Endophytic fungus Penicillium steckii DF33 promoted tanshinones biosynthesis in Salvia miltiorrhiza by regulating the expression of CYP450 genes.

Salvia miltiorrhiza, a prominent traditional Chinese medicinal resource, has been extensively employed in the management of cardiovascular and cerebrovascular ailments. Ensuring the consistency of S. miltiorrhiza raw materials revolves around the imperative task of maintaining stable tanshinones content and composition. An effective approach in this regard involves the utilization of endophytic fungi as inducers. Within this context, our study spotlights an endophytic fungus, Penicillium steckii DF33, isolated from the roots of S. miltiorrhiza. Remarkably, this fungus has demonstrated a significant capacity to boost the biosynthesis and accumulation of tanshinones. The primary objective of this investigation is to elucidate the underlying regulatory mechanism by which DF33 enhances and regulates the biosynthesis and accumulation of tanshinones. This is achieved through its influence on the differential expression of crucial CYP450 genes within the S. miltiorrhiza hairy roots system. The results revealed that the DF33 elicitor not only promotes the growth of hairy roots but also enhances the accumulation of tanshinones. Notably, the content of cryptotanshinone was reached 1.6452 ± 0.0925 mg g-1, a fourfold increase compared to the control group. Our qRT-PCR results further demonstrate that the DF33 elicitor significantly up-regulates the expression of most key enzyme genes (GGPPS, CPS1, KSL1, CYP76AH1, CYP76AH3, CYP76AK1, CYP71D411) involved in the tanshinone biosynthesis pathway. This effect is particularly pronounced in certain critical CYP450 genes and Tanshinone ⅡA synthase (SmTⅡAS), with their expression levels peaking at 7 days or 14 days, respectively. In summary, endophytic P. steckii DF33 primarily enhances tanshinone biosynthesis by elevating the expression levels of pivotal enzyme genes associated with the modification and transformation stages within the tanshinone biosynthesis pathway. These findings underscore the potential of employing plant probiotics, specifically endophytic and root-associated microbes, to facilitate the biosynthesis and transformation of vital constituents in medicinal plants, and this approach holds promise for enhancing the quality of traditional Chinese medicinal materials.

Antioxidative and Cytoprotective Effects of Ganoderma applanatum and Fomitopsis pinicola in PC12 Adrenal Phaeochromocytoma Cells.

Considering the impact of oxidative stress on the development of many diseases, together with the role of natural antioxidants in maintaining physiological balance in humans, medicinal mushrooms are potential sources of bioactive compounds against many diseases. In the present work, in vitro evaluation of the biological activities of the alcoholic extracts of two wild tree mushrooms, namely, Ganoderma applanatum and Fomitopsis pinicola, has been performed. Extraction of G. applanatum (GAE) and F. pinicola (FPE) was conducted with 60% ethanol and 100% ethanol sequentially. UPLC-MS/MS identification was conducted on the two mushrooms extracts. A total of 15 substances were identified in GAE, including 3 spiro meroterpenoids and 12 triterpenoids; a total of 14 chemical constituents were iden¬tified in FPE, including 8 triterpenoids, 4 triterpene glycosides, 1 lanosterol, and 1 lanostanoid. The resulting extracts were examined for their in vitro antioxidative and cytoprotective effects against AAPH-induced oxidative damage. Our results demonstrated that both extracts have potent antioxidative activities, when GAE was 0.2 mg/mL, the clearance rates of DPPH and ABTS have reached 93.34% and 99.93%, respectively. When FPE was 1.4 mg/mL and 0.6 mg/mL, the scavenging rates of DPPH and ABTS have reached 91.76% and 100%, respectively. Both the alcoholic extracts of G. applanatum and F. pinicola were able to protect the AAPH-induced damage and could effectively inhibit cell aging via ß-galactosidase (SA ß-gal) staining activity test and scanning electron microscopy analysis.

Endophytic fungus Cladosporium tenuissimum DF11, an efficient inducer of tanshinone biosynthesis in Salvia miltiorrhiza roots.

Salvia miltiorrhiza is a traditional medicinal plant mainly used for cardiovascular and cerebrovascular disease treatment. Tanshinones are the main bioactive constituents of S. miltiorrhiza, which mainly accumulate around its root periderm tissue. Endophytic fungi are important bioelicitors or probiotics that can promote the accumulation of secondary metabolites and sustainable cultivation of medicinal plants. Among them, endophytic Cladosporium spp., possessing a variety of biotransformation and metabolic abilities, is an ideal elicitor source. Here, we used a gnotobiotic system to investigate the effects of the endophytic fungus Cladosporium tenuissimum DF11 on tanshinone biosynthesis in S. miltiorrhiza roots. The results showed that C. tenuissimum DF11 mainly colonizes the intercellular space of the root tissues and promotes tanshinone biosynthesis and accumulation in S. miltiorrhiza roots by upregulating the expression of the genes encoding for key enzymes HMGR, DXS, DXR, GGPPS, CPS, KSL and CYP76AH1 of the tanshinone biosynthesis pathway. The expression levels of almost all genes encoding for key enzymes reached the response peak in the first or second week after DF11 colonization. Taken together, the endophytic fungus C. tenuissimum DF11 could promote secondary metabolite accumulation in S. miltiorrhiza roots. These results indicate that DF11 will be a potential biofertilizer fungus to regulate and stabilize the quality of cultivated S. miltiorrhiza medicinal materials.

Insight into the antioxidant activities of ten Fabaceae plant species that are medicinally used by the Aucan Tribal Peoples from the Republic of Suriname (South America).

Fabaceae are associated with a high antioxidant activity (AA) and a high total phenolic (TPC), total flavonoid (TFC), and selenium content (SeC). In this study, the aqueous extracts from ten Fabaceae species that are medicinally used by the Aucan Tribal Peoples from Suriname (South America), were evaluated for AA using a DPPH and a FRAP assay, and for TPC, TFC, and SeC using Folin-Ciocalteu's, an AlCl3 colorimetric, and an azure B-based method. Associations between pairs of these variables were determined by Pearson correlation coefficient. One-way ANOVA with post-hoc Tukey's test was used to evaluate the data for statistically significant differences (p < 0.05). The I. stipularis (bark), C. guyanensis (bark), A. jupunba (twigs), and M. urens (fruit) extracts had the highest DPPH IC50 values (36 - 70 µg/mL) and FRAP values (346 - 573 µM FeE/100 µg) and the highest TPC (25 - 41 GAEq/100 µg), TFC (21 - 39 REq/100 µg), and SeC (4 -17 µg/g). The values for the T. indica (leaf), P. macroloba (bark), M. pigra (whole plant), S. quinquangulata (leaf), A. sensitiva (whole plant), and L. leucocephala (leaf) extracts were > 10-fold lower. AA, TPC, TFC, and SeC correlated well with each other (correlation coefficient ≥ 0.83, p ≤ 0.0030). Thus, AA, TPC, TFC, and SeC may represent important determinants of the health benefits of the former four samples but not of the others. Future studies should focus on the precise contribution of AA, TPC, TFC, and SeC to the therapeutic value of medicinal Fabaceae.

Vaginal herb use and Chlamydia trachomatis infection: cross-sectional study among women of various ethnic groups in Suriname.

OBJECTIVE: Vaginal steam baths with herb leaves (herb use) is practised by some Surinamese women. We assessed herb use among women from the five most prevalent ethnic groups, and if herb use is associated with Chlamydia trachomatis infection. SETTING: Participants were recruited at a sexually transmitted infection (STI) clinic and a family planning clinic (FP) in Paramaribo, Suriname. PARTICIPANTS: 1040 women were included subsequently, comprising the following ethnic groups: Creole (26.7%), Hindustani (24.6%), Javanese (15.7%), Maroon (13.3%) and mixed descent (19.7%). METHODS: Nurses collected a questionnaire and vaginal swabs for nucleic acid amplification C. trachomatis testing. PRIMARY OUTCOMES: Determinants of vaginal herb use and C. trachomatis infection via univariable and multivariable logistic regression. RESULTS: Herb use was most common among Maroon (68.8%) and Creole women (25.2%). In multivariable analysis including only Maroon and Creole women, determinants significantly associated with vaginal herb use were (OR; 95% CI): Maroon ethnic descent (5.33; 3.26 to 8.71 vs Creole), recruitment at the STI clinic (2.04; 1.24 to 3.36 vs FP), lower education levels (3.80; 1.68 to 8.57 lower vs higher, and 2.02; 0.90 to 4.51 middle vs higher). Lower age and recruitment at the STI clinic were associated with C. trachomatis infection, but not vaginal herb use. CONCLUSION: In Suriname, vaginal herb use is common among Maroon and Creole women. Education, ethnic group and recruitment site were determinants for herb use. Vaginal herb use was not a determinant of C. trachomatis infection. Future research should focus on the effect of herb use on the vaginal microbiome and mucosal barrier.

Fifty years of primary health care in the rainforest: temporal trends in morbidity and mortality in indigenous Amerindian populations of Suriname.

BACKGROUND: The Amazonian Amerindian populations living in the southern and southwestern hinterlands of Suriname (South America) have come into contact with western health care since approximately fifty years ago. In this study, secondary data were used to assess the impact of Medical Mission's fifty-year old primary health care program on the health status of these populations. METHODS: Using data from the primary health care facilities of Medical Mission for 1965-1970, 1973-1977, 1982-1985, and 1997-2014, temporal trends in incidence and mortality of respiratory tract infections, gastroenteritis, and malaria; population composition; birth and death rates; and polyclinic consultations in these communities have been assessed over the period between 1965 and 2014. RESULTS: In the period covered by this study, the incidence of respiratory tract infections and gastroenteritis declined by about 75% and 53%, respectively, while malaria incidence rose sharply from the 1980s through 2005 but subsequently declined to levels approximating elimination. Crude death rates dropped by about 70% while birth rates declined by about 50% in the 1980s and since then remained at this level. The population doubled in size and increased in all age groups, particularly in the age group of ≥59 years. The infant mortality rate declined by 50%. In addition, the average yearly number of polyclinic visits per person decreased 6- to 7-fold during this period. CONCLUSIONS: The significant reduction of the infectious disease burden; the doubling of the population size and the growth of the proportion of elderly individuals due to the declining death rates; the declining infant mortality rates to levels comparable to the national average as well as the decline in average numbers of polyclinic consultations per person, indicate that Medical Missions health service provision achieved its goal of improving the health and survival of the indigenous people by providing free, accessible and permanent medical services. Building upon this successful experience Medical Mission could be instrumental in addressing potential contemporary life-style related health threats.

Beneficial effect of medicinal plants on the contractility of post-hypoxic isolated guinea pig atria - Potential implications for the treatment of ischemic-reperfusion injury.

Context Ischemic-reperfusion injury is accompanied by a decreased contractility of the myocardium. Positive-inotropic agents have proven useful for treating this condition but may exert serious side-effects. Objective In this study, aqueous preparations from Abelmoschus esculentus L. Moench (Malvaceae), Annona muricata L. (Annonaceae), Bixa orellana L. (Bixaceae), Cecropia peltata L. (Moraceae), Erythrina fusca Lour. (Fabaceae), Psidium guajava L. (Myrtaceae) and Terminalia catappa L. (Combretaceae) were evaluated for their ability to improve the decreased contractility of isolated guinea pig atria after hypoxic stress. Materials and methods Guinea pig atria isolated in Ringer-Locke buffer gassed with 100% O2 at 30 °C were exposed for 5 min to hypoxia, then allowed to recover in oxygenated buffer alone or containing a single plant extract (0.001-1 mg/mL). The contractility (g/s) and beating frequency (beats/min), as well as troponin C contents of the bathing solution (ng/mL), were determined and expressed as means ± SDs. Results The extracts of A. muricata, B. orellana, C. peltata and T. catappa caused an increase in the contractility compared to untreated atria of 340 ± 102%, 151 ± 13%, 141 ± 14% and 238 ± 44%, respectively. However, the latter two preparations increased the troponin C contents of the bathing solution to 36 ± 11 and 69 ± 33, compared to the value of 11 ± 3 ng/mL found with untreated atria. Conclusions Preparations from A. muricata and B. orellana may possess positive-inotropic properties which may improve the contractility of the post-hypoxic myocardium. Studies to assess their usefulness in ischemic-reperfusion injury are warranted.