Ethnomedical Applications and Conservation Status of Leguminosae- Caesalpinioideae Plants in Uttar Pradesh, India.

BACKGROUND: This review highlights the folklore, ethnomedicinal uses and conservation status of Caesalpinioideae in Uttar Pradesh (India). AIMS: It aims at compiling available data on traditional medicine, biological activity, phytochemical information and assessing the regional red list status of Caesalpinioideae in Uttar Pradesh. The information provided would help in formulating new drugs and medicines and addressing global conservation issues of such medicinally exploited species. METHODS: The current study included an extensive and systematic review of available literature, the study of previous collections of herbarium specimens, random interviews with locals and tribals, field surveys, and GeoCAT tool-based assessment during 2016-2020. The study reports that the majority of species of Caesalpinioideae are used for curing digestive problems (about 20 species) and skin diseases (19 species). RESULTS: Almost all the species have antimicrobial and antioxidant properties. These pharmacological activities can be attributed to the presence of various types of anthraquinones in plants. CONCLUSION: The regional conservation status reveals that eight species qualified for the status of regionally threatened category while two species fall under the near threatened category.

Morphologically favorable mutant of Trichoderma reesei for low viscosity cellulase production.

Metabolite production by filamentous fungi hampered because of high viscosity generated during growth. Low viscosity fermentation by mold is one of the preferred ways of large scale enzymes production. Cellulolytic enzymes play a key role during the process of lignocellulosic biomass conversion. In this study, a mutant RC-23-1 was isolated through mutagenesis (diethyl sulfate followed by UV) of Trichoderma reesei RUT-C30. RCRC-23-1 not only gave higher cellulase production but also generated lower viscosity during enzyme production. Viscosity of mutant growth was more than three times lower than parent strain. RC-23-1 shows unique, yeast-like colony morphology on solid media and small pellet-like growth in liquid media. This mutant did not spread like mold on solid media. This mutant produces cellulases constitutively when grown in sugars. Using only glucose, the cellulase production was 4.1 FPU/ml. Among polysaccharides (avicel, xylan, and pectin), avicel gave maximum of 6.2 FPU/ml and pretreated biomass (rice straw, wheat straw and sugarcane bagasse) produced 5.1-5.8 FPU/ml. At 7 L scale reactor, fed-batch process was designed for cellulase production using different carbon and nitrogen sources. Maximum yield of cellulases was 182 FPU/g of lactose consumed was observed in fed-batch process. The produced enzyme used for hydrolysis of acid pretreated rice straw (20% solid loading) and maximum of 60% glucan conversion was observed. RC-23-1 mutant is good candidate for large scale cellulase production and could be a model strain to study mold to yeast-like transformation.

Hydrothermal Pretreatment of Lignocellulosic Feedstocks to Facilitate Biochemical Conversion.

Biochemical conversion of lignocellulosic feedstocks to advanced biofuels and other bio-based commodities typically includes physical diminution, hydrothermal pretreatment, enzymatic saccharification, and valorization of sugars and hydrolysis lignin. This approach is also known as a sugar-platform process. The goal of the pretreatment is to facilitate the ensuing enzymatic saccharification of cellulose, which is otherwise impractical due to the recalcitrance of lignocellulosic feedstocks. This review focuses on hydrothermal pretreatment in comparison to alternative pretreatment methods, biomass properties and recalcitrance, reaction conditions and chemistry of hydrothermal pretreatment, methodology for characterization of pretreatment processes and pretreated materials, and how pretreatment affects subsequent process steps, such as enzymatic saccharification and microbial fermentation. Biochemical conversion based on hydrothermal pretreatment of lignocellulosic feedstocks has emerged as a technology of high industrial relevance and as an area where advances in modern industrial biotechnology become useful for reducing environmental problems and the dependence on fossil resources.

A Review of Chemistry and Pharmacology of Piperidine Alkaloids of Pinus and Related Genera.

BACKGROUND: Pinus belongs to the family Pinaceae, represented by several species across the globe. Various parts of the plant including needles are rich in biologically active compounds, such as thunbergol, 3-carene, cembrene, α-pinene, quercetin, xanthone. Of all the alkaloids, the piperidine group is one of the important component and holds considerable medicinal importance. METHODS: The group of alkaloids was initially identified from the genus Piper through which a large variety of piperidine molecules have been extracted. The planar structure of this heterocyclic nucleus enables acetamide groups to be added at various ring configurations. RESULTS: Piperidines have gained considerable importance. The broad range of its therapeutic application has paved a way for researchers to implant the nucleus from time to time in diversified pharmacophores and establish new profile. DISCUSSION: Biological functions of piperidine metabolites have been mostly examined on a limited scale, and that most of the findings are preliminary. We have tried to present various clinical applications of piperidine alkaloids in this study that researchers have already attempted to demystify with time. CONCLUSION: We have also illustrated different types of piperidine structures and their sources in different members of the family Pinaceae with special emphasis on Pinus. Given the importance of the piperidine nucleus, the study will enable the researchers to produce scaffolds of highest therapeutic efficacy.

Role and significance of lytic polysaccharide monooxygenases (LPMOs) in lignocellulose deconstruction.

Lytic polysaccharide monooxygenases (LPMOs) emerged a decade ago and have been described as biomass deconstruction boosters as they play an extremely important role in unravelling the enzymatic biomass hydrolysis scheme. These are oxidative enzymes requiring partners to donate electrons during catalytic action on cellulose backbone. Commercial cellulase preparations are mostly from the robust fungal sources, hence LPMOs from fungi (AA9) have been discussed. Characterisation of LPMOs suffers due to multiple complications which has been discussed and challenges in detection of LPMOs in secretomes has also been highlighted. This review focuses on the significance of LPMOs on biomass hydrolysis due to which it has become a key component of cellulolytic cocktail available commercially for biomass deconstruction and its routine analysis challenge has also been discussed. It has also outlined a few key points that help in expressing catalytic active recombinant AA9 LPMOs.

Ethnobotany and Pharmacology of Pinus Species Growing Naturally in Indian Himalayas: A Plant Review.

Pinus species are amongst the most commonly used medicinal plants in Indian Northwestern Himalayas. Nearly all parts of the plant especially needles are believed to possess medicinal and clinical values in traditional systems of medicine such as Ayurvedic, Unani or Homeopathic. They are used to heal numerous diseases, including afflictions of the skin, eyes, throats and ears. Various parts of the plant including needles are rich in biologically active compounds, such as thunbergol, 3-carene, cembrene, α-pinene, quercetin, xanthone, etc. The scientific data available for the biological potential of this plant and its constituents have been found to be scanty and also does not satisfy the basis of their age-old folklore and local uses. This review focuses on a detailed analysis of ethnopharmacognosical studies on the needles of different Pinus species growing naturally in northwestern region of Indian Himalayas including investigations and findings that have come up with a concrete view of the abilities of plant components for example phenolics, flavonoids, tannins and other constituents to treat oxidative, inflammatory and microbial responses. Careful evaluation of the data presented may be helpful for the researchers to discover and evaluate the specific chemical entities responsible for the traditional medicinal uses of P. roxburghii, P.wallichiana, P. merkusii, P.kesiya and P.thunbergii.

A screening approach for assessing lytic polysaccharide monooxygenase activity in fungal strains.

BACKGROUND: Efforts to develop efficient lignocellulose-degrading enzymatic preparations have led to the relatively recent discovery of a new class of novel cellulase boosters, termed lytic polysaccharide monoxygenases (LPMOs). These enzymes are copper-dependent metalloenzymes that initiate the biomass deconstruction process and subsequently work together with cellulases, hemicellulases, and other accessory enzymes to enhance their hydrolytic action. Given their wide distribution and diversity, screening and isolation of potent LPMOs from natural fungal diversity may provide an important avenue for increasing the efficiency of cellulases and thereby decreasing cellulosic ethanol production costs. However, methods for quick screening and detection are still not widely available. In this article, a simple and sensitive method is described by combining nonhydrolytic activity enhancement followed by LC-MS-based quantitation of LPMOs. RESULTS: In this study, a screening approach has been developed for the detection of nonhydrolytic cellulase-enhancing enzymes in crude fungal supernatants. With the application of a saturating benchmark cocktail of Celluclast 1.5L, fungal isolates were selected which had the capability of hydrolyzing pretreated rice straw by their synergistic enzyme fractions. Subsequently, these fungal extracts along with an LPMO-enriched commercial enzyme were investigated for their ability to produce Type I LPMO activity. An LC-MS-based methodology was applied to quantitate gluconic acid in enzymatic hydrolysates as an indirect measurement of Type I LPMO activity. CONCLUSION: The present study describes an LC-MS-based separation method to detect and quantitate LPMO activity in a commercial enzyme. This method was also applied to screen fungal extracts. The developed screening strategy has enabled detection of LPMO activity in two industrially important Penicillium strains.

Status of Selinum spp. L. a Himalayan Medicinal Plant in India: A Review of Its Pharmacology, Phytochemistry and Traditional Uses.

There is a growing trend of herbal medicines in India as well as other parts of the world. The use of herbal medicines alone or alongside prescription drugs for disease management is quite common now. We hereby carry out a review of dominant species of Selinum mainly S. vaginatum C.B. Clarke and S. wallichianum (DC.) Raizada & H.O. Saxena which are potent source of herbal medicine and whose potential is still not exploited. They are found in Northern Himalayas in the states of Kashmir, Himachal Pradesh and Uttarakhand in India, and also distributed in adjoining countries like China, Nepal, and West Pakistan. It is a perennial, primitive, high altitude, therapeutic herbal plant belonging to family Apiaceae (Umbelliferae) traditionally used for various diseases like epilepsy, seizures, and hysteria, etc. The aim of this review was to provide a summary on botanical characterization, traditional uses, phytochemistry and pharmacological activity of Selinum spp. The available information will be further tapped for commercial use of this plant without endangering its status. Based on the available evidence on the species pharmacology and chemistry, we highlight in which their therapeutic potential can be properly harnessed for possible integration into the country's healthcare system and be further passed on globally.

An assessment of benzimidazole resistance against caprine nematodes in Central India.

Current status of resistance to benzimidazole (BZ) group of anthelmintic drugs against caprine nematodes in Central India at Amanala goat farm, Jabalpur, Madhya Pradesh (M. P.), was systematically investigated using faecal egg count reduction (FECR) test and egg hatch test (EHT). Besides, allele-specific PCR (AS-PCR) was deployed to ascertain the susceptible genotype (alleles) especially of the Haemonchus contortus. Randomly selected 30 goats, irrespective of age and sex, were divided into three groups of 10 each, to serve as treated and untreated controls. It was ensured that the animals were not administered with an anthelmintic drug for the past 3 months prior to undertaking the study, and faecal egg counts were estimated. FECR test evidenced fenbendazole resistance by partial elimination (24.90%) copro-egg counts in the treated group of animals vis-à-vis controls with a lower confidence interval of -26%. Further, EHT revealed ED-50 value of 0.335 µg of thiabendazole/ml, confirming benzimidazole resistance in the animals of that farm. AS-PCR showed that 62% of H. contortus larvae were homozygous resistant (rr), 24% heterozygous (rS) and 14% homozygous susceptible (SS). The genotypic frequencies of three genotypes (rr, rS and SS) were significantly (P < 0.01) different. The prevalence of benzimidazole resistance allele (r) was also significantly (P < 0.01) higher (74%) as compared to susceptible allele (S) (26%). The resistance to benzimidazole has been discussed while emphasizing improved managemental practices designed to reduce exposure of the goat population to parasites, minimize frequency of anthelmintic use at optimum dose and rotational use of different chemical groups of medicines with different mode of action, so as to overcome and combat the upcoming problem in the field.

Ionic liquid pretreatment of biomass for sugars production: Driving factors with a plausible mechanism for higher enzymatic digestibility.

In this study, five ionic liquids (ILs) have been explored for biomass pretreatment for the production of fermentable sugar. We also investigated the driving factors responsible for improved enzymatic digestibility of various ILs treated biomass along with postulating the plausible mechanism thereof. Post pretreatment, mainly two factors impacted the enzymatic digestibility (i) structural deformation (cellulose I to II) along with xylan/lignin removal and (ii) properties of ILs; wherein, K-T parameters, viscosity and surface tension had a direct influence on pretreatment. A systematic investigation of these parameters and their impact on enzymatic digestibility is drawn. [C2mim][OAc] with ß-value 1.32 resulted 97.7% of glucose yield using 10 FPU/g of biomass. A closer insight into the cellulose structural transformation has prompted a plausible mechanism explaining the better digestibility. The impact of these parameters on the digestibility can pave the way to customize the process to make biomass vulnerable to enzymatic attack.

Evaluation of dipstick-ELISA using 28 kDa Fasciola gigantica cathepsin l cysteine proteinase (FgCL3) for serodiagnosis of fasciolosis in naturally infected goats.

In the present investigation 100 goats presented for slaughter were used to evaluate the 28kDa cathepsin l cysteine proteinase (FgCL3) dipstick-ELISA for the diagnosis of fasciolosis. Presence of Fasciola gigantica worms in liver at the time of slaughter was taken as a gold standard for the evaluation of the assay. Faeces, blood and liver were taken from all slaughtered goats. Biochemical parameters and coprological examinations using sedimentation technique were also taken into consideration. Total serum proteins, albumin, mean values of aspartate aminotransferase and alkaline phosphatase of positive group were not significantly different from those of the negative group. The diagnostic sensitivity of dipstick-ELISA was higher than coprological detection of fluke eggs. However, specificity was equal for both the tests. The accuracy of dipstick-ELISA in detecting circulatory antibodies during the course of disease was higher than coproscopic examination. In conclusion, the FgCL3 antigen dependent dipstick-ELISA was more reliable and has better scope for field application in the endemic areas.

Evidence of new pathogenic Theileria species in dogs.

Babesiosis, caused by tick-borne haematozoan organisms of the genus Babesia, is a parasitic disease of domestic and wild mammals. Canine Babesidae have historically been classified as "large Babesia" (Babesia canis) and "small Babesia" (Babesia gibsoni) based on the size of their intraerythrocytic forms. Recent publications, however, suggest that the diversity of piroplasm species infecting dogs might be greater than previously appreciated. Sequencing and phylogenetic analysis of the ssrRNA gene has revealed that canine piroplasms are found in three clades, 'true' Babesia sp. (B. canis and B. gibsoni), Theileria annae and Theileria-like group (B. conradae). This newly recognised piroplasm T. annae appears to be hyperendemic in northwest Spain. The vector for this emergent canine infection remains undescribed, although the Ixodeshexagonus is suspected based on their presence upon the dogs in NW Spain and the relative absence of others. These observations have introduced a major change in the approach to the epidemiology of babesiosis in dogs. More clinical samples and data will need to be collected and analyzed to understand the importance of Theileria species in dogs.

18F9 (4-(3,6-bis (ethoxycarbonyl)-4,5,6,7-tetrahydrothieno (2,3-c) pyridin-2-ylamino)-4-oxobutanoic acid) enhances insulin-mediated glucose uptake in vitro and exhibits antidiabetic activity in vivo in db/db mice.

Insulin resistance is central to the pathogenesis of type 2 diabetes mellitus. Previous studies have demonstrated that compounds that cause adipogenesis and improve glucose uptake in 3T3-L1 cells are potential insulin sensitizers. Therefore, we evaluated one such compound, 18F9, for (1) adipogenesis in human subcutaneous preadipocyte (SQ) cells, (2) glucose uptake in human skeletal muscle myotubes and SQ cells, and (3) antidiabetic activity in db/db mice. We also investigated its effect on ex vivo glucose uptake in soleus muscle isolated from continuously treated db/db mice. Gene expression profiling in soleus muscle and epididymal fat of db/db mice was performed to understand its effect on glucose metabolism, lipid metabolism, and thermogenesis. 18F9 enhanced adipogenesis in SQ cells and increased glucose uptake in SQ and human skeletal muscle myotubes cells. In db/db mice, 18F9 exhibited dose-dependent reduction in plasma glucose and insulin level. Interestingly, 18F9 was as efficacious as rosiglitazone but did not cause body weight gain and hepatic adverse effects. In addition, 18F9 demonstrated no change in plasma volume in Wistar rats. Furthermore, it enhanced ex vivo glucose uptake in soleus muscles in these mice, which substantiates our in vitro findings. Human peroxisome proliferator activated receptor-gamma transactivation assay revealed a weak peroxisome proliferator activated receptor-gamma transactivation potential (44% of rosiglitazone at 10 mumol/L) of 18F9. Gene expression profiling indicated that 18F9 increased insulin sensitivity mainly through a phosphoinositide 3-kinase-dependent mechanism. 18F9 also up-regulated genes involved in lipid transport and synthesis at par with rosiglitazone. Unlike rosiglitazone, 18F9 elevated the expression of Pdk4. In addition, 18F9 elevated the expression of glycogen synthase and adiponectin significantly higher than rosiglitazone. Taken together, these observations suggest that 18F9 is a safer and potent insulin sensitizer that demonstrates promising antidiabetic activity and is worth further development.

Immunodiagnostic/protective role of cathepsin L cysteine proteinases secreted by Fasciola species.

Cathepsin L, a major cysteine proteinase secreted by the parasite plays a pivotal role in various aspects of its pathogenecity. The enzyme takes part in nutrient acquisition by catabolizing host proteins to absorbable peptides, facilitates the migration of the parasite through the host intestine and liver by cleaving interstitial matrix proteins such as fibronectin, laminin and native collagen and is implicated in the inactivation of host immune defenses by cleaving immunoglobulins. Recently, Cathepsin L has been shown to suppress Th1 immune response in infected laboratory animals making them susceptible to concurrent bacterial infections. Accordingly, the protease has been recognized as an important target at which parasite intervention strategies should be directed. Fluke Cathepsin L cysteine proteinases are also reported as sensitive and specific markers for the immunodiagnosis of fasciolosis in ruminants. Further, vaccination of laboratory or large animals with these proteases resulted in a significant reduction in fluke burden and/or fecundity.