Highly sensitive analysis of trace germanium derived from the efficient electrosynthesis and spectral introduction of GeH4 on foam electrode.

BACKGROUND: The electrochemical hydride generation technology, which uses electrolysis instead of chemical reagents to generate reducing species to achieve gaseous transformation and sample introduction of the tested elements, has received widespread attention in the field of atomic spectroscopy due to its simple, economical, and green characteristics. However, limited by the effective area of the electrode, the introduction efficiency and spectral signal of most elements (e.g., germanium) in practical applications are lower than traditional chemical hydride generation. RESULTS: In this paper, an efficient electrochemical hydride generation (EHG) method based on metal foam electrode for µg L-1 level germanium was constructed. Systematic electrochemical and spectral tests showed that the low charge transfer resistance and the high electrochemical activity of nickel-based foam electrodes jointly promoted the efficient electroreduction of Ge(IV). Besides, the porous network structure of the metal foam material improves the contact probability of reactants while reducing the gas-evolution effect caused by bubble accumulation. Interestingly, adequate reaction sites are crucial for the conversion of germanium, but large foam electrodes are not always compatible with analytical performance. After coupling atomic fluorescence spectroscopy, this new electrolysis method has been proven to be suitable for efficient conversion and quantitative detection of Ge over a wide concentration range (5-150 µg L-1). SIGNIFICANCE: Our proposal to improve the electrosynthesis efficiency of germanane (GeH4) by using metal foam electrode is extremely effective for the detection of trace or ultra-trace germanium. The exploration of electrode material, structure, and especially effective area will also provide ideas for the establishment of highly sensitive analysis methods in the future.

Loureirin A Promotes Cell Differentiation and Suppresses Migration and Invasion of Melanoma Cells via WNT and AKT/mTOR Signaling Pathways.

Resina Draconis is a traditional Chinese medicine, with the in-depth research, its medicinal value in anti-tumor has been revealed. Loureirin A is extracted from Resina Draconis, however, research on the anti-tumor efficacy of Loureirin A is rare. Herein, we investigated the function of Loureirin A in melanoma. Our research demonstrated that Loureirin A inhibited the proliferation of and caused G0/G1 cell cycle arrest in melanoma cells in a concentration-dependent manner. Further study showed that the melanin content and tyrosinase activity was enhanced after Loureirin A treatment, demonstrated that Loureirin A promoted melanoma cell differentiation, which was accompanied with the reduce of WNT signaling pathway. Meanwhile, we found that Loureirin A suppressed the migration and invasion of melanoma cells through the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. Taken together, this study demonstrated for the first time the anti-tumor effects of Loureirin A in melanoma cells, which provided a novel therapeutic strategy against melanoma.

A novel electrooxidation vapor generation technique for the direct analysis of trace Os in ore/water samples.

All electrolytic vapor generation technologies are based on cathodic reduction, but this paper focuses on how to use anodic oxidation to realize the gaseous transformation of noble metal Os. Supported by RuO2-based dimensionally stable anode (DSA), we found that the conversion from trivalent/tetravalent Os to the OsO4 can be carried out continuously and stably, even at the µg L-1 level. Interestingly, there was a negative correlation between the conversion of OsO4 and the RuO2 content in the DSA. The decrease of oxygen absorption potential and the increase of current density suggest that the oxidation process of Os belongs to electrocatalytic behavior. The catalytic activity of the material has an obvious influence on the conversion of osmium while the formation of free radical may be crucial for the effective oxidation. Under the optimum conditions, this electrocatalytic synthesis of OsO4 combined with ICP-MS can realize the same effect of oxidation and detection of two osmium species [Os(III) and Os(IV)]. The proposed method exhibits a low limit of detection (5 pg kg-1), a wide linear range (0.1-100 µg L-1) and excellent anti-interference performance, which promotes the direct analysis of total Os in real ore samples without separation. Considering the catalytic activity of OsO4 in specific reactions, this green anodic electrosynthesis technology is also expected to provide more possibilities.

2B and 3C Proteins of Senecavirus A Antagonize the Antiviral Activity of DDX21 via the Caspase-Dependent Degradation of DDX21.

Senecavirus A (SVA), also known as Seneca Valley virus, is a recently discovered picornavirus that can cause swine vesicular disease, posing a great threat to the global swine industry. It can replicate efficiently in cells, but the molecular mechanism remains poorly understood. This study determined the host's differentially expressed proteins (DEPs) during SVA infection using dimethyl labeling based on quantitative proteomics. Among the DE proteins, DDX21, a member of the DEAD (Asp-Glu-Ala-Asp)-box RNA helicase (DDX) family, was downregulated and demonstrated inhibiting SVA replication by overexpression and knockdown experiment. To antagonize this antiviral effect of DDX21, SVA infection induces the degradation of DDX21 by 2B and 3C proteins. The Co-IP results showed that 2B and 3C did not interact with DDX21, suggesting that the degradation of DDX21 did not depend on their interaction. Moreover, the 3C protein protease activity was necessary for the degradation of DDX21. Furthermore, our study revealed that the degradation of DDX21 by 2B and 3C proteins of SVA was achieved through the caspase pathway. These findings suggest that DDX21 was an effective antiviral factor for suppressing SVA infection and that SVA antagonized its antiviral effect by degrading DDX21, which will be useful to guide further studies into the mechanism of mutual regulation between SVA and the host.

The Cellular and Viral circRNAs Induced by Fowl Adenovirus Serotype 4 Infection.

Circular RNAs (circRNAs) are a new class of noncoding RNAs that play vital roles in many biological processes. Virus infection induces modifications in cellular circRNA transcriptomes and expresses viral circRNAs. The outbreaks of Hydropericardium-hepatitis syndrome (HHS) caused by fowl adenovirus serotype 4 (FAdV-4) have resulted in huge economic losses to the poultry industry worldwide. To investigate the expression of circRNAs during FAdV-4 infection, we performed transcriptome analysis of FAdV-4-infected leghorn male hepatoma (LMH) cells. In total, 19,154 cellular circRNAs and 135 differentially expressed (DE) cellular circRNAs were identified. The characteristics of the DE cellular circRNAs were analyzed and most of them were related to multiple biological processes according to GO and KEGG enrichment analysis. The accuracy of 10 cellular circRNAs were verified by semiquantitative RT-PCR and sequencing. The change trend was consistent with the RNA sequencing results. Moreover, 2014 viral circRNAs were identified and 10 circRNAs were verified by the same methods. Our analysis showed that seven circRNAs with the same 3' terminal and variable 5' terminal regions were located at pTP protein and DNA pol protein of FAdV-4, which may be generated via alternative splicing events. Moreover, the expression level of viral circRNAs was closely related to the replication efficiency of the virus and partial of the viral circRNAs promoted the replication of FAdV-4. Competing endogenous RNA analysis further showed that the effects of cellular and viral circRNAs on host or viral genes may act via miRNAs. Collectively, our findings first indicate that FAdV-4 infection induced the differential expression of cellular circRNAs and FAdV-4 also expressed viral circRNAs, some of which affected FAdV-4 replication. These findings will provide new clues for further understanding FAdV-4 and provide a basis for investigating host-virus interactions.

Study on the Fracture Toughness of Softwood and Hardwood Estimated by Boundary Effect Model.

The tensile strength and fracture toughness of softwood and hardwood are measured by the Boundary Effect Model (BEM). The experimental results of single-edge notched three-point bending tests indicate that the BEM is an appropriate method to estimate the fracture toughness of the present fibrous and porous woods. In softwood with alternating earlywood and latewood layers, the variation in the volume percentage of different layers in a small range has no obvious influence on the mechanical properties of the materials. In contrast, the hardwood presents much higher tensile strength and fracture toughness simultaneously due to its complicated structure with crossed arrangement of the fibers and rays and big vessels diffused in the fibers. The present research findings are expected to provide a fundamental insight into the design of high-performance bionic materials with a highly fibrous and porous structure.

cGAS Restricts PRRSV Replication by Sensing the mtDNA to Increase the cGAMP Activity.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus that causes great economic losses globally to the swine industry. Innate immune RNA receptors mainly sense it during infection. As a DNA sensor, cyclic GMP-AMP synthase (cGAS) plays an important role in sensing cytosolic DNA and activating innate immunity to induce IFN-I and establish an antiviral cellular state. In contrast, the role of innate immune DNA sensors during PRRSV infection has not been elucidated. In this study, we found that cGAS facilitates the production of IFN-ß during PRRSV infection. Western blot and virus titer assays suggested that cGAS overexpression suppressed the replication of multiple PRRSV strains, while knockout of cGAS increased viral titer and nucleocapsid protein expression. Besides, our results indicated that the mitochondria were damaged during PRRSV infection and leaked mitochondrial DNA (mtDNA) into the cytoplasm. The mtDNA in the cytoplasm co-localizes with the cGAS, and the cGAMP activity was increased when the cGAS was overexpressed during PRRSV infection. Furthermore, the cGAMP also possesses an anti-PRRSV effect. These results indicate for the first time that cGAS restricts PRRSV replication by sensing the mtDNA in the cytoplasm to increase cGAMP activity, which not only explains the molecular mechanism by which cGAS inhibits PRRSV replication but also provides research ideas for studying the role of the cGAS-STING signaling pathway in the process of RNA virus infection.

Isolation of two new genes encoding heat shock protein 70 in Bemisia tabaci and analysis during thermal stress.

The heat shock protein 70 family (HSP70) is among the most varied HSP family with respect to structure and function. The phloem-feeding insect Bemisia tabaci (Gennadius) is an important pest of cotton, vegetables and ornamentals that transmits several plant viruses and causes enormous agricultural losses. In this study, two new HSP70 genes (Bthsp70-2 and Bthsp70-3) were isolated from the MED cryptic species B. tabaci, an important phloem-feeding pest of vegetables and ornamentals. Bthsp70-2 and Bthsp70-3 encoded proteins comprised of 652 and 676 amino acids, and the deduced proteins were closely related to other HSP70s in Hemiptera. Expression analyses using real-time quantitative PCR indicated that Bthsp70-2 and Bthsp70-3 were induced in B. tabaci pupae and adults during high and low thermal stress. Bthsp70-2 and Bthsp70-3 exhibited similar, but not identical, expression patterns when exposed to different durations of high temperature stress. Oral ingestion of dsBthsp70 reduced the expression level of Bthsp70-2 and Bthsp70-3 in B. tabaci and increased the mortality of B. tabaci during heat shock. In conclusion, Bthsp70-2 and Bthsp70-3 exhibit different expression patterns during thermal stress, thus expanding the roles of HSPs in B. tabaci.

Microniche geometry modulates the mechanical properties and calcium signaling of chondrocytes.

In articular cartilage, the function of chondrocytes is strongly related to their zone-specific microniche geometry defined by pericellular matrix. Microniche geometry is critical for regulating the phenotype and function of the chondrocyte in native cartilage and tissue engineering constructs. However the role of microniche geometry in the mechanical properties and calcium signaling of chondrocytes remains unknown. To recapitulate microniche geometry at single-cell level, we engineered three basic physiological-related polydimethylsiloxane (PDMS) microniches geometries fabricated using soft lithography. We cultured chondrocytes in these microniche geometries and quantified cell mechanical properties using atomic force microscopy (AFM). Fluorescent calcium indicator was used to record and quantify cytosolic Ca2+ oscillation of chondrocytes in different geometries. Our work showed that microniche geometry modulated the mechanical behavior and calcium signaling of chondrocytes. The ellipsoidal microniches significantly enhanced the mechanical properties of chondrocytes compared to spheroidal microniche. Additionally, ellipsoidal microniches can markedly improved the amplitude but weakened the frequency of cytosolic Ca2+ oscillation in chondrocytes than spheroidal microniche. Our work might reveal a novel understanding of chondrocyte mechanotransduction and therefore be useful for designing cell-instructive scaffolds for functional cartilage tissue engineering.

Multiple "Omics" data-based biomarker screening for hepatocellular carcinoma diagnosis.

The huge prognostic difference between early and late stage hepatocellular carcinoma (HCC) is a challenging diagnostic problem. Alpha-fetoprotein is the mostly widely used biomarker for HCC used in the clinic, however it's sensitivity and specificity of is not optimal. The development and application of multiple biotechnologies, including next generation sequencing, multiple "omics" data, that include genomics, epigenomics, transcriptomics, proteomics, metabolomics, metagenomics has been used for HCC diagnostic biomarker screening. Effective biomarkers/panels/models have been identified and validated at different clinical levels. A large proportion of these have a good diagnostic performance for HCC, especially for early HCC. In this article, we reviewed the various HCC biomarkers derived from "omics" data and discussed the advantages and disadvantages for diagnosis HCC.

MicroRNA-498 promotes proliferation, migration, and invasion of prostate cancer cells and decreases radiation sensitivity by targeting PTEN.

Prostate cancer (PCa) remains the secondary highest cause of cancer-related death in the United States in men. It has been reported that microRNAs can serve as key regulators in tumor development and progression in various cancers including PCa. In this study, we examined the effect of miR-498 on proliferation, radiosensitivity, invasion, and migration of PCa cells. The proliferation of LNCaP and DU-145 PCa cells with altered expression of miR-498 was evaluated by MTT assay. The invasion and migration of LNCaP and DU-145 PCa cells were assess by matrigel invasion assay and transwell migration assay. The protein expression level in PCa cells was examined by western blot. The function of miR-498 on radiation-induced apoptosis in LNCaP and DU-145 PCa cells was detected by Caspase-Glo3/7 assay. Forced expression of miR-498 improved the proliferation, invasion and migration in PCa cells. Furthermore, miR-498 decreased the sensitivity of PCa cells after ionizing radiation treatment. MiR-498 reduced the radiation-induced apoptosis in PCa cells by regulation of BAX and Bcl-2 expression. Meanwhile, miR-498 altered the expression of E-cadherin, N-cadherin, snail, and Vimentin in both LNCaP and DU-145 PCa cells and regulated epithelial to mesenchymal transition (EMT). Further study showed that aberrant expression of miR-498 changed the expression levels of phosphatase and tensin homolog and p-AKT in LNCaP and DU-145 PCa cells. In a summary, miR-498 displayed important roles in tumor development and progression in PCa cells, and might act as a potential prognostic biomarker and predict radiotherapy response in PCa.

Degradation of phenolic compounds with simultaneous bioelectricity generation in microbial fuel cells: Influence of the dynamic shift in anode microbial community.

This study evaluated the feasibility of microbial fuel cells (MFCs) for simultaneous electricity generation and degradation of phenolic compounds. The voltage generation was inhibited by 36.18-63.90%, but the degradation rate increased by 146.15-392.31% when the initial concentration of syringic acid (SA), vanillic acid (VA), and 4-hydroxybenzoic acid (HBA) increased from 0.3 to 3.0 g/L. The collaboration among the functional microbes significantly enhanced the degradation rate of parent compounds and their intermediates in MFCs systems, while the accumulated intermediates severely inhibited their complete mineralization in fermentative systems. High-throughput sequencing showed that the growth of fermentative bacteria prevailed, but electrogenic bacteria were inhibited in the anode microbial community (AMC) under high concentrations of phenolic compounds (3.0 g/L). These findings provide a better understanding of the dynamic shift and synergy effects of the AMC to evaluate its potential for the treatment of phenolic-containing wastewater.

Characterization of genes encoding small heat shock proteins from Bemisia tabaci and expression under thermal stress.

Small heat shock proteins (sHSPs) are probably the most diverse in structure and function among the various super-families of stress proteins, and they play essential roles in various biological processes. The sweet potato whitefly, Bemisia tabaci (Gennadius), feeds in the phloem, transmits several plant viruses, and is an important pest on cotton, vegetables and ornamentals. In this research, we isolated and characterized three α-crystallin/sHSP family genes (Bthsp19.5, Bthsp19.2, and Bthsp21.3) from Bemisia tabaci. The three cDNAs encoded proteins of 171, 169, and 189 amino acids with calculated molecular weights of 19.5, 19.2, and 21.3 kDa and isoelectric points of 6.1, 6.2, and 6.0, respectively. The deduced amino acid sequences of the three genes showed strong similarity to sHSPs identified in Hemiptera and Thysanoptera insects species. All three sHSPs genes from Bemisia tabaci lacked introns. Quantitative real-time PCR analyses revealed that the three BtsHSPs genes were significantly up-regulated in Bemisia tabaci adults and pupae during high temperature stress (39, 41, 43, and 45 °C) but not in response to cold temperature stress (-6, -8, -10, and -12 °C). The expression levels of Bthsp19.2 and Bthsp21.3 in pupae was higher than adults in response to heat stress, while the expression level of Bthsp19.5 in adults was higher than pupae. In conclusion, this research results show that the sHSP genes of Bemisia tabaci had shown differential expression changes under thermal stress.

Enhanced Bio-Electro-Fenton degradation of phenolic compounds based on a novel Fe-Mn/Graphite felt composite cathode.

Phenolic compounds are problematic byproducts generated from lignocellulose pretreatment. In this study, the feasibility degradation of syringic acid (SA), vanillic acid (VA), and 4-hydroxybenzoic acid (HBA) by Bio-Electro-Fenton (BEF) system with a novel Fe-Mn/graphite felt (Fe-Mn/GF) composite cathode were investigated. The nano-scale Fe-Mn multivalent composite catalyst with core shell structure distributed more evenly on GF surface to form a catalyst layer with higher oxygen reduction reaction performance. Accordingly, the maximum power density generated with Fe-Mn/GF cathode was 48.1% and 238.9% higher than Fe/GF and GF respectively, which further enhanced the in situ generation of H2O2 due to the superiority of nano-scale core shell structure and synergistic effect of Fe and Mn species. The degradation efficiency of the three phenolic compounds in the BEF system could reached 100% after optimization of influencing parameters. Furthermore, a possible SA degradation pathway by BEF process in the present system was proposed based on the detected intermediates. These results demonstrated an efficient approach for the degradation of phenolic compounds derived from lignocellulose hydrolysates.

Increased fermentative adenosine production by gene-targeted Bacillus subtilis mutation.

Adenosine, which is produced mainly by microbial fermentation, plays an important role in the therapy of cardiovascular disease and has been widely used as an antiarrhythmic agent. In this study, guanosine 5'-monophosphate (GMP) synthetase gene (guaA) was inactivated by gene-target manipulation to increase the metabolic flux from inosine 5'-monophosphate (IMP) to adenosine in B. subtilis A509. The resulted mutant M3-3 showed an increased adenosine production from 7.40 to 10.45 g/L, which was further enhanced to a maximum of 14.39 g/L by central composite design. As the synthesis of succinyladenosine monophosphate (sAMP) from IMP catalysed by adenylosuccinate synthetase (encoded by purA gene) is the rate-limiting step in adenosine synthesis, the up-regulated transcription level of purA was the potential underlying mechanism for the increased adenosine production. This work demonstrated a practical strategy for breeding B. subtilis strains for industrial nucleoside production.

Isoliquiritigenin Induces Mitochondrial Dysfunction and Apoptosis by Inhibiting mitoNEET in a Reactive Oxygen Species-Dependent Manner in A375 Human Melanoma Cells.

The mitochondrial protein mitoNEET is a type of iron-sulfur protein localized to the outer membrane of mitochondria and is involved in a variety of human pathologies including cystic fibrosis, diabetes, muscle atrophy, and neurodegeneration. In the current study, we found that isoliquiritigenin (ISL), one of the components of the root of Glycyrrhiza glabra L., could decrease the expression of mitoNEET in A375 melanoma cells. We also demonstrated that mitoNEET could regulate the content of reactive oxygen species (ROS), by showing that the ISL-mediated increase in the cellular ROS content could be mitigated by the mitoNEET overexpression. We also confirmed the important role of ROS in ISL-treated A375 cells. The increased apoptosis rate and the decreased mitochondrial membrane potential were mitigated by the overexpression of mitoNEET in A375 cells. These findings indicated that ISL could decrease the expression of mitoNEET, which regulated ROS content and subsequently induced mitochondrial dysfunction and apoptosis in A375 cells. Our findings also highlight mitoNEET as a promising mitochondrial target for cancer therapy.

MiR-27b promotes sheep skeletal muscle satellite cell proliferation by targeting myostatin gene.

To investigate the role of miR-27b in sheep skeletal muscle development, here we first cloned the sequence of sheep pre-miR-27b, then further investigated its expression pattern in sheep skeletal muscle in vivo, the relationship of miR-27b expression and sheep skeletal muscle satellite cell proliferation and differentiation in vitro, and then finally confirmed its target gene during this development process. MiR-27b sequence, especially its mature sequence, was conservative among different species. MiR-27b highly expressed in sheep skeletal muscle than other tissues. In skeletal muscle of Suffolk and Bashbay sheep, miR-27b was upregulated during foetal period and downregulated during postnatal period significantly (P<0.01), but it still kept a relatively higher expression level in skeletal muscle of postnatal Suffolk sheep than Bashbay. There is a potential target site of miR-27b on 3'-UTR of sheep myostatin (MSTN) mRNA, and the double luciferase reporter assay proved that miR-27b could successfully bind on this site. When sheep satellite cells were in the proliferation status, miR-27b was upregulated and MSTN was downregulated significantly (P<0.01). When miR-27b mimics was transfected into sheep satellite cells, the cell proliferation was promoted and the protein level of MSTN was significantly downregulated (P<0.01). Moreover, miR-27b regulated its target gene MSTN by translation repression at an early step, and followed by inducing mRNA degradation in sheep satellite cells. Based on these results, we confirm that miR-27b could promote sheep skeletal muscle satellite cell proliferation by targeting MSTN and suppressing its expression.

[Behavior and Mechanisms of Cd(â ¡) Adsorption from Water by Niobate-Modified Titanate Nanosheets].

Niobate-modified titanate nanosheets (Nb-TNS) were synthesized through a hydrothermal method and used to remove Cd(Ⅱ) from water. TEM and SEM characterizations indicated that the new nanocomposites were non-curled nanosheets. XRD showed that the material was composed of sodium tri-titanate and niobate, and titanate was the primary component. Ion exchange between Cd2+ and Na+ in the interlayers of the Nb-TNSs was the dominant mechanism for Cd(Ⅱ) adsorption, leading to good adsorption performance. The material exhibited rapid adsorption kinetics for Cd(Ⅱ), reaching equilibrium within 60 min, and the data fit well with the pseudo-second order model (R2=1). The maximum adsorption capacity of Cd(Ⅱ) was 287.9 mg·g-1, according to the Langmuir isotherm model, which was larger than that of most of traditional adsorbents. Higher pH promoted adsorption because the negatively charged material could capture Cd(Ⅱ) cations more easily. Co-existing inorganic ions (Na+and Ca2+) were unfavorable to the adsorption of Cd(Ⅱ) by Nb-TNS owing to the competition for adsorption sites. In addition, a slight inhibition effect on the adsorption in the presence of humic acid (HA) was found. Cd(Ⅱ) was efficiently desorbed from Nb-TNS after HNO3 treatment, and -ONa sites were restored with NaOH treatment. Considering its simple synthesis method, high removal efficiency for heavy metals, and good reusability, Nb-TNS is a promising material for remediation of areas contaminated by heavy metals.

A new insight into the theoretical design of highly dispersed and stable ceria supported metal nanoparticles.

How to design and develop ceria supported metal nanoparticles (M/CeO2) catalysts with high performance and sintering resistance is a great challenge in heterogeneous catalysis and surface science. In the present work, we propose two ways to improve the anti-sintering capability of M/CeO2 catalysts. One is to introduce Ti atom on CeO2 (1 1 1) to form monatomically dispersed Ti, TiOx or TiO2-like species on ceria. Density functional theory calculations show that the much stronger interactions between Au and Ti modified CeO2 (1 1 1) occur compared with that on CeO2 (1 1 1). According to the electronic analysis, the strong interactions are attributed to the electron transfer from the Ti modified ceria substrate to Au. The other is to dope Ti into CeO2 (1 1 1) to form TixCe1-xO2. This also leads to the interaction enhancement between Au and TixCe1-xO2 (1 1 1). Electronic analysis indicates that the charge protuberance of surface O atoms near Ti atom results in the strong interactions between metal and ceria. This work provides new ideas for preparing M/CeO2 catalysts with high dispersity and stability, and sheds light into the theoretical design of catalysts.

Advance in quality assessment of Chinese materia medica using microscopic and morphological methods.

Quality evaluation plays a vital role in ensuring safety and effectiveness of Chinese materia medica (CMM). Microscopic and morphological technologies can be used to distinguish CMM's characteristics, such as shape, size, texture, section, and smell, for authenticity and quality control of CMM. The microscopic and morphological applications of novel micro-technology, colorimeter, and texture analyzer for CMM identification are summarized and the future prospect is discussed in this paper. Various styles and complex sources of CMM are systemically reviewed, including cormophyte medicinal materials, fruit and seeds, pollen grain, and spore materials.