The Effects of Antofine on the Morphological and Physiological Characteristics of Phytophthora capsici.

Phytophthora capsici is an important plant pathogenic oomycete that causes great losses to vegetable production around the world. Antofine is an important alkaloid isolated from Cynanchum komarovii Al. Iljinski and exhibits significant antifungal activity. In this study, the effect of antofine on the mycelial growth, morphology, and physiological characteristics of P. capsici was investigated using colorimetry. Meanwhile, the activity of mitochondrial respiratory chain complexes of P. capsici was evaluated following treatment with a 30% effective concentration (EC30), as well as EC50 and EC70, of antofine for 0, 12, 24, and 48 h. The results showed that antofine had a significant inhibitory effect against P. capsici, with an EC50 of 5.0795 µg/mL. After treatment with antofine at EC50 and EC70, the mycelia were rough, less full, and had obvious depression; they had an irregular protrusion structure; and they had serious wrinkles. In P. capsici, oxalic acid and exopolysaccharide contents decreased significantly, while cell membrane permeability and glycerol content increased when treated with antofine. Reactive oxygen species (ROS) entered a burst state in P. capsici after incubation with antofine for 3 h, and fluorescence intensity was 2.43 times higher than that of the control. The activities of the mitochondrial respiratory chain complex II, III, I + III, II + III, V, and citrate synthase in P. capsici were significantly inhibited following treatment with antofine (EC50 and EC70) for 48 h compared to the control. This study revealed that antofine is likely to affect the pathways related to the energy metabolism of P. capsici and thus affect the activity of respiratory chain complexes. These results increase our understanding of the action mechanism of antofine against P. capsici.

Cell reprogramming in a predictable manner on the superhydrophobic microwell array chip.

Reprogramming of somatic cells into the pluripotent state is stochastic and inefficient using the conventional culture plates. Novel micro-culture systems employing precisely controlled biophysical cues can improve the reprogramming efficiencies dramatically. Here we perform iPSC induction on our previously developed superhydrophobic microwell array chip (SMAR-chip) where cells undergo distinctive morphology change, switching from 2D monolayers to 3D clumps, and develop into bona fide colonies in more than 90% of the microwells. The PDMS substrate, together with the microwell structure and the superhydrophobic layer constitute a well-controlled microenvironment favorable for the morphogenesis and pluripotency induction. Investigation of the molecular roadmap demonstrates that the SMAR-chip promotes the transition from the initiation phase to the maturation phase and overcomes the roadblocks for reprogramming. In addition, the SMAR-chip also promotes the reprogramming of human cells, opening our method for translational applications. In summary, our study provides a novel platform for efficient cell reprogramming and emphasizes the advantages of employing the insoluble microenvironmental cues for the precise control of cell fate conversion.

The impact of electricity-carbon market coupling on system marginal clearing price and power supply cost.

In this study, we assessed the impacts of the benchmark designs of emissions allowance allocation in China's national carbon emissions trading system with plant-level data and further estimated the marginal clearing price and power supply cost in Guangdong power market under electricity-carbon market coupling with unit commitment and economic dispatch model. We find that the existing allowances benchmark would result in a considerable surplus of allowances at about 222 Mt. But the benchmarking and exemplary levels on the heat rate of power supply would motivate thermal power units to reduce CO2 emissions. Under a tight balance of supply and demand in Guangdong, peaking thermal power plants will become the marginal clearing units and higher clearing prices will add to the revenue of lower cost inframarginal renewable energy power units. However, the combined impact of electricity-carbon market coupling would cause the marginal clearing price fluctuates obviously from 0 to 1159 CNY/MWh. Compared to the baseline scenario with free CO2 allowances allocation, the efficiency of thermal power utilization would decrease by 23%-59% and the net revenue per MWh power supply of coal-fired power units would decrease by 275%-325% under the stress scenario. Our study suggests that setting a more stringent allowances allocation benchmark for carbon price discovery is necessary. As electricity-carbon market coupling changes the role of coal-fired power plants to provide flexibility service and decrease their revenues, it calls for further market designs on proper reimbursement of flexible resources, under which the electricity market can effectively achieve the synergy among accommodating new energy, ensuring resource adequacy, and delivering cost efficiency. In addition, the synergy can be enhanced by formulating a tax program, which can promote renewable energy investment.

Mapping the economy of coal power plants retrofitted with post-combustion and biomass co-firing carbon capture in China.

As a major carbon-emitting country, China has considerable potential for carbon capture and storage (CCS) applications, but economics is critical for CCS deployment. In this study, the levelized cost of electricity (LCOE) at province level was calculated for operational coal-fired power plants after post-combustion carbon capture (CC) retrofitting. The costs of captured CO2 and avoided CO2 were then compared. The economics of CC-retrofitted coal-fired power plants co-firing with biomass (BECC) were also analyzed. CC retrofitting is economical in North and Northwest China (except Qinghai) where coal prices are low. However, BECC retrofit is more suitable for coal-fired power plants in Central China where biomass prices are low. Therefore, the variation in the resource conditions among provinces should be considered when promoting CC and BECC retrofits. Compared with other power sources, coal-fired power generation becomes much more expensive after retrofits. We calculated the carbon price and capacity subsidy intensity that could support the retrofit of coal units. Effective carbon price should be 142.44-406.06 CNY/t for retrofits, and the capacity subsidy intensity ranges from 175.14-721.52 CNY/kW. The current carbon market and capacity market are not sufficiently developed to support retrofitting. The development of the carbon market with effective carbon price signals and proper power market mechanisms is critical to the rollout of CCS in China.

Piezoelectric atomization of liquids with dynamic viscosities up to 175 cP at room temperature.

Piezoelectric atomization has been applied in the field of respiratory medicine delivery and chemistry. However, the wider application of this technique is limited by the viscosity of the liquid. High-viscosity liquid atomization has great potential for applications in aerospace, medicine, solid-state batteries and engines, but the actual development of atomization is behind expectations. In this study, instead of the traditional model of single-dimensional vibration as a power supply, we propose a novel atomization mechanism that uses two coupled vibrations to induce micro-amplitude elliptical motion of the particles on the surface of the liquid carrier, which produces a similar effect as localized traveling waves to push the liquid forward and induce cavitation to achieve atomization. To achieve this, a flow tube internal cavitation atomizer (FTICA) consisting of a vibration source, a connecting block and a liquid carrier is designed. The prototype can atomize liquids with dynamic viscosities up to 175 cP at room temperature with a driving frequency of 507 kHz and a voltage of 85 V. The maximum atomization rate in the experiment is 56.35 mg/min, and the average atomized particle diameter is 10 µm. Vibration models for the three parts of the proposed FTICA are established, and the vibration characteristics and atomization mechanism of the prototype were verified using the vibration displacement measurement experiment and the spectroscopic experiment. This study offers new possibilities for transpulmonary inhalation therapy, engine fuel supply, solid-state battery processing and other areas where high-viscosity microparticle atomization is needed.

Abundant fungi dominate the complexity of microbial networks in soil of contaminated site: High-precision community analysis by full-length sequencing.

During the past decade, the characterization of microbial community in soil of contaminated sites was primarily done by high-throughput short-read amplicon sequencing. However, due to the similarity of 16S rRNA and ITS genes amplicon sequences, the short-read approach often limits the microbial composition analysis at the species level. Here, we simultaneously performed full-length and short-read amplicon sequencing to clarify the community composition and ecological status of different microbial taxa in contaminated soil from a high-resolution perspective. We found that (1) full-length 16S rRNA gene sequencing gave better resolution for bacterial identification at all levels, while there were no significant differences between the two sequencing platforms for fungal identification in some samples. (2) Abundant taxa were vital for microbial co-occurrences network constructed by both full-length and short-read sequencing data, and abundant fungal species such as Mortierella alpine, Fusarium solani, Mrakia frigida, and Chaetomium homopilatum served as the keystone species. (3) Heavy metal correlated with the microbial community significantly, and bacterial community and its abundant taxa were assembled by deterministic process, while the other taxa were dominated by stochastic process. These findings contribute to the understanding of the ecological mechanisms and microbial interactions in site soil ecosystems and demonstrate that full-length sequencing has the potential to provide more details of microbial community.

The Regulatory Effect of Braided Silk Fiber Skeletons with Differential Porosities on In Vivo Vascular Tissue Regeneration and Long-Term Patency.

The development of small-diameter vascular grafts that can meet the long-term patency required for implementation in clinical practice presents a key challenge to the research field. Although techniques such as the braiding of scaffolds can offer a tunable platform for fabricating vascular grafts, the effects of braided silk fiber skeletons on the porosity, remodeling, and patency in vivo have not been thoroughly investigated. Here, we used finite element analysis of simulated deformation and compliance to design vascular grafts comprised of braided silk fiber skeletons with three different degrees of porosity. Following the synthesis of low-, medium-, and high-porosity silk fiber skeletons, we coated them with hemocompatible sulfated silk fibroin sponges and then evaluated the mechanical and biological functions of the resultant silk tubes with different porosities. Our data showed that high-porosity grafts exhibited higher elastic moduli and compliance but lower suture retention strength, which contrasted with low-porosity grafts. Medium-porosity grafts offered a favorable balance of mechanical properties. Short-term in vivo implantation in rats indicated that porosity served as an effective means to regulate blood leakage, cell infiltration, and neointima formation. High-porosity grafts were susceptible to blood leakage, while low-porosity grafts hindered graft cellularization and tended to induce intimal hyperplasia. Medium-porosity grafts closely mimicked the biomechanical behaviors of native blood vessels and facilitated vascular smooth muscle layer regeneration and polarization of infiltrated macrophages to the M2 phenotype. Due to their superior performance and lack of occlusion, the medium-porosity vascular grafts were evaluated in long-term (24-months) in vivo implantation. The medium-porosity grafts regenerated the vascular smooth muscle cell layers and collagen extracellular matrix, which were circumferentially aligned and resembled the native artery. Furthermore, the formed neoarteries pulsed synchronously with the adjacent native artery and demonstrated contractile function. Overall, our study underscores the importance of braided silk fiber skeleton porosity on long-term vascular graft performance and will help to guide the design of next-generation vascular grafts.

Automatic instance segmentation of orchard canopy in unmanned aerial vehicle imagery using deep learning.

The widespread use of unmanned aerial vehicles (UAV) is significant for the effective management of orchards in the context of precision agriculture. To reduce the traditional mode of continuous spraying, variable target spraying machines require detailed information about tree canopy. Although deep learning methods have been widely used in the fields of identifying individual trees, there are still phenomena of branches extending and shadows preventing segmenting edges of tree canopy precisely. Hence, a methodology (MPAPR R-CNN) for the high-precision segment method of apple trees in high-density cultivation orchards by low-altitude visible light images captured is proposed. Mask R-CNN with a path augmentation feature pyramid network (PAFPN) and PointRend algorithm was used as the base segmentation algorithm to output the precise boundaries of the apple tree canopy, which addresses the over- and under-sampling issues encountered in the pixel labeling tasks. The proposed method was tested on another miniature map of the orchard. The average precision (AP) was selected to evaluate the metric of the proposed model. The results showed that with the help of training with the PAFPN and PointRend backbone head that AP_seg and AP_box score improved by 8.96% and 8.37%, respectively. It can be concluded that our algorithm could better capture features of the canopy edges, it could improve the accuracy of the edges of canopy segmentation results.

Research on the Machinability of Micro-Tapered Hole Group in Piezoelectric Atomizer and the Improvement Method.

A metal atomizing sheet with a group of micro-tapered holes is the core constituent of a piezoelectric atomizer. However, the diameters of large-end and small-end micro-tapered holes in industrial applications deviate from the design values by 15.25% and 15.83%, respectively, which adversely impacts the effect of atomizers. In this study, two main factors that influence the machining quality of tapered holes, the external vibration disturbance and the internal system errors inside the laser processor, were explored; consequently, the vibration model of the machining device and the laser drilling model were established, respectively. Based on the models and the experimental results, it was found that the errors in diameter caused by these two factors accounted for 20% and 67.87% of the total deviation, respectively. Finally, an improved method was proposed, where a damping system was added to the machining device, and the diameter of the initial laser spot was corrected. The measurement results of tapered holes machined by the improved method showed that the deviation of the large diameters and the small diameters from the design values declined to 4.85% and 4.83%, respectively. This study lays a foundation for the high-precision and large-scale industry of atomizing sheets, and provides a new research direction for enhancing the performance of atomizers.

Design and Experimental Study of a Tetragonal Rotor Pump Based on Wankel Geometry.

Wankel pump designs have not been fully established, with existing designs limited to bicornous rotor pumps and triangular rotor pumps. Here, on the basis of Wankel geometry, we present a tetragonal rotor pump with a three-lobe epicycloid and its conjugate envelope as chamber and rotor profile. First, the design method and basic working principle of the pump are introduced. Four groups of prototypes with different shape factors were manufactured, and their flow and pressure characteristics were experimentally studied. Numerical study showed that the flow rate irregularity of the pump is lower than that of existing Wankel pumps. Finally, the feasibility the pump for mixing applications was verified by a flow field observation experiment. The work in this paper provides a new type of rotary displacement pump design, representing an study of reverse application of a Wankel engine structure.

The differences between surface degradation and bulk degradation of FEM on the prediction of the degradation time for poly (lactic-co-glycolic acid) stent.

The degradation time is a crucial factor in evaluating the performance of poly (lactic-co-glycolic acid) (PLGA) stents. Bulk degradation mode was commonly used to analyze the stent degradation behavior by finite element approach. However, the PLGA stents may present surface degradation more than bulk degradation under certain conditions, which will greatly affect the degradation time after implantation. In this study, the degradation processes of the poly (lactic-co-glycolic acid) stent were reproduced utilizing finite element analysis. Both bulk degradation and surface degradation modes were considered. The correlation between tensile stress and degradation rate was investigated. The degradation time was analyzed selectively. The stress distribution, fracture, and mass loss were also compared between bulk degradation mode and surface degradation mode. The simulation results showed that, in both evolution modes, the degradation began at the 'peak-valley' region and fracture occurred at the cross of links and rings. Additionally, high levels of Von-Mises stress were observed in these two regions. Compared with bulk degradation, the fracture time of the stent was delayed by 63% in the surface degradation mode. In conclusion, the mass loss rate and scaffolding period showed great differences between surface degradation and bulk degradation. Based on this study, it is suggested that bulk degradation mode is not applicable to the case of inadequate water uptake mode, such as the tracheal stent degradation process. More experimental research should be carried out to accurately predict the scaffolding period after implantation. The mechanical properties of the fracture zone should be strengthened.

The role of national carbon pricing in phasing out China's coal power.

As the country with the world's largest coal power capacity, China is launching a national carbon market. How the carbon pricing may contribute to phasing out China's coal power is a great concern. We collect full-sample data set of China's 4540 operating coal plant units and develop a stochastic Monte-Carlo financial model to assess the financial sustainability of the plant operation. Although China's coal plants have long residual technical lifetime, their operations are close to the break-even state. Even with low carbon price of 50 CNY/tCO2 growing at 4%/y and the permits being fully auctioned, the average residual lifetime of all the plants will be reduced by 5.43 years, and the cumulative CO2 emission from 2020 to 2050 will be reduced by 22.73 billion ton. The spatial disparity in the carbon pricing effect is significant, and the western regions are more vulnerable to the carbon pricing risk than the eastern regions.

Analysis of efficacy of intervention strategies for COVID-19 transmission: A case study of Hong Kong.

By the end of February 2021, COVID-19 had spread to over 230 countries, with more than 100 million confirmed cases and 2.5 million deaths. To control infection spread with the least disruption to economic and societal activities, it is crucial to implement the various interventions effectively. In this study, we developed an agent-based SEIR model, using real demographic and geographic data from Hong Kong, to analyse the efficiency of various intervention strategies in preventing infection by the SARS-CoV-2 virus. Close contact route including short-range airborne is considered as the main transmission routes for COVID-19 spread. Contact tracing is not that useful if all other interventions have been fully deployed. The number of infected individuals could be halved if people reduced their close contact rate by 25%. For reducing transmission, students should be prioritized for vaccination rather than retired older people and preschool aged children. Home isolation, and taking the nucleic acid test (NAT) as soon as possible after symptom onset, are much more effective interventions than wearing masks in public places. Temperature screening in public places only disrupted the infection spread by a small amount when other interventions have been fully implemented. Our results may be useful for other highly populated cities, when choosing their intervention strategies to prevent outbreaks of COVID-19 and similar diseases.

Evidence for lack of transmission by close contact and surface touch in a restaurant outbreak of COVID-19.

BACKGROUND: Coronavirus disease 2019 (COVID-19) is primarily a respiratory disease that has become a global pandemic. Close contact plays an important role in infection spread, while fomite may also be a possible transmission route. Research during the COVID-19 pandemic has identified long-range airborne transmission as one of the important transmission routes although lack solid evidence. METHODS: We examined video data related to a restaurant associated COVID-19 outbreak in Guangzhou. We observed more than 40,000 surface touches and 13,000 episodes of close contacts in the restaurant during the entire lunch duration. These data allowed us to analyse infection risk via both the fomite and close contact routes. RESULTS: There is no significant correlation between the infection risk via both fomite and close contact routes among those who were not family members of the index case. We can thus rule out virus transmission via fomite contact and interpersonal close contact routes in the Guangzhou restaurant outbreak. The absence of a fomite route agrees with the COVID-19 literature. CONCLUSIONS: These results provide indirect evidence for the long-range airborne route dominating SARS-CoV-2 transmission in the restaurant. We note that the restaurant was poorly ventilated, allowing for increasing airborne SARS-CoV-2 concentration.

A plant-by-plant strategy for high-ambition coal power phaseout in China.

More than half of current coal power capacity is in China. A key strategy for meeting China's 2060 carbon neutrality goal and the global 1.5 °C climate goal is to rapidly shift away from unabated coal use. Here we detail how to structure a high-ambition coal phaseout in China while balancing multiple national needs. We evaluate the 1037 currently operating coal plants based on comprehensive technical, economic and environmental criteria and develop a metric for prioritizing plants for early retirement. We find that 18% of plants consistently score poorly across all three criteria and are thus low-hanging fruits for rapid retirement. We develop plant-by-plant phaseout strategies for each province by combining our retirement algorithm with an integrated assessment model. With rapid retirement of the low-hanging fruits, other existing plants can operate with a 20- or 30-year minimum lifetime and gradually reduced utilization to achieve the 1.5 °C or well-below 2 °C climate goals, respectively, with complete phaseout by 2045 and 2055.

Na+ compartmentation strategy of Chinese cabbage in response to salt stress.

Na+/H+ antiporter (NHX), responsible for counter-transport of Na+ and H+ across membranes (Na+ compartmentalization), plays a central role in plant salt-tolerance. In order to explore the Na+ compartmentalization modes and salt tolerance strategy in Chinese cabbage (Brassica rapa L. ssp. pekinensis), the seedlings of a salt-susceptible cabbage cultivar (Kuaicai 38) and a salt-tolerant cabbage cultivar (Qingmaye) were exposed to 100-400 mM NaCl for 30 days. Both of these cultivars showed a gradual decrease in fresh weight and water content and an increase in root-shoot ratio with the increasing NaCl-treatment concentration. The distribution of Na+ in these two cultivars was similar, with the green leaves showing the highest Na+ content, followed by inflated midribs, stems, and roots. The Na+ concentration in the apoplast was higher than that in the protoplast of the leaves. The expression levels of BrNHX1-1 and BrNHX1-2 in the leaves of Qingmaye were the highest among all BrNHX members, and increased after salt treatment. However, only BrNHX1-1 was expressed in Kuaicai 38. These results indicate that Na+ compartmentation into vacuoles is the major salt-adaptation strategy in Chinese cabbage. Coordinated overexpression of BrNHX1-1 and BrNHX1-2 may confer greater salt-tolerance for Chinese cabbage.

An investigation of the PM2.5 and NO2 concentrations and their human health impacts in the metro subway system of Suzhou, China.

This study measured the particle concentrations with an aerodynamic diameter smaller than 2.5 µm (PM2.5), nitrogen dioxide (NO2), and relative humidity (RH) at five metro subway stations in Suzhou's subway system (Lines 1 and 2). The real-time monitoring campaign was conducted from March 30th to April 10th and August 4th to August 21st, 2015. The monitoring practice was carried out during rush (7:00-9:00 AM and 17:00-19:00 PM) and regular hours (other times) at the ground and underground levels under different weather conditions with a purpose of obtaining representative data. The monitored results show that the concentrations of PM2.5 in the train carriages were lower than the concentrations at the underground platforms during both spring and summer. The mean PM2.5 concentrations at all the underground platforms in all the sub-stations monitored were significantly higher than those at the ground level. The human health impact was calculated to be 6300 annual DALYs (or 375 deaths) due to exposure to the subway system in Suzhou according to the UNEP-SETAC toxicity (USEtox) model. Linear regression models were applied to evaluate the relationships between the PM2.5, NO2 concentrations, and RH. We found that a 10% increment in RH from the current average level of 50-60% can lead to a 9.8 µg m-3 concentration decrease in PM2.5. This further results in the total human health impact being reduced to 2451 DALYs (150-4753 DALYs), representing a 20% decrease (1.2-38%).

Effects of Viola yedoensis Makino anti-itching compound on degranulation and cytokine generation in RBL-2H3 mast cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The Chinese herb compound prescription Viola yedoensis Makino Anti-itching Compound (VYAC), which consists of Viola yedoensis Makino, herb, Sophora flavescens Aiton, root, and Dictamnus dasycarpus Turcz, root and rhizome, has been traditionally used to treat various skin allergic inflammatory diseases in clinic. AIM OF THE STUDY: The aim of this study is to investigate the effects of VYAC on degranulation and to determine its anti-inflammatory mechanism in RBL-2H3 mast cells. MATERIALS AND METHODS: VYAC was extracted with water-coction extraction (Shufen et al., 2012). The aqueous extracts were concentrated in vacuum under reduced pressure and lyophilized using a freeze dryer, and lyophilized powder was obtained. MTT was used to evaluate the cytotoxic of VYAC on RBL-2H3 cells. Degranulation was carried out with RBL-2H3 cell model, which was stimulated with A23187 plus PMA. ß-Hexosaminidase and histamine were measured to evaluate degranulation. The mRNA levels of inflammation cytokines (IL-1ß, TNF-α, IL-6, and iNOS) were investigated by RT-PCR to explain the anti-inflammatory mechanism of VYAC. RESULTS: VYAC did not show cytotoxic effect on RBL-2H3 cells in the range of 25-400µg/mL. A higher dose of VYAC (800µg/mL) showed significant cytotoxicity (P<0.05). VYAC could significantly inhibit ß-hexosaminidase and histamine release when treated with 100, 200, and 400µg/mL (P<0.05), but could not significantly inhibit ß-Hexosaminidase and histamine release when treated with 25 and 50µg/mL (p>0.05). The mRNA levels of inflammatory cytokines (TNF-α, IL-1ß, IL-6, and iNOS) could significantly decrease when treated with 200 and 400µg/mL (P<0.05) of VYAC, which were associated with the development of inflammation. CONCLUSIONS: Results showed that VYAC inhibited ß-hexosaminidase and histamine release, which was inhibit A23187 plus PMA stimulated RBL-2H3 cell degranulation and downregulated inflammatory cytokines (IL-1ß, TNF-α, IL-6, and iNOS) expression to block inflammatory development.

[Efficacy of routinely used patterns for schistosomiasis diagnosis in lake and marshland regions].

OBJECTIVE: To evaluate the efficacy of routinely used pattern for schistosomiasis diagnosis in lake and marshland regions. METHODS: A historically heavy endemic village of schistosomiasis named Jinggan Village from Jiangling County was selected for field survey. The residents aged 6-65 years were screened by indirect hemagglutination assay (IHA) and dipstick dye immunoassay (DDIA) in parallel. The serological positives were examined by Kato-Katz technique and miracidium hatching technique to determine the infection of schistosome. The consistency of the two serological methods was evaluated. In addition, the schistosome infection rates were estimated accordingto the 3 detection patterns namely IHA, DDIA, IHA+DDIA combined with the etiologic examination. RESULTS: A total of 530 individuals were examined by the serological tests. The positive rate of DDIA was 46.98% (249/530), significantly higher than that of IHA (28.49%, 151/530) (χ2 = 59.55, P < 0.01). Totally 279 individuals were serological positives determined by IHA or DDIA, while 252 of them were detected by stool examination, and 22 cases were determined as parasitological positives, while 7 and 3 cases were diagnosed as antibody negatives by IHA and DDIA, respectively. The estimated infection rates determined by IHA, DDIA, IHA plus DDIA combined with stool examination were 3.14%, 3.97%, 4.60%, respectively. CONCLUSIONS: Under the condition of endemic situation becoming more and more waning, the current routinely used pattern for schistosomiasis detection may lead to missed diagnosis. So, more sensitive and effective diagnostic tools or appropriate detection patterns need to be explored.