Evaluation of Safety of Stewart's Wood Fern (Dryopteris stewartii) and Its Anti-Hyperglycemic Potential in Alloxan-Induced Diabetic Mice.

Diabetes has become a critical challenge to the global health concerns. Cytotoxicity and development of resistance against available drugs for management of diabetes have shifted the focus of global scientific researchers from synthetic to herbal medications. Therefore, the current study was conducted to investigate the possible anti-hyperglycemic potential of Dryopteris stewartii using Swiss albino mice. To evaluate any possible toxic effect of the plant, acute oral toxicity test was performed while the anti-diabetic effects of aqueous and ethanol extracts at 500 mg/kg, positive, negative and normal control were assessed simultaneously. The anti-diabetic study revealed that aqueous extract has higher anti-diabetic potential than ethanol extract while lowered blood glucose level at second week reaching 150 mg/dL, exerting stronger anti-diabetic effects, compared to ethanol extract (190 mg/dL). Oral glucose tolerance findings revealed that aqueous extract decreased blood glucose level by -0.41-fold, compared to ethanol extract showing a decrease by only -0.29-folds. The histopathological evaluation of liver and pancreas of all groups revealed normal cell architecture with no morphological abnormalities. These results suggested the possible use of D. stewartii as anti-diabetic herbal drug in near future. However, these recommendations are conditioned by deep mechanistic studies.

Application of green silver nanoparticles synthesized from the red seaweeds Halymenia porphyriformis and Solieria robusta against oral pathogenic bacteria by using microscopic technique.

Aqueous extracts of two red seaweeds Halymenia porphyriformis and Solieria robusta were used to synthesize green silver nanoparticles. These biogenic nanoparticles were tested against four strains of oral pathogenic bacteria which cause tooth decay or cavities. Staphylococcus aureus (MT416445), Streptococcus viridans (MT416448), Lactobacillus acidophilus (MT416447) and Lactobacillus brevis (MT416446) were used. Characterization of AgNPs was done by UV-Visible spectroscopy, SEM, XRD and FTIR. XRD analysis revealed the crystalline nature of the particles. The size analysis by XRD of the green AgNPs by H. porphyriformis indicated it had smaller particles, 15.23 nm, when compared to AgNPs by S. robusta (17 nm). Both green synthesized silver nanoparticles showed moderate antibacterial activity against all strains of bacteria, except L. acidophilus. Both particles showed their maximum zone of inhibition against L. acidophilus at a lower concentration of 50 and 100 µg. However, it was concluded that silver nanoparticles of H. porphyriformis are more effective than that of S. robusta due to their smaller size.

Authentication of the therapeutic Lamiaceae taxa by using pollen traits observed under scanning electron microscopy.

Herbal medicines are gaining popularity worldwide for human healthcare because of their therapeutic potential. However, adulteration and use of unauthentic raw herbals as substitutes have become a major issue for the local communities and industry for reasons of safety and efficacy. Therefore, the authentication of medicinal plants before their use in herbal medicines is a need of time. Hence, the present study was designed with an aim, to authenticate the therapeutic Lamiaceous taxa by using pollen traits observed under scanning electron microscopy. Pollen micro-morphological studies solve the problem through discrimination and correct identification of the Lamiaceae species from the adulterants. Based on pollen features, Lamiaceae were further divided into two sub-families Lamioideae (tricolpate) and Nepetoideae (hexa-colpate). The pollen grains of Lamioideae were found as small to medium-sized, tricolpate, radially/bilateral symmetrical, sub-spheroidal and oblate shape. Besides, exine patterns bireticulate, reticulate and micro-reticulate, colpus surface sculpturing as psilate, gemmate, scabrate, and verrucate have also been reported. A significant variation was found in the pattern of the reticulum, thickness, and the number of secondary lumina per primary lumen. Similarly, Nepetoideae has a hexa zono-colpate pollen but tri and tetra zono-colpate pollen have also been observed. Hence, this study contributes to the authentication and correct identification of medicinally important Lamiaceae taxa by using scanning electron microscopic techniques and can help to solve the adulteration problem. Highlights Authentication of medicinally important Lamiceous taxa was carried out through scanning electron microscopic techniques. Chemotaxonomic characterization was used for the accurate identification of the therapeutic taxa. The medicinal, palynological and phytochemical significance of Lamiaceae taxa were evaluated. A significant variation was seen in the palynological traits that help in the determination and authentication of the therapeutic Lamiaceous species. Based on the chemotaxonomic characterization, our study can help to solve the adulteration problem for the reason of safety and efficacy.

Toxicological effects of the chemical and green ZnO NPs on Cyprinus carpio L. observed under light and scanning electron microscopy.

Nanoparticles in aquatic bodies cause serious harm to the aquatic organisms when accumulated in high amounts. However, green nanoparticles synthesized using plants can be less toxic as compared to chemical nanoparticles. Hence, we designed our study to investigate the toxicological effects of chemical and green zinc oxide nanoparticles (ZnO NPs) on the biological activity of juvenile Cyprinus carpio. The green ZnO NPs were synthesized from Solieria robusta, and chemical ZnO NPs were synthesized using zinc chloride solution and ammonium hydroxide. Characterization was done by using light microscopy, scanning electron microscope (SEM), Fourier transmission infrared radiation, and X-ray diffraction (XRD) techniques. The highest absorbance of nanoparticles was observed at 360 which confirmed the synthesis of ZnO. The SEM analysis showed that green nanoparticles were hexagonal while the chemical nanoparticles were spherical to cubic in shape. Definite peaks were observed in XRD of green and chemical NPs at 2θ angles 45.84° and 32.18°, respectively. Oxidative stress was determined by chemical analysis of catalase, glutathione S-transferase (GST), glutathione (GSH), and lipid peroxidation (LPO) activities. The toxicological effects of chemical ZnO NPs on the catalase, LPO, GST, and GSH activities were more than green ZnO NPs. The histopathological investigation proved that the effect of chemical nanoparticles was worse than green ZnO NPs. More tissue damage was found in chemical nanoparticles than green synthesized nanoparticles. It was concluded that chemical nanoparticles can be replaced by green nanoparticles, as green nanoparticles are eco-friendly with less toxicological effects. This replacement can limit the toxic effect of nanoparticles when they get accumulated in high amounts in water bodies.

Morphological and palynological assessment of some taxa of genus Echinochloa through light and scanning electron microscopy.

The present study was undertaken to gather information on selected species of Echinochloa based on their morphological and palyonological examination through light and scanning electron microscopy. As the Echinochloa species are most problematic grass weeds and exhibit high interspecific and intraspecific variability so due to their great morphological diversity it is difficult to recognize them. A lot of variation was observed in morphological features of studied taxa. In the present study, E. colona had more or less spreading dense racemes whereas in E. crus-galli racemes were linear to ovate. Upper glume surface was pubescent in E. colona, hispid in E. crus-galli and spinose in E. walteri. E. colona was comprised of five-nerved glumes whereas in E. crus-galli glumes were three-nerved. Although pollen basic characters in family Poaceae remains consistent but great variation was observed in pollen of studied taxa. The pollen size was quite variable among the species of genus Echinochloa. In polar view, the size of pollen grains ranged from 27.5 (20-35) µm to 40 (30-50) µm. E. crus-galli was appeared to be the smallest in size whereas E. colona was the largest.). P/E ratio ranged from 0.84 to 1.15 µm among the species. The highest value of pollen fertility was found in E. colona as 93.33% and lowest was in E. crus-galli as 87.50%. The morphological key of studied taxa is also given to study variations. In conclusion, it was observed that microscopy of morphological and palynological assessment can play a vital role in identification of taxonomically problematic taxa.

Morphological, microscopic, and physicochemical studies of Diospyros montana.

Adulteration is the root cause of producing not only a chemically and pharmacologically inferior but also in some instances hazardous or poisonous drug. Despite availability of several techniques, microscopy and physicochemical analyses are the most practical approaches for crude drug authentication. Hence, the present study aimed to evaluate morphological, microscopic, and physicochemical properties of root, bark, leaf, and fruit of Diospyros montana Morphological properties were determined by sensory organs, whereas microscopic features of cross-sections and powders were determined by light and scanning electron microscopy. The proximate and fluorescence analyses were performed using the standard guidelines. The physical examination of fresh, shade-dried, and powdered material showed no significant change in color. The identifying cellular structures included cuboidal cork, pitted tracheids, scalariform, reticulate and spiral xylary vessels, and rosettes, raphide, and cuboidal calcium oxalate crystals. The stomatal number, stomatal index, vein-islet and vein-termination number, and palisade ratio in the leaf were found to be 293.91 ± 32.68 mm-2 , 64.18 ± 3.42%, 22.00 ± 3.81 mm-2 and 38.40 ± 5.81 mm-2 , and 3.85 ± 0.60, respectively. Total ash, acid insoluble ash, water soluble ash and sulfated ash of leaf (9.00 ± 0.50%, 1.67 ± 0.23%, 2.00 ± 0.22% and 14.50 ± 0.99%, respectively), foaming index of bark and root (111.11 ± 2.11), and swelling index of fruit (19.00 ± 3.45) were higher than the other parts. The powder of different parts showed characteristic colors in the daylight and UV light upon treatment with various regents. The plant was found to be rich in saponins, fibers, and flavonoids. The results of the present study may serve as identifiers of different parts of Diospyros montana.

Antioxidant potential and chemical characterization of bioactive compounds from a medicinal plant Colebrokea oppositifolia Sm.

Colebrookea oppositifolia is a highly used medicinal plant and an enriched source of essential oils. Therefore, the present study was designed with the aim to extract the chemical constituents and to evaluate its antioxidant potential. Fresh plant parts were subjected to the extraction of volatile chemical constituents by maceration using n-hexane as the menstruum. The resulting n-hexane fractions were purified and then subjected to GC-MS and FTIR analysis. In-vitro antioxidant abilities were evaluated by, DPPH, total phenolic content (TPC), total flavonoid content (TFC) method against the standard solutions of (Gallic acid, Quercetin) as a positive control. The GC-MS analysis of leaves, stem and inflorescence showed a total of 100, 98 and 48 components out of which 47, 16 and 17 peaks were identified representing the 67.64 %, 73.16 % and 61.93 % of the total oily fractions, respectively. The FTIR spectrum indicated the presence of various functional groups. In-vitro antioxidant results exhibited that leaves showed the highest antioxidant potential by DPPH (3.365 ± 0.002), and the highest total phenolic content by FC method (203.00 ± 0.091). Foliar micromorphological features were found significant in the authentication of C. oppositifolia. Further pharmacognostic studies of this plant are recommended to evaluate its therapeutic potential.

Taxonomic study of subfamily Nepetoideae (Lamiaceae) by polynomorphological approach.

The present study is insight into pollen morphology for characterizing species and their utility in the taxonomic separation of certain taxa of subfamily Nepetoideae (Lamiaceae) from Pakistan. The pollen micromorphology of 11 species of the Nepetoideae was analyzed and documented using light microscopy and scanning electron microscopy (SEM) for both qualitative and quantitative characteristics. Most species have hexazonocolpate pollen grains but trizonocolpate and tetrazonocolpate pollen with circular and oval amb were also rarely observed in Mentha spicata. The basic pollen shape in most of the studied species was subspheroidal but prolate grains were also observed in M. spicata, S. coccinea, and S. plebeia. The exine sculpturing of Nepetoideae pollen was taxonomically very informative particularly at subfamily level. Observations of exine sculpturing with SEM revealed various types of pollen grains: reticulate, bireticulate, microreticulate, perforate, aerolate, and gammate. The bireticulate type further subdivided into three subtypes based on the number of secondary lumina in each primary lumen and is characterized by varying characteristics of the secondary reticulum and primary muri. A significant variation was observed in colpus surface ornamentation. The maximum polar diameter was found in O. americanum (58 ± 5.8 µm) and the maximum equatorial diameter observed in O. basilicum (50.25 ± 1.37 µm). Pollen features of the studied species were discussed and compared based on the current taxonomical concepts. The results showed that pollen traits of the subfamily Nepetoideae was found significant to classify the taxa. Furthermore, pollen features provide additional evidence to distinguish macromorphologically similar taxa from each other.

Morpho-palynological assessment of medicinal flora of district Lahore, Pakistan based on LM and SEM.

This study was carried out to identify 20 high valued medicinal plants of District Lahore, Punjab, Pakistan with the help of the palynological studies. The distinguishing characteristics for selected species were found by systematized methods using light microscopy and Scanning Electron Microscopy. In this study, important pollen qualitative and quantitative features of these medicinal species such as polar and equatorial outline and their diameter, P/E ratio, presence or absence of pore, sculpturing of pollen, exine thickness, intine thickness, intercellular distance of exine and intine, presence or absence of spines, length of spines, width of spines, presence or absence of colpi and length of colpi, were compared. Remarkable variations in these pollen characters have been observed. Pollen grains were usually circular-semicircular (Calendula officinalis), spheroidal (Convolvulus arvensis), bilobed and trilobed (Convolvulus arvensis), spiny, (Abutilon indicum), oblate (Terminalia chebula) and elongated (Euphorbia pilulifera). In polar view Conyza bonariensis exhibited the highest pollen size [195.9 (211.6-180.2) µm]. While in equatorial view Calendula officinalis and Euphorbia pilulifera had lowest value [22.5 (20-25) µm] whereas Carissa carandas [165 (140-179) µm] appeared to be the largest. Parthenium hysterophorus showed the highest P/E ratio, that is, 1.28. Colpi were reported in only 7 species. Most of the plants showed 100% fertility but Terminalia chebula showed the least value 35%. In the present, project the diversity in pollen morphology present pollen as a valuable taxonomic tool. It is also concluded that pollen features are helpful at the specific level as well as generic level and can provide us fruitful taxonomic solutions. RESEARCH HIGHLIGHTS: Palynological flora of the medicinal plant species of district Lahore, Pakistan is investigated first time on the basis of LM and SEM and the flora is found very significant for taxonomic investigations.

Acute and sub-acute toxicity study of a Pakistani polyherbal formulation.

BACKGROUND: Herbology is the prevailing system among the nationally-accepted alternative or complementary systems of medicine. The system is due to its general and patient-oriented methodology, is widely used in the general population exposing them to the risk of the side effects of the herbal medicines. METHOD: The aim of study was to assess the acute and sub-acute toxicity of the polyherbal formulation Hab-e-Kabad Noshadri tablets. In the acute arm of the study, a single dose of 2000 mg/kg was administered to Swiss Albino mice which were observed for physical symptoms and behavioral changes for 72 h. In sub-acute toxicity study repeated doses of the polyherbal preparation was administered to Wistar rats of both genders, separately. The animals received three doses of polyherbal product (50 mg/kg/day, 100 mg/kg/day and 200 mg/kg/day) for a period of 28 days. On 28th day of experiment, blood sampling of animals was done for hematological and biochemical analysis i.e. liver and renal function parameters, lipid profile and then sacrificed for histopathological examination of liver and kidney. RESULT: There was no morbidity and mortality noticed with single dose administration in acute toxicity study in mice. In sub-acute toxicity study, morphological changes with some damage in liver and kidney tissues of male and female animals were recorded at dose of 100 mg/kg/day and 200 mg/kg/day. CONCLUSIONS: It was found that prolonged use at higher dose i.e. 200 mg/kg/day of this polyherbal formulation should be avoided and practitioners should cautiously prescribe this formulation in patients with hepatic and renal impairment.

Pharmacognostic studies of stem, roots and leaves of Malva parviflora L.

OBJECTIVE: To establish quality control parameters of a locally occurring medicinal plant, Malva parviflora which is utilized as folk medicine in Sialkot area in Pakistan. METHODS: In pharmacognostic studies different types of evaluations were carried out that focus on microscopic, macroscopic, fluorescence analysis and organoleptic evaluations. RESULTS: The distinguishing characters of stem were the presence of parenchyma, cork cells, irregular shape calcium oxalate crystals, simple and compound starch granules and fusiform fibers with pits. Root microscopic characters were presence of simple and spherical starch granules with rounded or slit hilum, groups of lignified xylem fibers, reticulate vessels, and sieve tissues. Leaves microscopy indicated the presence of paracytic stomata, lignified fibers having pits, spiral and annular vessels, numerous sclereids while in fruit microscopy epicarp, thin walled cells endocarp, thin walled parenchyma and collenchyma of mesocarp and abundant thick walled endospermic cells containing aleurone grains and micro rosette crystals. Macroscopic study of leaves showed, 5-7 lobed reniform-shape, glabrous-surface, reticulate-venation in the leaves. Macroscopic features of roots showed type of root-taproot, surface-glabrous and stem was 1-10 dm tall simple to branched and may be prostrate or ascending. Similarly fruit was of schizocarp type. CONCLUSIONS: This study provides the scientific data for the proper identification and establishment of standards for the use of Malva parviflora.