Modulation in the enzymatic antioxidants, MDA level and elicitation in conessine biomolecule in Holarrhena pubescens (medicinal tree) cultures exposed to different heavy metals: Ni, Co, Cr and As.

Nodal explants of Holarrhena pubescens, an important medicinal tree, were cultured on Murashige and Skoog's medium (MS) containing 15 µM BA (control) alone and on medium supplemented with different concentrations (0, 1, 5, 25, 50, 100 and 200 mg/L) of heavy metals such as NiCl2, CoCl2, As2O3 and CrO3 to study their toxic effect. After 28 days of treatments, the nodal segments were harvested to assess the average number of shoots per explants, average shoot length, malondialdehyde content, proline content, conessine accumulation and antioxidant enzymatic activity. Among all the metals tried, best morphogenic response was achieved at 5 mg/L CrO3 where 80% culture differentiated an average of 3.21 ± 0.08 shoots per explant having 0.95 ± 0.018 cm average shoot length. Highest concentration (200 mg/L) of all the heavy metals proved lethal for morphogenesis. Maximum inhibition in average shoot number and average shoot length was observed in nodal explants treated with 25 mg/L As2O3 where an average of 0.49 ± 0.047 shoots having an average shoot length of 0.3 ± 0.02 cm. Contrarily, addition of heavy metals in culture medium proved strong elicitors, exhibiting significant enhancement in the biosynthesis of conessine, an important bioactive compound. HPLC analysis of the crude extract of in vitro grown untreated nodal cultures revealed an average of 117.06 ± 2.59 µg/g d. w. of conessine, whereas those treated with 100 mg/L of CoCl2 accounted for 297.1 ± 7.76 µg/g d. w. (an increase of 156% over control). Among the heavy metals tried, CoCl2 proved to be the best for conessine enhancement which was in the order of CoCl2 > Cr2O3 > NiCl2 > As2O3 in the nodal explants. Concomitantly, MDA content, the antioxidant enzymes activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GR) and ascorbate peroxidase (APX) were also observed to be differentially expressed with the increase in the heavy metals concentration from 1 to 200 mg/L. Free proline, too, increased up to 3.5-fold over control. The results obtained during the present investigation revealed that the overall response of the nodal explants in terms of morphogenesis, conessine content and antioxidant enzyme activities was metal specific as well as dose dependent.

Primary hyperparathyroidism presenting as acute hypercalcemic crisis: a case report.

BACKGROUND: Hyperparathyroid crisis, or "parathyroid storm" is a rare manifestation of primary hyperparathyroidism, characterized by sudden onset of symptomatic, severe hypercalcemia (> 3.5 mmol/L). Hemorrhage into a parathyroid adenoma has rarely been reported as an inciting or associated event. We present a case of hemorrhage into a longstanding adenoma presenting with acute onset of profound hypercalcemia and associated complications. CASE PRESENTATION: A 60-year-old male presented to hospital with sudden onset of confusion, muscle weakness, and ataxia. Initial labs showed serum calcium 4.79 mmol/L, parathyroid hormone 2043 ng/L; creatinine 364 µmol/L. Review of the patient's medical history indicated a 4-year history of recurrent nephrolithiasis, but no prior documented calcium levels. The hypercalcemia did not respond to 5 days of aggressive medical management with fluid resuscitation, denosumab and calcitonin, and later pamidronate and cinacalcet. He continued to deteriorate, requiring intubation and continuous renal replacement therapy. Imaging demonstrated 4.8 cm cystic right paratracheal mass; Technetium (Tc99m) Sestamibi scintigraphy was non-localizing. Urgent parathyroidectomy was completed, revealing a 5 × 3.3 × 1.8 cm hemorrhagic, atypical hypercellular parathyroid. Unfortunately, the patient died from complications from anticoagulation therapy for treatment of deep vein thrombosis 4 weeks after admission. His renal function had not recovered at the time of his death. CONCLUSION: This case gives potential insight into the etiology of hyperparathyroid crisis, and the difficulty in achieving control of hypercalcemia with medical means. Surgical intervention is the definitive management in these cases and should be considered urgently.

Arabian Primrose leaf extract mediated synthesis of silver nanoparticles: their industrial and biomedical applications.

The present study encompasses green synthesis of silver nanoparticles (AgNPs) using aqueous leaf extract of Arabian Primrose within 6 min of reaction at 60 °C, pH 7 and their characterisation using physico-chemical analytical techniques. UV-Visible spectroscopy elucidated the surface plasmon resonance around 420 nm. FESEM and TEM images revealed that AgNPs were spherical with average diameter 10-60 nm. XRD pattern confirmed their crystalline nature. The leaf extract rich in phenolics and flavonoids was subjected to GC-MS analysis that identified bioactive compounds helping in reduction and stabilisation of AgNPs. The synthesised AgNPs possessed high anti-oxidant potential against DPPH and H2O2 radicals. Incidentally, the AgNPs acted as excellent nanocatalyst towards borohydride reduction and degradation of structurally different organic dyes. The AgNPs also exhibited selective colorimetric sensing of hazardous mercuric, ferric ions and ammonia. Such AgNPs were cytotoxic against HeLa cells (IC50 7.18 µg/mL) and compatible towards normal L20B cells. These AgNPs showed effective anti-microbial activity against different human pathogens tested (bacterial and fungal). This is probably the first report of AgNPs synthesis using Arabian Primrose leaf extract showing strong anti-oxidant, catalytic, biosensing, anti-cancer and anti-microbial activities and find remarkable applications in medical, industrial and ecological sectors.

Biocontrol of mosquito vectors through herbal-derived silver nanoparticles: prospects and challenges.

Mosquitoes spread several life-threatening diseases such as malaria, filaria, dengue, Japanese encephalitis, West Nile fever, chikungunya, and yellow fever and are associated with millions of deaths every year across the world. However, insecticides of synthetic origin are conventionally used for controlling various vector-borne diseases but they have various associated drawbacks like impact on non-targeted species, negative effects on the environment, and development of resistance in vector species by alteration of the target site. Plant extracts, phytochemicals, and their nanoformulations can serve as ovipositional attractants, insect growth regulators, larvicides, and repellents with least effects on the environment. Such plant-derived products exhibit broad-spectrum resistance against various mosquito species and are relatively cheaper, environmentally safer, biodegradable, easily accessible, and are non-toxic to non-targeted organisms. Therefore, in this review article, the current knowledge of phytochemical sources exhibiting larvicidal activity and their variations in response to solvents used for their extraction is underlined. Also, different methods such as physical, chemical, and biological for silver nanoparticle (AgNPs) synthesis, their mechanism of synthesis using plant extract, their potent larvicidal activity, and the possible mechanism by which these particles kill mosquito larvae are discussed. In addition, constraints related to commercialization of nanoherbal products at government and academic or research level and barriers from laboratory experiments to field trial have also been discussed. This comprehensive information can be gainfully employed for the development of herbal larvicidal formulations and nanopesticides against insecticide-resistant vector species in the near future. Graphical abstract.

Green synthesis of silver nanoparticles using Indian Belladonna extract and their potential antioxidant, anti-inflammatory, anticancer and larvicidal activities.

KEY MESSAGE: Atropa acuminata aqueous leaf extract biosynthesized silver nanoparticles showed strong antioxidant, anticancerous (HeLa cells) and anti-inflammatory activities. Besides, this bio syn-AgNP also proved effective against mosquito vectors causing malaria, dengue and filariasis. Present study highlights eco-friendly and sustainable approach for the synthesis of silver nanoparticles (AgNP) using aqueous leaf extract of A. acuminata, a critically endangered medicinal herb. The addition of 1 mM silver nitrate to aqueous leaf extract resulted in the synthesis of AgNP when solution was heated at 60 °C for 30 min at pH 7. Absorption band at 428 nm, as shown by UV-Vis spectroscopy confirmed the synthesis of AgNP. XRD patterns revealed the crystalline nature of AgNP and TEM analysis showed that most of the nanoparticles were spherical in shape. Zeta potential of AgNP was found to be - 33.5 mV which confirmed their high stability. FT-IR investigations confirmed the presence of different functional groups involved in the reduction and capping of AgNP. The synthesized AgNP showed effective DPPH (IC50-16.08 µg/mL), H2O2 (IC50-25.40 µg/mL), and superoxide (IC50-21.12 µg/mL) radical scavenging activities. These plant-AgNP showed significant inhibition of albumin denaturation (IC50-12.98 µg/mL) and antiproteinase activity (IC50-18.401 µg/mL). Besides, biosynthesized AgNP were found to have strong inhibitory effect against a cervical cancer (HeLa) cell line (IC50-5.418 µg/mL) as well as larvicidal activity against 3rd instar larvae of Anopheles stephensi (LC50-18.9 ppm, LC90-40.18 ppm), Aedes aegypti (LC50-12.395 ppm, LC90-36.34 ppm) and Culex quinquefasciatus (LC50-17.76 ppm, LC90-30.82 ppm) and were found to be non-toxic against normal cell line (HEK 293), and a non-target organism (Mesocyclops thermocyclopoides). This is the first report on the synthesis of AgNP using aqueous leaf extract of A. acuminata, validating their strong therapeutic potential.

Bioengineering of Piper longum L. extract mediated silver nanoparticles and their potential biomedical applications.

The present investigation highlights the strong antioxidant, anticancer and larvicidal potential of green synthesized silver nanoparticles (AgNPs) using aqueous leaf extract of Piper longum L. for their diverse therapeutic applications. The optimum conditions for the synthesis of AgNPs were recorded as 1 mM AgNO3, 60 ±â€¯2 °C at pH 6 for 120 min. Synthesized AgNPs proved to be highly stable and monodispersed as characterized through various techniques. UV-Vis spectrum of biosynthesized AgNPs showed a maximum absorption peak at 420 nm. Field emission-Scanning electron microscopy (FE-SEM) and High resolution-Transmission electron microscopy (HR-TEM) micrographs showed the spherical shape of AgNPs with mean diameter size of 28.8 nm. Existence of crystallographic AgNPs was proved by X-ray diffraction (XRD) pattern analysis. Presence of phenolics, terpenoids and flavonoids compounds which act as bioreducing agents were confirmed by Fourier-transform infrared spectroscopy (FTIR) analysis. Furthermore, the AgNPs and leaf extracts prepared individually in different solvents such as methanol, ethyl acetate, chloroform, hexane and aqueous were assessed for their bio-efficacies. AgNPs showed the enhanced antioxidant (IC50 67.56 µg) and radical-scavenging activities (IC50 196.8 µg) as compared to the crude leaf extracts. Anticancer activity revealed the strong and dose-dependent cytotoxic effect of AgNPs against the HeLa cells showing maximum IC50 value being 5.27 µg/mL after 24 h and was also found to be non-toxic to normal cells (HEK). The AgNPs induced the fragmentation of DNA in the cells, indicating the occurrence of apoptosis and necrosis. Subsequently, an efficient larvae mortality was also recorded against Anopheles stephensi having LC50 and LC90 values being 8.969 and 16.102 ppm, followed by Aedes aegypti (LC50;14.791 and LC90;28.526 ppm) and Culex quinquefasciatus (LC50;18.662 and LC90;40.903 ppm) after 72 h of exposure. Besides, they showed no toxicity against Mesocyclops thermocyclopoides (non-target organism). This is the first report showing strong anti-tumorous and larvicidal activity of AgNPs synthesized using P. longum leaf extract against cervical cancer cell line and mosquito vectors causing dengue, malaria and filariasis. Based on our findings, we suggest that AgNPs derived using P. longum leaf extract possessed excellent anti-cancerous and mosquito larvicidal potential and therefore, can be bioprospected further for the management of these hazardous health diseases. This study has given a new insight for the novel drug designing after conducting experiments on the in vivo models.

Assessment of genetic diversity, population structure and sex identification in dioecious crop, Trichosanthes dioica employing ISSR, SCoT and SRAP markers.

Twenty inter-simple sequence repeat (ISSR) and twenty two start codon targeted (SCoT) primers were employed to analyze genetic diversity and population structure among 52 Trichosanthes dioica Roxb. accessions collected from nine different eco-geographical regions of India. ISSR markers proved to be more informative in genetic diversity assessment and produced higher mean number of polymorphic bands (15.25 with 95.96% polymorphism) and polymorphic information content (PIC) value (0.47) compared to SCoT markers (12.55 polymorphic bands with 92.20% polymorphism and PIC: 0.45). Total genetic diversity (Ht) and genetic diversity within populations (Hs) in T. dioica accessions was found to be very high (0.45 and 0.43, respectively). AMOVA analysis also revealed higher genetic variation within populations (81%) than among them (19%). Among different T. dioica populations, very low genetic differentiation (Gst: 0.05) and high gene flow (Nm: 9.32) were observed. T. dioica populations of Bihar state were found to be highly diverse and Kolkata and Cuttack populations were least diverse. T. dioica male plants were more variable than females. UPGMA, Neighbor-Joining and population structure analyses divided T. dioica populations into three main clusters. First cluster comprised of Meerut population, second cluster included of Cuttack and Kolkata populations and populations of Bihar, Delhi and Kanpur occurred in third cluster. Genetic diversity was found to be strongly positively correlated with the latitude and strongly negatively correlated with annual mean rainfall of different T. dioica cultivated regions. For sex identification, one SRAP primer combination, 'Em-6/Me-4' amplified two molecular markers of around 230 and 290 bp specific to male T. dioica plants of Bihar, Kanpur, North Delhi and Meerut populations and were completely absent from female plants.

Fabrication of silver nanoparticles using Arnebia hispidissima (Lehm.) A. DC. root extract and unravelling their potential biomedical applications.

The present study reports the biosynthesis of silver nanoparticles using aqueous root extract of Arnebia hispidissima. They were prepared by adding 10 mL root extract in 90 mL silver nitrate (0.5 mM) solution and heating at 60 ± 2 °C for 12 min at pH 7.5. Characterization of the biosynthesized silver nanoparticles was done using UV-Visible spectroscopy, field emission scanning electron microscopy, energy dispersive X-ray analysis, transmission electron microscopy, X-ray diffraction analysis, dynamic light scattering measurements and Fourier-transform infrared spectroscopy. The synthesized silver nanoparticles were crystalline in nature exhibiting different shapes like sphere, rod, triangle, hexagon and polygon. Their zeta potential was -23.6 mV confirming their high stability. Fourier-transform infrared spectroscopy analysis showed the presence of phenolics, flavonoids and proteins as reducing, capping and stabilizing agents. The synthesized nanoparticles showed effective in vitro anti-oxidant activity against DPPH (IC50 = 9.86 µg/mL) and H2O2 (IC50 = 53.78 µg/mL) radicals. The nanoparticles showed dose-dependent cytotoxicity against HeLa (cervical cancer, IC50 = 4.44 µg/mL) cells and were non-toxic towards normal L20 B cells (non-malignant mouse cell line). They also exhibited strong anti-microbial activity against Candida albicans, Candida tropicalis, Geotrichum candidum (fungal strains); Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae (bacterial strains). This is the first report of synthesis of silver nanoparticles using A. hispidissima root extract validating their bioefficacy against HeLa cancer cells and diverse microorganisms.

Piriformospora indica, an excellent system for heavy metal sequestration and amelioration of oxidative stress and DNA damage in Cassia angustifolia Vahl under copper stress.

Present investigation reveals copper induced phytotoxicity, oxidative stress and DNA damage in Cassia angustifolia Vahl and its amelioration by employing a symbiotic fungus, Piriformospora indica. Seeds were germinated on Knop's medium containing five Cu levels (0, 1, 10, 50, 100 and 200 mg L-1), with and without P. indica. Colonization with P. indica significantly (P < 0.05) ameliorated Cu induced oxidative stress. However, maximum amelioration was observed at 50 mg L-1 Cu with P. indica. Atomic absorption spectroscopy revealed that P. indica colonization significantly inhibited Cu accumulation in shoots. Maximum decline in Cu accumulation in shoots was observed at 50 mg L-1 (27.27%) with P. indica over Cu alone. Besides, P. indica colonized seedlings stored 16.86% higher Cu in roots as compared to Cu alone at 200 mg L-1. Similarly, maximum proline accumulation increased up to 19.32% over Cu alone at 50 mg L-1 Cu with P. indica. Significant elevation in antioxidant enzyme levels of superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase and glutathione reductase was seen with P. indica. Contrary to increase in antioxidant level, toxic parameters such as lipid peroxidation and hydrogen peroxide decreased significantly with P. indica. Maximum decline in lipid peroxidation (13.76%) and hydrogen peroxide (18.58%) was observed at 50 mg L-1 with P. indica over Cu alone. P. indica significantly reduced DNA damage as well as changed the protein profile in C. angustifolia seedlings. Thus, P. indica proved to be an excellent system to alleviate Cu induced oxidative stress and might be useful as a phytostabilization tool.

In vitro elicitation, isolation, and characterization of conessine biomolecule from Holarrhena antidysenterica (L.) Wall. callus and its larvicidal activity against malaria vector, Anopheles stephensi Liston.

In vitro elicitation of an important compound conessine has been done in the bark-derived callus culture of Holarrhena antidysenterica (L.) Wall. employing different elicitors. For induction of callus, green bark explants excised from field-grown plants were cultured on MS medium augmented with different concentrations (0, 1, 2.5, 5, and 10 µM) of various growth regulators such as BA, IBA, NAA, and 2,4-D either alone or in combinations. The maximum amount of conessine (458.18 ± 0.89d µg/g dry wt.) was achieved in callus developed on MS medium supplemented with 5 µM BA and 5 µM 2,4-D through HPLC analysis. Elicitation in conessine content in the above callus was achieved employing a variety of organic (phenylalanine, tyrosine, chitosan, tryptophan, casein hydrolysate, proline, sucrose, and yeast extract) as well as inorganic elicitors (Pb(NO3)2, As2O3, CuSO4, NaCl, and CdCl2) in different concentrations. The optimum enhancement in conessine content (3518.58 ± 0.28g µg/g dry wt.) was seen at the highest concentration (200 mg/L) of phenylalanine. The enhancement was elicitor specific and dose dependent. The overall increment of the conessine content was seen in the order of phenylalanine > tryptophan > Pb(NO3)2 > sucrose > NaCl > As2O3 > casein hydrolysate > CdCl2 > chitosan > proline > yeast extract > CuSO4 > tyrosine. The isolation and purification of conessine was done using methanol as a solvent system through column chromatography (CC) and TLC. The isolated compound was characterized by FT-IR, 1H-NMR, and HRMS which confirmed with the structure of conessine. The bioassays conducted with the isolated compound revealed a strong larvicidal activity against Anopheles stephensi Liston with LC50 and LC90 values being 1.93 and 5.67 ppm, respectively, without harming the nontarget organism, Mesocyclops thermocyclopoides Harada, after 48 h of treatment. This is our first report for the isolation and elicitation of conessine in the callus culture of H. antidysenterica.

Green synthesis of silver nanoparticles using Holarrhena antidysenterica (L.) Wall.bark extract and their larvicidal activity against dengue and filariasis vectors.

The present study was carried out to evaluate the larvicidal potential of methanol, hexane, acetone, chloroform, and aqueous bark extracts of Holarrhena antidysenterica (L.) Wall. and silver nanoparticles (AgNPs) synthesized using aqueous bark extract against the third instar larvae of Aedes aegypti L. and Culex quinquefasciatus Say. AgNPs were prepared by adding 10 ml of aqueous bark extract in 90 ml of 1 mM silver nitrate (AgNO3) solution. After 5 min of mixing, a change in color from yellow to dark brown occurred indicating the synthesis of AgNPs. Their further characterization was done through ultraviolet-visible spectroscopy (UV-Vis), X-ray diffraction analysis (XRD), field emission scanning electron microscope (FE-SEM), transmission electron microscopy (TEM), and Fourier transform infrared spectroscopy (FT-IR). UV-Vis spectrum of synthesized AgNPs showed a maximum absorption peak at 420 nm wavelength. Crystalline nature of AgNPs was confirmed by the presence of characteristic Bragg reflection peaks in XRD pattern. TEM images have shown that most of the AgNPs were spherical in shape with an average size of 32 nm. FT-IR spectrum of AgNPs showed prominent absorbance peaks at 1012.2 (C-O) and 3439.44 cm-1 (O-H) which represent the major constituents of phenolics, terpenoids, and flavonoids compounds. LC-MS analysis of the bark extract confirmed the presence of carbonyl and hydroxyl functional groups which were directly correlated with FT-IR results. These AgNPs were assayed against different mosquito vectors, and the maximum mortality was recorded against the larvae of A. aegypti with LC50 and LC90 values being 5.53 and 12.01 ppm, respectively. For C. quinquefasciatus, LC50 and LC90 values were 9.3 and 19.24 ppm, respectively, after 72 h of exposure. Bark extracts prepared in different solvents such as methanol, chloroform, hexane, acetone, and water showed moderate larvicidal activity against A. aegypti their respective LC50 values being 71.74, 94.25, 102.25, 618.82, and 353.65 ppm and LC90 values being 217.36, 222.24, 277.82, 1056.36, and 609.37 ppm. For C. quinquefasciatus, their LC50 values were 69.43, 112.39, 73.73, 597.74, and 334.75 ppm and LC90 values of 170.58, 299.76, 227.48, 1576.98, and 861.45 ppm, respectively, after 72 h of treatment. AgNPs proved to be nontoxic against the non-target aquatic organism, Mesocyclops thermocyclopoides Harada when exposed for 24, 48, and 72 h. The results showed that bark extract-derived AgNPs have extremely high larvicidal potential compared to other organic solvents as well as aqueous bark extract alone. These AgNPs, therefore, can be used safely for the control of dengue and filarial vectors that cause severe human health hazards.

Effects of salinity and drought on growth, ionic relations, compatible solutes and activation of antioxidant systems in oleander (Nerium oleander L.).

Nerium oleander is an ornamental species of high aesthetic value, grown in arid and semi-arid regions because of its drought tolerance, which is also considered as relatively resistant to salt; yet the biochemical and molecular mechanisms underlying oleander's stress tolerance remain largely unknown. To investigate these mechanisms, one-year-old oleander seedlings were exposed to 15 and 30 days of treatment with increasing salt concentrations, up to 800 mM NaCl, and to complete withholding of irrigation; growth parameters and biochemical markers characteristic of conserved stress-response pathways were then determined in stressed and control plants. Strong water deficit and salt stress both caused inhibition of growth, degradation of photosynthetic pigments, a slight (but statistically significant) increase in the leaf levels of specific osmolytes, and induction of oxidative stress-as indicated by the accumulation of malondialdehyde (MDA), a reliable oxidative stress marker-accompanied by increases in the levels of total phenolic compounds and antioxidant flavonoids and in the specific activities of ascorbate peroxidase (APX) and glutathione reductase (GR). High salinity, in addition, induced accumulation of Na+ and Cl- in roots and leaves and the activation of superoxide dismutase (SOD) and catalase (CAT) activities. Apart from anatomical adaptations that protect oleander from leaf dehydration at moderate levels of stress, our results indicate that tolerance of this species to salinity and water deficit is based on the constitutive accumulation in leaves of high concentrations of soluble carbohydrates and, to a lesser extent, of glycine betaine, and in the activation of the aforementioned antioxidant systems. Moreover, regarding specifically salt stress, mechanisms efficiently blocking transport of toxic ions from the roots to the aerial parts of the plant appear to contribute to a large extent to tolerance in Nerium oleander.

Strong Anti-tumorous Potential of Nardostachys jatamansi Rhizome Extract on Glioblastoma and In Silico Analysis of its Molecular Drug Targets.

BACKGROUND: Glioblastoma has been reckoned as the prime cause of death due to brain tumours, being the most invasive and lethal. Available treatment options, i.e. surgery, radiotherapy, chemotherapy and targeted therapies are not effective in improving prognosis, so an alternate therapy is insistent. Plant based drugs are efficient due to their synergistic action, multi-targeted approach and least side effects. METHODS: The anti-tumorous potential of Nardostachys jatamansi rhizome extract (NJRE) on U87 MG cell line was evaluated through various in vitro and in silico bio-analytical tools. RESULTS: NJRE had a strong anti-proliferative effect on U87 MG cells, Its IC50 was 33.73±3.5, 30.59±3.4 and 28.39±2.9 µg/mL, respectively after 24, 48 and 72 h. NJRE at 30 µg/mL induced DNA fragmentation, indicating apoptosis, early apoptosis began in the cells at 20 µg/mL, whereas higher doses exhibited late apoptosis as revealed by dual fluorescence staining. NJRE at 60 and 80 µg /mL caused a G0/G1 arrest and at 20 and 40 µg/mL showed excessive nucleation and mitotic catastrophe in the cells. Immuno-blotting validated the apoptotic mode of cell death through intrinsic pathway. NJRE was harmless to normal cells. In silico docking of NJRE marker compounds: oroselol, jatamansinol, nardostachysin, jatamansinone and nardosinone have revealed their synergistic and multi-targeted interactions with Vestigial endothelial growth factor receptor 2 (VEGFR2), Cyclin dependent kinase 2 (CDK2), B-cell lymphoma 2 (BCL2) and Epidermal growth factor receptor (EGFR). CONCLUSION: A strong dose specific and time dependent anti-tumorous potential of NJRE on U87 MG cells was seen. The extract can be used for the development of safe and multi-targeted therapy to manage glioblastoma, which has not been reported earlier.

Insulinoma or non-insulinoma pancreatogenous hypoglycemia? A diagnostic dilemma.

Insulinoma is the most common cause of endogenous hyperinsulinemic hypoglycemia in adults. An alternate etiology, non-insulinoma pancreatogenous hypoglycemia (NIPH), is rare. Clinically, NIPH is characterized by postprandial hyperinsulinemic hypoglycemia, negative 72-h fasts, negative preoperative localization studies for insulinoma and positive selective arterial calcium infusion tests. Histologically, diffuse islet hyperplasia with increased number and size of islet cells is present and confirms the diagnosis. Differentiating NIPH from occult insulinoma preoperatively is challenging. Partial pancreatectomy is the procedure of choice; however, recurrence of symptoms, although less debilitating, occurs commonly. Medical management with diazoxide, verapamil and octreotide can be used for persistent symptoms. Ultimately, near-total or total pancreatectomy may be necessary. We report a case of a 67-year-old male with hypoglycemia in whom preoperative workup, including computerized tomography abdomen, suggested insulinoma, but whose final diagnosis on pathology was NIPH instead.

Assessment of Antioxidant Enzyme Activity and Mineral Nutrients in Response to NaCl Stress and its Amelioration Through Glutathione in Chickpea.

Salinity stress has been reckoned as one of the major threat towards crop productivity as it causes significant decline in the yield. The impact of NaCl stress (0, 1, 10, 50, 100 and 200 mg L(-1)) as well as glutathione (10 mg L(-1)) either alone or in combination has been evaluated on the induction of multiple shoots, antioxidant enzymes' activity, lipid peroxidation, relative permeability, concentration of nutrients, photosynthetic pigments, protein and proline content of nodal segments of chickpea after 14 days of culture. The antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) were found to be increased under salt stress as well as glutathione-supplemented medium. A significant decrease in the concentrations of chlorophylls a, b, total chlorophyll and carotenoid was observed under salt stress. Concentrations of nitrogen, phosphorus, potassium, calcium, carbon, magnesium and sulphur showed an initial increase up to 10 mg L(-1) NaCl, but a decline was seen at higher NaCl levels. Proline content and malondialdehyde concentration were found to be increased under salt stress. Three isoforms of SOD, one of CAT and four of GPX were expressed during native polyacrylamide gel electrophoresis (PAGE) analysis. However, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the stressed nodal explants revealed the over-expression of several polypeptide bands related to NaCl stress. These findings for the first time suggest that glutathione (GSH) helps in ameliorating NaCl stress in nodal explants of chickpea by manipulating various biochemical and physiological responses of plants.

Genetic diversity analysis among male and female Jojoba genotypes employing gene targeted molecular markers, start codon targeted (SCoT) polymorphism and CAAT box-derived polymorphism (CBDP) markers.

To detect genetic variations among different Simmondsia chinensis genotypes, two gene targeted markers, start codon targeted (SCoT) polymorphism and CAAT box-derived polymorphism (CBDP) were employed in terms of their informativeness and efficiency in analyzing genetic relationships among different genotypes. A total of 15 SCoT and 17 CBDP primers detected genetic polymorphism among 39 Jojoba genotypes (22 females and 17 males). Comparatively, CBDP markers proved to be more effective than SCoT markers in terms of percentage polymorphism as the former detecting an average of 53.4% and the latter as 49.4%. The Polymorphic information content (PIC) value and marker index (MI) of CBPD were 0.43 and 1.10, respectively which were higher than those of SCoT where the respective values of PIC and MI were 0.38 and 1.09. While comparing male and female genotype populations, the former showed higher variation in respect of polymorphic percentage and PIC, MI and Rp values over female populations. Nei's diversity (h) and Shannon index (I) were calculated for each genotype and found that the genotype "MS F" (in both markers) was highly diverse and genotypes "Q104 F" (SCoT) and "82-18 F" (CBDP) were least diverse among the female genotype populations. Among male genotypes, "32 M" (CBDP) and "MS M" (SCoT) revealed highest h and I values while "58-5 M" (both markers) was the least diverse. Jaccard's similarity co-efficient of SCoT markers ranged from 0.733 to 0.922 in female genotypes and 0.941 to 0.746 in male genotype population. Likewise, CBDP data analysis also revealed similarity ranging from 0.751 to 0.958 within female genotypes and 0.754 to 0.976 within male genotype populations thereby, indicating genetically diverse Jojoba population. Employing the NTSYS (Numerical taxonomy and multivariate analysis system) Version 2.1 software, both the markers generated dendrograms which revealed that all the Jojoba genotypes were clustered into two major groups, one group consisting of all female genotypes and another group comprising of all male genotypes. During the present investigation, CBDP markers proved more informative in studying genetic diversity among Jojoba. Such genetically diverse genotypes would thus be of great significance for breeding, management and conservation of elite (high yielding) Jojoba germplasm.

In vitro enhancement of psoralen as an important anticancer compound in Psoralea corylifolia through precursor feeding.

CONTEXT: Psoralea corylifolia L. (Fabacese) is rich source of bioactive compounds, which endows the plant with immense value for its use in pharmaceuticals, health, and body-care products. OBJECTIVE: The current study was designed (i) for the determination of psoralen from callus derived from different plant parts, and (ii) for the enhancement of psoralen in in vitro condition with the treatment of various psoralen pathway precursors. MATERIALS AND METHODS: B5 media were employed for raising the cultures from different plant parts such as leaf, node, root, and green seeds. Cotyledons' calluses were derived from cotyledon of green seeds that were elicited on MS + 10 µM BA + 5 µM IBA medium supplemented at 0.1, 1, 2.5, 5, 25, and 50 mg/L of various psoralen pathway precursors such as umbelliferone, cinnamic acid, and NADPH. The method for extraction of psoralen was modified from the Singh method and the content of psoralen was measured using HPLC. RESULTS: HPLC analysis of callus derived from different parts of P. corylifolia revealed that a maximum of psoralen (2601.8 µg/g fresh wt.) was recorded in cotyledons' callus. Cotyledonary callus was chosen for the enhancement of psoralens because of higher amount of psoralen in it. In vitro evaluation showed that all the precursors enhanced the psoralen amount dramatically so that the optimum amount of psoralen (2518.8 µg/g fresh wt.) was obtained at 2.5 mg/L cinnamic acid. DISCUSSION AND CONCLUSION: The results obtained indicate that cinnamic acid is one of the important precursors of psoralen pathway that induced a maximum amount of psoralen with in vitro conditions.

Micropropagation and validation of genetic and biochemical fidelity amongst regenerants of Cassia angustifolia Vahl employing RAPD marker and HPLC.

In vitro protocol has been established for clonal propagation of Cassia angustifolia Vahl which is an important source of anticancerous bioactive compounds, sennoside A and B. Nodal explants excised from field raised elite plant (showing optimum level of sennoside A and B) of C. angustifolia when reared on Murashige and Skoog's medium augmented with different cytokinins, viz. N(6)-benzyladenine (BA), N(6)-(2-isopentenyl) adenine (2iP) and 6-furfuryl aminopurine (Kn) differentiated multiple shoots in their axils. Of the three cytokinins, BA at 5 µM proved optimum for differentiating multiple shoots in 95 % cultures with an average of 9.14 shoots per explant within 8 weeks of culture. Nearly, 95 % of the excised in vitro shoots rooted on half strength MS medium supplemented with 10 µM indole-3-butyric acid (IBA). The phenotypically similar micropropagated plants were evaluated for their genetic fidelity employing random amplified polymorphic DNA (RAPD) markers. Eleven individuals, randomly chosen amongst a population of 120 regenerants were compared with the donor plant. A total of 36 scorable bands, ranging in size from 100 to 1,000 bp were generated amongst them by the RAPD primers. All banding profiles from micropropagated plants were monomorphic and similar to those of mother plant proving their true to the type nature. Besides, high performance liquid chromatography evaluation of the sennoside A and B content amongst leaves of the mature regenerants and the elite mother plant too revealed consistency in their content.

Strong larvicidal potential of Artemisia annua leaf extract against malaria (Anopheles stephensi Liston) and dengue (Aedes aegypti L.) vectors and bioassay-driven isolation of the marker compounds.

Malaria and dengue are the two most important vector-borne human diseases caused by mosquito vectors Anopheles stephensi and Aedes aegypti, respectively. Of the various strategies adopted for eliminating these diseases, controlling of vectors through herbs has been reckoned as one of the important measures for preventing their resurgence. Artemisia annua leaf chloroform extract when tried against larvae of A. stephensi and A. aegypti has shown a strong larvicidal activity against both of these vectors, their respective LC50 and LC90 values being 0.84 and 4.91 ppm for A. stephensi and 0.67 and 5.84 ppm for A. aegypti. The crude extract when separated through column chromatography using petroleum ether-ethyl acetate gradient (0-100%) yielded 76 fractions which were pooled into three different active fractions A, B and C on the basis of same or nearly similar R f values. The aforesaid pooled fractions when assayed against the larvae of A. stephensi too reported a strong larvicidal activity. The respective marker compound purified from the individual fractions A, B and C, were Artemisinin, Arteannuin B and Artemisinic acid, as confirmed and characterized through FT-IR and NMR. This is our first report of strong mortality of A. annua leaf chloroform extract against vectors of two deadly diseases. This technology can be scaled up for commercial exploitation.

Marked enhancement in the artemisinin content and biomass productivity in Artemisia annua L. shoots co-cultivated with Piriformospora indica.

A significant enhancement in artemisinin content, an important anti-malarial compound, has been achieved in Artemisia annua L. shoots by co-cultivating with Piriformospora indica, a mycorrhiza-like fungus. The in vitro shoots derived from nodal cultures of A. annua were implanted on four different culture media namely, (i) Murashige & Skoog (MS) basal, (ii) MS + 5 µM indole-3-butyric acid (IBA), (iii) MS + P. indica and, (iv) MS + 5 µM IBA + P. indica. After 2 months, it was observed that the cultures reared on MS + 5 µM IBA + P. indica showed optimum growth in terms of shoot and root proliferation over those cultured without P. indica. The average shoot number on MS + 5 µM IBA + P. indica was 17.83 ± 1.01 and on MS + P. indica alone was 12.75 ± 1.10. A drastic decline in shoot number was observed without P. indica which was 2.0 ± 0.12 on basal and 4.9 ± 1.52 on 5 µM IBA. Similarly, a maximum average of 16.83 ± 0.82 roots were achieved on MS + 5 µM IBA + P. indica which declined to 10.75 ± 1.02 on MS + P. indica. A further decrease in root number occurred in shoots without P. indica, their average being 2.5 ± 0.12 on basal and 8.91 ± 1.57 on 5 µM IBA. HPLC analysis of the aforesaid cultures revealed that the quantity of artemisinin was significantly higher (1.30 ± 0.03 %) in shoots cultured on 5 µM IBA + P. indica compared to those of control (0.80 ± 0.01 %).