Search

Microfluidic systems aim to detect sample matter quickly with high sensitivity and resolution, on a small scale. With its increased use in medicine, the field is showing significant promise in prostate cancer diagnosis and management due, in part, to its ability to offer point-of-care testing. This review highlights some of the research that has been undertaken in respect of prostate cancer and microfluidics. Firstly, this review considers the diagnosis of prostate cancer through use of microfluidic systems and analyses the detection of prostate specific antigen, proteins, and circulating tumor cells to highlight the scope of current advancements. Secondly, this review analyses progressions in the understanding of prostate cancer physiology and considers techniques used to aid treatment of prostate cancer, such as the creation of a micro-environment. Finally, this review highlights potential future roles of microfluidics in assisting prostate cancer, such as in exosomal analysis. In conclusion, this review shows the vast scope and application of microfluidic systems and how these systems will ensure advancements to future prostate cancer management.

The objective of this research was to examine the response of different X-ray doses to two okra genotypes, Hassawi okra and the commercial Clemenson genotype on germination and biochemical analysis...

Aedes mosquitoes are implicated in the transmission of several viruses, including Dengue virus (DENV) to millions of people worldwide. The global expansion of Aedes mosquitos'habitats creates a desperate need for control mechanisms with minimum negative effects. Deciphering the molecular interactions between DENV and its vector is a promising field to develop such efficient control strategies. As soon as the viremic blood is ingested by the mosquito, DENV is encountered by different innate immunity responses. During the past three decades, different pathways of innate immunity have been identified in Aedes spp. Recognition of viral molecular patterns, including viral RNA, and vector attempts to resist DENV infection are the most important defense mechanisms. Crosstalk between innate immune pathways and redundancy of anti-DENV responses become more evident as research progresses. The viral evasion and repression of vector immune response are increasingly being discovered. Such viral strategies are potential targets to be disrupted in order to limit DENV infection and spread. Vector-related non-immune factors such as gut microbiota can also be tapped for efficient control of DENV infection in Aedes mosquito's populations without affecting their fitness. Current trends in controlling DENV in its vector are exploring the potentials of using genetically engineered mosquitoes via RNA-based systems to degrade DENV genome once released into the midgut cells cytoplasm at the early phase of the infection.

MVs are released in response to several stress agents, in an attempt to prevent continued cellular damage. After an initial stimulus of prostate cancer cells with sublytic C5b-9 and activation of MV release through PKC, cells take at least 20 min to fully recover their ability to microvesiculate. This release of MVs through activation of sublytic C5b-9 was inhibited by the PKC inhibitor bisindoylmaleimide I but not the Rho kinase inhibitor, Y27632. After stimulus there is a rise of 79 nMs(-1) over 11 s, reaching a peak [Ca(2+)]i of 920 nM. The concentration of cytosolic calcium then falls steadily at 2.4 nMs(-1) over 109 s reaching baseline levels (50-100 nM) within 10-15 min. In PC3 cells the rate of release of MVs from stimulated cells also reaches a minimum within 10-15 min. Using fura-2 AM-loaded cells, upon stimulation, cells were found to release MVs with a concentration of intravesicular calcium estimated at ∼ 430 nM.

Using a Quartz Crystal Microbalance with dissipation monitoring, QCM-D (label-free system) measuring changes in resonant frequency (Δf) that equate to mass deposited on a sensor, we showed the attachment, over a 60 min period, of a monolayer of PC3 cells to the gold electrodes of the quartz crystal sensor, which had been rendered hydrophilic. That MVs were released upon BzATP stimulation of cells was confirmed by NTA analysis (average 250 nm diameter), flow cytometry, showing high phosphatidylserine exposition and by fluorescent (Annexin V Alexa Fluor® 488-positive) and electron microscopy. Over a period of 1000s (16.7 min) during which early apoptosis increased from 4% plateauing at 10% and late apoptosis rose to 2%, the Δf increased 20 Hz, thereupon remaining constant for the last 1000s of the experiment. Using the Sauerbrey equation, the loss in mass, which corresponded to the release of 2.36 × 106 MVs, was calculated to be 23 ng. We therefore estimated the mass of an MV to be 0.24 pg. With the deposition on the QCM-D of 3.5 × 107 MVs over 200s, the decrease in Δf (Hz) gave an estimate of 0.235 pg per MV.

A reverse phase HPLC method is developed for the simultaneous analysis of Trifluoper- azine (TFP) and prochlorperazine (PCP) in pharmaceutical preparations. HPLC was carried out on a C18 column using acetonitrile as a mobile phase at 1 mL min � 1 flow rate and the effluent was monitored at 250 nm. Chlorperazine hydrochloride (CPZ) was used as an internal standard. The retention time of the drugs was 10.879 and 13.708 min, respectively. This method produced a linear response in the concentration range between 5-200 l gm l � 1 of trifluoperazine and 10-500 l gm l � 1 of prochlorperazine. In this study, a HPLC method was successfully applied for the quantitative assay of trifluoperazine and prochlorperazine in tablets and ampule of commercial preparations which is simple, rapid and does not require any separation step for each drug.

Complex organic additives are known to improve growth and differentiation of in vitro plant cultures. The present investigation was conducted to determine the effect of various concentrations of yeast extract (YE) and casein hydrolysate (CH) on callus growth and somatic embryogenesis in date palm cultivar Nabout Saif. Callus induced from shoot tip explants was grown on callus multiplication medium supplemented with either YE or CH at 0.0, 0.1, 0.25, 0.5, and 1 g l−1. To induce somatic embryogenesis, callus was transferred to a hormone-free medium containing the corresponding concentration of additives. The results have shown that callus weight and the number of somatic embryos were directly proportional to increases in the concentration of organic additives tested. Callus growth was best achieved when 1 g l−1 of either YE or CH was added to the culture medium. At this concentration of YE, callus growth was double that of the control medium. On CH-containing media growth was 2.3 times that of the control. This indicates that CH is more effective in enhancing callus growth. However, YE was more effective in enhancing somatic embryogenesis. The data show that the best somatic embryo formation was obtained on either 1 g l−1 YE or 0.5 g l−1 CH which produced 45 and 30 embryos per culture, respectively, as compared to 20 embryos produced in the control treatment. Resultant somatic embryos successfully rooted and regenerated plantlets which exhibited normal growth in the greenhouse. Enhanced plant regeneration, an essential criterion for commercial micropropagation, was achieved.

The methods of Plasma Membrane-derived Vesicle (PMV) isolation and quantification vary considerably in the literature and a new standard needs to be defined. This study describes a novel filtration method to isolate PMVs in plasma, which avoids high speed centrifugation, and to quantify them using a Becton Dickinson(BD) FACS Calibur (TM) flow cytometer, as annexin V-positive vesicles, larger than 0.2 mu m in diameter. Essentially microvesicles (which comprise a mixture of PMVs and exosomes) from citrate plasma were sonicated to break up clumped exosomes, and filtered using Millipore 0.1 mu m pore size Hydrophilic Durapore membranes in Swinnex 13 mm filter holders. Phosphatidylserine-positive PMVs detected with annexin V-PE were quantified using combined labelling and gating strategies in conjunction with Polysciences Polybead Microspheres (0.2 mu m) and BDTrucount tubes. The PMV absolute count was calculated on the analysis template using the Trucount tube lot number information and expressed in PMV count/ml. Having estimated a normal reference range (0.51 x 10(5)-2.82 x 10(5) PMVs/ml) from a small sample of human donors, using the developed method, the effect of certain variables was investigated. Variations such as freezing of samples and gender status did not significantly alter the PMV absolute count, and with age plasma PMV levels were only marginally reduced. Smokers appeared to have reduced PMV levels. Nicotine, as for calpeptin was shown to dose-dependently (from 10 up to 50 mu m) reduce levels of early apoptosis in THP-1 monocytes and to decrease the level of PMV release. Fasting individuals had 2-3 fold higher PMV absolute counts compared to non-fasting subjects.

Plasma membrane-derived vesicles (PMVs) or microparticles are vesicles (0.1-1mum in diameter) released from the plasma membrane of all blood cell types under a variety of biochemical and pathological conditions. PMVs contain cytoskeletal elements and some surface markers from the parent cell but lack a nucleus and are unable to synthesise macromolecules. They are also defined on the basis that in most cases PMVs express varying amounts of the cytosolic leaflet lipid phosphatidylserine, which is externalised during activation on their surface. This marks the PMV as a biologically distinct entity from that of its parent cell, despite containing surface markers from the original cell, and also explains its role in events such as phagocytosis and thrombosis. There is currently a large amount of variation between investigators with regard to the pre-analytical steps employed in isolating red cell PMVs or RPMVs (which are slightly smaller than most PMVs), with key differences being centrifugation and sample storage conditions, which often leads to result variability. Unfortunately, standardization of preparation and detection methods has not yet been achieved. This review highlights and critically discusses the variables contributing to differences in results obtained by investigators, bringing to light numerous studies of which RPMVs have been analysed but have not yet been the subject of a review.

A benzopyranopyridine inhibitor of mitogen-activated protein kinase-activated protein kinase 2 (MK2) is prepared rapidly and efficiently in one step using microwave dielectric heating, whereas a substrate-selective p38 MAPK inhibitor was prepared using conventional heating techniques. The former had MK2 inhibitory activity above 2.5 microM concentration, whereas the latter showed no MK2 inhibition at 10 microM. However, rather than rescuing the reduced cellular growth rate and aged morphology of hTERT-immortalised WS dermal fibroblasts, both induce a state resembling stress-induced cellular senescence, suggesting that these inhibitors may have limited therapeutic use.

Complement C2 receptor inhibitor trispanning (CRIT) inhibits the classical pathway (CP) C3 convertase formation by competing with C4b for the binding of C2. The C-terminal 11-amino-acid of the first CRIT-extracellular domain (CRIT-H17) has a strong homology with a sequence in the C4beta chain, which is responsible for the binding of C2. Since the CP and alternative pathway (AP) C3 convertases have many functional and structural similarities, we further investigated the effects of CRIT-H17 on the AP. The factor D-mediated cleavage of factor B (FB) was blocked by CRIT-H17. By ELISA and immunoblot, CRIT-H17 was shown to bind FB. CRIT-H17 had no decay activity on the C3bBb complex as compared to decay-accelerating factor. Binding of CRIT-H17 to FB did not interfere with the assembly of C3bB complex. In a haemolytic assay using C2-deficient serum, CRIT-H17 interfered with AP complement activation.

Complement C2 receptor inhibitor trispanning (CRIT) is a receptor for the second component of complement and is found in various tissues and hemopoietic cells. On binding to CRIT, C2 cannot be activated to potentially form a variant-C3 convertase as it is rendered non-cleavable by C1s. CRIT thus limits the amount of C3 convertase formed on the cell surface. In this study we have shown, using flow cytometry and immunofluorescence microscopy, that human CRIT undergoes endocytosis from the plasma membrane. The endocytosis, possibly ligand mediated, occurs via clathrin-coated pits as it can be inhibited by prior incubation of cells in hypertonic medium or with chlorpromazine, at 37 degrees C. However, inhibition of endocytosis was not possible after treatment with nystatin, or filipin, inhibitors of caveolae/raft-dependent endocytosis. In the presence of C2 alone, CRIT associates with the adapter protein, beta-arrestin-2, and whether in association with C2 or not, then appears in the perinuclear region, but does not appear to be translocated into the nucleus. Apart from the C3aR and C5aR that internalize the anaphylatoxic peptides, this is the first report of the internalization via the clathrin pathway of a receptor for a complement serum protein.

Variability in callus growth and somatic embryogenesis in response to silver nitrate (AgNO3) among genotypes of date palm (Phoenix dactylifera L.) was investigated. Callus was cultured on MS medium containing 53.7 μM naphthaleneacetic acid (NAA) and 7.4 μM 2-isopentenyladenine (2iP) and supplemented with AgNO3 at 0, 12.5, 25, 37.5, 50, 62.5, 75, 87.5, or 100 μM. Subsequently, somatic embryogenesis was induced by transferring callus to hormone-free MS medium containing corresponding AgNO3 concentrations. Callus growth of cvs. Barhee, Naboot Saif, Ruzaiz, and Hillali was significantly promoted in response to 37.5 μM AgNO3 but optimum growth occurred at 50 μM in the former three cultivars, whereas cv. Hillali grew best at 62.5 μM. In contrast, cv. Khusab exhibited significant increase in callus weight at 12.5 μM but maximum growth occurred at 62.5 μM. Callus proliferation, expressed in increased callus fresh weight, which precedes the formation of somatic embryos during the regeneration stage, also varied depending upon AgNO3 concentration and cultivars. Regeneration percentage was significantly enhanced in all genotypes except cv. Hillali that was unaffected. Optimum AgNO3 concentrations were 62.5, 50, 37.5 μM for cvs. Barhee, Hillali, and Ruzaiz, respectively, whereas 12.5 μM was optimum for cvs. Naboot Saif and Khusab. Significant increase in number of somatic embryos was observed in cvs. Barhee, Naboot Saif, and Ruzaiz in response to 75, 12.5, and 37.5 μM AgNO3 but 75, 87.5, and 75 μM, respectively, were optimum. Embryo numbers in cvs. Hillali and Khusab were unchanged by AgNO3 and at concentrations higher than 50 μM numbers decreased. This study identified optimum AgNO3 levels for somatic embryo production of some commercial date palm cultivars.

Embryogenic callus derived from offshoot tip of date palm ( Phoenix dactylifera L.) was transferred to MS medium containing 0, 25, 50, 75, or 100 μM AgNO 3 combined with 0 or 0.5 μM 2iP. Embryogenic callus weight, number of embryos developed and embryo elongation were significantly influenced by the interaction between silver nitrate (AgNO 3 ) and 2iP. In the absence of 2iP, callus weight was greatest with 75 μM AgNO 3 , but in the presence of 2iP omitting silver nitrate resulted in the highest callus proliferation. The number of embryos increased in response to increasing silver nitrate concentration in the absence of 2iP, but in the presence of 2iP increasing the concentration of silver nitrate gave the opposite trend. The number of resultant embryos was the highest on 25 μM AgNO 3 in the presence of 0.5 μM 2iP. This treatment also caused maximum embryo elongation. The results have shown that silver nitrate promoted callus proliferation and enhanced the formation and elongation of somatic embryos of date palm. Furthermore, the action of silver nitrate was clearly modified by the addition of 2iP. Depending on the response, 2iP modification ranged from slight alteration to complete inversion of the general trend associated with increasing silver nitrate concentration. The observed stimulatory action of AgNO 3 on date palm somatic embryogenesis may contribute to improve existing regeneration systems particularly for recalcitrant date palm genotypes.

Shoots regenerated from leaf-derived callus of spinach (Spinacia oleracea L. cultivar ‘High Pack’) were induced to flower by exposure to 14-h photoperiods (50 μmol m−2 s−1) and day/night temperatures of 20° C 16° C . Leaves from male and female plants served as sources for explants. Shoots regenerated from callus were induced to flower on a hormone-free medium. The majority of regenerated plants exhibited sex corresponding to their respective explant donors; a proportion of regenerants of male origin, however, shifted to monoecious. These sex-modified plantlets developed viable seeds in vitro. This phenomenon can be useful in producing seeds from elite selected male plants.

Purpose Abnonnalities in pulmonary function have been reported in association with chronic liver disease of varied etiology. The aim of this study was to better define the frequency and nature of these abnormalities in patients who were being evaluated for liver transplantation. patients and methods We performed a battery of pulmonary function tests and chest radiographs in 116 consecutive patients (50 men, 66 women; aged 19 to 70 years, mean 44.6 years) with severe advanced liver disease who were hospitalized specifically for evaluation for possible orthotopic liver transplantation and were able to perform technically satisfactory tests. In 17 patients, quantitative whole-body technetium-99m macroaggregated albumin perfusion scanning was also performed for assessment of possible right-to-left shunting through intrapulmonary vascular dilatations. Results The most commonly affected test of lung function was the single-breath diffusing capacity for carbon monoxide (DL CO ), which was abnormal in 48%, 45%, and 71% of patients who never smoked, former smokers, and current smokers, respectively. Ventilatory restriction was noted in 25% of all patients, airflow obstruction (reduced ratio of forced expiratory volume in 1 second to forced vital capacity) in only 3%, and a widened alveolar-arterial oxygen gradient in 45%. Diffusion impairment was accompanied by a restrictive defect in only 35% of the patients and by an abnormally widened alveolar-arterial oxygen gradient in 60%. When diffusion impairment was accompanied by an oxygenation defect, it was also associated with a significantly increased right-to-left shunt fraction (mean 24.9%) assessed from quantitative whole-body perfusion imaging. On the other hand, isolated diffusion impairment unaccompanied by significant hypoxemia (noted in approximately a third of the patients with a reduced DL CO ) was not associated with evidence of significant intrapulmonary shunting (mean rightto-left shunt fraction 6.7%). Conclusions Most patients with advanced liver disease have one or more types of abnormality in lung function, a reduced DL CO being the single most common functional defect Mechanisms accounting for the abnormality in gas transfer may include intrapulmonary vascular dilatations, diffuse interstitial lung disease, pulmonary vaso-occlusive disease, and/or ventilation-perfusion unbalance.

Callus was induced from leaf explants of four spinach ( Spinacia oleracea L.) cultivars on Murashige-Skoog (MS) medium supplemented with 2 mg 1 −1 6-(furfurylamino)purine (kinetin) and 0.1, 0.5 or 1 mg 1 −1 2,4-dichlorophenoxyacetic acid (2,4-D) either in darkness or exposed to light (50 μE m −2 s −1 ). Callus production was cultivar dependent. Maximum callus proliferation was obtained with 0.5 mg 1 −1 2,4-D, which also gave rise to the highest percentage of shoot formation. Callus induced in darkness exhibited higher regeneration capacity than callus induced in the light. Regeneration capacity of the callus varied among cultivars and was enhanced by the addition of ascorbic acid. Gibberellic acid was required for shoot regeneration. Complete plants were obtained by inducing the shoots to root with 1 mg 1 −1 indole butyric acid (IBA).

The regulatory receptor complement receptor inhibitor trispanning (CRIT), on binding complement C2, is able to limit the amount of C3 convertase formed on the surface of cells. Recently CRIT was shown to be expressed in highly differentiated cells including glomerular podocytes in the kidney, keratinocytes in oesophagus and sertoli cells in the testis. The pattern of CRIT mRNA transcription in adult kidney corresponds with expression of CRIT protein in glomerular podocytes, and in a mature podocyte cell line. CRIT mRNA and protein are expressed in fetal kidney from 14 weeks’ gestation with increasing expression levels at gestational week 21. Some expression of CRIT was noted in parietal epithelial cells in the Bowman's capsule where podocyte precursor cells form, but only in the more immature glomerulus. Our results indicate that besides complement regulation, CRIT may play a role in podocyte maturation.