A Vaccine Based on the A/ASIA/G-VII Lineage of Foot-and-Mouth Disease Virus Offers Low Levels of Protection against Circulating Viruses from the A/ASIA/Iran-05 lineage

The recent emergence and circulation of the A/ASIA/G-VII (A/G-VII) lineage of foot-and-mouth disease virus (FMDV) in the Middle East has resulted in the development of homologous vaccines to ensure susceptible animals are sufficiently protected against clinical disease. However, a second serotype A lineage called A/ASIA/Iran-05 (A/IRN/05) continues to circulate in the region and it is therefore imperative to ensure vaccine strains used will protect against both lineages. In addition, for FMDV vaccine banks that usually hold a limited number of strains, it is necessary to include strains with a broad antigenic coverage. To assess the cross protective ability of an A/G-VII emergency vaccine (formulated at 43 (95% CI 8–230) PD50/dose as determined during homologous challenge), we performed a heterologous potency test according to the European Pharmacopoeia design using a field isolate from the A/IRN/05 lineage as the challenge virus. The estimated heterologous potency in this study was 2.0 (95% CI 0.4–6.0) PD50/dose, which is below the minimum potency recommended by the World Organisation for Animal Health (OIE). Furthermore, the cross-reactive antibody titres against the heterologous challenge virus were poor (≤log10 0.9), even in those cattle that had received the full dose of vaccine. The geometric mean r1-value was 0.2 (95% CI 0.03–0.8), similar to the potency ratio of 0.04 (95% CI 0.004–0.3). Vaccination decreased viraemia and virus excretion compared to the unvaccinated controls. Our results indicate that this A/G-VII vaccine does not provide sufficient protection against viruses belonging to the A/IRN/05 lineage and therefore the A/G-VII vaccine strain cannot replace the A/IRN/05 vaccine strain but could be considered an additional strain for use in vaccines and antigen banks.

Anti-bacterial activity of Rosela Flower Extract (Hibiscus sabdariffa L.) against Extended-Spectrum Beta-Lactamase (ESBL) Eschericia coli

Roselle (Hibiscus sabdariffa L.) contains cyanidin-3-rutinoside, delphinidin, delphinidin-3-monoglucoside, cyanidin-3-monoglucoside, cyanidin-3-sambubioside, cyanidin-3,5-diglucoside may inhibit the growth of Extended-Spectrum-Beta-Lactamase (ESBL) Escherichia coli, but there is no research reported the determination of MIC of Roselle (Hibiscus sabdariffa L.) extract powder against ESBL E.coli ATCC 6110 and ATCC 5949 and their potency ratio compared to meropenem. This study aimed to determine the MIC of Roselle flower extract powder on the growth of ESBL E. coli ATCC 6110 and ATCC 5949 and determine the potency ratio compared to meropenem. This study used two methods in determining MIC, namely the agar diffusion method and the dilution method with Nutrient Agar media which was incubated at 37 ±1°C for 24 hours, while the determination of the potency ratio was carried out by diffusion method with the same media which was incubated at 37 ±1°C for 24 hours. The results obtained were diameter of inhibition zone (mm) which were then observed and analyzed to calculate the potency ratio. The results showed that the MIC of Roselle flower extract powder was obtained at a concentration of 12,500 ppm by diffusion method and at a concentration of 3,125 ppm by dilution method with 24 hours incubation at 37 ± 1o C and the potency ratio of Roselle flower extract powder compared to meropenem was 89.7% and 97.97% against ESBL E.coli ATCC 6110 and ESBL E.coli ATCC 5949 respectively.

Benefit and Risk Evaluation of Biased μ-Receptor Agonist Oliceridine versus Morphine

Background To improve understanding of the respiratory behavior of oliceridine, a μ-opioid receptor agonist that selectively engages the G-protein–coupled signaling pathway with reduced activation of the β-arrestin pathway, the authors compared its utility function with that of morphine. It was hypothesized that at equianalgesia, oliceridine will produce less respiratory depression than morphine and that this is reflected in a superior utility. Methods Data from a previous trial that compared the respiratory and analgesic effects of oliceridine and morphine in healthy male volunteers (n = 30) were reanalyzed. A population pharmacokinetic–pharmacodynamic analysis was performed and served as basis for construction of utility functions, which are objective functions of probability of analgesia, P(analgesia), and probability of respiratory depression, P(respiratory depression). The utility function = P(analgesia ≥ 0.5) – P(respiratory depression ≥ 0.25), where analgesia ≥ 0.5 is the increase in hand withdrawal latency in the cold pressor test by at least 50%, and respiratory depression ≥ 0.25 is the decrease of the hypercapnic ventilatory response by at least 25%. Values are median ± standard error of the estimate. Results The two drugs were equianalgesic with similar potency values (oliceridine: 27.9 ± 4.9 ng/ml; morphine 34.3 ± 9.7 ng/ml; potency ratio, 0.81; 95% CI, 0.39 to 1.56). A 50% reduction of the hypercapnic ventilatory response by morphine occurred at an effect-site concentration of 33.7 ± 4.8 ng/ml, while a 25% reduction by oliceridine occurred at 27.4 ± 3.5 ng/ml (potency ratio, 2.48; 95% CI, 1.65 to 3.72; P < 0.01). Over the clinically relevant concentration range of 0 to 35 ng/ml, the oliceridine utility function was positive, indicating that the probability of analgesia exceeds the probability of respiratory depression. In contrast, the morphine function was negative, indicative of a greater probability of respiratory depression than analgesia. Conclusions These data indicate a favorable oliceridine safety profile over morphine when considering analgesia and respiratory depression over the clinical concentration range. Editor’s Perspective What We Already Know about This Topic What This Article Tells Us That Is New

Validated TLC-Contact Bioautography Method for Identification of Kanamycin Sulfate in Injection Preparation

Background: TLC-contact bioautography is one of an effective method for identification antibiotics, by which many antibiotics could be identification and determination simultaneously. Objective: To evaluate kanamycin sulfate in injection preparations based on its inhibitory activity against Escherichia coli ATCC 8739 as test organism. Methods: Sample and standard solutions were spotted onto TLC silica gel 60 F254 plate and developed in 10% potassium dihydrogen phosphate solutionas as mobile phase. The TLC-contact bioautography method was validated according to USP guidelines by considering specificity, LOD, LOQ, linearity, accuracy and precision parameters. Results: The TLC-contact bioautography method was found to be high sensitivity with LOD of 0.75 µg and LOQ 2.31 µg. Linearity range of 100-350 µg/mL with r = 0.9993 and linear regression equation was y = 0.0019x + 0.0338. The recovery obtained from addition of blank samples by three different concentrations of kanamycin sulfate standard was 101.40% ¬+ 2.02%. The precision of the method was good with coefficient of variation 0.080%. The TLC-contact bioautography method was supported by determination of kanamycin sulfate potency ratio in the injection preparation and kanamycin sulfate standard using 3-3 design. Random block design obtained the potential for kanamycin sulfate in injection preparations compared to kanamycin sulfate standard was 100.6%. Conclusion: The TLC-contact bioautography for kanamycin sulfate in injection preparations could be applied to the quality control analysis of the investigated drugs.

The Intriguing Effects of Substituents in the N-Phenethyl Moiety of Norhydromorphone: A Bifunctional Opioid from a Set of “Tail Wags Dog” Experiments

(−)-N-Phenethyl analogs of optically pure N-norhydromorphone were synthesized and pharmacologically evaluated in several in vitro assays (opioid receptor binding, stimulation of [35S]GTPγS binding, forskolin-induced cAMP accumulation assay, and MOR-mediated β-arrestin recruitment assays). “Body” and “tail” interactions with opioid receptors (a subset of Portoghese’s message-address theory) were used for molecular modeling and simulations, where the “address” can be considered the “body” of the hydromorphone molecule and the “message” delivered by the substituent (tail) on the aromatic ring of the N-phenethyl moiety. One compound, N-p-chloro-phenethynorhydromorphone ((7aR,12bS)-3-(4-chlorophenethyl)-9-hydroxy-2,3,4,4a,5,6-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-7(7aH)-one, 2i), was found to have nanomolar binding affinity at MOR and DOR. It was a potent partial agonist at MOR and a full potent agonist at DOR with a δ/μ potency ratio of 1.2 in the ([35S]GTPγS) assay. Bifunctional opioids that interact with MOR and DOR, the latter as agonists or antagonists, have been reported to have fewer side-effects than MOR agonists. The p-chlorophenethyl compound 2i was evaluated for its effect on respiration in both mice and squirrel monkeys. Compound 2i did not depress respiration (using normal air) in mice or squirrel monkeys. However, under conditions of hypercapnia (using air mixed with 5% CO2), respiration was depressed in squirrel monkeys.

Intravenous oxycodone compared to morphine for post-operative opioid-related adverse events in opioid naïve patients: a prospective randomized controlled trial

Bacground: Intravenous oxycodone compared to morphine for postoperative pain relief is controversial. The purpose of this study was to assess opioid-related adverse events of oxycodone versus morphine in opioid-naive patients after orthopaedic surgery. Methods: Patients scheduled for total hip arthroplasty under general anesthesia combined with a multimodal analgesia (acetamnophen, nonsteroidal anti-inflammatory) were randomized in a triple-blinded trial to postopertaive pain treatment with either intravenous oxycodone or morphine (potency ratio 1:1). After surgery, patients received similar drug regimen for titration in the postoperative care unit (bolus 2-3 mg, 5 min period, when pain score was >3/10) followed by an intravenous patient controlled analgesia (bolus 1 mg, lockout time 7 min) postoperatively. The primary outcome was number of patients with ≥1 opioid-related adverse events within the first 24 hours (at least one of the following complications: nausea, vomiting, respiratory depression, pruritus, urinary retention requiring evacuation, allergy, hallucination). Secondary outcomes included pain scores, opioid consumption. Patients were followed up to 4 months. Results: The intention-to-treat analysis included 241 patients with similar characteristics. There were 55 patients with at least one opioid-related adverse events in oxycodone group versus 46 in morphine group (48% vs 40%, p=0.19; relative risk= 1.22 [0.91; 1.63]). Oxycodone versus morphine requirements were respectively: 6 [0-11] versus 8 [0-12] mg (p=0.06) for titration, 15 [8-26] versus 8 [5-16] mg (p=0.001) for PCA dose, and 22 [12-37] mg versus 19 [11-28] mg for the titration and PCA accumulated consumption (p=0.048). During the first 24 hours, there were no other differences in secondary outcomes between both drugs for (respectively oxycodone versus morphine in %): nausea (15 versus 13), vomiting (5 versus 5 ), urinary retention (20 versus 12 ) and pain scores. Conclusion: This study demonstrated that oxycodone required lower doses for titration in postoperative care unit, but did not significantly reduce opioid-related adverse events within the first 24 hours compared to morphine.

In vitro equivalence of generic and branded amoxicillin tablet by microbiological assay method

BackgroundIndonesian Ministry of Health advocate doctors, especially in government-owned healthcare facility, to prescribe generic drugs including amoxicillin. Although BPOM (the National Agency of Drug and Food Control) already guarantees that the generic amoxicillin and the branded one were interchangeable, lack of confidence in generic drugs still remains among patients, pharmacists, and doctors. This issue supported by lack of publication confirmed the therapeutic equivalence of branded and generic drugs. This study aims to evaluate and compare the in vitro microbiological assay of different generic and branded amoxicillin that are available in Indonesian market, especially those used in government-owned healthcare facilities.MethodsMicrobiological assays for five samples of amoxicillin tablet containing 500 mg amoxicillin available in Indonesia were determined using a method from Indonesia Pharmacopeia. Samples were coded as Products A to E. The assay was carried out by measuring the diameter of the inhibition zones in the plate agar incubated with Escherichia coli and Staphylococcus aureus. The obtained data were evaluated to determine the sample potency and compared with the amoxicillin reference standard.ResultsMinor and insignificant differences (p > 0.05) were found in the diameters of the inhibition zones. Potency ratio measured both in E. coli and S. aureus were all between 95% and 105%. The lowest of the tested samples were from Product C, which resulted to ratio potencies of 96.3% and 95.5% in E. coli and S. aureus, respectively.ConclusionsAll five samples were in the range of the acceptance criteria. Therefore, from the view of the microbiological assay, these products are in equivalence in quality and are interchangeable.

Minimal Local Analgesic Dose of Intrathecal Bupivacaine and Ropivacaine in Patients Undergoing Cesarean Section: A Comparative Study

Introduction: Spinal anesthesia is a reasonable option for cesarean section. Bupivacaine and ropivacaine have been used as intrathecal drugs alone or in combination with various opiods. Ropivacaine is considered a valid and safe alternative to bupivacaine for intrathecal anesthesia. This study aims to determine the median effective dose (ED50) of intrathecal bupivacaine and ropivacaine for cesarean section and defines this as the minimum local anesthetic dose (MLAD). Methods: Forty pregnant women undergoing elective cesarean section were allocated and randomized into two groups. The initial dose was 13mg for both ropivacaine and bupivacaine groups and was increased or decreased of 0.3mg, using the up-down sequential allocation technique. Efficacy was accepted if adequate sensory dermatomal anesthesia to pinprick to T6 was attained within 20 minutes after intrathecal injection and required no supplemental epidural injection for procedure until at least 50 minutes after the intrathecal injection of test drugs. The MLAD for both bupivacaine and ropivacaine was calculated with 95% confidence interval using the formula of Dixon and Massey. Comparison of different variables between the groups was done using t-test with significant p value at < 0.05. Results: The two groups were comparable in terms of demographic profile and clinical characteristics. The MLAD of ropivacaine and bupivacaine were 11.63 mg (95% CI, 11.5-12.92) and 10.459 mg (95% CI, 10.12-10.87) respectively. The potency ratio between spinal ropivacaine and bupivacaine was 0.89. Conclusion: Ropivacaine provided clinically surgical anaesthesia of shorter duration without compromising neonatal outcome and can be used as a safe alternative to bupivacaine.

Synthesis and biological evaluation of some novel 1-substituted fentanyl analogs in Swiss albino mice

ABSTRACT Fentanyl [N-(1-phenethyl-4-piperidinyl)propionanilide] is a potent opioid analgesic agent, but a has narrow therapeutic index. We reported earlier on the synthesis and bioefficacy of fentanyl and its 1-substituted analogs (1-4) in mice. Here we report the synthesis and biological evaluation of four additional analogs, viz. N-isopropyl-3-(4-(N-phenylpropionamido)piperidin-1-yl)propanamide (5), N-tbutyl- 3-(4-(N-phenylpropionamido)piperidin-1-yl)propanamide (6), isopropyl 2-[4-(N-phenylpropionamido)piperidin-1-yl]propionate (7) and t-butyl 2-[4-(N-phenylpropionamido)piperidin-1-yl]propionate (8). The median lethal dose (LD50) determined by intravenous, intraperitoneal and oral routes suggests these analogs to be comparatively less toxic than fentanyl. On the basis of observational assessment on spontaneous activities of the central, peripheral, and autonomic nervous systems, all the analogs were found to be similar to fentanyl. Naloxone hydrochloride abolished the neurotoxic effects of these analogs, thereby ascertaining their opioid receptor-mediated effects. All the analogs displayed significant analgesic effects, measured by formalin-induced hind paw licking and tail immersion tests at their respective median effective dose (ED50). They also exhibited 8-12 fold increase in therapeutic index over fentanyl. However, 5 and 6 alone produced lower ED50 (20.5 and 21.0 μg/kg, respectively) and higher potency ratio (1.37 and 1.33, respectively) compared to fentanyl. They could thus be considered for further studies on pain management