These findings verify that the coculture model system was functio

These findings verify that the coculture model system was functional and particles that were applied apically (on top of the filter membrane) and able to diffuse through the collagen-1 coated filter membrane and reach the endothelial monolayer. Under coculture Gemcitabine solubility dmso conditions with H441 on the upper-side of the filter membrane and apical exposure of NPs, no uptake could be observed in ISO-HAS-1 for both NPs (Fig. 7, right column), although a detectable uptake was seen after 48 h exposure on the apical side of the filter membrane. The barrier properties were also evaluated following the apical (H441) exposure to Sicastar Red and AmOrSil. TER (Fig. 8A) was measured after exposure to Sicastar Red

(60–300 μg/ml) for 4 h and 4 h/20 h (4 h exposure and 20 h further JNJ-26481585 molecular weight cultivation in fresh serum-containing medium). Very high concentrations (300 μg/ml) resulted in a dramatic decrease of TER after 4 h (11.5 ± 6.6% of t0) and remained significant reduced during the 20 h recovery period (24 ± 21% of t0). Furthermore, TER was also checked for the permanent incubation for 48 h to Sicastar Red (60 μg/ml) and AmOrSil (300 μg/ml). No significant alterations to the TER occurred during the 48 h exposure compared to the untreated control, which demonstrated that a functional barrier was present during coculture transport experiments. The untreated control showed reduced TER values

after 24 h (91 ± 8% of t0), and these further decreased after 48 h (76 ± 11% of t0). But, even with the reduction Adenosine of TER, a functional barrier could be maintained after 48 h with 390 ± 83 Ω cm2. IL-8 and sICAM released from cells was determined after Sicastar Red exposure for 4 h/20 h (60–300 μg/ml). As control groups, transwell-monocultures (H441, seeded on the top and ISO-HAS-1 seeded on the bottom side of the

filter membrane) were evaluated along with the coculture under the same culture conditions with Sicastar Red applied apically (on the H441 side). A concentration of 300 μg/ml in the CC resulted in a dramatic IL-8 release into the upper compartment (27 ± 9-fold of untreated control uc) but not into the lower compartment, which was on the contrary observed for the H441 transwell-monoculture without ISO-HAS-1 in the lower chamber (4 ± 1.2-fold of uc). However, a significant increase of sICAM (1.76 ± 0.4% of uc) could be detected in the lower compartment of the CC (ISO-HAS-1 side) after exposure to 300 μg/ml Sicastar Red. The monoculture with ISO-HAS-1 showed higher levels of sICAM (60 μg/ml: 2.25 ± 1.3%, 100 μg/ml: 2.3 ± 0.6%, 300 μg/ml: 3.3 ± 1.1% of uc) in the apical (upper) compartment (the stimulated side and basolateral side of the ISO-HAS-1). A concentration of 60 μg/ml Sicastar Red did not cause an IL-8 elevation after 4 h/20 h but after 48 h continuous exposure (7.5 ± 3.5% of uc).

The presence and functionality of P-gp (mdr1) proteins were probe

The presence and functionality of P-gp (mdr1) proteins were probed respectively by immunocytochemistry and bi-directional permeability studies with the two established substrates, 3H-digoxin and Rh123. A positive immunocytochemical signal was obtained on the apical surface of RL-65 cell layers cultured in both media for 8 days while no green fluorescence was detected when cells were only incubated with the FITC-labelled secondary antibody (Fig. 6). However, no statistical difference (p > 0.05) between AB and BA transport across 8-day old RL-65 layers was observed for any of the two P-gp substrates investigated ( Fig. 7), suggesting

negligible transporter-mediated drug trafficking ZD6474 in the cell culture model. In vivo and ex vivo absorption studies are frequently conducted in rats to predict the pharmacokinetics of inhaled drug candidates in humans ( Tronde et al., 2003). However, variations in drug disposition in human and rat lungs have yet to be fully appraised. A rat respiratory epithelial cell culture model suitable for permeability screening would aid better understanding of interspecies differences in pulmonary drug absorption, including the role of drug transporters, in addition to providing an ethical alternative to animal testing. This

study evaluates the potential of layers of the bronchial/bronchiolar epithelial rat cell line, RL-65, as an in vitro permeability screening tool. It demonstrates that RL-65 cells

cultured at an AL interface on Transwell® supports formed layers morphologically Venetoclax solubility dmso similar to the upper airway epithelium with a TEER and 14C-mannitol paracellular permeability values in agreement with those in established human bronchial epithelial cell models. Expression of the drug transporters P-gp Oxalosuccinic acid and octn2 was confirmed in the cell layers, although no vectorial transport of widely used P-gp probes was observed. This preliminary characterisation of air-interfaced RL-65 cell layers identifies a potentially useful tool for investigating differences in drug permeability between the human and rat airway epithelia. Morphological analysis of RL-65 cells grown in presence of serum revealed multilayered cultures with an uppermost layer of non-viable cells (Fig. 4), thus providing a poor representation of the native epithelium. This indicated that a serum containing medium is unsuitable for the development of RL-65 cells into polarised layers mimicking the airway epithelium. Likewise, sub-optimal growth of the cell line had previously been described in presence of serum (Roberts et al., 1990). Our study also demonstrated that the sole consideration of markers of epithelial barrier formation such as TEER and paracellular permeability values is potentially misleading for a reliable assessment of cell-based absorption screens, and highlights the importance of morphological examinations in the characterisation of those models.

5 days—median, 312 0 days (259 0–458 0 days) The overall antibod

5 days—median, 312.0 days (259.0–458.0 days). The overall antibody response in HIV-infected patients receiving one or two doses of the meningococcal serogroup C conjugate vaccine was 81.4% (35 of the 43 patients evaluated). Of the 35 responders, 31 had received a single dose and 4 had received two doses. As shown in Table

1, after the first dose of the vaccine, side effects were observed in 16.3% of the HIV+ group patients and in 44% of the HIV− group patients (p = 0.004). The reported side effects are shown in Table 2. No side effects were reported among the 10 patients who received a second (booster) HTS assay dose of the vaccine. In the present study, 72.1% of the HIV-infected patients evaluated were responsive to a single dose of meningococcal serogroup C conjugate vaccine (as is usually recommended), and this rate increased to 81.4% when those receiving a second dose were included. However, 100% of the

non-HIV-infected patients achieved protective levels after receiving the first dose, a result that is consistent with those of other studies involving healthy children or adolescents [35], [36], [37] and [38]. The magnitude of the antibody response obtained was significantly smaller in the HIV-infected patients than in the non-HIV-infected patients. The differences found were expected, given the results of studies of the use of other vaccines in HIV-infected patients. In general, the response to vaccination was weaker in HIV-infected patients than in those not so infected. However, the response obtained in the present study was significant for the prevention of meningococcal disease in such a susceptible Selleckchem LY2109761 population. It is of note that Digestive enzyme two doses provided better results than did a single dose. These results are in accordance with a recent publication of the Advisory Committee on Immunization Practices of the Centers for Disease Control and Prevention and the American Academy of Pediatrics, which recommends an immunization

schedule with two doses of the quadrivalent conjugate vaccine (serogroups A, C, Y, and W135) for HIV-infected patients [39]. Nevertheless, in our study only 40% of the HIV-infected patients who were revaccinated responded to the booster dose, possibly due to immune system dysfunction caused by the HIV. It is debatable whether the interval between the two doses influenced the response in those patients. Further studies, with shorter intervals between doses, are needed in order to evaluate such aspect. We found that the antibody response in HIV-infected patients did not correlate with clinical variables or with the results of viral and immunological tests. Therefore, the responders and non-responders presented the same profile: CDC classification B and C; absolute CD4 count >350 cells/mm3 (with a proportion >25%); and viral loads below the detection limit in most cases. The HIV+ group showed very similar characteristics since the beginning, but no sample was calculated to determine the associations between those variables.

As HSV-2 infection is often subclinical, measurement of clinical

As HSV-2 infection is often subclinical, measurement of clinical disease as a primary endpoint is problematic. click here An important feature of candidate vaccines will be modification of the construct so that an antibody assay can distinguish between vaccinated and infected persons. Secondary endpoints should include frequency of clinically apparent HSV genital disease, and in those who seroconvert, frequency of genital viral shedding. Mathematical modeling suggests that even low efficacy preventative vaccine could impact the HSV-2 epidemic

by decreasing shedding and reducing viral transmission [90]. Such a vaccine would have the highest impact in high-prevalence populations [91]; for instance, a vaccine which marginally decreases HSV-2 susceptibility but reduces shedding frequency by 75% could reduce HSV-2 incidence by 30% over a 10 year period [92]. Thus, it is important to study both acquisition, and in those who acquire, frequency of viral shedding. An effective therapeutic HSV-2 vaccine could both improve the clinical course in individual patients,

and decrease this website HSV transmission through reduction in shedding, for a public health benefit. The approach to efficiently evaluate such vaccines relies on evaluation of viral shedding in a cohort of highly adherent persons with clinically apparent genital HSV-2; we have found that this population is highly motivated to participate in daily genital shedding studies [93]. The participants obtain genital swabs for detection of viral shedding before and after vaccination in a one-way crossover study design. These studies are ideal for proof-of-concept, Thalidomide as they can rapidly provide an answer to whether the vaccine has efficacy and can be efficiently performed with fewer than 100 persons [94]. Reduction in viral shedding is the more sensitive primary endpoint for therapeutic vaccine trials, and serves as a useful surrogate endpoint for recurrence rate

and transmission likelihood. As initial therapeutic vaccine trials should target persons with symptomatic infection, important secondary endpoints include frequency of genital lesions and prodromal symptoms. These are the clinical endpoints that have been requested in the past by FDA for licensure studies. In addition, the density of HIV receptor-positive cells in the genital mucosal following therapeutic immunization will need to be evaluated. Although prior vaccines that have been tested in human clinical trials have almost exclusively targeted glycoproteins, the HSV vaccine pipeline is rich with novel platforms that have shown efficacy in animal models (Table 1). The challenge will be quickly moving these candidate vaccines into human clinical trials. There has been concern about safety of replication-competent vaccines due to possibility of recombination with clinical strains or the establishment of latency.

two-dose boys = $256 million over 70 years of vaccination in a po

two-dose boys = $256 million over 70 years of vaccination in a population of 10 million, results not shown). Compared to no vaccination, all two- and three-dose girls-only and girls & boys HPV vaccination strategies investigated produce cost-effectiveness ratios below the $40,000/QALY-gained cost-effectiveness threshold ( Fig. 2, and see Supplementary Table 3 for detailed results). In the base-case, two-dose girls-only vaccination (vs. no vaccination) consistently produces the lowest incremental cost-effectiveness ratio with cost/QALY-gained varying between $7900 [IQR: 7000;9700] and $10,400 [IQR: 8800;13,400] ( Fig. 2b–f). The only

exception is when two-dose duration of protection is assumed to be 10 years ( Fig. 2a). In the sensitivity analysis, two-dose girls-only vaccination Selleck Autophagy Compound Library cost-effectiveness ratios remained below $40,000/QALY-gained ( Fig. 3a). The maximum cost per dose for two-dose girls-only vaccination to remain cost-effective (vs. no vaccination) is predicted to be $128, $218 and $252 assuming two-dose vaccine protection lasts 10, 20 and 30 years, PD0332991 nmr respectively (see Supplementary Fig. 4 and Table 4). The incremental cost-effectiveness ratio of

giving the third dose of vaccine to girls (i.e., of three-dose girls-only vs. two-dose girls-only) is estimated to be below $40,000/QALY-gained if: (i) three doses provide longer protection than two doses (i.e., more than 5 years), and ii) two-dose protection is less than 30 years ( Fig. 2 and Fig. 3). Under most scenarios, two-dose girls & boys vaccination (vs. two-dose girls-only) provides fewer or similar QALYs-gained and is more expensive than three-dose girls-only vaccination (i.e., is dominated; Fig. 2 and Fig. 3). The mafosfamide only exceptions are: (i) if the third dose provides little or no additional protection to two doses, (ii) when extreme scenarios for burden of HPV-disease among MSM are assumed (e.g., 7% males are MSM, the relative risk of disease among

MSM vs. male heterosexuals is 17, and girls-only vaccination is assumed to have no effect on HPV-related disease incidence in MSM) or (iii) when vaccine cost for boys is 10–40% of the cost for girls ( Fig. 3b, Supplementary Fig. 3). Finally, the incremental cost-effectiveness ratio of three-dose girls & boys vaccination (vs. three-dose girls-only) is greater than $100,000/QALY-gained under all base-case scenarios and most scenarios investigated in sensitivity analysis ( Fig. 2 and Fig. 3). In the sensitivity analysis, three-dose girls & boys vaccination is estimated to be less than $40,000/QALY-gained if the cost per dose for girls and boys is substantially reduced (Supplementary Fig. 4c).

They used the Assessment of Quality of Life questionnaire, which

They used the Assessment of Quality of Life questionnaire, which ranges from 0 (death) to 1 (full health). The two exercise groups did not differ significantly (mean between-group difference 0.05 points in favour of supervised exercise, 95% CI −0.15 to 0.25). This study pooled data from five eligible papers to conclude that post-discharge physiotherapy does provide better patient outcomes after total hip replacement, in

terms of strength of hip abductor muscles of the operated leg, gait speed, and cadence. Outpatient supervised rehabilitation provided no better results than unsupervised home exercise programs for most outcome measures, with the exception of the Timed Up and Go test, which was faster in the physiotherapist-supervised group. The studies included in our review found similar results

to other published studies in this area. A non-randomised, controlled selleck trial (Sashika et al 1996) showed that a six-week selleck compound home program including hip range of motion exercises, isometric exercises, and eccentric strengthening increased strength of hip abductors, walking speed, and cadence. Unlu et al (2007) evaluated a six-week program including the same exercises as Sashika et al (1996), though with two comparison groups: one home based and one supervised by a physiotherapist. Both treatment groups showed an improvement in isometric hip abductor torque, gait speed, and cadence. Di Monaco et al (2009) performed a systematic review of controlled trials of physical exercise programs after total hip replacement, which also supported the usefulness of rehabilitation from late phase (> 8wks post-operative). This review included some of the studies in our review (Jan et al 2004, Trudelle-Jackson and Smith 2004, and Unlu et al 2007), 17-DMAG (Alvespimycin) HCl and concluded that for these programs to be effective they should comprise weight bearing exercises with hip abductor eccentric strengthening. In our systematic

review, functional outcomes were measured using a wide range of tools. As a consequence meta-analysis of these data was not possible. The review by Minns Lowe (2009) was also unable to meta-analyse these data and concluded it was not possible to determine whether post-discharge physiotherapy is effective due to insufficient evidence. In the absence of meta-analysis, it is worth considering some details of the trials that demonstrated good outcomes in a range of diverse measures, such as the Timed Up and Go test and self-perceived function. Jan et al (2004) showed that a 12-week home exercise program performed for 60 min daily increased bilateral hip muscle strength, walking speed, and functional score (Harris Hip Score). These improvements were significant in a highly compliant patient group (practice ratio > 50%) and patients from a low-compliance group compared to the controls.

GABA-T activity assay was expressed as the concentration that cau

GABA-T activity assay was expressed as the concentration that caused 50% inhibition of the GABA-T (γ-aminobutyric acid transaminase) is the major inhibitory neurotransmitter in the mice brain (IC50). The GABA-T assay was used to examine the newly synthesized compounds. Vigabatrin was used as a standard anticonvulsant drug for comparison. The results of our preliminary screening indicated that compounds 5d and 5h showed strong where as compound 5l showed moderate activity. The other compounds 5a–c showed weak activity (Table 1). The compounds 5e–g, 5i–k, 5m, 5n far active compare to standard Vigabatrin. Subsequently, we may conclude the following structure activity relationship’s (SAR’s).

(i) The presence of basic skeleton (maleic imide moiety) is necessary for the development of active anticonvulsant derivates. (ii) Introducing napthol PI3K Inhibitor Library cell line to form Michael adduct with maleic imide moiety MK-8776 purchase GABA-T activity was observed far from standard drug (compound 5a–b). (iii) Introducing one bromine atom (electron withdrawing group) in position 4 of phenol to form Michael adduct with maleic imide moiety potent GABA-T activity with IC50 (100.5 ± 5.2 μM) as compare to standard drug (compound 5d) While instead of bromine atom if we replace position 4 of phenol by chlorine to

form Michael adduct with maleic imide moiety GABA-T activity was drop down from strong to weak. (iv) According to the above findings the presence of one −NO2 electron withdrawing group in position 4 of phenol showed very weak activity toward GABA-T (compound 5e) while increases number of –NO2 electron withdrawing group in position 2, 4, 6 of phenol little beat increased Methisazone GABA-T activity (compound 5l). (v) In compound

5m the presence of phenyl group as well as Introducing heterocyclic phenol (N-methyl-4-hydroxy Quinoline, 5n) in position 4 of phenol to form Michael adduct with maleic imide moiety decreased activity toward GABA-T. (vi) In compounds 5i–d, −CH3 group at respective o, m and p- position of phenol showed weakly active toward GABA-T. Incorporation of weak electron donating group irrespective of their position on phenol was not effect cytotoxically on GABA-T. (vii) In compound 5h, –CHO group at O-position of phenol to form maleimide have potent activity as IC50 (160.4 ± 6.2 μM) toward GABA-T than standard drug IC50 (250 ± 6.4 μM). We have demonstrated that the reaction of 1-(4-acetylphenyl)-pyrrole-2,5-diones with phenols via Michael addition reaction resulting 1-(4-acetylphenyl)-3-aryloxypyrrolidine-2,5-diones, enables the efficient synthesis of Michael adduct in single step in satisfactory overall yields. The products of this reaction are of potential medicinal interest. Moreover, we have shown from the biological evaluation part that 5d and 5h derivatives have excellent inhibition against GABA-T with respect to standard Vigabatrin. All authors have none to declare. The authors thank the S. V.

n ) administration of mice with c-di-GMP induces recruitment of m

n.) administration of mice with c-di-GMP induces recruitment of monocytes and granulocytes [20] and activates the host immune response [21] and [22]. In one study, the lungs and draining lymph nodes from mice intranasally Gefitinib treated either with c-di-GMP

or phosphate buffered saline (PBS) were examined 24 or 48 h after treatment for differences in cell number or composition. Results showed that the draining lymph nodes of c-di-GMP-treated mice had significantly higher total cell numbers as well as higher percentages of CD44low cells and CD86 positive cells. In the lung, however, the picture was less clear with no difference in total numbers of monocytes or neutrophils or pulmonary DCs as determined by flow cytometry [21]. However, there was some indication that c-di-GMP did affect lung parenchymal cells in that the lungs from c-di-GMP-treated mice had a larger proportion of alveolar macrophages which were newly recruited (CD11chiMHCIIlowCD11b+). Also, DCs (CD11chiMHCIIhi), although not significantly increased in number, expressed higher levels of CD40 and CD86 than PBS-treated control mice [21]. Work from our own laboratories has indicated that 24 h after a single i.n. administration of c-di-GMP, there is a significant increase in the number of pulmonary DCs with higher expression

of CD40 and CD80 but not CD86 or MHCII [23]. The treatment also induced a rapid but transient recruitment of neutrophils and other inflammatory new cells into the bronchoalveolar space [23] and increased levels of selleck compound proinflammatory cytokines and chemokines IL-12p40, IL-1β, IL-6, keratinocyte derived chemokine (KC), MCP-1, macrophage inflammatory protein (MIP)-1β, RANTES and tumor necrosis factor (TNF)-α in a dose-dependent manner [22]. A number of recent studies have shown that the innate immune response elicited by c-di-GMP is a potent immunomodulator for the treatment of bacterial infections. In this regard, studies of the effect of c-di-GMP on the course of bacterial infection have clearly shown a striking protective

effect of c-di-GMP administration against a number of serious bacterial infections. Using a mouse model of mastitis, Karaolis and co-workers showed that, despite no direct bactericidal activity, c-di-GMP co-administered with S. aureus directly into the mammary glands significantly decreases bacterial burdens [24]. Previous work by the same group had shown that c-di-GMP inhibits biofilm formation of the same S. aureus strain as well as its adherence to HeLa cells [25]. To rule out the possibility that the c-di-GMP-mediated protection is solely due to its role in the inhibition of biofilm formation, subsequent work showed that pretreatment with c-di-GMP 12 and 6 h before intramammary infection with S. aureus also results in a 1.5 log and a 3.

They were ready to transfer of this knowledge to the outside worl

They were ready to transfer of this knowledge to the outside world only on the basis of substantial payment. Recent trend shows a decline in the number of traditional herbal healers in the tribal areas since the younger generation is not interested to continue this tradition. Hence, there is an urgent need to record and preserve all information on plants used

by different tribal communities for various purposes before it is completely lost. Tribal herbal healers should also be encouraged by some means so that their knowledge is FRAX597 sustained for future generations. In Kodagu district, the tribal populations living far away from urban area still rely on traditional herbal medicine for their primary health care needs. The unfortunate part is that due to forest fire and forest cutting for coffee and cardamom plantations, ginger cultivation, etc. many species are facing threat of extinction. There is immediate need for their conservation. Ibrutinib manufacturer And also there is need for phytochemical analysis and pharmacological investigations of these important disappearing plants to strengthen the documentation of ethnic drugs. It would help in developing novel drug(s) to treat chronic diseases. All authors have none to declare. The authors are grateful to Dr. Halesh for help in the identification of some plants. Thanks are also due to the tribal people of Kodagu district, especially

Raju, Dobi, Thamma and Era who have provided the valuable information and co-operated during field work. “
“Prolonged antibiotic treatment is the main cause of fungal infections, especially candidiasis. Candida

albicans, causative agent of candidiasis is a yeast and one of the constituents of regular flora of the skin, gastro-intestinal tract, mouth, rectum and vagina. Although Candida is an endosymbiont of the human body, it can cause problems if there is an overgrowth, resulting in candidiasis. 1 Candidiasis usually occurs when there is an imbalance in the regular flora of the body, and in people who have compromised immune systems. Different factors can lead to Candidal overgrowth such as a person’s diet, immune suppression and prolonged antibiotic treatment, however, CYTH4 research findings support that the prolonged use of antibiotics can also play a major role in the development of candidiasis. Prolonged dose of antibiotics can lead to an imbalance in the essential gut flora, an imbalance that wipe out beneficial microflora and allows harmful bacteria, yeasts and parasites to overgrow in the stomach.2, 3, 4 and 5 Steroids and some cancer medications also weaken the immune system and can allow yeast to flourish. If this condition is not treated and controlled, the affected person can begin to suffer a slew of negative side effects such as oropharyngeal candidiasis, Intertrigo, Candida vulvovaginitis (Vaginitis), Systemic yeast infections (IDSA).

The availability of a fast and automated analytics platform will

The availability of a fast and automated analytics platform will expand the scope, robustness, and evolution of Design of Experiment (DOE) studies. It is envisaged

that this will lead to expanded use of Quality by Design (QbD) approaches PI3K Inhibitor Library supplier in vaccine process development. Currently, the development of purification processes for vaccine polysaccharides is exceedingly complex, time-consuming, and laborious. HTPD of polysaccharides has lagged significantly behind current developmental archetypes for other biologicals such as monoclonal antibodies. The lack of simple, high throughput analytical tools has played a role in hindering the evolution of HTPD for polysaccharides. Purification process development does not require the exquisite accuracy demanded of release

assays. Instead, speed, simplicity, and precision are paramount. Especially in the context of high throughput process development, the desire to find the best conditions on a microplate, relative to the other wells, is critical. Excluding affinity separations, the maximum purification factor that can be achieved in a single-stage equilibrium experiment is typically 2 logs, obviating the need for extremely sensitive analytics. Accuracy is more important in the subsequent scale-up and demonstration of promising purification conditions. Polysaccharides, endotoxin, proteins, and nucleic acids are the MLN8237 major components found in harvested bacterial fermentation broths employed in industrial polysaccharide vaccine manufacturing. Their critical importance is underscored by the old inclusion of the respective assays in the batch release package for product characterization. In the current work, analytical techniques for quantifying polysaccharides, endotoxin, and proteins were qualified. In selecting methods, emphasis was given to procedural simplicity, amenability to automation, robustness,

and precision over accuracy. In addition, the qualification process included evaluating the impact of impurities commonly encountered alongside the carbohydrate product as well as a diverse library of polysaccharides. Novel procedures were described to simplify methods and facilitate automation. A phenol sulphuric acid assay was optimized for high throughput quantitation of mono-, di-, and poly-saccharides. The assay requires only 25 μL of sample and involves no heating steps that can stymie automation. The described procedure also reduces the quantities of hazardous chemicals such as phenol and sulphuric acid, requiring only 150 μL total per sample. A linear range of approximately 2 logs (e.g. glucose: 8–1000 μg/mL) was observed for every tested carbohydrate, with the actual range derived from the specific composition of reactive sugars present. The precision of the described assay was found to be 10%. The PyroGene™ assay was simplified to a single measurement while removing a heated incubation step.