In vitro induction and selection of mutants obtained through gamma irradiation with improved processing traits in potato (Solanum tuberosum L.).

PURPOSE: This manuscript aimed for the generation of γ-irradiation derived mutants of potato genotype PAU/RR-1501 possessing desirable processing traits. MATERIALS AND METHODS: Nodal cuttings from virus-free explants were established on basal MS medium and irradiated with different doses (0, 5, 10 and 20 Gy) of γ-irradiation. The 5 and 10 Gy treated plantlets were multiplied and used for micro-tuber induction. Harvested micro-tubers were planted in pots for the selection and evaluation of mutants in M1V2 generation. RESULTS: Four weeks post-treatment, plantlets (5 Gy) showed enhanced growth as compared to the control while 20 Gy treatment exhibited completely ceased shoot growth. The highest number and weight of mini-tubers per plant was recorded for 10 Gy followed by 5 Gy treatment as compared to control. The γ-irradiation treatments caused changes in the skin color and shape of M1V2 tubers. CONCLUSION: Under the 5 Gy treatment 49.9% of clones produced exhibited cream and 8.53% brown skin color. Nine putative mutants were identified in genotype PAU/RR-1501 exhibiting promising processing traits.

Phytochemicals: Alternative for Infertility Treatment and Associated Conditions.

Infertility and obstetric complications have become global health issues in the past few years. Infertility is defined as the inability of a couple to conceive even after twelve months or more of regular and unprotected intercourse. According to WHO data published in the year 2020, 186 million people have infertility globally. Factors leading to infertility are variable in both males and females. But some common factors include smoking, alcohol consumption, obesity, and stress. Various synthetic drugs and treatment options are available that are effective in treating infertility, but their prolonged usage produces various unwanted adverse effects like hot flashes, mood swings, headaches, and weight gain. In extreme cases, these may also lead to the development of anxiety and depression. Herbal remedies have gained a lot of popularity over the years, and people's inclination toward them has increased all over the world. The prime reason is that these show significant therapeutic efficacy and have fewer side effects. The therapeutic efficacy of plants can be attributed to the presence of diverse phytochemical classes of constituents like alkaloids, flavonoids, and volatile oils. These secondary metabolites, or phytomolecules, can be used to develop herbal formulations. The review highlights the applications and mechanisms of action of various phytochemicals for treating infertility. Also, it focuses on the various future prospects associated with it.

Immunomodulatory effects of ß-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer.

BACKGROUND: Macrophages are mononuclear CD34+ antigen-presenting cells of defense mechanism and play dual roles in tumor burden. The immunomodulatory and their antitumor function of ß-defensin 2 is still unclear, despite the accumulating evidence of the response in infection. So, the aim of present study is to elucidate the role of ß-defensin 2 on the level of ROS, cytokines, chemokine expression in macrophages and antitumor function in breast cancer. METHOD: Swiss albino mice were used to harvest PEC macrophages and C127i breast cancer cells line for tumor model was used in this study. Macrophages were harvested and characterized by flow-cytometry using F4/80 and CD11c antibodies. MTT was performed to estimate cytotoxicity and dose optimization of ß-defensin 2. Oxidative stress was analyzed by H2O2 and NO estimation followed by iNOS quantified by q-PCR. Cytokines and chemokines estimation was done using q-PCR. Co-culture experiment was performed to study anti-tumor function using PI for cell cycle, Annexin -V and CFSE analysis for cell proliferation. RESULTS: PEC harvested macrophages were characterized by flow-cytometry using F4/80 and CD11c antibodies with the purity of 8% pure population of macrophages. It was found that 99% of cells viable at the maximum dose of 100 ng/ml of ß-defensin 2 in MTT. Levels of NO and H2O2 were found to be decreased in ß-defensin 2 as compared to control. Expression of cytokines of IFN-γ, IL-1α, TNF-α, TGF-ßwas found to be increased while IL-3 was decreased in ß-defensin 2 group as compared to control. Levels of chemokines CXCL-1, CXCL-5 and CCL5 increased in treated macrophages while CCL24 and CXCL-15 expression decreased. Adhesion receptor (CD32) and fusion receptor (CD204) were decreased in the ß-defensin 2 group as compared to control. Anti-tumor experiment was performed using co-culture experiment apoptosis (Annexin-V) was induced, cell cycle arrest in phage and cell proliferation of C127i cells was decreased. CONCLUSION: This is the first report of ß-defensin 2 modulates macrophage immunomodulatory and their antitumor function in breast cancer. ß-defensin 2 as a new therapeutic target for immunotherapy as an adjuvant in vaccines.

Amelioration of Chromium-Induced Oxidative Stress by Combined Treatment of Selected Plant-Growth-Promoting Rhizobacteria and Earthworms via Modulating the Expression of Genes Related to Reactive Oxygen Species Metabolism in Brassica juncea.

Chromium (Cr) toxicity leads to the enhanced production of reactive oxygen species (ROS), which are extremely toxic to the plant and must be minimized to protect the plant from oxidative stress. The potential of plant-growth-promoting rhizobacteria (PGPR) and earthworms in plant growth and development has been extensively studied. The present study was aimed at investigating the effect of two PGPR (Pseudomonas aeruginosa and Burkholderia gladioli) along with earthworms (Eisenia fetida) on the antioxidant defense system in Brassica juncea seedlings under Cr stress. The Cr toxicity reduced the fresh and dry weights of seedlings, enhanced the levels of superoxide anion (O2•-), hydrogen peroxide (H2O2), malondialdehyde (MDA), and electrolyte leakage (EL), which lead to membrane as well as the nuclear damage and reduced cellular viability in B. juncea seedlings. The activities of the antioxidant enzymes, viz., superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate peroxidase (APOX), glutathione peroxidase (GPOX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) were increased; however, a reduction was observed in the activity of catalase (CAT) in the seedlings under Cr stress. Inoculation of the PGPR and the addition of earthworms enhanced the activities of all other antioxidant enzymes except GPOX, in which a reduction of the activity was observed. For total lipid- and water-soluble antioxidants and the non-enzymatic antioxidants, viz., ascorbic acid and glutathione, an enhance accumulation was observed upon the inoculation with PGPR and earthworms. The supplementation of PGPR with earthworms (combined treatment) reduced both the reactive oxygen species (ROS) and the MDA content by modulating the defense system of the plant. The histochemical studies also corroborated that the combined application of PGPR and earthworms reduced O2•-, H2O2, lipid peroxidation, and membrane and nuclear damage and improved cell viability. The expression of key antioxidant enzyme genes, viz., SOD, CAT, POD, APOX, GR, DHAR, and GST showed the upregulation of these genes at post-transcriptional level upon the combined treatment of the PGPR and earthworms, thereby corresponding to the improved plant biomass. However, a reduced expression of RBOH1 gene was noticed in seedlings supplemented under the effect of PGPR and earthworms grown under Cr stress. The results provided sufficient evidence regarding the role of PGPR and earthworms in the amelioration of Cr-induced oxidative stress in B. juncea.

Reviving mitochondrial bioenergetics: A relevant approach in epilepsy.

Epileptogenesis is most commonly associated with neurodegeneration and a bioenergetic defect attributing to the fact that mitochondrial dysfunction plays a key precursor for neuronal death. Mitochondria are the essential organelle of neuronal cells necessary for certain neurophysiological processes like neuronal action potential activity and synaptic transmission. The mitochondrial dysfunction disrupts calcium homeostasis leading to inhibitory interneuron dysfunction and increasing the excitatory postsynaptic potential. In epilepsy, the prolonged repetitive neuronal activity increases the excessive demand for energy and acidosis in the brain further increasing the intracellular calcium causing neuronal death. Similarly, the mitochondrial damage also leads to the decline of energy by dysfunction of the electron transport chain and abnormal production of the ROS triggering the apoptotic neuronal death. Thus, the elevated level of cytosolic calcium causes the mitochondria DNA damage coinciding with mtROS and releasing the cytochrome c binding to Apaf protein further initiating the apoptosis resulting in epileptic encephalopathies. The various genetic and mRNA studies of epilepsy have explored the various pathogenic mutations of genes affecting the mitochondria functioning further initiating the neuronal excitotoxicity. Based on the results of previous studies, the recent therapeutic approaches are targeting basic mitochondrial processes, such as energy metabolism or free-radical generation, or specific interactions of disease-related proteins with mitochondria and hold great promise to attenuate epileptogenesis. Therefore, the current review emphasizes the emerging insights to uncover the relation between mitochondrial dysfunction and ROS generation contributing to mechanisms underlying epileptic seizures.

Vim stereotactic radiosurgical thalamotomy for drug-resistant idiopathic Holmes tremor: a case report.

We share our experience with stereotactic gamma knife thalamotomy (GKT) for medically refractory Holmes tremor (HT). A 22-year-old patient underwent gamma knife thalamotomy at ventrointermediate nucleus for disabling HT of the right upper limb. A single 4-mm isocenter was used to target the ventral intermediate nucleus with 130 Gy radiation. At 4 months follow up, we observed 84% improvement in his Fahn-Tolosa-Marin (FTM) rating scale with significant improvement in the right upper limb dystonic tremor. There was only subtle improvement in the ataxic component of the right lower limb. At 1 year after stereotactic GKT, there was sustained neurological improvement with no side effect, We present the stereotactic GKT as a treatment modality for drug-resistant HT. Moreover, it may be considered an alternate treatment modality especially in patients reluctant or contraindicated for any invasive surgical technique. CLINICAL TRIAL REGISTRATION NUMBER: Not required.

Protective efficacy of Chlorophytum borivilianum root extract against murine visceral leishmaniasis by immunomodulating the host responses.

BACKGROUND: The available drugs for treating visceral leishmaniasis are limited. Moreover, the disease is associated with suppression of immune function. Therefore, therapies with effective immunomodulatory agents are needed which can decrease parasitic burden and enhance adaptive immunity. OBJECTIVES: The present study was planned to evaluate the antileishmanial efficacy of crude ethanolic extract of roots of Chlorophytum borivilianum (CBREE) against murine visceral leishmaniasis through immunomodulation. MATERIALS AND METHODS: The in vitro studies were carried out to check leishmanicidal activity against promastigote form and cytotoxicity against HeLa cells. The parasite load in liver smears, immunological and biochemical changes induced by 500 and 1000 mg/kg b.wt. of CBREE were assessed on 1, 7, 14 and 21 post treatment days in infected and treated BALB/c mice. RESULTS: CBREE showed inhibitory effect on growth of promastigotes with IC50 of 28.25 µg/mL and negligible cytotoxicity. The extract was toxicologically safe in BALB/c mice when administered orally with 5 g/kg b.wt. of extract. A significant reduction in parasite load was observed along with active immunomodulation through enhanced Th1 type of immune responses and suppressed Th2 type of immune responses. CONCLUSION: The treatment with both doses showed no toxic effect as evidenced by normal liver and kidney function tests and normal histological observations of liver and kidney. Therefore, it should be further explored for its active components in pursuit of the new effective antileishmanial agents in the plant kingdom.

Evaluation of in vitro and in vivo antileishmanial potential of bergenin rich Bergenia ligulata (Wall.) Engl. root extract against visceral leishmaniasis in inbred BALB/c mice through immunomodulation.

BACKGROUND: Medicinal plants with immunomodulatory properties can provide good alternative therapeutics for curing visceral leishmaniasis. Bergenia ligulata (Wall.) Engl. is an interesting plant with strong antioxidant, antimicrobial, immunomodulatory and hepatoprotective properties. AIM: The present study was planned to determine the antileishmanial activity of plant extract by modulating the immune responses of inbred BALB/c mice. METHODOLOGY: Bergenin, the principle active component of B. ligulata, was quantitated in crude extract by performing RP-HPLC. The therapeutic potential was assessed through in vitro antileishmanial activity and in mice model through parasite load, cytokine assays, IgG antibody levels, DTH responses, histopathology and biochemical enzyme assays. RESULTS: B. ligulata showed the presence of glycosides, saponins, carbohydrates, tannins, flavonoids and bergenin which contributed to the antileishmanial activity of extract with IC50 of 22.70 µg/mL. Furthermore, the higher dose significantly reduced the parasite load by 95.56 %. The reduction was further associated with significant enhancement of IL-12 and IFN-γ levels in comparison to IL-10 and IL-4 cytokines. The switching towards Th1 type of immune response was also confirmed by elevated antibody levels of IgG2a isotype as compared to IgG1 as well as increased DTH responses. The histology of liver and kidney further complimented the non toxic nature of plant extract in addition to its negligible toxicity on HeLa cells. CONCLUSIONS: The current study revealed the significant antileishmanial and immunomodulatory properties of this plant extract against murine visceral leishmaniasis. Further, the bioactive components will be explored to assess their efficacy for the development of safe and cost effective drug.

Antileishmanial and immunomodulatory potential of Ocimumsanctum Linn. and Cocosnucifera Linn. in murine visceral leishmaniasis.

The role of immunomodulation in the therapeutic treatment of visceral leishmaniasis has gained eminence in view of moderate to severe drawbacks of the currently available drugs like toxicity, drug resistance and prohibitive costs. The potential for modulation of the immune system of many herbal plants can be tapped to address these problems. We conducted the present research study to investigate the antileishmanial and immunomodulatory effects of Ocimum sanctum Linn. and Cocos nucifera Linn. during the progression of visceral leishmaniasis in BALB/c mouse model. The IC50 values of the ethanolic leaf extract of O. sanctum and that of the aqueous husk-fibre extract of C. nucifera against the sodium stibogluconate (SSG) susceptible strain (MHOM/IN/80/Dd8) were found to be 73.3 and 62 µg/ml respectively. On treatment of infected BALB/c mice with the extracts, we observed a reduction in hepatic parasite load by 43.63 % (O. sanctum), 65.42 % (C. nucifera) and 75.61 % (O. sanctum + C. nucifera) at 1st post treatment day (p.t.d.), while at 15th p.t.d., the reduction was 73.61 % (O. sanctum), 76.59 % (C. nucifera) and 94.12 % (O. sanctum + C. nucifera). This was accompanied by an up-scaling of the DTH response, skewing of the humoral response towards Th1 type and hepatoprotection in the form of normalization of liver function tests. Overall, administration of the extracts of these two plants in combination as compared to their administration alone rescued the affected mice from the disease greatly, which can be attributed to their antileishmanial and immunomodulatory activities.

Anticonvulsant activity of ethanol extracts of Vetiveria zizanioides roots in experimental mice.

CONTEXT: Vetiveria zizanioides Linn. (Gramineae), an aromatic plant commonly known as vetiver, is traditionally used for various ailments. Ethanol and aqueous extract of this plant found extensive use in Indian folklore medicine and used in treatment of a wide range of disorders including seizure. However, the anticonvulsant activity of this plant has not been studied. OBJECTIVE: To evaluate anticonvulsant activity of ethanol extract of V. zizanioides (EEVZ) in experimental mice. MATERIALS AND METHODS: Anticonvulsant activity of EEVZ was determined by maximal electroshock stimulation (MES) and pentylenetetrazole (PTZ) in mice for 8 d experimental protocol. The extract at a dose of 100, 200 and 400 mg/kg body weight was administered once by oral route. RESULTS: LD50 value of EEVZ in mice was found at a dose of 600 mg/kg body weight. EEVZ at a dose of 400 mg/kg significantly (p < 0.001) reduced flexion (l5.98 to 3.73 s), extension (13.73 to 0.96 s), clonus (14.07 to 4.93 s), stupor (6.29 to 1.22 s) in the MES model. Further, it increases onset of clonic (88.25 to 708.32 s/30 min) and tonic (139.52 to 1126.39 s/30 min) in the PTZ model. In the PTZ model, 33% normal control and 83% EEVZ (100 mg/kg) animals were alive, while 100% protection was achieved in standard drug phenobarbital (20 mg/kg), EEVZ (200 mg/kg) and EEVZ (400 mg/kg) animals. DISCUSSION AND CONCLUSION: Findings demonstrate that V. zizanioides shows significant anticonvulsant activity in mice.

Comparison of protective and curative potential of Daucus carota root extract on renal ischemia reperfusion injury in rats.

CONTEXT: Daucus carota Linn (Apiaceae), a useful vegetable, is traditionally used in treating kidney and hepatic dysfunctions. OBJECTIVE: To evaluate the protective and curative potential of D. carota root extract on renal ischemia reperfusion injury in rats. MATERIALS AND METHODS: Wistar rats were selected with 8 + 8 groups (n = 6). Renal pedicles of rats were occluded for 45 min and allowed for reperfusion period. In protective and curative studies, 14 days prior and 14 days after the induction of ischemia/reperfusion (I/R), rats received petroleum ether extract (PEE 250 and 500 mg/kg), fractional methanol extract (FME 250 and 500 mg/kg) and direct methanol extract (DME 250 and 500 mg/kg) of Daucus carota root, orally, once daily. RESULTS: PEE at a dose of 500 mg/kg significantly (p < 0.001) reduced the levels of serum creatinine (0.853-3.090 mg/dl), uric acid (1.300-3.500 mg/dl) and urea (58.26-132.00 mg/dl) compared to disease control. FME at a dose of 500 mg/kg body weight significantly (p < 0.001) reduced the levels of serum creatinine (0.960-3.090 mg/dl), uric acid (1.700-3.500 mg/dl) and urea (77.17-132.00 mg/dl) compared to disease control. DME at a dose of 500 mg/kg body weight significantly (p < 0.001) reduced the levels of serum creatinine (1.173-3.090 mg/dl), uric acid (2.267-3.500 mg/dl) and urea (84.75-132.00 mg/dl) compared to disease control. DISCUSSION AND CONCLUSION: Findings demonstrate that postconditioning with the D. carota root extract significantly improves kidney function in I/R rats.

Protective effect of Lannea coromandelica Houtt. Merrill. against three common pathogens.

BACKGROUND: Ayurvedic text reports suggested Lannea coromandelica is used in various microbial origin disorders like dysentery, sore eyes and leprosy, genital wounds. OBJECTIVE: The present study was designed to investigate the antimicrobial effect of L. coromandelica Houtt. Merrill. (Anacardiaceae) on microbes which cause female reproductive tract infection. MATERIALS AND METHODS: Ethanolic and aqueous bark extract (Ext.) of L. coromandelica were screened against strains of Streptococcus pyogens, Staphylococcus aureus, and Candida albicans. Antimicrobial assay had been done with agar well diffusion method. RESULTS: Ethanolic extracts [100% (16 mg), 75% (12 mg) and 50% (8 mg)] of L. coromandelica exhibited zone of inhibition (ZI) 19.21 mm, 18.45 mm, 16.41 mm and 18.12 mm, 17.35 mm, 16.35 mm against S. aureus and S. pyogens, respectively. However, only 100% and 75% ethanolic extract showed (ZI-19.18 mm, 16.29 mm) activity against C. albicans. Nevertheless, aqueous extract (100%) showed higher antifungal activity (ZI-16.97 mm). Ciprofloxacin and amphotericin B were used as a standard drugs in the present study. CONCLUSION: The results demonstrated that L. coromandelica Houtt. Merrill. have antibacterial activity against S. pyogens, S. aureus and antifungal property against C. albicans. Our findings corroborate the ethnobotanical use of L. coromandelica in traditional medicine system (Ayurveda) of India.

Polypharmacy (herbal and synthetic drug combination): a novel approach in the treatment of type-2 diabetes and its complications in rats.

The aim of present study was to evaluate the effect of aqueous leaf extract of Annona squamosa with Glipizide in a high fat diet and streptozocin-induced type-2 diabetes. Nine groups (n = 6) of male Sprague-Dawley rats were used for the study, with. Basal blood glucose, urine volume, and body weights were measured and the rate were kept on a high fat diet. After 15 days, streptozocin in sub-diabetic dose (35 mg/kg) was administered to the animals to induce diabetes. With 1 week of consistent hyperglycemia, treatment was initiated. Aqueous extract of Annona squamosa was administered orally at 350 mg/kg body weight alone and in combination with reduced and reducing dose combinations of Glipizide. Blood glucose, body weight, urine volume were measured every 10th day. The elevated blood-glucose level in diabetic rats was controlled better with combination therapy compared with the synthetic drug alone or the herbal stand-alone drug. All the results were statistically significant (P < 0.001). A combination of Annona squamosa along with Glipizide may be helpful in dose reduction of Glipizide up to 50%, reducing the risk of the onset of insulin therapy.

An updated review on the Oenothera genus.

Oenothera genus (Onagraceae) has been used as a folk remedy since ancient times for the treatment of asthma, gastrointestinal disorders, neuralgia, skin diseases, and hepatic and kidney diseases. Different chemical constituents like lipids, flavonoids, tannins, steroids and triterpenes have been isolated from this genus. The various notable pharmacological activities reported from the genus are antioxidant, cytotoxic, antibacterial, antiviral, anti-inflammatory, antihyperlipidaemic, thrombolytic and antidiarrhoeal. The present paper is to summarize the worldwide reported biological activities and phytoconstituents associated with this genus for about 50 years and highlight the medicinally important species belonging to this genus so that these species can be further explored and used as therapeutic agents for various diseases.