispensing. This paper simulates the droplet jetting process which has been divided into 5 stages named backflow, growth, droplet extension, breakage, and separation, and analyses the combined effects of system parameters, such as pressure, viscosity, needle stroke, and nozzle diameter, on the changes of morphologies of ejected droplets, which is verified by experiments. The simulation and experiment results show that a higher driving pressure is quite suitable for the high-viscosity liquid to form normal droplets by avoiding adhesion. When increasing the needle stroke, the pressure should also be lowered properly to prevent the flow-stream. Besides, the nozzle with a large diameter is much more likely to cause sputtering or satellite-droplet problems. The results have a great significance for guiding the parameter settings of the needle-type dispensing approach.

Do surgeons and surgical facilities disturb the clean air distribution close to a surgical patient in an orthopedic operating room with laminar airflow?

BACKGROUND: Airflow distribution in the operating room plays an important role in ensuring a clean operating microenvironment and preventing surgical site infections (SSIs) caused by airborne contaminations. The objective of this study was to characterize the airflow distribution in proximity to a patient in an orthopedic operating room. METHODS: Experimental measurements were conducted in a real operating room at St. Olav's Hospital, Norway, with a laminar airflow system. Omnidirectional anemometers were used to investigate the air distribution in the operating zone, and 4 different cases were examined with a real person and a thermal manikin. RESULTS: This study showed that the downward airflow from the laminar airflow system varies in each case with different surgical arrangement, such as the position of the operating lamp. The results indicate that the interaction of thermal plumes from a patient and the downward laminar airflow may dominate the operating microenvironment. CONCLUSIONS: The airflow distribution in proximity to a patient is influenced by both the surgical facility and the presence of medical staff. A thermal manikin may be an economical and practical way to study the interaction of thermal plumes and downward laminar airflow. The provision of higher clean airflow rate in the operating microenvironment may be an effective way to prevent the development of SSIs caused by indoor airborne contamination.

Indoor air pollutants, ventilation rate determinants and potential control strategies in Chinese dwellings: A literature review.

After nearly twenty years of rapid modernization and urbanization in China, huge achievements have transformed the daily lives of the Chinese people. However, unprecedented environmental consequences in both indoor and outdoor environments have accompanied this progress and have triggered public awareness and demands for improved living standards, especially in residential environments. Indoor pollution data measured for >7000 dwellings (approximately 1/3 were newly decorated and were tested for volatile organic compound (VOC) measurements, while the rest were tested for particles, phthalates and other semi-volatile organic compounds (SVOCs), moisture/mold, inorganic gases and radon) in China within the last ten years were reviewed, summarized and compared with indoor concentration recommendations based on sensory or health end-points. Ubiquitous pollutants that exceed the concentration recommendations, including particulate matter, formaldehyde, benzene and other VOCs, moisture/mold, inorganic gases and radon, were found, indicating a common indoor air quality (IAQ) issue in Chinese dwellings. With very little prevention, oral, inhalation and dermal exposure to those pollutants at unhealthy concentration levels is almost inevitable. CO2, VOCs, humidity and radon can serve as ventilation determinants, each with different ventilation demands and strategies, at typical occupant densities in China; and particle reduction should be a prerequisite for determining ventilation requirements. Two directional ventilation modes would have profound impacts on improving IAQ for Chinese residences are: 1) natural (or window) ventilation with an air cleaner and 2) mechanical ventilation with an air filtration unit, these two modes were reviewed and compared for their applicability and advantages and disadvantages for reducing human exposure to indoor air pollutants. In general, mode 2 can more reliably ensure good IAQ for occupants; while mode 1 is more applicable due to its low cost and low energy consumption. However, besides a roadmap, substantial efforts are still needed to develop affordable, applicable and general ventilation solutions to improve the IAQ of residential buildings in China.

TRANSLUCENT GREEN, an ERF family transcription factor, controls water balance in Arabidopsis by activating the expression of aquaporin genes.

Water is the most abundant molecule in almost all living organisms. Aquaporins are channel proteins that play critical roles in controlling the water content of cells. Here, we report the identification of an AP2/EREBP family transcription factor in Arabidopsis thaliana, TRANSLUCENT GREEN (TG), whose overexpression in transgenic plants gave enhanced drought tolerance and vitrified leaves. TG protein is localized in the nucleus, binds DRE and GCC elements in vitro, and acts as a transcriptional activator in yeast cells. Microarray analysis revealed a total of 330 genes regulated by TG, among which five genes encode aquaporins. A transient expression assay showed that TG directly binds to the promoters of three aquaporin genes, such as AtTIP1;1, AtTIP2;3, and AtPIP2;2, indicating that TG directly regulates the expression of these genes. Moreover, overexpression of AtTIP1;1 resulted in vitrified phenotypes in transgenic Arabidopsis plants, similar to those observed in TG overexpression lines. Water injection into wild-type leaves recapitulated the vitrified leaf phenotypes, which was reversed by cutting off the water supply from vascular bundles. Taken together, our data support that TG controls water balance in Arabidopsis through directly activating the expression of aquaporin genes.

Association of circulating endothelial progenitor cells (CD14+-EPC) with renal function in patients with coronary artery disease.

BACKGROUND: The relationships between the endothelial progenitor cells (EPCs)-CD34(+) and CD14(+) and coronary artery disease (CAD) were reported and the association of CD34(+) cells with renal function was studied previously. Another kind EPC-CD14(+) cell and its association with renal function in patients with CAD have not been reported yet. Our aim was to assess CD14(+) cell counts versus renal function in CAD. METHODS AND RESULTS: We studied 242 patients with severe angiographic CAD and 30 healthy control participants. The CD14(+) cells were enumerated by flow cytometry. With lowering glomerular filtration rate (GFR), CD14(+) cell numbers (percentage of lymphocytes, median and interquartile range) decreased: 0.04 (0.03-0.06), 0.03 (0.02-0.05), 0.02 (0.01-0.03) for estimated glomerular filtration rate (eGFR) ≥90, 60 to 89, and 30 to 89 mL/min per 1.73 m(2), respectively (P < .001 for trend). The CD14(+) cell counts correlated with eGFR (r = .27, P = .03). By multivariate liner regression analysis, the difference remains significant (P = .02). CONCLUSIONS: The CD14(+) cell depletion is associated with renal dysfunction in CAD.

Characterization of Arabidopsis MYB transcription factor gene AtMYB17 and its possible regulation by LEAFY and AGL15.

MYB transcription factors compose one of the largest transcription factor families in Arabidopsis, which play important roles in various developmental processes as well as defense responses against environmental stresses. In this study, we report the characterization of AtMYB17 gene, a putative R2R3 type MYB gene family member in Arabidopsis. AtMYB17 was found exclusively localized in nuclear, with an activation domain at its C-terminus. AtMYB17 was highly expressed in inflorescences and siliques, especially at early flower developmental stages. The level of AtMYB17 transcripts was also found to increase after imbibition during seed germination and gradually concentrate to the shoot apex. Bioinformatics analysis identified several binding sites of LEAFY (LFY) and AGL15 in the promoter region of AtMYB17. Promoter-GUS fusion analysis showed that the LFY binding sites were important in fine-tuning regulation of the spatio-temporal expression of AtMYB17 in transgenic plants. Moreover, AtMYB17 was up-regulated in 35S::AGL15 plants. Taken together, our data suggest that LFY may be involved in the regulation of AtMYB17, possibly together with AGL15, and thereafter in early inflorescence development and seed germination.