Shining insights: Deciphering the biogenic synthesis of Ajuga bracteosa-mediated gold nanoparticles with advanced microscopy techniques.

In this study, gold nanoparticles (AuNPs) were bioreduced from Ajuga bracteosa, a medicinal herb known for its therapeutic properties against various diseases. Different fractions of the plant extract were used, including the methanolic fraction (ABMF), the n-hexane fraction (ABHF), the chloroform fraction (ABCF), and the aqueous extract for AuNPs synthesis. The characterization of AuNPs was performed using UV-Vis spectrophotometry, FT-IR, XRD, EDX, and TEM. UV-Vis spectroscopy confirmed the formation of AuNPs, with peaks observed at 555 nm. FT-IR analysis indicated strong capping of phytochemicals on the surface of AuNPs, which was supported by higher total phenolic contents (TPC) and total flavonoid contents (TFC) in AuNPs. XRD results showed high crystallinity and a smaller size distribution of AuNPs. TEM analysis revealed the spherical shape of AuNPs, with an average size of 29 ± 10 nm. The biologically synthesized AuNPs exhibited superior antibacterial, antioxidant, and cytotoxic activities compared to the plant extract fractions. The presence of active biomolecules in A. bracteosa, such as neoclerodan flavonol glycosides, diterpenoids, phytoecdysone, and iridoid glycosides, contributed to the enhanced biological activities of AuNPs. Overall, this research highlights the potential of A. bracteosa-derived AuNPs for various biomedical applications due to their remarkable therapeutic properties and effective capping by phytochemicals. RESEARCH HIGHLIGHTS: This research underscores the growing significance of herbal medicine in contemporary healthcare by exploring the therapeutic potential of Ajuga bracteosa and gold nanoparticles (AuNPs). The study highlights the notable efficacy of A. bracteosa leaf extracts and AuNPs in treating bacterial infections, demonstrating their bactericidal effects on a range of strains. The anti-inflammatory properties of plant extracts and nanoparticles are evidenced through paw edema method suggesting their applicability in managing inflammatory conditions. These findings position A. bracteosa and AuNPs as potential candidates for alternative and effective approaches to modern medication.

Knowledge and Behavior of Primary Care Physicians Regarding Utilization of Standardized Tools in Screening and Assessment of Anxiety, Depression, and Mood Disorders at a Large Integrated Health System.

INTRODUCTION: Standardized screening, objective evaluation, and management of behavioral health conditions are major challenges in primary care. The Generalized Anxiety Disorder Scale (GAD-7), Patient Health Questionnaire (PHQ-9), and Mood Disorder Questionnaire (MDQ) provide standardized screening and symptom management tools for generalized anxiety disorder (GAD), major depressive disorder (MDD), and Mood Disorders (MD), respectively. This study explores family physicians' knowledge, attitudes, and practices regarding the utilization of GAD-7, PHQ-9, and MDQ in outpatient primary care offices. METHODS: The study method was a cross-sectional electronic and paper survey utilizing a self-administered questionnaire that assessed primary care physicians' demographics, knowledge, attitudes, and practices in rural and urban outpatient clinical settings regarding GAD-7, PHQ-9, and MDQ. Statistical software SAS 9.4 was used for descriptive and Chi-Square statistics. RESULTS: Out of 320 total participants,145 responded (45.3%). Responding family physicians demonstrated a high level of familiarity with the GAD-7 (97.9%), PHQ-9 (97.9%), and MDQ (81.3%) assessment tools. However, the reported utilization rates were relatively lower than knowledge, with 62.7%, 73.1%, and 31.9% extremely likely or likely to utilize the GAD-7, PHQ-9, and MDQ as screening and monitoring tools, respectively. Less than a quarter of the total respondents use the objective score for the future management of GAD, with significantly more residents utilizing the score for GAD-7 compared to attendings (P < .05). There was no statistical significance difference between residents and attendings for the objective evaluation of Major Depressive Disorder (P = .26) and Mood Disorders (P = .05). CONCLUSIONS: Despite being knowledgeable of the utility of GAD-7, PHQ-9, and MDQ, the primary care physicians in a large integrated health system in Central Pennsylvania and Northern Maryland report inconsistent utilization in their practice. Further studies are needed to determine the underlying factors contributing to the suboptimal usage of these screening tools and ways to increase it.

Phytochemical-Mediated Biosynthesis of Silver Nanoparticles from Strobilanthes glutinosus: Exploring Biological Applications.

The application of green synthesis for silver nanoparticles in nanomedicine has experienced significant growth. Strobilanthes glutinosus, a plant primarily located in the Himalayas, remains largely unexplored. Considering the biomedical value of S. glutinosus, phytochemicals from this plant were used for the biosynthesis of silver nanoparticles. Silver nanoparticles were synthesized from aqueous extract of root and leaves of Strobilanthes glutinosus. The synthesized silver nanoparticles were characterized using UV-Vis spectrophotometry, Fourier-transform infrared spectroscopy, transmission electron microscopy, and X-ray diffraction. Total phenolic and flavonoid contents of plants were determined and compared with nanoparticles. The biomedical efficacy of plant extracts and silver nanoparticles was assessed using antioxidant and antibacterial assays. The UV-Vis spectra of leaf- and root-extract-mediated AgNPs showed characteristic peaks at 428 nm and 429 nm, respectively. TEM images revealed the polycrystalline and spherical shapes of leaf- and root-extract-mediated AgNPs with size ranges of 15-60 nm and 20-52 nm, respectively. FTIR findings shown the involvement of phytochemicals of root and leaf extracts in the reduction of silver ions into silver nanoparticles. The crystalline face-centered cubic structure of nanoparticles is depicted by the XRD spectra of leaf and root AgNPs. The plant has an ample amount of total phenolic content (TPC) and total flavonoid content (TFC), which enhance the scavenging activity of plant samples and their respective AgNPs. Leaf and root AgNPs have also shown good antibacterial activity, which may enhance the medicinal value of AgNPs.

Biofabrication of Fe3O4 Nanoparticles from Spirogyra hyalina and Ajuga bracteosa and Their Antibacterial Applications.

Iron oxide nanoparticles (NPs) have attracted substantial interest due to their superparamagnetic features, biocompatibility, and nontoxicity. The latest progress in the biological production of Fe3O4 NPs by green methods has improved their quality and biological applications significantly. In this study, the fabrication of iron oxide NPs from Spirogyra hyalina and Ajuga bracteosa was conducted via an easy, environmentally friendly, and cost-effective process. The fabricated Fe3O4 NPs were characterized using various analytical methods to study their unique properties. UV-Vis absorption peaks were observed in algal and plant-based Fe3O4 NPs at 289 nm and 306 nm, respectively. Fourier transform infrared (FTIR) spectroscopy analyzed diverse bioactive phytochemicals present in algal and plant extracts that functioned as stabilizing and capping agents in the fabrication of algal and plant-based Fe3O4 NPs. X-ray diffraction of NPs revealed the crystalline nature of both biofabricated Fe3O4 NPs and their small size. Scanning electron microscopy (SEM) revealed that algae and plant-based Fe3O4 NPs are spherical and rod-shaped, averaging 52 nm and 75 nm in size. Energy dispersive X-ray spectroscopy showed that the green-synthesized Fe3O4 NPs require a high mass percentage of iron and oxygen to ensure their synthesis. The fabricated plant-based Fe3O4 NPs exhibited stronger antioxidant properties than algal-based Fe3O4 NPs. The algal-based NPs showed efficient antibacterial potential against E. coli, while the plant-based Fe3O4 NPs displayed a higher zone of inhibition against S. aureus. Moreover, plant-based Fe3O4 NPs exhibited superior scavenging and antibacterial potential compared to the algal-based Fe3O4 NPs. This might be due to the greater number of phytochemicals in plants that surround the NPs during their green fabrication. Hence, the capping of bioactive agents over iron oxide NPs improves antibacterial applications.

Comparable quality performance between telemedicine and office-based care for abnormal BMI screening and management.

Background: Despite widespread adoption during COVID-19, there is limited evidence supporting the quality of telemedicine care in managing patients with abnormal BMI. Objective: To evaluate the comparability of telemedicine and in-person (office) quality performance for abnormal body mass index (BMI kg/m2) screening and management in primary care. Methods: This retrospective cohort study measured Healthcare Effectiveness Data and Information Set (HEDIS) quality performance for abnormal BMI screening (patients with BMIs <18.5 or >25 kg/m2 and a qualifying documented follow up plan) across an 8-hospital integrated health system seen via primary care from 4/1/20 - 9/30/21. Encounters were divided into three exposure groups: office (excluding telemedicine), telemedicine (excluding office), and blended telemedicine (office + telemedicine). Demographic stratification compared group composition. Chi squared tests determined statistical differences in quality performance (p = <0.05). Results: Demographics of sub-groups for the 287,387 patients (office: 222,333; telemedicine: 1,556; blended-telemedicine: 63,489) revealed a modest female predominance, majority ages 26-70, mostly White non-Hispanics of low health risk, and the majority BMI representation was overweight, followed closely by class 1 obesity. In both HEDIS specified and HEDIS modified performance, blended-telemedicine performed better than office (12.56%, 95% CI 12.29%-13.01%; 11.16%, 95% CI: 10.85%-11.48%; p < 0.0001); office performed better than telemedicine (4.29%, 95% CI 2.84%-5.54%; 4.79%, 95% CI 3.99%-5.35%; p < 0.0001). Conclusion: Quality performance was highest for blended-telemedicine, followed by office-only, then telemedicine-only. Given the known cost savings, adding telemedicine as a care venue might promote value within health systems without negatively impacting HEDIS performance.

An efficient and simple strategy for fabricating a polypyrrole decorated ceramic-polymeric porous membrane for purification of a variety of oily wastewater streams.

A ceramic-polymeric membrane was fabricated through in-situ oxidative polymerization of pyrrole (Py) on alumina (Al2O3) ceramic ultrafiltration support. The establishment of polypyrrole (PPy) active layer on the ceramic support led to a new PPy coated ceramic-polymeric membrane. Various salient features such as surface wettability, surface morphology, composition and functional goups of PPy coated ceramic-polymeric membrane were determined by various characterization techniques water contact angle (WCA), scanning electron microscopy (SEM), energy dispersive x-ray (EDX) analysis and attenuated total reflectance fourier transform infrared (ATR-FTIR). The PPy coated ceramic-polymeric membrane showed superhydrophilic nature owing to its under water oil contact angle of ≥160° (superoleophobic). Thanks to stable deposition of PPy active layer on ceramic support, the membrane retained a separation efficiency of >99% for O/W emulsions at varied transmembrane pressures ranging from 0.5 bar to 2 bar with a feed concentration of 125 ppm of oil in water. Moreover, the PPy coated ceramic-polymeric membrane exhibited an ideal behaviour to the applied transmembrane pressure with a linear increase from 380 LMH to 2112 LMH in permeate flux as the pressure increased from 0.5 bar to 2 bar. As the concentration of oil was raised from 50 ppm to 250 ppm, the separation efficeincy separation remained at >99%. From among the different types of oils (Motor oil, Diesel oil and Crude oil) to mimic the oily waste water streams, the permeate flux was found to be highest in case of motor oil with a value reaching to 1690 LMH at 1 bar. The stability test revealed that the PPy coated ceramic-polymeric membrane was able to separate >99% of 125 ppm O/W surfactant stabilized emulsion for a period of 420 min.

Comparison of Quality Performance Measures for Patients Receiving In-Person vs Telemedicine Primary Care in a Large Integrated Health System.

Importance: Despite its rapid adoption during the COVID-19 pandemic, it is unknown how telemedicine augmentation of in-person office visits has affected quality of patient care. Objective: To examine whether quality of care among patients exposed to telemedicine differs from patients with only in-person office-based care. Design, Setting, and Participants: In this retrospective cohort study, standardized quality measures were compared between patients with office-only (in-person) visits vs telemedicine visits from March 1, 2020, to November 30, 2021, across more than 200 outpatient care sites in Pennsylvania and Maryland. Exposures: Patients completing telemedicine (video) visits. Main Outcomes and Measures: χ2 tests determined statistically significant differences in Health Care Effectiveness Data and Information Set (HEDIS) quality performance measures between office-only and telemedicine-exposed groups. Multivariable logistic regression controlled for sociodemographic factors and comorbidities. Results: The study included 526 874 patients (409 732 office-only; 117 142 telemedicine exposed) with a comparable distribution of sex (196 285 [49.7%] and 74 878 [63.9%] women), predominance of non-Hispanic (348 127 [85.0%] and 105 408 [90.0%]) and White individuals (334 215 [81.6%] and 100 586 [85.9%]), aged 18 to 65 years (239 938 [58.6%] and 91 100 [77.8%]), with low overall health risk scores (373 176 [91.1%] and 100 076 [85.4%]) and commercial (227 259 [55.5%] and 81 552 [69.6%]) or Medicare or Medicaid (176 671 [43.1%] and 52 513 [44.8%]) insurance. For medication-based measures, patients with office-only visits had better performance, but only 3 of 5 measures had significant differences: patients with cardiovascular disease (CVD) receiving antiplatelets (absolute percentage difference [APD], 6.71%; 95% CI, 5.45%-7.98%; P < .001), patients with CVD receiving statins (APD, 1.79%; 95% CI, 0.88%-2.71%; P = .001), and avoiding antibiotics for patients with upper respiratory infections (APD, 2.05%; 95% CI, 1.17%-2.96%; P < .001); there were insignificant differences for patients with heart failure receiving ß-blockers and those with diabetes receiving statins. For all 4 testing-based measures, patients with telemedicine exposure had significantly better performance differences: patients with CVD with lipid panels (APD, 7.04%; 95% CI, 5.95%-8.10%; P < .001), patients with diabetes with hemoglobin A1c testing (APD, 5.14%; 95% CI, 4.25%-6.01%; P < .001), patients with diabetes with nephropathy testing (APD, 9.28%; 95% CI, 8.22%-10.32%; P < .001), and blood pressure control (APD, 3.55%; 95% CI, 3.25%-3.85%; P < .001); this was also true for all 7 counseling-based measures: cervical cancer screening (APD, 12.33%; 95% CI, 11.80%-12.85%; P < .001), breast cancer screening (APD, 16.90%; 95% CI, 16.07%-17.71%; P < .001), colon cancer screening (APD, 8.20%; 95% CI, 7.65%-8.75%; P < .001), tobacco counseling and intervention (APD, 12.67%; 95% CI, 11.84%-13.50%; P < .001), influenza vaccination (APD, 9.76%; 95% CI, 9.47%-10.05%; P < .001), pneumococcal vaccination (APD, 5.41%; 95% CI, 4.85%-6.00%; P < .001), and depression screening (APD, 4.85%; 95% CI, 4.66%-5.04%; P < .001). Conclusions and Relevance: In this cohort study of patients with telemedicine exposure, there was a largely favorable association with quality of primary care. This supports telemedicine's value potential for augmenting care capacity, especially in chronic disease management and preventive care. This study also identifies a need for understanding relationships between the optimal blend of telemedicine and in-office care.

PD-L1 Over-Expression Varies in Different Subtypes of Lung Cancer: Will This Affect Future Therapies?

Programmed death-ligand (PD-L) 1 and 2 are ligands of programmed cell death 1 (PD-1) receptor. They are members of the B7/CD28 ligand-receptor family and the most investigated inhibitory immune checkpoints at present. PD-L1 is the main effector in PD-1-reliant immunosuppression, as the PD-1/PD-L pathway is a key regulator for T-cell activation. Activation of T-cells warrants the upregulation of PD-1 and production of cytokines which also upregulate PD-L1 expression, creating a positive feedback mechanism that has an important role in the prevention of tissue destruction and development of autoimmunity. In the context of inadequate immune response, the prolonged antigen stimulation leads to chronic PD-1 upregulation and T-cell exhaustion. In lung cancer patients, PD-L1 expression levels have been of special interest since patients with non-small cell lung cancer (NSCLC) demonstrate higher levels of expression and tend to respond more favorably to the evolving PD-1 and PD-L1 inhibitors. The Food and Drug Administration (FDA) has approved the PD-1 inhibitor, pembrolizumab, alone as front-line single-agent therapy instead of chemotherapy in patients with NSCLC and PD-L1 ≥1% expression and chemoimmunotherapy regimens are available for lower stage disease. The National Comprehensive Cancer Network (NCCN) guidelines also delineate treatment by low and high expression of PD-L1 in NSCLC. Thus, studying PD-L1 overexpression levels in the different histological subtypes of lung cancer can affect our approach to treating these patients. There is an evolving role of immunotherapy in the other sub-types of lung cancer, especially small cell lung cancer (SCLC). In addition, within the NSCLC category, squamous cell carcinomas and non-G12C KRAS mutant NSCLC have no specific targetable therapies to date. Therefore, assessment of the PD-L1 expression level among these subtypes of lung cancer is required, since lung cancer is one of the few malignances wherein PD-L1 expression levels is so crucial in determining the role of immunotherapy. In this study, we compared PD-L1 expression in lung cancer according to the histological subtype of the tumor.

MicroRNA Mediated Plant Responses to Nutrient Stress.

To complete their life cycles, plants require several minerals that are found in soil. Plant growth and development can be affected by nutrient shortages or high nutrient availability. Several adaptations and evolutionary changes have enabled plants to cope with inappropriate growth conditions and low or high nutrient levels. MicroRNAs (miRNAs) have been recognized for transcript cleavage and translational reduction, and can be used for post-transcriptional regulation. Aside from regulating plant growth and development, miRNAs play a crucial role in regulating plant's adaptations to adverse environmental conditions. Additionally, miRNAs are involved in plants' sensory functions, nutrient uptake, long-distance root transport, and physiological functions related to nutrients. It may be possible to develop crops that can be cultivated in soils that are either deficient in nutrients or have extreme nutrient supplies by understanding how plant miRNAs are associated with nutrient stress. In this review, an overview is presented regarding recent advances in the understanding of plants' responses to nitrogen, phosphorus, potassium, sulfur, copper, iron, boron, magnesium, manganese, zinc, and calcium deficiencies via miRNA regulation. We conclude with future research directions emphasizing the modification of crops for improving future food security.

Sewage waste water application improves the productivity of diverse wheat (Triticum aestivum L.) cultivars on a sandy loam soil.

Water stress due to climate change is an emerging threat to wheat (Triticum aestivum L.) productivity in the arid regions of the world which will impact the future food security. In this scenario, the investigations are needed to check the feasibility of alternate sources of irrigation water to fulfill the irrigation demands of the crops in the arid regions. This 2-year study was aimed to investigate the influence of three irrigation sources (sewage water, canal water, and underground water) on the productivity of 10 wheat cultivars under an arid climate of Layyah, Pakistan. The results indicated that the number of fertile tillers, grains per spike, 1000-grain weight, and grain yield varied from 114 to 168 m-2, 34.8 to 53.3, 33.4 to 38.4 g, and 2.68 to 4.05 Mg ha-1, respectively in various wheat cultivars. The highest fertile tillers (168 m-2) were recorded in cultivar Gold-2016 followed by Aas-2011 (155 cm), AARI-2011 (153 m-2), and Ujala-2016 (150 m-2). The highest 1000-grain weight of 38.4 g was recorded in cultivar NARC-2016. The grains per spike (53.3) were the highest in cultivar Ujala-2016. The grain yields were the highest in cultivars Ujala-2016 (4.05 Mg ha-1) and Gold-2016 (3.91 Mg ha-1). The highest grain yield of 3.71 Mg ha-1 was recorded with sewage water irrigation against the grain yield of 3.18 and 2.91 Mg ha-1 in canal and underground water irrigation, respectively. There existed a strong co-relation of fertile tillers and grains per spike with the grain yield of wheat. Application of sewage water also enhanced the total nitrogen, extractable potassium, and available phosphorous in soil. In crux, the cultivation of recently bread wheat cultivars (viz. Ujala-2016, Gold-2016) and the irrigation of field with sewage water in the absence of canal water might be a viable option to boost wheat productivity under arid regions. A range of genetic variability existed for different traits in the cultivars; therefore, these can be used to breed wheat cultivars to be used for sewage water cultivation.

Antihyperglycemic effect of Conocarpus erectus leaf extract in alloxan-induced diabetic mice.

Synthetic drugs have widely been helpful in management of diabetes mellitus type 2. However, side effects associated with synthetic drugs serve as an impetus to explore plants as alternate mode of treatment. The hydroethanolic leaf extracts of Conocarpus erectus were evaluated for phenolic contents, flavonoid distribution, antioxidant activity and antidiabetic potential. The maximum extract yield, total phenolic and flavonoid contents were exposed by 60% ethanolic extract. The Antioxidant and anti α-glucosidase tendency of 60% ethanolic extract was the most promising and complemented by in-vivo antihyperglycemic impact on mice. The findings were substantial regarding suppression of blood glucose levels in alloxan induced diabetic mice establishing the Conocarpus erectus as proficient pool of nutraceuticals for diabetes mellitus type 2 management.

Cytotoxic activity of extracts and crude saponins from Zanthoxylum armatum DC. against human breast (MCF-7, MDA-MB-468) and colorectal (Caco-2) cancer cell lines.

BACKGROUND: Zanthoxylum armatum DC has been an important traditional plant known for its medicinal properties. It is well known for its antimicrobial, larvicidal and cytotoxic activities. METHODS: The potential anticancer effects of the methanol extract and the crude saponins from fruit, bark and leaves of Z. armatum on breast (MDA-MB-468 and MCF-7) and colorectal (Caco-2) cancer cell lines using MTT, neutral red uptake(NRU) and DAPI stain assays were evaluated. RESULTS: In MTT assay the methanol extract of fruit (Zf), bark (Zb) and leaves (Zl) of Zanthoxylum armatum, showed significant and dose dependent growth inhibition of MCF-7, MDA MB-468 and Caco-2 cancer cell lines in a dose of 200 µg/ml and above. The saponins (Zf.Sa, Zb.Sa and Zl.Sa) showed significant activity against MDA MB-468 (95, 94.5 and 85.3%) as compared to MCF-7 (79.8, 9.43, 49.08%) and Caco-2 (75.8, 61.8, 68.62%) respectively. The extracts were further tested in more sensitive NRU assay and its was found that Zf extract showed higher cytotoxic activity as compared to Zb and Zl extracts with 100 µg/ml concentration. The breast cancer cell lines showed more sensitivity toward the crude saponins from fruit and bark with maximum inhibition of up to 93.81(±2.32) % with respect to 71.19(± 2.76) of Actinomycin-D. DAPI staining experiment showed that saponins from fruit induced apoptosis mode of cell death in all three types of cell lines while saponins form leaves and bark showed similar results against MDA MB-468 indicated by nuclear fragmentation and chromatin condensation. The effect of saponins from fruit, bark and leaves (Zf.Sa, Zb.Sa and Zl.Sa) against Caco-2 cell lines inhibited the growth of Caco-2 by 53.16 (±3.31) %, 66.43 (± 3.24) and 45.96 (± 10.67) respectively with respect to Actinomycin-D (4 µM) which showed the growth inhibition of 65.40(±4.29) %. CONCLUSION: The current study clearly demonstrates that the extract and crude saponins from fruit, bark and leaves of traditional medicinal plant Zanthoxyllum armatum DC., has the potential to exert its cytotoxic effect on cancer cell lines isolated form human by a mechanism involving apoptosis. The overall finding demonstrate that this plant specially fruits, could be potential source of new anticancer compounds for possible drug development against cancer.

Evaluation of anti-oxidative, antimicrobial and anti-diabetic potential of Adiantum venustum and identification of its phytochemicals through GC-MS.

The present study aimed to explore antioxidant, antimicrobial and anti-diabetic properties of the fern Adiantum venustum. The TPC (total phenolic content) of methanolic extract of the plant was 247.95±0.0007µg of Gallic acid equivalents per gram of dried extract (mg GAE/g). The highest TPC was in n-butanolic fraction, which was 981.45±0.1562mg GAE/g. Hexane fraction showed lowest TPC (256.95±0.0420mg GAE/g). Ethyl acetate fraction exhibited highest total flavonoid content (TFC), i.e., 62.0±0.050mg of Rutin equivalents per gram of sample. DPPH (1,1-diphenyl-2- picryl hydrazyl) radical scavenging activity of the plant was significant. n-Butanolic fraction was most potent with IC50 being 1.06mg/mL. The IC50 of methanolic extract was 1.50mg/mL, that of aqueous fraction was 2.51 mg/mL, and that of chloroform fraction was 2.65mg/mL. Antibacterial potential of the fern was tested against two Gram-positive bacterial strains, Pseudomonas aeruginosa and Escherichia coli and two Gram-negative bacterial strains, Bacillus subtilis and Staphylococcus aureus. n-Butanolic fraction showed highest zone of inhibition (ZOI, 25.13±1.237 mm) against P. aeruginosa. Ethyl acetate fraction was most active (ZOI, 15.75±1.060 mm) against S. aureus. Against E. coli, aqueous and n-butanolic fractions were most active (ZOI, 14.75±0.353 and 14.50±0.707mm, respectively). Minimum inhibitory concentration (MIC) of methanolic extract against B. subtilis was as low as 1.98mg/mL. The hexane fraction was least toxic against all the fractions. The plant displayed significant alpha-amylase inhibitory activity. Chloroform fraction was most effective with lowest IC50 (1.10mg/mL) followed by ethyl acetate fraction (1.92mg/mL). A. venustum is rich in phenolics and has substantial antioxidant, antimicrobial and alpha-amylase inhibitory potential.

Anti-inflammatory constituents from Perovskia atriplicifolia.

CONTEXT: Perovskia atriplicifolia Benth (Labiantae) has long been used as a traditional herbal medicine for anti-inflammation in Pakistan; this prompted us to isolate anti-inflammatory compounds from this plant. OBJECTIVE: The objective of this study was to isolate and characterize the anti-inflammatory principles from Perovskia atriplicifolia. MATERIALS AND METHODS: The CHCl3-soluble fraction of the methanol extract of the whole plant on column chromatography yielded compounds 1-6. The anti-inflammatory potential of the compounds 1-6 was evaluated by Leukotriene C4 (LTC4) Release Assay which was performed according to the established protocol. LTC4 in the supernatant of each well was measured using an ELISA kit (Cayman Chemical Co., Ann Arbor, MI). RESULTS: The bioassay-guided phytochemical investigation of the CHCl3 soluble fraction of the methanol extract of Perovskia atriplicifolia furnished six compounds, abrotanone (1), abrotandiol (2), (+)-pinoresinol (3), (+)-syringaresinol (4), (+)-lariciresinol (5), and (+)-taxiresinol (6). The compounds (1-6) were evaluated for their inhibitory activities on LTC4 release. Among the tested compounds, (+)-taxiresinol (6) exhibited the most potent inhibition of LTC4 release with an IC50 value of 3.4 ± 0.09 µM followed by compounds 4, 5, 3, and 2 with an IC50 value ranging from 7.9 ± 0.04 to 17.2 ± 0.07 µM. Abrotanone (1) showed the lowest inhibition of LTC4 release with an IC50 value of 35.1 ± 0.05 µM (the positive control, zileuton, 0.77 ± 0.05 µM). CONCLUSION: Compounds 1-6 were found to possess inhibitory activity and seem to have potential therapeutic effect on inflammatory diseases.

Anticancer investigations on Carissa opaca and Toona ciliata extracts against human breast carcinoma cell line.

This study was aimed to determine the effectiveness of two ethnobotanically important plant species Carissa opaca and Toona ciliata against cancer cells. Antiproliferative activity of the plant extracts and their fractions was tested against MCF-7 breast cancer cell line using MTT assay. A concentration dependent inhibition was observed for both crude extracts. C. opaca crude extract showed 78.5% inhibition while T. ciliata showed 57% activity against cancer cells at 500 µg/mL. Fractions were tested at 200 µg/mL concentration and were more active than crude extracts. Chloroform fraction of C. opaca showed maximum inhibition 99% followed by ethyl acetate and methanol fraction of C. opaca exhibiting 96% and 94% inhibition, respectively. Ethyl acetate fraction of T. ciliata showed 78% inhibition of cancer cells at the same concentration. Preliminary phytochemical screening revealed the chemical composition of C. opaca extract containing alkaloids, flavonoids, tannins and saponins while T. ciliata had tannins and coumarins. Present investigation suggests that tested plant species possess potent anticancer compounds specially chloroform, ethyl acetate and methanol fractions of C. opaca and ethyl acetate fraction of T. ciliata can be an important source of anticancer drugs.

Targeted mutagenesis of mitochondrial carbonic anhydrases VA and VB implicates both enzymes in ammonia detoxification and glucose metabolism.

Prior studies with carbonic anhydrase (CA) inhibitors implicated mitochondrial CA in ureagenesis and gluconeogenesis. Subsequent studies identified two mitochondrial CAs. To distinguish the contribution of each enzyme, we studied the effects of targeted disruption of the murine CA genes, called Car5A and Car5B. The Car5A mutation had several deleterious consequences. Car5A null mice were smaller than wild-type littermates and bred poorly. However, on sodium-potassium citrate-supplemented water, they produced offspring in expected numbers. Their blood ammonia concentrations were markedly elevated, but their fasting blood sugars were normal. By contrast, Car5B null mice showed normal growth and normal blood ammonia levels. They too had normal fasting blood sugars. Car5A/B double-knockout (DKO) mice showed additional abnormalities. Impaired growth was more severe than for Car5A null mice. Hyperammonemia was even greater as well. Although fertile, DKO animals were produced in less-than-predicted numbers even when supplemented with sodium-potassium citrate in their drinking water. Survival after weaning was also reduced, especially for males. In addition, fasting blood glucose levels for DKO mice were significantly lower than for controls (153 ± 33 vs. 230 ± 24 mg/dL). The enhanced hyperammonemia and lower fasting blood sugar, which are both seen in the DKO mice, indicate that both Car5A and Car5B contribute to both ammonia detoxification (ureagenesis) and regulation of fasting blood sugar (gluconeogenesis). Car5A, which is expressed mainly in liver, clearly has the predominant role in ammonia detoxification. The contribution of Car5B to ureagenesis and gluconeogenesis was evident only on a Car5A null background.

A novel steroidal saponin glycoside from Fagonia indica induces cell-selective apoptosis or necrosis in cancer cells.

Fagonia indica is a small spiny shrub of great ethnopharmacological importance in folk medicine. The aqueous decoction of aerial parts is a popular remedy against various skin lesions, including cancer. We used a biological activity-guided fractionation approach to isolate the most potent fraction of the crude extract on three cancer cell lines: MCF-7 oestrogen-dependent breast cancer, MDA-MB-468 oestrogen-independent breast cancer, and Caco-2 colon cancer cells. A series of chromatographic and spectroscopic procedures were utilised on the EtOAc fraction, which resulted in the isolation of a new steroidal saponin glycoside. The cytotoxic activity of the saponin glycoside was determined in cancer cells using the MTT and neutral red uptake assays. After 24h treatment, the observed IC(50) values of the saponin glycoside were 12.5 µM on MDA-MB-468 and Caco-2 cells, but 100 µM on MCF-7 cells. Several lines of evidence: PARP cleavage, caspase-3 cleavage, DNA ladder assays, and reversal of growth inhibition with the pan-caspase inhibitor Z-VAD-fmk, suggested stimulation of apoptosis in MDA-MB-468 and Caco-2 cells, but not in MCF-7 cells, which do not express caspase-3. The haemolytic activity of the saponin glycoside was confirmed in sheep red blood cells, with cell lysis observed at >100 µM, suggesting that, at this concentration, the saponin glycoside caused necrosis through cell lysis in MCF-7 cells. Using the DNA ladder assay, the saponin glycoside (12.5 µM) was not toxic to HUVEC (human umbilical vein endothelial cells) or U937 cells, indicating some selectivity between malignant and normal cells. We conclude that the steroidal saponin glycoside isolated from F. indica is able to induce apoptosis or necrosis in cancer cells depending on the cell type.

In vitro cytotoxic activity of Aesculus indica against breast adenocarcinoma cell line (MCF-7) and phytochemical analysis.

Aesculus indica (Linn.) (Sapindaceae) is an ethanobotanically important plant specie traditionally used against rheumatism, skin and vein complaints. Cytotoxic potential of Aesculus indica crude leaf extract and its fractions was investigated against MCF-7 cell line. Crude extract of Aesculus indica was prepared in methanol by maceration technique. Crude extract was fractionated into four organic and one aqueous fraction on polarity basis. MTT assay was used to evaluate the reduction of viability of MCF-7 breast cancer cell line. Cell viability was inhibited by Aesculus indica crude extract in a dose dependent manner ranging from 34.2% at 10 µg/ml to 94% at 500µg/ml. Activity was found in an ascending order from hexane showing 29.8% inhibition to aqueous fraction indicating maximum inhibition, 60%. Phytochemical analysis of crude and fractionated extracts revealed presence of flavonoids, saponins, coumarins and tannins upto varying degrees. Methanol and aqueous fraction of methanol extract of Aesculus indica can be good source of cytotoxic compounds.

Novel acylated steroidal glycosides from Caralluma tuberculata induce caspase-dependent apoptosis in cancer cells.

AIM OF THE STUDY: Pregnane glycosides are potent cytotoxic agents which may represent new leads in the development of anti-tumour drugs, particularly in the treatment of breast cancer, because of the structural similarity to estrogenic agonists. Caralluma species are natural sources of a wide variety of pregnane glycosides. The aim of the study was to isolate, using an activity-guided fractionation approach, novel pregnane glycosides for testing on breast cancer and other tumour lines. MATERIALS AND METHODS: The effect of crude extracts, specific organic fractions and isolated compounds from Caralluma tuberculata was tested on the growth and viability of MCF-7 estrogen-dependent, and MDA-MB-468 estrogen-independent breast cancer cells, Caco-2 human colonic cells, HUVECs and U937 cells. Neutral red uptake and MTT assays were used. Apoptosis was detected by Western blot of poly-(ADP ribose) polymerase (PARP) as were other markers of nuclear fragmentation (DNA ladder assay, staining of cells with nuclear dye DAPI). The involvement of caspases was investigated using the pan-caspase inhibitor Z-VAD-FMK. RESULTS: The ethyl acetate fraction of Caralluma tuberculata was found to be the most potent anti-proliferative fraction against all three cancer cell lines. Two novel steroidal glycosides were isolated from the active fraction after a series of chromatographic experiments. The structure of the isolated compounds was elucidated solely based on 2D-NMR (HMBC, HETCOR, DQF-COSY) and MS spectral analysis as compound 1: 12-O-benzoyl-20-O-acetyl-3ß,12ß,14ß,20ß-tetrahydroxy-pregnan-3-ylO-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1 → 4)-3-methoxy-ß-D-ribopyranoside, and as compound 2: 7-O-acetyl-12-O-benzoyl-3ß,7ß,12ß,14ß-tetrahydroxy-17ß-(3-methylbutyl-O-acetyl-1-yl)-androstan-3-ylO-ß-D-glucopyranosyl-(1 → 4)-6-deoxy-ß-D-allopyranosyl-(1 → 4)-ß-D-cymaropyranosyl-(1 → 4)-ß-D-cymapyranosyl-(1→ 4)-ß-D-cymaropyranoside. Compound 1 (pregnane glycoside) and compound 2 (androstan glycoside) induced apoptosis at <25 µM after 48 h as assessed by cell shrinkage, PARP cleavage, DNA fragmentation, and reversal with the caspase inhibitor. CONCLUSIONS: Two novel steroid glycosides isolated from Caralluma tuberculata possess moderate, micromolar cytotoxic activity on breast cancer and other cells in vitro, which may indicate a source of activity in vivo of interest to future drug design.

Electrically wired mitochondrial electrodes for measuring mitochondrial function for drug screening.

In the continual search of new therapeutics, many possible drug candidates are excluded, because they are found to negatively affect mitochondrial function. We have developed an approach for directly, electrochemically assaying mitochondrial metabolic activity as a function of metabolic substrate to determine drug toxicity. By wiring mouse mitochondria to a carbon electrode surface, electrons can be intercepted before they reach Complex IV, the terminal step of electron transport chain. The electrons are rerouted, to a separate electrode of the electrochemical cell, the cathode. This allows for the direct measurement of electrical current and potential of the mitochondria during their oxidation of substrates such as pyruvate and fatty acids when there are different concentrations of drug present. This analytical technique has been shown to reliably assay several classical mitochondrial toxins and exhibits potential for the further development of a drug candidate screening technique, as well as other applications where the quantitative study of mitochondrial dysfunction is important.