Fulltext search in archive

Ginger and turmeric rhizomes are used in folk medicine for the treatment of several cerebrovascular diseases with limited scientific basis for their action. Hence, in this study, we investigate the effects of two Zingiberaceae varieties (ginger and turmeric) on ectonucleotidases (NTPDase and 5'-nucleotidase), adenosine deaminase (ADA) and acetylcholinesterase (AChE) activities in synaptosomes of cerebral cortex from l-NAME induced hypertensive rats. The animals were divided into seven groups (n = 10): normotensive control rats; hypertensive rats; hypertensive rats treated with atenolol; normotensive and hypertensive rats treated with 4% supplementation of turmeric and ginger rhizomes, respectively. After 14 days of pre-treatment with both rhizomes the animals were induced with hypertension by oral administration of l-NAME. The results revealed an increase of ATP and AMP hydrolysis as well as ADA and AChE activities of cerebral cortex synaptosomes in induced rats when compared with the control. The supplementation of both rhizomes prevented these alterations by decreasing ATP and AMP hydrolysis and ADA and AChE activities in cerebral cortex. In conclusion, this study demonstrated that both rhizomes interfere with the purinergic and cholinergic neurotransmission in cerebral cortex of hypertensive rats. Therefore, we can suggest that both rhizomes exert neuroprotective potential under hypertensive state.

Cytostatic treatment is often negatively affected by dose-limited toxicities. Novel agents, including nanoparticle-based drug delivery systems (DDS), are becoming available to overcome this problem. Despite achieving a lesser toxicity in exchange for more favorable pharmacokinetic profiles, the use of DDS is often associated with a particular toxicity profile. The accumulation of DDS in tumor tissue is much faster than in normal tissues where toxic events occur. While only a small amount of DDS is delivered to the target tissue, and accumulated there, most of the administered dose remains in circulation. The removal of this fraction, which is no longer effective, is thought to reduce toxicity. Pegylated liposomal doxorubicin (PLD) has been proven to be effective in platinum-resistant ovarian carcinoma with the reduced risk for cardiotoxicity. Once saturation in tumor tissue is achieved, prolonged circulation seems ineffective, whereas other toxicity risks (palmar-plantar erythrody sesthesia and mucositis) have been reported. Therefore, extracorporeal elimination of circulating nanoparticles using plasma filtration would probably reduce this risk of toxicity. The elimination rate could be kinetically regulated, i.e. based on individual doxorubicin pharmacokinetic variables. Plasma filtration can significantly influence the exposure to PLD (plasma concentration-time profile-AUC of PLD) and would be a suitable, well tolerated method enabling individualized, more effective and safer chemotherapy.

Sulfate-reducing bacteria (SRB) are most likely involved in both the initiation and maintenance of inflammatory bowel disease (IBD); unfortunately present antibacterial chemotherapeutics used in the treatment of IBD have been ineffective. Thus, the antimicrobial activity of salicylamide derivatives against two different genera of intestinal SRB, Desulfovibrio and Desulfomicrobium, was investigated. Six 2-(phenylcarbamoyl)phenyl N-[(benzyloxy)carbonyl]alkanoates and three 2-hydroxy-N-[(2S)-1-oxo-1-(phenylamino)alkan-2-yl]benzamides showed MIC values in the range from 0.22 to 0.35 μM against Desulfovibrio piger Vib-7 and in the range from 0.27 to 8.52 μM against Desulfomicrobium sp. Rod-9, while MIC values of ciprofloxacin were 41.2 μM and 39.3 μM. The highest potency against the two strains was observed for 4-chloro-N-{(2S)-1-[(3,4-dichlorophenyl)amino]-3-methyl-1-oxobutan-2-yl}-2-hydroxybenzamide (MIC 0.22 μM and 0.27 μM). 4-Chloro-2-[(4-nitrophenyl)carbamoyl]phenyl (2S)-2-{[(benzyloxy)carbonyl]amino}-3-methylbutanoate showed high activity against D. piger Vib-7 (MIC = 0.26 μM), while 4-chloro-2-[(4-methylphenyl)carbamoyl]phenyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(1H-indol-2-yl)propanoate expressed high activity against Desulfomicrobium sp. Rod-9 (MIC = 0.31 μM). Structure-activity relationships are discussed.

Phenazine-1-carboxylic acid has extensive pharmacological activity, including antibiotic and immunomodulatory, but the anticancer activity remains unknown. Treatment of prostate cancer cell line (DU145) with phenazine-1-carboxylic acid stimulated inhibition of cell proliferation in concentration- and time-dependent manner. Dual staining confirmed phenazine-1-carboxylic acid stimulated prostate cancer programmed cell death in time-dependent manner. To investigate the exact mechanism, phenazine-1-carboxylic acid-stimulated oxidative stress and mitochondrial-related apoptotic pathway in human prostate cancer cells were examined in this study. Phenazine-1-carboxylic acid increased the generation of reactive oxygen species (ROS) in prostate cancer cell lines, which triggered the pro-apoptotic JNK signaling. Phosphorylated JNK stimulated the depolarization of mitochondrial membrane potential (ΔΨm) and downregulation of anti-apoptotic protein Bcl-2 related with the upregulation of pro-apoptotic protein Bax. Downregulation of anti-apoptotic Bcl-2 family protein in corresponding with loss of ΔΨm, stimulate the increased production of cytochrome c and programmed cell death inducing factor (AIF) from mitochondria, and ultimately induced the caspase-dependent and caspase-independent programmed cell death. Altogether, the present study suggests that phenazine-1-carboxylic acid showed an antitumor activity in prostate cancer cells by reactive oxygen species production and mitochondrial-related apoptotic pathway. The results of the present study offered an insight into the prospective of phenazine-1-carboxylic acid for prostate cancer therapy.

Developing plant derived chemopreventive agents that can divert the carcinogenic process and inhibit tumour progression may greatly reduce serious health consequences of cancer. This study investigated the antiproliferative and apoptosis inducing ability of Rubus fairholmianus root acetone (RFRA) on MCF-7 cells. RFRA treatments showed an increase in the in vivo antioxidant levels such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) in mice. The extract showed significant antitumour activity with 76.57% increase in life span of ascites tumour bearing mice and 1.94 cm³ reductions in solid tumour volume at 250 mg/kg. The cytotoxicity spectrum analysis indicated that RFRA is a potent cytocidal agent with varying levels of toxicity (IC₅₀: 57.14-29.36 μg/ml). A significant dose dependent decrease in viability, proliferation and increase in cytotoxicity, caspase 3/7 activity were observed in MCF-7 cells. RFRA (20 μg/ml) induced apoptosis in MCF-7 cells, which was observed by morphology; DNA ladder formation and increased apoptotic population in flow cytometry analysis. These findings strongly suggest the use of R. fairholmianus as natural antioxidant with profound antitumour activities. It can act as a potent cytotoxic agent, which is able to induce apoptosis in MCF-7 cells via caspase 3/7 upregulation.

Betahistine dihydrochloride, which is widely prescribed for the treatment of symptoms associated with Meniere's syndrome, is generally administered orally in solid or liquid formulations. There is a strong need of profound investigation of alternative routes of administration of betahistine to overcome difficulties related to oral administration. The aim of this study was to evaluate betahistine cytotoxicity and permeability in vitro and to assess the drug's relevance for incorporation in drug delivery systems for nasal administration. RPMI epithelial model was used to evaluate drug permeability in vitro. The cytotoxicity of betahistine was assessed by MTT test. Chitosan microspheres were used as a betahistine delivery system. RPMI 2650 formed a thick, impermeable cell layer on the apical side of the filter inserts and developed enough TEER values to confirm confluence. According to the obtained results, BET showed high permeability coefficients (Papp values in the range 2.3 × 10^-5 to 19 × 10^-5) and could, therefore, be successfully used in nasal drug delivery formulations. Also, BET exhibited a good safety profile regarding nasal epithelium toxicity. A dose-dependent reduction in cell viability was observed. The microspheres as drug delivery systems affected BET permeation profiles due to the presence of chitosan as an absorption enhancer.

Oxidative stress and NF-κB signaling plays a major role in pathogenesis of osteoarthritis. In the present study, we analyzed the potent role of carnosol against osteoarthritis in cells treated using monosodium iodoacetate (MIA) model through in vitro studies. MIA caused dose-dependent cell death and induced programmed cell death by increasing subG1 accumulation and caspase-3 expressions. MIA caused oxidative stress by increasing reactive oxygen species, lipid peroxidation and further induced NF-κB expression and down regulated Nrf-2 levels. Pre-treatment with carnosol significantly protected the cells by reducing the oxidative stress markers and improved the cell viability up to 98%. Further, carnosol down regulated NF-κB nuclear expression with a concomitant increase in Nrf-2 nuclear localization and up regulated the nuclear Nrf-2 levels. Carnosol also inhibited MIA-induced subG1 accumulation and caspase-3 activation. This study demonstrates that, carnosol might act as potent antioxidant and regulate MIA-induced oxidative stress, NF-κB signaling and programmed cell death by up regulating the Nrf-2 levels.

The protective and anticonvulsive efficacy of two prophylactic mixtures (PANPAL consisting of pyridostigmine, benactyzine, and trihexyphenidyle and pyridostigmine plus biperiden) administered prior to the administration of soman in a lethal dose (1.5 LD₅₀) with or without antidotal treatment (atropine + HI-6) was evaluated using rats as experimental animals. The pretreatment was applied 30 and 60 min before intoxication and the antidotal therapy was administered 1 min after soman poisoning. The anticonvulsive efficacy of both prophylactic combinations was determined using a seven degree scale. Non-treated soman-poisoned rats died within 10 min after soman challenge showing severe tremor, subconvulsions and generalized convulsions. More than 90% of pretreated animals survived for 24 hrs following soman poisoning and they were observed to be free from soman-induced toxic signs 24 hrs after soman administration. Our findings confirm that both prophylactic mixtures are able not only to protect experimental animals from the lethal effects of soman but also to eliminate most soman-induced toxic signs. Our results confirm that both prophylactic mixtures should be considered as a pretreatment for nerve agent poisoning, especially in the case of the threat of exposure to soman.

The purpose of our study was to examine the early expression of p21 and activated transcription factors ATF-2, CREB, Elk-1, p53 after soman and VX poisoning, to throw light on the pathogenetic mechanism of nerve agent-induced non-specific effects. Male Wistar rats were i.m. poisoned by soman (60 μg.kg^-1 - 70% LD₅₀) or VX (8 μg.kg^-1 - 70% LD₅₀). Samples were taken 4, 24, and 72 hours after poisoning, immunohistochemically stained and phospho-ATF-2^Thr-69/71, phospho-CREB^Ser-133, phospho-Elk-1^Ser-383, phospho-p53^Ser-15, and protein p21 expressions were measured using computer Image analysis in apical and cryptal enterocytes of the colon transversum. After soman poisoning, we observed an increased phospho-CREB in cryptal enterocytes 4, 24, and 72 h after poisoning, while apical enterocytes expressed increased phospho-CREB only 72 h after intoxication. Phospho-Elk-1 significantly dropped 4 and 24 h after soman poisoning in the cryptal compartment. Activation of ATF-2 and p53 and expression of p21 were not changed 4, 24, and 72 h after soman poisoning. VX poisoning did not change any of measured parameters. Soman and VX showed a different effect on cellular signalling. Soman seems to cause additional effects, which are not related to the basic mechanism of nerve agent-induced toxicity and which temporarily suppress promitotic pathways of proliferating cells and persist in cells during the differentiation process.

Tong Luo Jiu Nao (TLJN), a modern formula of traditional Chinese medicine, has proven to be clinically efficacious in several vascular cerebral diseases but its specific effect is not known. In the present study we investigated the acute and persisting effects of TLJN on anxiety model in Wistar male rats. TLJN was administered intragastrically during three successive days (Days 1-3) and then, the double TLJN dose was given on Day 7. For the evaluation of anxiety-related behavior of animals, we used the open field (OF) and elevated plus maze (EPM) paradigms. Testing in the OF was performed on Days 1, 2, 3, 4, 8, 14 and 22; in the EPM on the Day 23.
Two-way repeated-measures ANOVA revealed significant differences between control and TLJN treated animals in the open field model where TLJN induced increased exploratory activity, indicating reduced anxiety-like behavior. The effect persisted for several days after the treatment and was still present on Days 14 and 22. Reduced anxiety-like behavior was also observed on EPM 16 days after the last TLJN administration.
The results demonstrate behavioral effects of intragastric administered TLJN, which indicate reduced anxiety. Persistence of the induced behavioral changes suggests prolonged duration of action.

The ability of two newly developed oximes (K361, K378) to reduce tabun-induced acute neurotoxic signs and symptoms was compared with the oxime K203 and trimedoxime using a functional observational battery. The neuroprotective effects of the oximes studied combined with atropine on rats poisoned with tabun at a sublethal dose (310 μg/kg i.m.; 90% of LD₅₀ value) were evaluated. Tabun-induced neurotoxicity was monitored by functional observational battery at 2 h following tabun challenge. The results indicate that all tested oximes combined with atropine enable tabun-poisoned rats to survive till the end of experiment. Both newly developed oximes (K361, K378) combined with atropine were able to decrease tabun-induced neurotoxicity in the case of sublethal poisonings although they did not eliminate all tabun-induced acute neurotoxic signs and symptoms. Their ability to decrease tabun-induced acute neurotoxicity was slightly lower than that of trimedoxime and the oxime K203. Therefore, the newly developed oximes are not suitable for the replacement of commonly used oximes (especially trimedoxime and obidoxime) in the treatment of acute tabun poisonings.

Current biomedical research requires quantitative and qualitative studies of tissue metabolites. In this review, we discuss recent advances in multinuclear magnetic resonance (MR) technology that enhance biomedical research. The advances presented herein clearly show that MR is a diagnostic tool for the study of tumor tissue metabolism associated with changes in small molecule concentrations. Biomedical MR offers a non-invasive view of cancer cell metabolism by detection of resonance nuclei such as hydrogen-1, carbon-13, phosphorus-31 and fluorine-19. Due to current progress in MR technology, it is now possible to monitor changes in cancer cell metabolism before and after therapy.
Our criteria for the selection of research papers for this review were focused on those that show progress in biomedicine achieved by using MR. Our review demonstrates that small molecule detection in cancer tissue by MR has advanced biomedical research allowing for significant improvements in tumor detection and treatment.

This study investigated the effects of sericin-derived oligopeptides on natural killer (NK) activity. In vitro exposure of human peripheral blood mononuclear cells with sericin-derived oligopeptides resulted in an augmentation of NK cell activity against K562 target cells and the effects appeared to be dose-related. Experiments designed to examine whether enhanced NK activity was due to direct or indirect activation of NK cells revealed that sericin oligopeptides did not induce activity of purified NK cells, and that sericin oligopeptides augmented NK activity indirectly by inducing the production of IL-2 and IFN-γ cytokines. In in vivo experimentation where mice were orally administered with sericin oligopeptides and splenic mononuclear cells tested against YAC-1 target cells, significant increase in NK activity was obtained compared to control mice. Elevated levels of IL-2 were also evident in all oligopeptides-treated groups. As demonstrated both in vitro and in vivo, these results indicate that sericin-derived oligopeptides have efficient NK-enhancing activity and suggest the potential therapeutic applications of such oligopeptides for functional improvement of NK cells, and possibly for treatment of tumor and infectious diseases in which NK activity contributes to host defense.

MSCs produce CD4(+)CD25(+)FOX3(+) regulatory T (Treg) cells from activated peripheral blood mononuclear cells (PBMC), T-CD4+ and T-CD8+ cells in vitro and in vivo. Here we investigated whether the deficiency of androgen/AR in MSCs influence Treg induction from total PBMC, splenocytes, CD4+CD25-through AR/TGF-β interaction. Eight to 12-week-old wild type and general androgen receptor knockout (ARKO) mice were used. MSCs were collected, characterized and function of Treg cells was studied. Our result showed that depletion of AR suppressed the immunosuppressive effect of MSCs, and demonstrated that WT-MSC-induced Treg cell expansion was partially impaired by blocking androgen receptor signal. Furthermore, the levels of TGF-β were lower in the T cell coculture with ARKO-MSC compared to WT-MSC. Exposure of ARKO-MSC cells to exogenous active TGF-β partially restored the induction of Treg cell expansion by ARKO-MSC cells. Our data suggest that ARKO-MSC hampers Treg cell expansion and function via androgen/AR and TGF-β signal pathways interaction. To the best of our knowledge, this study is the first investigating the interaction of MSCs from ARKO mice and WT Tregs in an allogeneic co-culture model. Together, these results might provide great insight into treatment of inflammatory and autoimmune diseases.

The radiosensitizing potential of Plumbagin (PLB) against chemo- and radioresistant B16F1 melanoma cells growing in vitro was investigated. Clonogenic assay revealed a sensitization enhancement ratio (SER) of 1.5 for PLB treatment in combination with radiation. PLB pretreatment for 1 h prior to radiation resulted in elevated intracellular ROS levels compared to the group treated with radiation alone. Alkaline comet assay analysis revealed PLB's potential to enhance the radiation induced DNA damage. Cell cycle studies have shown enhanced G₂/M arrest for combination treatment of PLB with radiation. Cell death exerted by PLB combination was mainly through programmed cell death, involving the depletion of mitochondrial membrane potential, increase in the expression of p53, Bax, Cytochrome c, PARP and Caspase 3 cleavage. In conclusion, this study demonstrates the radiosensitizing potential of PLB to inhibit the growth of melanoma cells in vitro, which may be attributed to the oxidative stress and DNA damage leading to enhanced mitochondria-mediated programmed cell death. Also, this study demonstrate the ability of PLB to augment ionizing radiation induced tumor cell kill which further warrant the avenue for the development of a clinically useful radiosensitizer.

The potency of two novel oximes (K920, K923) to reactivate tabun-inhibited acetylcholinesterase and to reduce acute toxicity of tabun was compared with the oxime K203 and trimedoxime using in vivo methods. The study determining percentage of reactivation of tabun-inhibited peripheral acetylcholinesterase (diaphragm) and central acetylcholinesterase (brain) in tabun-poisoned rats showed that the reactivating efficacy of both newly developed oximes is lower than the reactivating potency of the oxime K203 and trimedoxime. The therapeutic efficacy of both newly developed oximes roughly corresponds to their weak reactivating efficacy. Their potency to reduce acute toxicity of tabun in mice was lower compared to the oxime K203 and trimedoxime. All differences in reactivating efficacy of oximes and different protective ratios were found for selected doses of oximes used in this study. Based on the results obtained, we can conclude that the reactivating and therapeutic potency of both newly developed oximes does not prevail the effectiveness of the oxime K203 and trimedoxime and, therefore, they are not suitable for their replacement of commonly used oximes for the treatment of acute tabun poisoning. The conclusion is only relevant for the experimental animals used in this study because of remarkable species differences in reactivating properties of oximes.

The ability to detect pathogenic and physiologically relevant molecules in the body with high sensitivity and specificity offers a powerful opportunity in the early diagnosis and treatment of diseases. Early detection and diagnosis can be used to greatly reduce the cost of patient care associated with the advanced stages of many diseases. However, despite their widespread clinical use, these techniques have a number of potential limitations. For example, a number of diagnostic devices have slow response times and are burdensome to patients. Furthermore, these assays are expensive and cost the health care industry billions of dollars every year. Therefore, there is a need to develop more efficient and reliable sensing and detection technologies. A biosensor is commonly defined as an analytical device that uses a biological recognition system to target molecules or macromolecules. Biosensors can be coupled to a physiochemical transducer that converts this recognition into a detectable output signal. Typically biosensors are comprised of three components: (1) the detector, which identifies the stimulus; (2) the transducer, which converts this stimulus to a useful output; and (3) the signal processing system, which involves amplification and display of the output in an appropriate format. The goal of this combination is to utilize the high sensitivity and selectivity of biological sensing for analytical purposes in various fields of research and technology. We review here some of the main advances in this field over the past few years, explore the application prospects, and discuss the issues, approaches, and challenges, with the aim of stimulating a broader interest in developing biosensors and improving their applications in medical diagnosis.

Antiepileptic drugs may have varying toxicity or efficacy depending on administration time. VPA administration could be associated with a great deal of haematological toxicity and can cause aplasic anaemia or peripheral cytopenia affecting one or more cell lines. The objective of this study is to experimentally verify if VPA-induced haematological toxicity in mice varies according to drug administration time in the 24 h scale. Different groups of mice received 620 mg/kg of VPA by i.p. route at four different circadian stages. The results obtained showed that VPA treatment induced a significant decrease in the haematological parameter rates (cytopenia) depending on the circadian time; otherwise the Cosinor analysis showed that each haematological variable followed a significant blood type circadian physiological rhythm in controls and in treated mice. The highest significant haematological toxicity illustrated by the leucopenia index and thrombocytopenia was observed in the middle of the dark-activity phase (19 HALO). Chronotherapy may play an important role in improving the control of seizures and limiting the adverse effects of VPA treatment. Indeed, the data obtained indicate that the optimal haematological tolerance is observed when VPA was injected in the middle of the light-rest span of mice, which is physiologically analogous to the end of the activity of the diurnal phase in human patients.

The aim of this study was to evaluate the effectiveness of individual (inactive) proenzymes and mixtures thereof in cancer treatment and to compare this treatment with more frequently used therapy based on active proteases. Experiments focused on explanation of possible mechanisms of proenzyme action against tumours are included.
Proenzyme therapy of sarcoma S-180 significantly reduced tumour growth and prolonged survival of mice. The effect of trypsinogen and chymotrypsinogen was synergistic. Proenzyme therapy of melanoma B16-F10 bearing mice reduced both tumour growth and prevalence of metastases. Active enzyme based therapy of melanoma B16-F10 was less effective. Severe combined immunodeficiency (SCID) mice bearing sarcoma S-180 did not respond to the proenzyme therapy, indicating that the effect of this therapy is dependent on fully developed acquired immunity. Measured decreased levels of TGF-β and an increased amount of alpha-2 macroglobulin in serum contributed to the elucidation of the cancer treatment mechanism.
Proenzyme therapy based on administration of a mixture of trypsinogen and chymotrypsinogen is effective in cancer treatment.

Elephantopus scaber Linn., family Asteraceae, is a small herb found in Neotropics, Europe, Asia, Africa and Australia. The plant parts of this herb have been used traditionally for the treatment of a number of diseases in many countries. Sesquiterpene lactones, triterpenoids, steroids, flavonoids and essential oil constituents have been reported from various parts of the plant. The plant has been extensively screened for anticancer activity. Sesquiterpene lactones such as deoxyelephantopin, isodeoxyelephantopin, scabertopin, and isoscabertopin have been found to be prominent anticancer constituents. Many other biological activities such as antimicrobial, hepatoprotective, antioxidant, antidiabetic, anti-inflammatory, analgesic, antiasthamatic, antiplatelet, and wound healing have been reported in various research papers. The present review has been envisaged with an intension to provide scientific information about the ethnomedical, phytochemical and pharmacological profile of E. scaber.

Epidemiological data indicate that selenium status is inversely connected with cancer risk. Animal and human studies have demonstrated that most inorganic and organic forms of selenium compounds have an anticancer action. This work investigated the impact of organic selenium on the multiple signalling pathways involved in the inhibition of the viability of prostate cancer cells. Prostate adenocarcinoma cells (PC-3) were incubated with seleno-l-methionine (SeMet) at four concentrations and cell viability and programmed cell death were determined by the WST-1, BrdU assays and Tali image based cytometer. The expression of chosen cell-cycle regulatory genes was determined by real-time RT-PCR analysis and confirmed at the protein level. SeMet treatment of PC-3 cells resulted in an inhibition of cell proliferation in a dose- and time-dependent manner. The inhibition of proliferation correlated with the up-regulation of gene expression and the protein levels of CCNG1, CHEK1, CDKN1C and GADD45A, whereas SeMet down-regulated the expression of CCNA1 and CDK6 genes. Therefore SeMet inhibits the proliferative activity of prostate cancer cells by a direct influence on the expression of genes involved in the regulation of cell cycle progression.

The neuroprotective effects of a novel oxime KR-22934, the oxime K203 and obidoxime in combination with atropine in rats poisoned with tabun at a sublethal dose (200 μg/kg i.m.; 80% LD₅₀) were studied. The tabun-induced neurotoxicity was monitored at 24 h following tabun challenge using a functional observational battery and an automatic measurement of motor activity. The results indicate that all tabun-poisoned rats treated with oximes in combination with atropine were able to survive within 24 h following tabun poisoning. One tabun-poisoned rat without antidotal treatment died within 24 h. The oximes KR-22934 and K203 combined with atropine showed a similar potency to decrease tabun-induced neurotoxicity at 24 h after tabun administration while the neuroprotective efficacy of obidoxime was slightly higher. However, no oxime was able to eliminate tabun-induced neurotoxicity completely. When atropine was administered alone, negligible neuroprotective efficacy was observed. Based on the results, a novel oxime KR-22934 did not bring any improvement of the neuroprotective efficacy of antidotal treatment of acute tabun poisonings.

Recombinant human bone morphogenetic protein-2 (rhBMP-2) is an osteo-inductive reagent that can be used therapeutically to promote spinal arthrodesis and fracture healing. Despite its clinical use, the effect of rhBMP-2 on expression of endogenous osteogenic growth factors is poorly described. The goal of this study was to determine if local rhBMP-2 treatment modulates expression of pro-osteogenic growth factors during fracture healing. Rat femur fractures were wrapped with control or rhBMP-2 collagen soaked sponge. Radiographic observations demonstrated that rhBMP-2 treatment enhanced fracture callus formation and decreased bridging time. Expression of fracture site growth factor mRNAs was determined by quantitative polymerase chain reaction. The gene expression data demonstrated that rhBMP-2 treatment significantly increased mRNA levels of BMP-2 at day 4, BMP-4 at days 2, 4, 7, and 10, BMP-7 at day 2, and TNF-α at day 7. Treatment with rhBMP-2 was shown to significantly reduce mRNA levels of BMP-2 on day 2 and 10, TNF-α on day 14, and BMP-6 on days 2, 10, 14, and 21. TGFβ-1 was also significantly reduced on days 2 and 21. In conclusion, treatment with rhBMP-2 appears to enhance expression of other osteogenic factors that could contribute to bone formation.

The ability of two novel oximes K378 and K348 and currently available oximes (HI-6, obidoxime) to reactivate sarin-inhibited acetylcholinesterase and to reduce acute toxicity of sarin was evaluated. Both new potential oxime reactivators were chosen for this study based on the data obtained during the extensive work on oximes development, from structure-activity relationship studies and in vitro evaluation of their ability to reactivate acetylcholinesterase inhibited by organophosphorus compounds. In vivo determined percentage of reactivation of sarin-inhibited rat blood and brain acetylcholinesterase showed that the potency of both novel oximes K378 and K458 to reactivate sarin-inhibited acetylcholinesterase roughly corresponds to low reactivating efficacy of obidoxime. On the other hand, the oxime HI-6 was found to be efficient reactivator of sarin-inhibited acetylcholinesterase in the peripheral compartment. While the oxime HI-6 was able to reduce the acute toxicity of sarin more than three times, both novel oximes decreased the acute toxicity of sarin less than two times. Based on the results, we can conclude that reactivating and therapeutic efficacy of both novel oximes K378 and K458 are significantly lower compared to the oxime HI-6 and, therefore, it is not suitable to replace the oxime HI-6 for the antidotal treatment of acute sarin poisoning.

Maggot debridement therapy (MDT) is increasingly being used as a fast and effective treatment approach for healing chronic wounds. The removal of necrotic tissue from wounds is a fundamental step in successful treatment. Larvae of Lucilia sericata produce a cocktail of proteolytic and antimicrobial substances in the gut as well as salivary glands called excretion/secretion (ES) products. Three groups of proteolytic enzymes (serine, aspartyl and metalloproteinases) were identified in the larval ES products to date. We prepared cDNA library from the salivary glands and identified novel putative serine proteases, metalloproteinase and signal peptide protease. In situ hybridisation revealed following expression profiles in all the three larval instars during the feeding stage: serine protease 1 - frontgut and fat body including grease coupler of salivary glands; serine protease 2 - salivary glands and Malpighian tubes; serine protease 3 - salivary glands and anterior midgut; prenyl metalloproteinase and signal peptide protease - salivary glands and fat body. We also investigated the expression of chymotrypsin, previously identified protease, and found that it is produced only in the anterior part of the midgut. In conclusion, we identified five novel putative proteases of medicinal maggots and demonstrated that they could be secreted into the wound during the MDT.

Hermetia illucens larvae have been used in Europe and America as a medical resource and medicinal insect for the treatment of skin damage such as burns and wound healing. This study was carried out to evaluate the effect of a new substance causing antibacterial activity against various pathogenic bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The antibacterial activity of hexanedioic acid was determined using different antimicrobial indicators such as turbidometric assay, resazurin assay, and agar disk diffusion. Hexanedioic acid showed the selective-growth inhibitory effects against the growth and proliferation of Staphylococcus aureus, MRSA, Klebsiella pneumonia, and Shigella dysenteriae in a concentration dependent manner. The growth inhibitory zones of bacteria treated with 80 μg/ml of hexanedioic acid for 24 h were measured as 18.27 ± 0.18, 23.35 ± 0.15, 16.62 ± 0.18, and 12.96 ± 0.24 mm, respectively. The minimum inhibitory concentration (MIC) values of hexanedioic acid against the viability of these bacteria for 24 h were measured as 140.377, 137.369, 139.117, and 139.704 μg/ml, respectively. These results demonstrate that hexanedioic acid has antibacterial properties that effectively inhibit the growth/proliferation of pathogenic bacteria. Furthermore, this study suggests novel aspects for the utilization of hexanedioic acid as a medicinal substance.

The aim of the study was to investigate clock gene expression in Bos taurus and the alteration of that during two pathological conditions, evaluating the daily expression pattern of four clock genes (Per2, Cry2, Bmal1, Clock) in peripheral blood cells. Five healthy cows, five affected by Brucellosis (BR) and five affected by Bovine Viral Diarrhoea-Mucosal Disease (BVD-MD) were housed in indoor stalls under natural spring conditions, blood samples were collected at 4 h intervals over a 24 h period. Statistical analysis showed rhythmic expression of clock genes mRNAs in healthy cows. Cows affected by BR did not show any rhythmic expression of clock genes mRNAs, cows affected by BDV mRNA levels of Bmal1, Clock and Cry2 changed during the day. These findings highlighted that circadian system could be involved in homeostasis alteration and that clock genes could be considerate as regulatory genes or early response genes during inflammation, so, their regulation should be evaluated in health research and treatment.

Avidity is an important feature of antibodies associated with their pathogenic effects. Anticardiolipin antibodies (aCLs) are the most commonly examined antiphospholipid antibodies, however, their avidity has been investigated only marginally. The aim of the study was to compare the avidity of the antibodies specifically bound to the cardiolipin-coated microtitrate wells with those bound to the wells bearing no cardiolipin in various conditions. We analysed 22 serum samples with high, medium and low levels of aCL IgG. The avidity of aCL IgG was determined in serially diluted sera by the ELISA method in the presence of increasing urea concentrations (from 2 to 8 mol/L) as a chaotropic agent. The serum dilution 1:50 and 1:100 and the concentrations of urea 6 mol/L and 8 mol/L seemed to be suitable for the determination of aCL avidity. The avidity of antibodies specifically bound to the cardiolipin and those bound to the cardiolipin-free wells significantly differed in their avidity. The simultaneous determination of higher-avidity aCL and lower-avidity polyspecific antibodies, whose presence complicates the ELISA methods by an increase of the background signal, might limit the shortcomings of some ELISA methods.

Epigallocatechin-3-gallate (EGCG) is widely used as a weight controlling supplement. Concerns about its safety evoked after cases of hepatotoxicity occurred upon its use. The underlying factors that could be involved in EGCG associated hepatotoxicity are not fully studied. In this study, we investigated the possible impact of lipopolysaccharide (LPS), as an inflammagen, on the effect of EGCG on hepatocytes. HepG2 cells were treated with different concentrations of EGCG (100, 200, 500 μM), with and without LPS (10 nM)-presensitization of the cells. Viability of HepG2 cells decreased with the increased concentrations of EGCG; the viability was even lesser in LPS-presensitized cells. Oxidative stress (Ox.LDL and CXCL16), the expression of nuclear retinoic receptors (RAR, RXR) and the biomarkers of hepatocellular injury (TNFα, TGFβ1) were all relatively higher in LPS-presensitized cells compared to non-sensitized cells upon treatment with EGCG. Sensitization of HepG2 cells with LPS alone did not affect the viability or any of the other biomarkers considered in this study. In conclusion, EGCG alone can be harmful to liver at high concentrations and this effect may become more pronounced under the influence of an inflammagen.