Cryopreservation and assessment of genetic fidelity Acorus calamus Linn., an endangered medicinal plant.

BACKGROUND: Acorus calamus Linn. is a medicinally valuable monocot plant belonging to the family Acoraceae. Over-exploitation and unscientific approach towards harvesting to fulfill an ever-increasing demand have placed it in the endangered list of species. OBJECTIVE: To develop vitrification-based cryopreservation protocols for A. calamus shoot tips, using conventional vitrification and V cryo-plate. MATERIALS AND METHODS: Shoot tips (2 mm in size) were cryopreserved with the above techniques by optimizing various parameters such as preculture duration, sucrose concentration in the preculture medium, and PVS2 dehydration time. Regenerated plantlets obtained post-cryopreservation were evaluated by random amplified polymorphic DNA (RAPD) to test their genetic fidelity. RESULTS: The highest regrowth of 88.3% after PVS2 exposure of 60 min was achieved with V cryo-plate as compared to 75% after 90 min of PVS2 exposure using conventional vitrification. After cryopreservation, shoot tips developed into complete plantlets in 28 days on regrowth medium (0.5 mg/L BAP, 0.3 mg/L GA3, and 0.3 mg/L ascorbic acid). RAPD analysis revealed 100% monomorphism in all cryo-storage derived regenerants and in vitro donor (120-days-old) plants. CONCLUSION: Shoot tips of A. calamus that were cryopreserved had 88.3% regrowth using V cryo-plate technique and the regerants retained genetic fidelity. https://doi.org/10.54680/fr24210110412.

Phytocannabinoids for Cancer Therapeutics: Recent Updates and Future Prospects.

Phytocannabinoids (pCBs) are lipid-soluble phytochemicals present in the plant, Cannabis sativa L. and non-cannabis plants which have a long history in recreation and traditional medicine. The plant and the constituents isolated were central in the discovery of the endocannabinoid system (ECS), the most new target for drug discovery. The ECS includes two G-protein-coupled receptors; the cannabinoid receptors-1 and -2 (CB1 and CB2) for marijuana's psychoactive principle Δ(9)-tetrahydrocannabinol (Δ(9)-THC), their endogenous small lipid ligands; namely anandamide (AEA) and 2-arachidonoylglycerol (2-AG), also known as endocannabinoids and the enzymes for endocannabinoid biosynthesis and degradation such as fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL). The ECS has been suggested as a pro-homeostatic and pleiotropic signaling system activated in a time- and tissue-specific way during pathological conditions including cancer. Targeting the CB1 receptors becomes a concern because of adverse psychotropic reactions. Hence, targeting the CB2 receptors or the endocannabinoid metabolizing enzymes by pCBs obtained from plants lacking psychotropic adverse reactions has garnered interest in drug discovery. These pCBs derived from plants appear safe and effective with a wider access and availability. In the recent years, several pCBs derived other than non-cannabinoid plants have been reported to bind to and functionally interact with cannabinoid receptors and appear promising candidate for drug development including cancer therapeutics. Several of them also targets the endocannabinoid metabolizing enzymes that control endocannabinoid levels. In this article, we summarize and critically discuss the updates and future prospects of the pCBs as novel and promising candidates for cancer therapeutics.

Growth inhibitory and adjuvant therapeutic potential of aqueous extract of Triticum aestivum on MCF-7 and HeLa cells.

AIM: The purpose of the present study is to evaluate the potent growth inhibitory effects of aqueous wheatgrass extract (AWE) alone and in combination with cisplatin on human breast and cervical cancer cells. MATERIALS AND METHODS: The cytotoxic potential of AWE alone and in combination with cisplatin was evaluated on human breast and cervical cancer cells (MCF-7 and HeLa) by cell viability assay. Further, the mode of cell death induced by AWE was determined by nuclear morphological examination and cell cycle analysis. These effects were then correlated with the expression of genes involved in apoptosis and proliferation (cyclin D1 and Bax) by RT-PCR. RESULTS: AWE showed dose- and time dependent selective cytotoxicity towards the cancer highlighting its safe profile. Lower dose combinations of AWE and cisplatin induced increased growth inhibition compared with the individual drugs on both cell lines (combination index < 1) indicating strong synergistic interactions. AWE was found to induce apoptosis and arrested the cells at G0-G1 phase of the cell cycle which correlated with the modulation of expression of bax and cyclin D1 in a time-dependent manner in MCF-7 and HeLa cells. CONCLUSION: These results suggest that the anti-cancer potential of AWE may be due to apoptosis induction and its anti-proliferative properties. This study also provides the first evidence demonstrating synergism between AWE and cisplatin, which may enhance the therapeutic index of prevention and/or treatment of human breast and cervical cancer.

Annual variation in flowering phenology, pollination, mating system, and pollen yield in two natural populations of Schima wallichii (DC.) Korth.

BACKGROUND: Schima wallichii is a highly valuable tree of tropical forest in north-east Himalaya region that grows naturally in a wide range of altitudes between 750 and 2400 m asl with varying environments. Flowering phenology of tropical tree species at population level is generally ignored and therefore a detailed knowledge of flowering and fruiting patterns of important multipurpose tree species is critical to the successful management of forest genetic resources. MATERIALS AND METHODS: The study was conducted at two different altitudes (i.e., 750 m and 900 m asl) in the tropical semideciduous forest of north-east Himalaya. The floral phenology including flowering synchrony in the populations, anthesis, anther dehiscence, stigma receptivity, pollinators visitation frequency, and mating system including index of self-incompatibility were worked out in Schima wallichii according to the ear-marked standard methods given by various scientists for each parameter. RESULTS: The flowering period in Schima wallichii varied from 33 to 42 days with mean synchrony of 0.54 to 0.68 between the populations. The stigma was receptive up to 2.5 days only and showed slightly protandrous type of dichogamy. Average pollen production ranged between 6.90 × 10(7) pollen per tree in 2007 and 15.49 × 10(8) pollen per tree in 2011. A three-year masting cycle was noticed in this species. The frequency of visitation of honey bees was fairly high (5.2 ± 1.12 visits/flower/hour) as compared to other pollinators. The hand pollination revealed maximum fruit (74.2 ± 5.72%) and seed (70.8 ± 7.46%) settings. CONCLUSIONS: The variation in flowering phenology and pollen yield individually and annually along with temporal separation in anther dehiscence and pollinator's visitation cause pollen limited reproduction, which ultimately influences the reproductive success in Schima wallichii.

Fluorescent gold clusters as nanosensors for copper ions in live cells.

This paper reports the use of fluorescent gold nanoclusters synthesized using bovine serum albumin (Au-BSA) for the sensing of copper ions in live cells. The fluorescence of the clusters was found to be quenched by Cu(2+) enabling its detection in cells. The selectivity of the nanosensor was demonstrated in the presence of several cations excluding Hg(2+). We did not study the effect of Hg(2+) since it was reported earlier. The present study suggests that Cu(2+) induced fluorescence quenching is due to its binding to BSA rather than the fluorescence quenching by metal-metal interaction as in the case of Hg(2+). The Au-BSA showed excellent selectivity to Cu(2+) at various pH conditions. The 'turn off' of fluorescence can be retrieved by a Cu(2+) chelator glycine. Our results showed that gold clusters can be used as a 'turn off' sensor for copper and a 'turn on' sensor for glycine. Under the experimental conditions, the probe showed a response for Cu(2+) over a range of 100 µM to 5 mM with a detection limit of 50 µM. The role of Cu(2+) in the misfolding and disassembly of Prion Protein (PrP) leading to various maladies is well ascertained. The methodology we reported here seems to be useful in supplementing other techniques in predicting disease conditions involving Cu(2+).

Trends of greenhouse gas emissions from the road transport sector in India.

The road transport sector is the largest consumer of commercial fuel energy within the transportation system in India and accounts for nearly 35% of the total liquid commercial fuel consumption by all sectors. Gasoline and diesel consumption for road transportation have quadrupled between 1980 and 2000 due to about nine times increase in the number of vehicles and four-fold increase in freight and passenger travel demands. The paper elaborates the trends of energy consumption and consequent emissions of greenhouse gases such as CO(2), CH(4) and N(2)O and ozone precursor gases like CO, NO(x) and NMVOC in the road transport sector in India for the period from 1980 to 2000. For the first time, efforts have been made to apportion the fuels, both diesel and gasoline, across different categories of vehicles operating on the Indian roads. In order to generate more comprehensive and complete emission estimates, additionally, other minor fuel types like light diesel oil and fuel oil along with lubricants have also been taken into account. Emission estimates have revealed that nearly 27 Mt of CO(2) were emitted in 1980, increasing to about 105 Mt in 2000. Similar trends have also been observed for other gases. Further scope for improvements in emission estimation is possible by generating country specific emission factors for different vehicle categories and improvement in documentation of fuel consumption at segregated levels by fuel types and vehicle types.

Wilson's disease presenting as isolated obsessive-compulsive disorder.

Wilson's disease (WD) is a genetic neurodegenerative disorder; it exhibits wide heterogeneity in symptoms and usually presents with liver disease and/ or neuropsychiatric manifestations. The common neurological manifestations observed are dysarthria, gait disturbance, dystonia, rigidity, tremor, dysphagia and chorea. The frequent psychiatric manifestations reported are personality and mood changes, depression, phobias, cognitive impairment, psychosis, anxiety, compulsive and impulsive behavior. Isolated obsessive-compulsive disorder (OCD) is a rare presentation of WD. Reported herein is a case of a 17-year-old boy with isolated OCD. He presented to the psychiatrist with symptoms of contamination obsessions and washing compulsions, along with compulsion of repeated feet tapping and was treated with adequate doses of fluoxetine for 6 months but did not improve. Later on, he was diagnosed as a case of WD and showed improvement with chelating and behavior therapy. This implies the importance of the occurrence of isolated psychological symptoms in WD.

Localized attachment loss in Pendred syndrome: incidental?

BACKGROUND: Pendred syndrome is a rare, inherited, autosomal recessive disorder with an iodine organification defect of thyroxin produced by the thyroid gland. Its clinical features include sensorineural hearing loss, classically congenital and prelingual, and goiter. METHODS: This is the first case report of Pendred syndrome in the dental literature with oral findings that include localized extensive attachment loss involving mandibular incisor teeth and idiopathic hypercementosis involving multiple teeth in addition to other oral manifestations, suggesting hypothyroidism-like macroglossia and macrocheilia. Furthermore, serum alkaline phosphatase along with inorganic calcium and phosphate levels were also elevated. Peripheral neutrophil function test suggested a defective function of neutrophils. RESULTS: Management of the case included augmenting thyroxin supplementation, in consultation with an endocrinologist, and extraction of hopeless mandibular central incisors followed by placement of immediate transitional dentures. CONCLUSION: A comprehensive medical history and systemic and laboratory evaluations should be considered a prerequisite to identify, manage, and report such rare conditions in routine clinical practice.

Anti-neutrophil cytoplasmic autoantibodies: a renewed paradigm in periodontal disease pathogenesis?

In the past, various models including the autoimmunity model have been proposed to explain the pathogenesis of periodontal diseases. The aim of this review is to introduce the pathogenic role of anti-neutrophil cytoplasmic autoantibodies (ANCAs) in various autoimmune diseases and compare these conditions with periodontal disease to elucidate common pathogenic mechanisms. Also, a novel model to explain the pathogenesis of periodontal disease based on the concept of ANCA-associated autoimmunity is proposed. This encompasses a wide array of biochemical mediators that range from direct and indirect initiators of ANCA production and eventual release of proinflammatory mediators and free radicals, all of which have been implicated in periodontal tissue destruction in the past. In addition, specific links between the typical ANCA-associated diseases and periodontal disease are discussed. Finally, a new paradigm in the periodontal disease-associated destruction is proposed that includes the currently accepted mechanism, namely, the genetic-microbial-host interactions.

Pollen production, microsporangium dehiscence and pollen flow in Himalayan cedar (Cedrus deodara Roxb. ex d. Don).

Microsporangium dehiscence, pollen production and dispersal were studied in Himalayan cedar (Cedrus deodara) during 1998 and 1999. Microsporangium dehiscence showed diurnal periodicity and was found to be related to air temperature and relative air humidity, with a strobilus taking 2 d to dehisce completely in warmer conditions and 3 d in cooler ones. The frequency of flowering in C. deodara was highly variable during the two successive years; however, cyclical production of pollen grains was observed in 50% of the trees. The maximum concentration of pollen grains in the air was found between 1200 and 1600 h, and this period was also noted to be the best time for pollination. Studying migration of pollen grains from isolated single trees in three directions showed that migration was not uniform in all directions. Long-distance transport of pollen grains was observed in the downhill direction. However, in the uphill and horizontal directions grains could travel only up to 97.5 and l95.1 m, respectively, and the frequency of pollen grains to the source frequency at these distances was only 1.9 and 2-5%, respectively. The results suggest that an isolation barrier of 190 m may be considered as a minimum for the management of deodar seed orchards.

Saw palmetto berry extract inhibits cell growth and Cox-2 expression in prostatic cancer cells.

The cytotoxicity of a commonly used material to alleviate the symptoms of benign prostatic hyperplasia (BPH), Saw Palmetto Berry Extract (SPBE), was examined as neat oil using a set of prostatic cell lines; 267B-1, BRFF-41T and LNCaP. Proliferation of these prostatic derived cell lines is inhibited to different degrees when dosed for 3 days with SPBE. The amount of SPBE required to inhibit 50% growth (IC50) of these cell lines was 20-30 nl equivalents of SPBE per ml of medium for cell lines 267B-1 and BRFF-41T and approximately 10-fold more for the LNCaP cell line. The effect of SPBE dosing on these cell lines is not irreversible, since a 30 min treatment with SPBE at an IC50 concentration does not inhibit their growth. Normal prostate cells were inhibited by 20-25% when grown in the presence of 200 nl SPBE equivalent per ml media. Growth of other non-prostatic cancer cell lines, i.e. Jurkat and HT-29, was affected by approx. 50% and 40%, respectively. When LNCaP cells were grown in the presence of dihydrotestosterone and SPBE, the IC50 concentration decreased significantly compared to LNCaP cells grown in the presence of serum and SPBE. Reduced cellular growth after SPBE treatment of these cell lines may relate to decreased expression of Cox-2 and may be due to changes observed in the expression of Bcl-2. Expression of Cox-1 under similar conditions is not affected because of its constitutive expression. Since increased Cox-2 expression is associated with an increased incidence of prostate cancer, and decrease in its expression by SPBE would provide a basis for further investigation of its use against BPH and in prostatic cancer chemoprevention.

The anticalcification effect of polyethylene glycol-immobilized on hexamethylene diisocyanate treated pericardium.

Pathologic calcification is thought to be the main cause of failure in the present generation tissue valves fabricated from glutaraldehyde pretreated bovine pericardium (BP). The present investigation describes the in vitro calcification and enzymatic degradation of bovine pericardia after hexamethylene diisocyanate (HMDIC) crosslinking and subsequent modification with polyethylene glycol. The enzymatic degradation of these treated surfaces were monitored by scanning electron micrography and tensile strength measurements. Various proteases, such as alpha-chymotrypsin, bromelain, esterase, trypsin and collagenase were investigated for tissue stability. Incubation of these enzymes with crosslinked pericardia had variably reduced their tensile strength. Among these treated surfaces, polyethylene glycol (PEG) grafted BP via isocyanate functionalities had retained maximum strength. The PEG modified tissues had also indicated a substantial reduction in calcification, when compared to other treated tissues. Further, the biocompatibility of various pericardial tissues were established by platelet adhesion and octane contact angle measurements. It is assumed that the PEG modification of pericardium may interfere with the cellular activation of injury (platelets) to reduce tissue associated calcification. In conclusion, it seems the PEG modification of bovine pericardium via HMDIC may provide new ways of controlling tissue biodegradation and calcification. However, more in vivo studies are needed to develop applications.

Lymphatic filariasis in two distinct communities of upper Assam.

Current status of bancroftian filariasis among two distinct communities of Upper Assam was evaluated. High prevalence (10.3% mf rate) in tea workers descendents of tribals migrated from endemic states such as Orissa, Bengal, Uttar Pradesh and southern states of India, were recorded. Among non tea workers (indigenous population) significantly low mf rate was found in spite of the fact that both the communities are living in close vicinity and in similar ecological and mosquitogenic conditions. Culex quinquefasciatus was incriminated as vector in the areas of both the communities. A group of recent migrants of Bihar and Bengal living in the locality of non tea worker population were also found microfilaraemic. Significantly lower infection rate in non tea worker population appears to be associated with host susceptibility to the infection. Familial clustering in microfilaraemic subjects noticed in tea workers group also suggests involvement of racial susceptibility to infection.

Effect of fabrication, sterilization and mediators--blood compatibility of polyurethanes.

The possible changes in the surface and physical properties of polyether urethane urea (PEUU) implants, including their interaction with blood, due to the different preparation methods, sterilization techniques and long term storage in different environmental conditions have been investigated by conducting the studies of mechanical properties, contact angle, platelet adhesion and protein adsorption. Considerable variations in the mechanical properties have been observed for the PEUU grafts stored in different conditions. Changes in platelet adhesion and albumin adsorption have also been observed in the case of samples that underwent different sterilization methods. The effect of mediators like bromelain, an enzyme present in pineapple juice, on albumin adsorption and platelet adhesion on PEUU surfaces have been investigated. It seems the presence of pineapple juice increases the adsorption of albumin and reduces the adhesion of platelets on PEUU surfaces.

Effect of plasma glow, glutaraldehyde and carbodiimide treatments on the enzymic degradation of poly (L-lactic acid) and poly (gamma-benzyl-L-glutamate) films.

The hydrolytic and enzymic degradation of poly(L-lactic acid) (PLA) and poly(gamma-benzyl L-glutamate) (PBGA) films, together with a series of surface treatments, were studied, as a function of exposure time. The degradation of these polymers was monitored by weight loss, contact angle, pH changes and tensile strength studies. Glutaraldehyde treatment retained the maximum strength of PLA in buffer, followed by carbodiimide, compared with control films. On the other hand, plasma glow reversed the effect. The ability of alpha-chymotrypsin, carboxypeptidase, ficin, esterase, bromelain and leucine aminopeptidase to modulate the degradation of PLA and PBGA was also investigated. Addition of these enzymes to the polymer-buffer system reduced the tensile strength of these polymers variably. Among the six enzymes studied, leucine aminopeptidase showed the highest enzymic effect on the degradation of the glutaraldehyde-treated and bare PLA or bare PBGA films. However, glutaraldehyde-cross-linked PLA demonstrated maximum stability in buffers or in all other enzyme systems studied compared with bare PLA. It is conceivable that surface treatments on these polymers might have altered their physical and chemical configuration and the subsequent degradation properties. Surface modifications may provide new ways of controlling the biodegradation of polymers for a variety of biomedical applications.

Albumin adsorption on to aluminium oxide and polyurethane surfaces.

The changes in protein adsorption onto aluminium surfaces coated with different thicknesses of oxide layers were examined. The oxide layers on aluminium substrates were derived by the anodizing technique. Protein adsorption studies were conducted using 125I-labelled albumin and the amount of albumin adsorbed was estimated with the help of a gamma counter. An increase in albumin adsorption was observed on oxide layer coated aluminium surfaces. The effect of anti-Hageman factor on albumin and fibrinogen adsorption on to bare aluminium, oxide layer coated aluminium and bare polyether urethane urea surfaces was also investigated. It was observed that the presence of anti-Hageman factor increased the adsorption of albumin and fibrinogen on to all these substrates.

Effect of vitamin B6 on the synthesis and degradation of aspartate aminotransferase in chicken embryo fibroblasts.

The effect of pyridoxal depletion and supplementation on the intracellular level of mitochondrial and cytosolic aspartate aminotransferase in cultured chicken embryo fibroblasts was examined. No apoenzyme was detected in cells grown in the presence of pyridoxal, and the specific activity of total enzyme did not vary profoundly from primary to quaternary cultures. Under pyridoxal depletion, up to 40% apoenzyme was found in tertiary cultures which was entirely due to the mitochondrial isoenzyme. Cytosolic apoenzyme was never detected. Total aspartate aminotransferase relative to total protein was increased 2-fold in secondary cultures; only the mitochondrial isoenzyme contributed to the increased specific activity. The cytosolic isoenzyme decreased steadily and was below the limit of detection in quaternary cultures. The changes are attributed to an increased and decreased synthesis of mitochondrial and cytosolic isoenzyme, respectively. No induction of either isoenzyme was observed after incubating the cells with different hormones and substrates. In secondary cultures, no degradation of mitochondrial isoenzyme could be detected under pyridoxal deficiency or supplementation during 4.4 days, an interpassage duration. The cytosolic aspartate aminotransferase was degraded initially with an apparent half-life of approximately 0.9 day under both sets of conditions. The pronounced stability of mitochondrial aspartate aminotransferase, even though one-third of it was present as apoenzyme, excludes the formation of the apoform to be the rate-limiting step in its degradation. The present results show that pyridoxal affects the synthesis of mitochondrial and cytosolic aspartate aminotransferase, but differently.