Miracle-dz/UniFAR-FTUs huggingface dataset:
CYP267A1 and CYP267B1 from Sorangium cellulosum So ce56 are Highly Versatile Drug Metabolizers

The guidelines of the Food and Drug Administration and International Conference on Harmonization have highlighted the importance of drug metabolites in clinical trials. As a result, an authentic source for their production is of great interest, both for their potential application as analytical standards and for required toxicological testing. Since we have previously shown promising biotechnological potential of cytochromes P450 from the soil bacterium Sorangium cellulosum So ce56, herein we investigated the CYP267 family and its application for the conversion of commercially available drugs including nonsteroidal anti-inflammatory, antitumor, and antihypotensive drugs. The CYP267 family, especially CYP267B1, revealed the interesting ability to convert a broad range of substrates. We established substrate-dependent extraction protocols and also optimized the reaction conditions for the in vitro experiments and Escherichia coli–based whole-cell bioconversions. We were able to detect activity of CYP267A1 toward seven out of 22 drugs and the ability of CYP267B1 to convert 14 out of 22 drugs. Moderate to high conversions (up to 85% yield) were observed in our established whole-cell system using CYP267B1 and expressing the autologous redox partners, ferredoxin 8 and ferredoxin-NADP+ reductase B. With our existing setup, we present a system capable of producing reasonable quantities of the human drug metabolites 4′-hydroxydiclofenac, 2-hydroxyibuprofen, and omeprazole sulfone. Due to the great potential of converting a broad range of substrates, wild-type CYP267B1 offers a wide scope for the screening of further substrates, which will draw further attention to future biotechnological usage of CYP267B1 from S. cellulosum So ce56.
In Vitro Screening of Cytotoxic, Antimicrobial and Antioxidant Activities of Clinacanthus nutans (Acanthaceae) leaf extracts

Purpose: To evaluate the in vitro cytotoxic, antioxidant and antimicrobial activities of Clinacanthus nutans extracts and semi-fractions. Method: The plant was subjected to cold solvent extraction to produce petroleum ether, ethyl acetate and methanol crude extracts, followed by isolation using bioassay-guided fractionation. The crude extracts (0.2 to 10.0 mg/ml) were tested against HeLa and K-562 cell lines using 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and antioxidant activity using 1, 1-diphenyl-2picrylhydrazyl (DPPH) assay. Furthermore, fractions isolated from ethyl acetate leaf extract (0.02 to 10.0 mg/ml) were tested against Bacillus cereus (ATCC11778), Escherichia coli (ATCC25922), Salmonella enterica Typhimurium (ATCC14028) and Candida albicans (ATCC10231) using minimum inhibitory concentration (MIC) and minimum bactericidal or fungicidal (MBC/MFC) assays. Results: Petroleum ether extracts demonstrated the strongest cytotoxic activity against HeLa and K562 cells with IC50 of 18.0 and 20.0 µg/mL, respectively. Petroleum ether extracts also displayed the highest radical scavenging activity of 82.00 ± 0.02 %, compared with ascorbic acid and α-tocopherol corresponding values of 88.7 ± 0.0 and 86.6 ± 0.0 %, respectively. In MIC assay, all the crude extracts and fractions showed inhibition against all tested microorganisms. Fraction 7 displayed the lowest MIC and MBC/MFC values against B. cereus and C. albicans at 1.39 mg/ml, respectively. Ampicillin and amphotericin B displayed MIC value of 1.3 mg/ml. Conclusion: The bioactive compounds from C. nutans are potential cytotoxic, antimicrobial and antioxidant agents.
Assessing the impact of abuse-deterrent opioids (ADOs): identifying epidemiologic factors related to new entrants with low population exposure

ABSTRACT Objective: Prescription opioid misuse and abuse is a serious public health concern that can lead to overdose, addiction, and death. The development of opioid formulations designed to deter misuse and abuse is considered an important step towards addressing the problem, but the extent to which abuse-deterrent opioids (ADOs) actually deter abuse in the community requires confirmation through epidemiologic studies. Epidemiologic evaluations of misuse and abuse present several unique challenges related to study design, sources of data, and methodology, particularly for new entrant ADOs with low population exposure. The purpose of this article is to review the critical methodologic issues that must be considered when designing an epidemiologic evaluation of prescription opioid misuse and abuse, and deterrence for new entrant ADOs. Methods: A systematic feasibility assessment was conducted by critically evaluating and applying epidemiologic principles to combinations of epidemiology study design, study directionality, population, data source, clinical and patient-reported endpoints, type of comparator, effect size, and the ability to control for confounding variables. Results: Ten epidemiologic factors were identified that are considered key to an epidemiologic evaluation, and the issues associated with each are summarized. Conclusion: Given the low population exposure of new entrant ADOs, there is limited feasibility in conducting the epidemiologic studies necessary to evaluate the effectiveness of these products in deterring abuse. Clear regulatory guidance is needed.
Effect of Desensitizing Agents on the Bond Strength of Mild and Strong Self-etching Adhesives.

BACKGROUND Desensitizing agents are used, almost as routine practice, in many adhesive restorative procedures. There is still debate as to their effect in dentin bonding, particularly with self-etching adhesives. The present study aimed to evaluate the effect of different desensitizing agents on the bond strength of mild and strong self-etching adhesive systems to dentin. MATERIALS AND METHODS One hundred twenty recently extracted, noncarious human molars were used to obtain superficial dentin substrate for bonding. No desensitizer was used in the control groups. The experimental groups were pretreated with Gluma Desensitizer, MicroPrime B, and Dentin Desensitizer immediately prior to bonding with self-etching adhesives Optibond XTR, Xeno IV, and iBond. A bonding jig was used to fabricate composite cylinders, which were stored for either 24 hours or three months, after which the shear bond strength (SBS) was evaluated using a notched-edge testing device at a crosshead speed of 1 mm/min. Failure mode distribution was also evaluated at 24 hours and three months. A two-way analysis of variance, Tukey test, and Student t-test, with a significance level of p<0.05, were used for data analysis. RESULTS At 24 hours, there was no significant difference in SBS when the same adhesive was used with any of the experimental desensitizing agents compared with the control group without desensitizer. However, at three months, Dentin Desensitizer bonded with Optibond XTR demonstrated significantly lower SBS (p<0.001), while Gluma bonded with iBond showed significantly higher SBS values (p=0.034) relative to their corresponding control group. Only MicroPrime B bonded with Xeno IV and iBond with no desensitizer demonstrated a significant reduction in SBS after three months (p=0.034 and p=0.002, respectively). The most prevalent type of failure was adhesive. CONCLUSION Desensitizing agents can be used in combination with self-etching adhesives to control hypersensitivity without adversely affecting their bond strength to dentin.
Immunoreactivity for the group III metabotropic glutamate receptor subtype mGluR4a in the superficial laminae of the rat spinal dorsal horn

Studies indicate that metabotropic glutamate receptors (mGluRs) may play a role in spinal sensory transmission. We examined the cellular and subcellular distribution of the mGluR subtype 4a in spinal tissue by means of a specific antiserum and immunocytochemical techniques for light and electron microscopy. A dense plexus of mGluR4a‐immunoreactive elements was seen in the dorsal horn, with an apparent accumulation in lamina II. The immunostaining was composed of sparse immunoreactive fibres and punctate elements. No perikaryal staining was seen. Immunostaining for mGluR4a was detected in small to medium‐sized cells but not in large cells in dorsal root ganglia. At the electron microscopic level, superficial dorsal horn laminae demonstrated numerous immunoreactive vesicle‐containing profiles. Labelling was present in the cytoplasmic matrix, but accretion of immunoreaction product to presynaptic specialisations was commonly observed. Axolemmal labelling was confirmed by using a preembedding immunogold technique, which revealed distinctive deposits of gold immunoparticles along presynaptic thickenings with an average centre‐to‐centre distance of 41 nm (41.145 ± 13.59). Immunoreactive terminals often formed synaptic contacts with dendritic profiles immunonegative for mGluR4a. Immunonegative dendritic profiles were observed in apposition to both mGluR4a‐immunoreactive and immunonegative terminals. Diffuse immunoperoxidase reaction product was also detected in dendritic profiles, some of which were contacted by mGluR4a‐immunoreactive endings, but only occassionally were they observed to accumulate immunoreaction product along the postsynaptic density. Terminals immunoreactive for mGluR4a also formed axosomatic contacts. The present results reveal that mGluR4a subserves a complex spinal circuitry to which the primary afferent system seems to be a major contributor. J. Comp. Neurol. 430:448–457, 2001. © 2001 Wiley‐Liss, Inc.
Stromal uptake and transmission of acid is a pathway for venting cancer cell-generated acid

Significance Metabolism energizes cancer growth, but only if its end product, acid, is removed effectively. A bottleneck for acid handling is slow diffusion across the underperfused extracellular milieu of hypoxic tumors. Here, we characterize the acid-handling mechanisms operating in stromal myofibroblasts that can improve the flow of acid through tumors. We show that myofibroblasts are high-capacity reservoirs that absorb excess extracellular acidity, via the AE2 transporter, and transmit this acid load across a syncytium fused by channels, such as connexin-43. Furthermore, the cytokine TGFβ1, which orchestrates many cancer–stromal interactions, can stimulate acid uptake and transmission in stromal cells with lower baseline activities. Because many colorectal cancer cells do not express AE2 and connexin-43, acid traffic would be routed preferentially through the stromal compartment of tumors. Proliferation and invasion of cancer cells require favorable pH, yet potentially toxic quantities of acid are produced metabolically. Membrane-bound transporters extrude acid from cancer cells, but little is known about the mechanisms that handle acid once it is released into the poorly perfused extracellular space. Here, we studied acid handling by myofibroblasts (colon cancer-derived Hs675.T, intestinal InMyoFib, embryonic colon-derived CCD-112-CoN), skin fibroblasts (NHDF-Ad), and colorectal cancer (CRC) cells (HCT116, HT29) grown in monoculture or coculture. Expression of the acid-loading transporter anion exchanger 2 (AE2) (SLC4A2 product) was detected in myofibroblasts and fibroblasts, but not in CRC cells. Compared with CRC cells, Hs675.T and InMyoFib myofibroblasts had very high capacity to absorb extracellular acid. Acid uptake into CCD-112-CoN and NHDF-Ad cells was slower and comparable to levels in CRC cells, but increased alongside SLC4A2 expression under stimulation with transforming growth factor β1 (TGFβ1), a cytokine involved in cancer–stroma interplay. Myofibroblasts and fibroblasts are connected by gap junctions formed by proteins such as connexin-43, which allows the absorbed acid load to be transmitted across the stromal syncytium. To match the stimulatory effect on acid uptake, cell-to-cell coupling in NHDF-Ad and CCD-112-CoN cells was strengthened with TGFβ1. In contrast, acid transmission was absent between CRC cells, even after treatment with TGFβ1. Thus, stromal cells have the necessary molecular apparatus for assembling an acid-venting route that can improve the flow of metabolic acid through tumors. Importantly, the activities of stromal AE2 and connexin-43 do not place an energetic burden on cancer cells, allowing resources to be diverted for other activities.
Improved informed consent documents for biomedical research do not increase patients' understanding but reduce enrolment: a study in real settings.

AIMS The aim was to evaluate the comprehension of participants of an improved informed consent document (ICD). METHOD This was a randomized controlled French multicentre study performed in real conditions. Participants were adult patients undergoing screening for enrolment in biomedical research studies, who agreed to answer a validated questionnaire evaluating objective and subjective comprehension scored from 0 (no comprehension) to 100 (excellent comprehension). Patients were provided either the original ICD or an ICD modified in terms of structure and readability. The primary end point was the score of objective comprehension. The secondary end-points were the enrolment rate in the clinical study and patient characteristics associated with the score of objective comprehension. RESULTS Four hundred and eighty-one patients were included, 241 patients in the original ICD group and 240 patients in the modified ICD group. There was no difference between the two groups for the score of objective comprehension (original ICD 72.7 (95% CI 71.3, 74.1) vs. modified ICD 72.5 (95% CI 71.0, 74.0); P = 0.81). However, the rate of enrolment in the clinical study was lower in the group who received the modified ICD (64.4% (95% CI 58.3, 70.5)) than for the original ICD (73.0% (95% CI 67.4, 78.7)) (P = 0.042). Only female gender and high educational level were associated with a better objective comprehension. CONCLUSIONS Improving ICDs had no effect on participants' understanding, whereas the rate of enrolment was lower in this group. In attempts at improving potential participants' understanding of clinical research information, efforts and future trials should focus on other ways to improve comprehension.
Elastic Deformation in Orthotropic Vessels: Theoretical and Experimental Results

Excursions of the semiaxes were used to define the distortion and shape of the vessel cross section considered for conic and almost conic models. Some describing functions and mensuration expressions closely approximate, yet greatly simplify, calculations of perimeter and cross sections. Theory and experiments with models were in good agreement, but preliminary experiments showed the real case to be more complicated. Physical properties of the pulmonary trunk varied along its axis, growing stiffer toward the periphery. Furthermore, the dimension variations with transmural loading alter concepts of properties derived from measurements of a single diameter. The influence of the pericardium on the deformation of the main pulmonary artery under transmural stress was also explored. It was seen that the pericardium exerts significant restraint on the distortion and volume variations of this vessel. Over normal pressure ranges, removal of the pericardium can result in trunk volumes up to 100% greater than realized with the pericardium intact.
Analysis of geolocation accuracy by GPS: dedicated support signal integration and collaborative network in location-based services

Abstract: Currently, mobile devices, tablets and smartphones, among others, integrate geolocation systems supported by GPS or A-GPS sensors that have digital mapping functionalities, with the possibility of storage, routing and location information systems, among those, called location-based services (LBS). This technology allows the user to integrate approximate apparent position in real time between 3 and 6 meters. This approximation to the real position is valid for the LBS in applications of routing and tourism among others, but not for cadastral type surveys, urban inventories, cartographic updating or topographic support points among others. The article presents the integration of the processes required to obtain sub metric precisions in geolocation with mobile devices, in order to make accessible to all the professional use and the integration of new services.
A Study of Complications in Preterm Deliveries After Prolonged Premature Rupture of the Membranes

The risk of infection for mother and baby after spontaneous rupture of the membranes was evaluated in a prospective study of 24 patients with ruptured membranes before the 36th week of pregnancy. The mean length of pregnancy after rupture was 10 days and 2 hours. Only patients harboring Group B streptococci or E. coli in the urogenital tract were treated with antibiotics (during delivery). With the exception of 1 woman, all patients harbored one or more pathogens in the urogenital tract. Four mothers were infected but all recovered. One of 26 infants died from infection. The study did not confirm any association between prolonged rupture of the membranes and the frequency of idiopathic respiratory distress syndrome, nor did it contradict attempts to actively prolong pregnancy after rupture of the membranes.
Single daily amikacin versus cefotaxime in the short-course treatment of spontaneous bacterial peritonitis in cirrhotics.

AIM To compare the efficacy and safety of single daily amikacin vs. cefotaxime in the 5-d treatment of spontaneous bacterial peritonitis (SBP). METHODS Thirty-seven cirrhotic patients with SBP, 19 in group A and 18 in group B, were studied. Group A received 1 g of cefotaxime every 6 h, and group B received 500 mg of amikacin qd. Both antibiotics were administered up to 5 d and the responses were compared. RESULTS Infection was cured in 15 of 19 patients (78.9%) treated with cefotaxime and in 11 of 18 (61.1%) treated with amikacin. Four patients of the Cefotaxime group (21.1%) and five patients of the Amikacin group (27.8%) died. Two in each group (10.5% vs 11.1%) had renal impairment during study period. One in each group (5.3% vs 5.6%) may be considered to suffer from nephrotoxicity due to increased urinary beta(2)-microglobulin concentration. CONCLUSION In this study, single daily doses of amikacin in the treatment of SBP in cirrhotics were not associated with an increased incidence of renal impairment or nephrotoxicity. However, a 5-d regimen of amikacin is less effective than a 5-d regimen of cefotaxime in the SBP treatment.
Prolonged inhibition of neutral endopeptidase 24.11 by sinorphan in stroke-prone spontaneously hypertensive rats.

The cardiovascular consequences of inhibition of the neutral endopeptidase 24.11 (NEP) with the orally active NEP inhibitor sinorphan were evaluated by determining long-term effects of the drug on hemodynamic, hormonal and structural parameters in stroke-prone spontaneously hypertensive rats (SHR-SP). Systolic blood pressure increased in young SHR-SP from 194 +/- 2 to 266 +/- 7 mmHg, whereas in sinorphan (30 mg/kg p.o. bid) treated animals systolic blood pressure increased only from 193 +/- 4 to 229 +/- 4 mmHg during the treatment period of 9 weeks. The increase in relative heart weight was also delayed. Plasma ANP was higher in the sinorphan group than in the controls. The results of a second study demonstrate a substantial improvement of cardiac pump function and ventricular hypertrophy in old SHR-SP with compromised cardiac function by long-term inhibition of NEP. Thirteen-month-old SHR-SP were treated with sinorphan (30 mg/kg p.o. bid) for two weeks. At the end of experiment, the increase in ANP plasma levels did not reach statistical significance, whereas plasma cGMP was higher in sinorphan treated animals than in controls. Left ventricular end-diastolic pressure was markedly elevated in controls and significantly lower in sinorphan treated animals. In addition, sinorphan reduced cardiac hypertrophy in these old SHR-SP. In conclusion, the results of the present studies demonstrate that long-term NEP inhibition with sinorphan has inhibitory effects on malignant hypertension and associated cardiac hypertrophy in young SHR-SP on a high-sodium diet. NEP inhibition substantially improves cardiac pump function and reduces ventricular hypertrophy of old SHR-SP with compromised cardiac function.
The effect of end-stage renal disease on innate and adaptive immunity

The progressive loss of kidney function is associated with an inflammatory state and functional defects of the innate and adaptive immune system. The combined effects of increased activation and immune dysfunction could explain the susceptibility of patients in End Stage Renal Disease (ESRD) to viral and bacterial infections, their weak responses to vaccination and increased risk of malignant tumors and cardiovascular disease. It is important to underline that these immune complications mainly affect patients on dialysis. The mechanisms of these immunological disturbances observed in ESRD patients are still unclear. Early identification of chronic kidney disease allows implementing interventions to slow the progression to ESRD. Therefore, measures aimed at attenuating immune abnormalities in ESRD should be a main research area as this could lead to increased survival and better quality of life in HD patients.
The association between bedding material and the bacterial counts of Staphylococcus aureus, Streptococcus uberis and coliform bacteria on teat skin and in teat canals in lactating dairy cattle

Several mastitis-causing pathogens are able to colonize the bovine teat canal. The objective of this study was to investigate the association between the treatment of sawdust bedding with a commercial alkaline conditioner and the bacterial counts on teat skin and in the teat canal. The study used a crossover design. Ten lactating Holstein cows that were free of udder infections and mastitis were included in the study. The animals were bedded on either untreated sawdust or sawdust that had been treated with a hydrated lime-based conditioner. Once a day, fresh bedding material was added. After 3 weeks, the bedding material was removed from the cubicles, fresh bedding material was provided, and the cows were rotated between the two bedding material groups. Teat skin and teat canals were sampled using the wet and dry swab technique after weeks 1, 2, 3, 4, 5 and 6. Staphylococcus aureus, Streptococcus uberis, Escherichia coli and other coliform bacteria were detected in the resulting agar plate cultures. The treatment of the bedding material was associated with the teat skin bacterial counts of Str. uberis, Esch. coli and other coliform bacteria. An association was also found between the bedding material and the teat canal bacterial counts of coliform bacteria other than Esch. coli. For Staph. aureus, no associations with the bedding material were found. In general, the addition of a hydrated lime-based conditioner to sawdust reduces the population sizes of environmental pathogens on teat skin and in teat canals.