Nitroimidazole derivatives potentiated against tumor hypoxia: Design, synthesis, antitumor activity, molecular docking study, and QSAR study.

A hypoxic environment occurs predominantly in tumors. During the growth phase of a tumor, it grows until it exceeds its blood supply, leaving regions of the tumor in which the oxygen pressure is dramatically low. They are virtually absent in normal tissues, thus creating perfect conditions for selective bioreductive therapy of tumors. To this aim, a novel series of cytotoxic radiosensitizer agents were synthesized by linking the nitroimidazole scaffold with oxadiazole or triazole rings. The majority of the compounds exhibited moderate to excellent antiproliferative activities toward HCT116 cell line under normoxic and hypoxic conditions. The structure-activity relationship study revealed that compounds containing the free thiol group either in the oxadiazoles 11a,b or the triazoles 21a,b-23a,b demonstrated the strongest antiproliferative activity, which proves that the free thiol group plays a crucial role in the antiproliferative activity of our compounds under both normoxic (half-maximal inhibitory concentration [IC50 ] = 12.50-24.39 µM) and hypoxic conditions (IC50 = 4.69-11.56 µM). Radiosensitizing assay of the four most active cytotoxic compounds 11b and 21-23b assured the capability of the compounds to enhance the sensitivity of the tumor cells to the DNA damaging activity of γ-radiation (IC50 = 2.23-5.18 µM). To further investigate if the cytotoxicity of our most active compounds was due to a specific signaling pathway, the online software SwissTargetPrediction was exploited and a molecular docking study was done that proposed cyclin-dependent kinase 2 (CDK2) enzyme to be the most promising target. The CDK2 inhibitory assay assured this assumption as five out of six compounds demonstrated a comparable inhibitory activity with roscovitine, among which compound 21b showed threefold more potent inhibitory activity in comparison with the reference compound. A further biological evaluation proved compound 21b to have an apoptotic activity and cell cycle arrest activity at the G1 and S phases. During the AutoQSAR analysis, the model demonstrated excellent regression between the predicted and experimental activity with r2 = 0.86. Subsequently, we used the model to predict the activity of the test set compounds that came with r2 = 0.95.

MHD dissipative Powell-Eyring fluid flow due to a stretching sheet with convective boundary conditions and slip velocity.

The novelty and motivation of this research can be emphasized by examining how the heat transfer mechanism of a non-Newtonian Powell-Eyring fluid, which flows because of a stretched sheet, is affected by factors like viscous dissipation, the slip velocity phenomenon, and Joule heating. In addition, the investigation delves into the heat transfer behavior of the fluid flow when it comes into contact with a convectively heated stretched surface that is influenced by varying fluid properties. This analysis also takes into account the influence of changing fluid characteristics and the presence of magnetic field. The numerical solutions of modelled equations that governing the problem are detected using the shooting technique. Also, in order to confirm the validity of the present investigation, a proper comparison with certain published works as a particular case of the present model is presented, and a perfect agreement is noted. With the use of diagrams and tables, the flow problem's effective parameters are thoroughly discussed. Likewise, through a tabular representation, the values of the local Nusselt number and the skin-friction coefficient are computed and analyzed. Many significant conclusions can be drawn from numerical results. Most importantly, the local Nusselt number rises monotonically with both the surface convection parameter and the slip velocity parameter, but the local skin-friction coefficient has the opposite trend. The results indicate that the nanofluid temperature is enhanced by factors such as the surface convection parameter, magnetic field, and viscous dissipation. On the other hand, the slip velocity phenomenon leads to the opposite effect.

Efficient application of newly synthesized green Bi2O3 nanoparticles for sustainable biodiesel production via membrane reactor.

Introduction of waste and non-edible oil seeds coupled with green nanotechnology offered a pushover to sustainable and economical biofuels and bio refinery production globally. The current study encompasses the synthesis and application of novel green, highly reactive and recyclable bismuth oxide nanocatalyst derived from Euphorbia royealeana (Falc.) Boiss. leaves extract via biological method for sustainable biofuel synthesis from highly potent Cannabis sativa seed oil (34% w/w) via membrane reactors. Advanced techniques such as X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Energy Diffraction X-Ray (EDX), and FT-IR were employed to illustrate the newly synthesized green bismuth oxide nanoparticles. 92% of FAMEs were produced under optimal reaction conditions such as a 1.5% w/w catalyst weight, 1:12 oil to methanol molar ratio, and a reaction temperature of 92 ⸰C for 3.5 h via membrane reactor. The synthesized Cannabis biodiesel was identified using the FT-IR and GC-MS techniques. The fuel properties of synthesized biofuels (acid number 0.203 mg KOH/g, density 0.8623 kg/L, kinematic viscosity 5.32 cSt, flash point 80 °C, pour point -11 °C, cloud point -11 °C, and Sulfur 0.00047 wt %, and carbon residues 0.2) were studied and established to be comparable with internationally set parameters. The experimental data (R2 = 0.997) shows that this reaction follow pseudo first-order kinetics. These findings affirm the application of green bismuth oxide nanoparticles as economical, highly reactive and eco-friendly candidate for industrial scale biodiesel production from non-edible oil seeds.

Outcomes of traumatic brain injury in the patient of 60 years and above: a single centre retrospective study.

INTRODUCTION: The elderly is at risk for traumatic brain injury (TBI), but local data on their morbidity and mortality outcomes was lacking. This study aims to assess the outcome in mortality and functional outcome, Glasgow Outcome Scale (GOS) and factors associated with poor outcomes in patients with TBI more than 60 years old. MATERIALS AND METHODS: This single centre retrospective cohort study was carried out involving patients age 60 years old and above with TBI between June 2018 to May 2021. The mortality and GOS at hospital discharge, 30th day, and 90th day of trauma were analysed. The simple logistic regression (SLR) and multiple logistic regression (MLR) were performed to determine factors associated with poor outcomes and mortality. RESULTS: A total of 248 patients were analysed. The mean age was 67.5 ± 6.31 years. 156 (62.9%), 26 (10.5%), and 66 (26.6%) had mild, moderate, and severe TBI, respectively. The overall mortality rate was 9.7% and the median(IQR) GOS score were 4(2); p<0.001 at hospital discharge, 30th day and 90th day. There was significant difference in GOS outcomes after 90 days χ2(2) = 136.76 p<0.001. Upon MLR, there was a significant association of polytrauma, Adj. OR 11.04 (2.503-48.711); p < 0.002 and TBI severity: moderate TBI, Adj. OR 71.44(13.028-391.782); p < 0.001 and severe TBI, Adj OR 2533.51 (213.050-30127.644); p<0.001 towards poor outcome. However, only severity of TBI: moderate TBI, Adj. OR 19.48 (1.899-199.094); p=0.012 and severe TBI, Adj OR 26.42 (2.864-243.722); p=0.004 is associated with mortality. CONCLUSION: Polytrauma and moderate-severe head injury are associated with poor outcomes and moderate-severe head injury is associated with high mortality.

Schistosomiasis as causative organism for chronic osteomyelitis. Case report.

INTRODUCTION AND IMPORTANCE: Osteomyelitis is an infection of the bone mainly caused by bacteria and rarely by fungi and parasites. Parasitic osteomyelitis (schistosoma) is a very rare and unique condition with a few literatures. CASE PRESENTATION: A 14-year- old boy presented with right upper arm sinus for 4 years and exposed bone for 1 year. He has a history of hematuria. Blood tests were normal, urine general was normal and no Ova was seen. In Radiological assessment, X-ray showed Sequestrum in the anterior part of the upper humerus with Involucrum, MRI showed abnormal expansion, cortical thickening and diffuse altered marrow signal in the shaft of humerus with multiple cortical defects, sinus tracts, peri-osseus enhancing sheets and collections. The patient underwent Sequestrectomy and samples were collected for culture & sensitivity and showed no growth, Acid Fast Bacilli was negative, Histopathology test showed marked mixed inflammatory infiltrate composed mainly of eosinophils surrounding numerous Ova of Schistosoma haematobium, the patient was shifted to Praziquantel, wound care and regular follow-up. Long term clinical & radiological follow up showed good healing and the patient was satisfied. CLINICAL DISCUSSION: Parasitic osteomyelitis caused by Schistosoma is a very rare, and unique condition with a limited published cases in literature. Janet.T.Scott et al. stated that Schistosoma Haematobium is associated with chronic Osteomyelitis during investigation about potential risk factors of Buruli ulcer, which recognized as emerging public health problem by WHO in 1997 in West Africa, that lead to severe complications like amputations. CONCLUSION: Schistosomiasis can cause chronic osteomyelitis, good history taking and examination, high index of suspicious, collection of adequate tissue samples and sending them to a reliable laboratory are the corner stone of diagnosis.

Polymeric Insulator Conditions Estimation by Using Leakage Current Characteristics Based on Simulation and Experimental Investigation.

The current work contributes an estimate of the time-frequency characteristics of a leakage current in assessing the health condition of a polluted polymeric insulator. A 33 kV polymer insulator string was subjected to a series of laboratory tests under a range of environmental conditions, including pollution, wetting rate (WR), non-soluble deposit density (NSDD), and non-uniform distribution pollution (FT/B). The temporal and frequency features of the leakage current were then extracted and used as assessment indicators for insulator conditions based on laboratory test findings. Two indices were generated from the leakage current waveform in the time domain: the curve slope index (F1), which is determined by measuring the inclination of the curve between two successive time peaks of the leakage current, and the crest factor indicator (F2). The frequency domain of the leakage current signal was used to calculate the other two indices. These are the odd harmonic indicators derived from the odd frequency harmonics of the leakage current up to the 9th component (F3) and the 5th to 3rd harmonics ratio (F4). The findings showed that the suggested indicators were capable of evaluating insulator conditions. Finally, the confusion matrix for the experimental and prediction results obtained with the proposed indices was used to assess which indicator performed the best. Therefore, the analysis suggests an alternative and effective method for estimating the health condition of a polluted insulator through leakage current characteristics obtained in the time and frequency domains.

Thiamine deficiency and oxidative stress induced by prolonged metronidazole therapy can explain its side effects of neurotoxicity and infertility in experimental animals: Effect of grapefruit co-therapy.

We aimed to explore the possible neurotoxicity and infertility mechanisms of prolonged metronidazole (MTZ) use and the effects of antioxidant grapefruit (GP) co-therapy on MTZ-induced complications. Sixty male albino Wistar rats were divided into four groups (n = 15 each). Group I (control group) received 1% dimethyl sulfoxide (27 ml/ kg/day), group II (MTZ group) received MTZ (400 mg/kg/day), group III (MTZ + GP) received MTZ (400 mg/kg/ day) plus GP juice (27 ml/kg/ day) and group IV (GP group) received GP juice (27 ml/kg) for 60 days. Semen analyses were performed. Free testosterone, gonadotrophin (follicle-stimulating hormone (FSH) and luteinizing hormone) and thiamine levels were measured. Samples of cerebellar, testicular and epididymal tissues were used for both colorimetric assays of oxidative stress markers and histopathological examinations. Significant decreases in the sperm count, percent total sperm motility, serum thiamine levels, free testosterone levels and FSH levels were observed in the MTZ and MTZ + GP groups (p < 0.05 for all parameters). Significantly higher oxidative stress levels (p < 0.05) were observed in the cerebellar and testicular tissue homogenates of these groups than in those of the control group, and associated disruptions in the cerebellar, testicular and epididymal structures were apparent compared to those of the control group. Although the GP group showed a significantly higher sperm count and significantly lower oxidative stress than the control group (p < 0.05), with histological similarity to the control group, the GP group exhibited significantly higher prolactin levels and lower free testosterone and FSH levels than the control group (p < 0.05). Oxidative stress and decreased thiamine levels could explain the MTZ-induced neurotoxicity and infertility side effects that aggravated by GP co-administration.

Extreme environments: microbiology leading to specialized metabolites.

The prevalence of multidrug-resistant microbial pathogens due to the continued misuse and overuse of antibiotics in agriculture and medicine is raising the prospect of a return to the preantibiotic days of medicine at the time of diminishing numbers of drug leads. The good news is that an increased understanding of the nature and extent of microbial diversity in natural habitats coupled with the application of new technologies in microbiology and chemistry is opening up new strategies in the search for new specialized products with therapeutic properties. This review explores the premise that harsh environmental conditions in extreme biomes, notably in deserts, permafrost soils and deep-sea sediments select for micro-organisms, especially actinobacteria, cyanobacteria and fungi, with the potential to synthesize new druggable molecules. There is evidence over the past decade that micro-organisms adapted to life in extreme habitats are a rich source of new specialized metabolites. Extreme habitats by their very nature tend to be fragile hence there is a need to conserve those known to be hot-spots of novel gifted micro-organisms needed to drive drug discovery campaigns and innovative biotechnology. This review also provides an overview of microbial-derived molecules and their biological activities focusing on the period from 2010 until 2018, over this time 186 novel structures were isolated from 129 representatives of microbial taxa recovered from extreme habitats.

Hybrid PSO-FLC for dynamic global peak extraction of the partially shaded photovoltaic system.

Particle Swarm Optimization (PSO) is widely used in maximum power point tracking (MPPT) of photovoltaic (PV) energy systems. Nevertheless, this technique suffers from two main problems in the case of partial shading conditions (PSCs). The first problem is that PSO is a time invariant optimization technique that cannot follow the dynamic global peak (GP) under time variant shading patterns (SPs) and sticks to the first GP that occurs at the beginning. This problem can be solved by dispersing the PSO particles using two new techniques introduced in this paper. The two new proposed PSO re-initialization techniques are to disperse the particles upon the SP changes and the other one is upon a predefined time (PDT). The second problem is regarding the high oscillations around steady state, which can be solved by using fuzzy logic controller (FLC) to fine-tune the output power and voltage from the PV system. The new contribution of this paper is the hybrid PSO-FLC with two PSO particles dispersing techniques that is able to solve the two previous mentioned problems effectively and improve the performance of the PV system in both normal and PSCs. A detailed list of comparisons between hybrid PSO-FLC and original PSO using the two proposed methodologies are achieved. The results prove the superior performance of hybrid PSO-FLC compared to PSO in terms of efficiency, accuracy, oscillations reduction around steady state and soft tuning of the GP tracked.

Low risk of recurrence following artesunate-Sulphadoxine-pyrimethamine plus primaquine for uncomplicated Plasmodium falciparum and Plasmodium vivax infections in the Republic of the Sudan.

BACKGROUND: First-line schizontocidal treatment for uncomplicated malaria in the Republic of the Sudan is artesunate (total dose 12 mg/kg) plus Sulphadoxine/pyrimethamine (25/1.25 mg/kg) (AS/SP). Patients with Plasmodium vivax are also treated with 14 days primaquine (total dose 3.5 mg/kg) (PQ). The aim of this study was to assess the efficacy of the national policy. METHODS: Patients above 1 year, with microscopy-confirmed, Plasmodium falciparum and/or P. vivax malaria were treated with AS/SP. Patients with P. falciparum were randomized to no primaquine (Pf-noPQ) or a single 0.25 mg/kg dose of PQ (Pf-PQ1). Patients with P. vivax received 14 days unsupervised 3.5 mg/kg PQ (Pv-PQ14) on day 2 or at the end of follow up (Pv-noPQ). Primary endpoint was the risk of recurrent parasitaemia at day 42. G6PD activity was measured by spectrophotometry and the Accessbio Biosensor™. RESULTS: 231 patients with P. falciparum (74.8%), 77 (24.9%) with P. vivax and 1 (0.3%) patient with mixed infection were enrolled. The PCR corrected cumulative risk of recurrent parasitaemia on day 42 was 3.8% (95% CI 1.2-11.2%) in the Pf-noPQ arm compared to 0.9% (95% CI 0.1-6.0%) in the Pf-PQ1 arm; (HR = 0.25 [95% CI 0.03-2.38], p = 0.189). The corresponding risks of recurrence were 13.4% (95% CI 5.2-31.9%) in the Pv-noPQ arm and 5.3% (95% CI 1.3-19.4%) in the Pv-PQ14 arm (HR 0.36 [95% CI 0.1-2.0], p = 0.212). Two (0.9%) patients had G6PD enzyme activity below 10%, 19 (8.9%) patients below 60% of the adjusted male median. Correlation between spectrophotometry and Biosensor™ was low (rs = 0.330, p < 0.001). CONCLUSION: AS/SP remains effective for the treatment of P. falciparum and P. vivax. The addition of PQ reduced the risk of recurrent P. falciparum and P. vivax by day 42, although this did not reach statistical significance. The version of the Biosensor™ assessed is not suitable for routine use. Trial registration https://clinicaltrials.gov/ct2/show/NCT02592408.

MCNP5 evaluation of photoneutron production from the Alexandria University 15 MV Elekta Precise medical LINAC.

Photoneutron production, and the dose equivalent, in the head assembly of the 15 MV Elekta Precise medical linac; operating in the faculty of Medicine at Alexandria University were estimated with the MCNP5 code. Photoneutron spectra were calculated in air and inside a water phantom to different depths as a function of the radiation field sizes. The maximum neutron fluence is 3.346×10-9 n/cm2-e for a 30×30 cm2 field size to 2-4 cm-depth in the phantom. The dose equivalent due to fast neutron increases as the field size increases, being a maximum of 0.912 ± 0.05 mSv/Gy at depth between 2 and 4 cm in the water phantom for 40×40 cm2 field size. Photoneutron fluence and dose equivalent are larger to 100 cm from the isocenter than to 35 cm from the treatment room wall.

Design, synthesis and biological evaluation studies of novel quinazoline derivatives as cytotoxic agents.

Some novel quinazoline derivatives 6a-h were designed, synthesized and evaluated for their in vitro cytotoxic activity against lymphoma cell line compared to etoposide as a reference drug. Compounds (S)-2-(6,8-diiodo-2-phenylquinazolin-4-ylamino)-3-phenylpropanoic acid (6 f), (S)-2-(6,8-diiodo-2-phenylquinazolin-4-ylamino)-3-(1H-imidazol-4-yl)propanoic acid (6 g) and (S)-2-(6,8-diiodo-2-phenylquinazolin-4-ylamino)-3-(1H-indol-3-yl) propanoic acid (6 h) had comparable higher cytotoxic activity than the reference drug. Compound 6 f, the most active compound, had IC50=13.2 µM. In an attempt to interpret such anti-cancer activity of these derivatives, their anti-inflammatory action was examined using the carrageenan induced rat paw edema method. The most active compounds showed moderate anti-inflammatory activity compared to the reference drug. In order to identify the most relevant physicochemical features important for high antitumor activity of the target compounds, specific 2D descriptors were calculated and correlated with the antileukemic activity.

Treatment of hepatitis C virus infection in patients on maintenance hemodialysis: a single United Arab Emirates center experience.

BACKGROUND: Hepatitis C is prevalent among hemodialysis patients. In patients with normal kidney function, treatment with pegylated interferon and ribavirin can lead to eradication of HCV (hepatitis C virus). But the treatment is more problematic in patients with impaired kidney function, in part due to the altered pharmacokinetics of these medications. Despite recent guidelines, the optimal strategy in this group of patients is not well defined. METHOD: In a retrospective study, we reviewed all patients with chronic hepatitis C on hemodialysis treated at Sheikh Khalifa Medical City, in the United Arab Emirates between 2003 and 2009. The aim of our study was to determine the rate of sustained viral response (SVR) and to establish the safety and rate of dropouts in the different treatment regimens used (patients treated with peginterferon only and patients treated with peginterferon and low dose ribavirin). RESULTS: 22 patients were treated during this period. 5 patients received monotherapy with a reduced dose of peginterferon alfa 2b s.c. once weekly while 17 patients were treated with a combination of reduced dose of peginterferon alfa 2a or 2b s.c. once weekly and a low dose ribavirin (200mg/day). A SVR was achieved in 73% (16/22 patients) of the total patient population and in 76% (13/17 patients) in the sub-group of patients treated with a combination therapy. The tolerability was high. No patients had to discontinue their treatment. The use of ESA (erythropoietin stimulating agents) and G-CSF was common in the combination therapy (94% and 53% respectively). CONCLUSION: In our study of patients on hemodialysis with chronic hepatitis C, the use of peginterferon or a combination of peginterferon with a low daily dose of ribavirin achieved a high rate of SVR and the rate of dropout was low after pursuing an aggressive management of side effects.

Towards non-surgical therapy for uterine fibroids: catechol-O-methyl transferase inhibitor shrinks uterine fibroid lesions in the Eker rat model.

BACKGROUND: Uterine leiomyomas (fibroids) are the most common pelvic tumors in women. We assessed the potential therapeutic utility of Ro 41-0960, a synthetic catechol-O-methyl transferase inhibitor (COMTI), in the Eker rat. METHODS: We randomized uterine fibroid-bearing Eker rats for treatment with Ro 41-0960 (150 mg/kg/12 h) versus vehicle for 2 and 4 weeks. The fibroids were measured by caliper and subjected to histological evaluation. Urinary levels of 2-hydroxy estrogen (E(2)), 16-hydroxy E2 and DPD (osteoporosis marker) and serum liver enzymes were evaluated. Expressions of Cyclin D1, proliferating cell nuclear antigen (PCNA), Poly [ADP-ribose] polymerase1 (PARP1), tumor suppressor gene (P53) and transforming growth factor (TGFß3) were assessed in fibroids using immunohistochemical analysis or RT-PCR. Apoptosis was confirmed using terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL). RESULTS: Ro 41-0960-treated rats exhibited fibroid volumes of 86 ± 7% and 105 ± 12% of initial burden, at 2 and 4 weeks post-treatment, respectively, significantly lower than control group (240 ± 15% and 300 ± 18%; P< 0.01). Ro 41-0960 increased the urinary 2-hydroxy E2/16-hydroxy E(2) ratio, level of p53 mRNA and TUNEL positivity (P< 0.05) and decreased PARP1, PCNA and cyclin D1 proteins and TGFß3 mRNA (P< 0.05). Ro 41-0960 did not change normal tissue histology, liver functions or urinary DPD level. CONCLUSIONS: Ro 41-0960 (COMTI) arrested growth/shrunk uterine fibroids in Eker rats. This result may be related to modulation of estrogen-dependent genes involved in apoptosis, proliferation and extracellular matrix deposition via accumulation of 2-hydroxy estrogen. The efficacy and safety of Ro 41-0960 in rats suggest its candidacy for treatment of uterine fibroids.

CMOS Conductometric System for Growth Monitoring and Sensing of Bacteria.

We present the design and implementation of a prototype complementary metal-oxide semiconductor (CMOS) conductometric integrated circuit (IC) for colony growth monitoring and specific sensing of Escherichia coli (E. coli) bacteria. The detection of E. coli is done by employing T4 bacteriophages as receptor organisms. The conductometric system operates by measuring the resistance of the test sample between the electrodes of a two-electrode electrochemical system (reference electrode and working electrode). The CMOS IC is fabricated in a TSMC 0.35-µm process and uses a current-to-frequency (I to F) conversion circuit to convert the test sample resistance into a digital output modulated in frequency. Pulsewidth control (one-shot circuit) is implemented on-chip to control the pulsewidth of the output digital signal. The novelty in the current work lies in the ability of the CMOS sensor system to monitor very low initial concentrations of bacteria (4×10(2) to 4×10(4) colony forming unit (CFU)/mL). The CMOS system is also used to record the interaction between E. coli and its specific receptor T4 bacteriophage. The prototype CMOS IC consumes an average power of 1.85 mW with a 3.3-V dc power supply.

Decolorization of Mordant red 73 azo dye in water using H2O2/UV and photo-Fenton treatment.

Decolorization of the Mordant red 73 (MR73) azo dye in water was investigated in laboratory-scale experiments using UV/H(2)O(2) and photo-Fenton treatments. Photodegradation experiments were carried out in a stirred batch photoreactor equipped with a low-pressure mercury lamp as UV source at 254 nm. The effect of operating parameters such as pH, [H(2)O(2)](,) [dye] and the presence of inorganic salts (NaNO(3), NaCl and Na(2)CO(3)) were also investigated. The results indicated that complete dye decolorization was obtained in less than 60 min under optimum conditions. Furthermore, results showed that dye degradation was dependent upon pH, [H(2)O(2)] and initial dye concentration. The presence of chloride ion led to large decreases in the photodegradation rate of MR73 while both nitrate and carbonate ions have a slight effect. The photo-Fenton treatment, in the presence of Fe powder as a source of Fe(2+) ions, was highly efficient and resulted in 99% decolorization of the dye in 15 min. Mineralization of MR73 dye was investigated by determining chemical oxygen demand (COD). In a 3h photoperiod "65%" of the dye was mineralized by the H(2)O(2)/UV process, while the photo-Fenton treatment was more efficient producing 85% mineralization over the same 3-h period.

Toward gene therapy of uterine fibroids: targeting modified adenovirus to human leiomyoma cells.

BACKGROUND: To circumvent the paucity of the primary adenovirus (Ad5) receptor and the non-specific Ad5 tropism in the context of uterine leiomyoma cells, Ad5 modification strategies would be beneficial. METHODS: We screened several modified adenoviruses to identify the most efficient and selective virus toward human leiomyoma cells to be used as candidate for delivering therapeutic genes. We propagated: wild-type Ad5-luc, fiber-modified viruses: ad5 RGD-luc, Ad5-Sigma-luc, Ad5/3-luc and Ad5-CAV2-luc, as well as transcriptional targeted viruses: ad5 survivin-luc, Ad5-heparanase-luc, Ad5-MSLN-CRAD-luc and Ad5-SLPI-luc, on 293 cells and purified them by double CsCL density centrifugation. Then we transfected primary cultures of human leiomyoma cells derived from fibroids of four different patients, telomerase-immortalized human leiomyoma cell line (huLM), telomerase-immortalized normal human myometrial cell line (HM9) and immortalized normal human liver cells (THLE3) with the viruses at 5, 10 and 50 plaque-forming units (PFU)/cell. After 48 h, luciferase activities were measured and normalized to the total cellular protein content. RESULTS: Ad5-RGD-luc and Ad5-CAV2-luc, Ad5-SLPI-luc and Ad5-MSLN-CRAD-luc at 5, 10 and 50 pfu/cell showed significantly higher expression levels of luciferase activity in both primary and immortalized human leiomyoma cells when compared with Ad5-Luc. Additionally, these modified viruses demonstrated selectivity toward leiomyoma cells, compared with myometrial cells and exhibited lower liver cell transduction, compared with Ad5-luc, at the same dose levels. CONCLUSIONS: Ad5-CAV2-luc, Ad5-RGD-luc, Ad5-SLPI-luc and Ad5-MSLN-CRAD-luc are promising delivery vehicles in the context of leiomyoma gene therapy.

Effects of various nonopioid receptor antagonists on the antinociceptive activity of Muntingia calabura extracts in mice.

This study was carried out in mice to determine the nonopioid receptor signaling pathway(s) that might modulate the antinociceptive activity of the aqueous and chloroform extracts of Muntingia calabura (M. calabura) leaves, using the hot-plate test. The leaves of M. calabura were sequentially soaked [1:2 (w/v); 72 h] in distilled water (dH(2)O) and chloroform. The 50% concentration extracts were selected for this study based on the plant's previously established antinociceptive profiles. The mice (n = 7) were pretreated (s.c.) for 10 min with the selected nonopioid receptor antagonists, followed by the (s.c.) administration of the respective extract. The latency of discomfort was recorded at the interval time of 0.5, 1, 2, 3, 4 and 5 h after the extract administration. The 5 mg/kg atropine, 10 mg/kg phenoxybenzamine, 10 mg/kg yohimbine, 10 mg/kg pindolol, 1 mg/kg haloperidol and 10 mg/kg bicuculline caused significant (p < 0.05) reduction in the aqueous extract-induced antinociceptive activity. The 10 mg/kg phenoxybenzamine, 10 mg/kg yohimbine, 10 mg/kg pindolol and 10 mg/kg bicuculline caused significant (p < 0.05) reduction in the chloroform extract-induced antinociceptive activity. In conclusion, the central antinociceptive activity of M. calabura leaves appears to be involved in the modulation of various nonopioid receptor signaling pathways. Its aqueous extract antinociceptive activity is mediated via modulation of the muscarinic, alpha(1)-adrenergic, alpha(2)-adrenergic, beta-adrenergic, dopaminergic and GABAergic receptors, while its chloroform extract activity is mediated via modulation of the alpha(1)-adrenergic, alpha(2)-adrenergic, beta-adrenergic and GABAergic receptors.