Comparative Evaluation of LAMP and Nested PCR for the Rapid Diagnosis of Mycobacterium marinum Infection.

Purpose: Culture of Mycobacterium marinum is very time-consuming, taking several weeks to produce positive results. Seeking rapid and sensitive diagnostic methods for diagnosis can greatly improve patient treatment. Our study aimed to compare the rapid diagnostic abilities of polymerase chain reaction (PCR), nested PCR and loop mediated isothermal amplification (LAMP) of detecting M. marinum in skin samples from patients with M. marinum infection. Methods: A total of 6 M. marinum strains and 6 skin samples with definite diagnosis of M. marinum infection were included in the study. We optimized LAMP performance for detection of M. marinum genomic DNA and confirmed the specificity of the primers. Then, the sensitivity of the LAMP and nested PCR assays were assessed by M. marinum strains and clinical samples. Results: Nested PCR was 10-fold more sensitive than the LAMP assay by serial dilution of M. marinum DNA. PCR positive samples were all positive by LAMP detection of 6 clinical M. marinum strains. Out of 6 clinical skin specimens confirmed as M. marinum infection, 0 (0%), 3 (50%), 3 (50%), and 4 (66.6%) were positive by PCR, nested PCR, LAMP and culture. The LAMP shared the same sensitivity than nested PCR in M. marinum strains and clinical samples, but it was easy to perform and faster than nested PCR assay. Conclusion: Compared with conventional PCR, LAMP and nested PCR are more sensitive and have a higher detection rate of M. marinum in clinical skin specimens. The LAMP assay proved to be more suitable for rapid diagnosis of M. marinum infection in a shorter time, especially in resource-limited settings.

Preparation, characterization, and pharmacokinetics of rivaroxaban cocrystals with enhanced in vitro and in vivo properties in beagle dogs.

Rivaroxaban (RIV) is a direct Factor Xa inhibitor anticoagulant, but the oral bioavailability of RIV is estimated to be only 60% due to its poor solubility. The aim of the present study was to improve the solubility and bioavailability of RIV. Five cocrystals-p-hydroxybenzoic acid (HBA), 2,4-dihydroxybenzoic acid (DBA), nicotinamide (NA), isonicotinamide (IA), and succinic acid (SA)-were used as cofomers and were successfully obtained and characterized by powder X-ray diffraction, thermal analysis, and Fourier transform infrared spectra. RIV-DBA and RIV-HBA cocrystals showed obvious improvements in solubility, dissolution (under sink conditions), and intrinsic dissolution rates versus RIV. Moreover, the dissolution of RIV-HBA, RIV-DBA, and RIV-SA cocrystals under non-sink conditions showed obvious "spring and parachute" patterns. The in vitro permeability levels in a Caco-2 cell model of RIV-DBA and RIV-IA cocrystals were significantly improved versus RIV. Pharmacokinetic studies in beagle dogs showed that RIV-DBA and RIV-HBA cocrystals had higher bioavailability than RIV. The enhancements in solubility and bioavailability indicate the potential of RIV cocrystals as a better candidate for the treatment of thrombosis versus RIV.

Optimization of Nanostructured Lipid Carriers of Fenofibrate Using a Box-Behnken Design for Oral Bioavailability Enhancement.

BACKGROUND: Fenofibrate (FNB) is a commonly used hypolipidemic agent. However, the oral bioavailability of FNB is limited by slow dissolution due to its low solubility. Thus, investigations on novel FNB formulations are necessary for their use. OBJECTIVE: The objective of this study is to enhance the oral bioavailability of FNB using optimized Nanostructured Lipid Carrier (NLC) formulations. METHODS: Hot homogenization followed by ultrasonication was used to prepare FNB-NLCs. These formulations were optimized using a Box-Behnken design, where the amount of FNB (X1), a ratio of solid lipid/liquid lipid (X2), and the percentage of emulsifier (X3) were set as independent variables, while the particle size (Y1), and Entrapment Efficiency (EE%) (Y2), were used as dependent factors. An in vitro dissolution test was then performed using a paddle method, while an in vivo pharmacokinetic study of FNB-NLC formulation was performed in rats. RESULTS: FNB-NLCs were successfully prepared and optimized using a Box-Behnken design. The particle size and EE% of the FNB-NLC had less than 5% difference from predicted values. The in vitro dissolution and oral bioavailability of the FNB-NLC were both higher than those of raw FNB. CONCLUSION: A Box-Behnken design was successfully applied to optimize FNB-NLC formulation for the enhancement of the dissolution and bioavailability of FNB, a poorly water-soluble drug.

Bile acid sequestrants: a review of mechanism and design.

OBJECTIVE: Bile acid sequestrants (BAS) are used extensively in the treatment of hypercholesterolaemia. This brief review aimed to describe the design and evaluation of three types of BAS: amphiphilic copolymers, cyclodextrin/poly-cyclodextrin and molecular imprinted polymers. The mechanisms underlying the action of BAS are also discussed. KEY FINDINGS: BAS could lower plasma cholesterol, improve glycemic control in patients with type 2 diabetes and regulate balance energy metabolism via receptors or receptor-independent mediated mechanisms. Different types of BAS have different levels of ability to bind to bile acids, different stability and different in-vivo activity. CONCLUSIONS: A growing amount of evidence suggests that bile acids play important roles not only in lipid metabolism but also in glucose metabolism. The higher selectivity, specificity, stability and in-vivo activity of BAS show considerable potential for lipid-lowering therapy.

Effect of Surfactants and Polymers on the Dissolution Behavior of Supersaturable Tecovirimat-4-Hydroxybenzoic Acid Cocrystals.

(1) Background: Pharmaceutical cocrystals have attracted remarkable interest and have been successfully used to enhance the absorption of poorly water-soluble drugs. However, supersaturable cocrystals are sometimes thermodynamically unstable, and the solubility advantages present a risk of precipitation because of the solution-mediated phase transformation (SMPT). Additives such as surfactants and polymers could sustain the supersaturation state successfully, but the effect needs insightful understanding. The aim of the present study was to investigate the roles of surfactants and polymers in the dissolution-supersaturation-precipitation (DSP) behavior of cocrystals. (2) Methods: Five surfactants (SDS, Poloxamer 188, Poloxamer 407, Cremophor RH 40, polysorbate 80) and five polymers (PVP K30, PVPVA 64, HPC, HPMC E5, CMC-Na) were selected as additives. Tecovirimat-4-hydroxybenzoic (TEC-HBA) cocrystals were chosen as a model cocrystal. The TEC-HBA cocrystals were first designed and verified by PXRD, DSC, SEM, and FTIR. The effects of surfactants and polymers on the solubility and dissolution of TEC-HBA cocrystals under sink and nonsink conditions were then investigated. (3) Results: Both the surfactants and polymers showed significant dissolution enhancement effects, and most of the polymers were more effective than the surfactants, according to the longer Tmax and higher Cmax. These results demonstrate that the dissolution behavior of cocrystals might be achieved by the maintained supersaturation effect of the additives. Interestingly, we found a linear relationship between the solubility and Cmax of the dissolution curve for surfactants, while no similar phenomena were found in solutions with polymer. (4) Conclusions: The present study provides a basis for additive selection and a framework for understanding the behavior of supersaturable cocrystals in solution.

Hair removal using a new intense pulsed light source in Chinese patients.

BACKGROUND: Unwanted hair is a widespread cosmetic problem. Several lasers and intense pulsed light (IPL) have been utilized for this purpose. A new IPL device (Lumenis One) with OPT is one of the newer modalities to be studied in Chinese patients. OBJECTIVE: This study evaluates the short-term efficacy and side effects of the new IPL device for epilation in Chinese patients. METHODS: Eighteen Chinese women with Fitzpatrick skin types III-V and black hair, were treated four times at 4 to 6-week intervals using IPL (Lumenis One) on the axillae (n=13) and the upper lip (n=5). The energy density for treatment ranged from 14 to 22 J/cm(2). Parameters utilized were 695-/755-nm filters, triple pulse for patients on the axillae, and 640-/695-nm filters, double pulse for patients on the upper lip (3.5- to 7-ms pulse, 30- to 90-ms pulse delay, 15x35 mm spot size). Hair reduction was assessed at baseline, immediately before each treatment session, and at 4 weeks after the fourth treatment. Patient's satisfaction on a 5-point scale was also evaluated. RESULTS: The average hair reduction for all sites was 49.9% after one session, 58.6% after two sessions, 79.3% after three sessions, and 83.8% after four sessions (p=0.001). The hair reduction of 44.1%, 52.1%, 81.1%, and 86.0% were achieved after each treatment for axillae, with 65.1%, 75.7%, 74.6%, and 78.0% for upper lip. Patients got more satisfaction after four sessions (score 3.1) than that after two sessions (2.0) (p=0.001). In both the assessments, upper lip appeared to show a better response than axillae after two IPL treatments, which reversed after four treatments. No significant complications or adverse events were reported. CONCLUSION: The new IPL device provides a safe and effective means of hair removal for Chinese patients. Treatment efficacy varies with the anatomic location and number of treatments. However, further study is necessary to determine the long-term clinical efficacy in Chinese patients.

The impact of immunosenescence on the efficacy of immune checkpoint inhibitors in melanoma patients: a meta-analysis.

BACKGROUND: Immunosenescence, the age-related decline of immunity, affects the immune responses of melanoma patients. Through immune responses, immune checkpoint inhibitors (ICIs) exert their antitumor robustness. In different ages of melanoma patients, especially the older patients, the effectiveness of ICIs remains unclear. It is still controversial whether ICIs should be used in treating older patients. MATERIALS AND METHODS: The authors included clinical trials of ICIs in older and younger patients. The authors used hazard ratio (HR) and 95% CI of overall survival (OS). RESULTS: From four phase III randomized clinical trials 2,251 melanoma patients were included. We found that ICIs significantly prolonged the OS for melanoma patients in both younger (HR, 0.71; 95% CI, 0.60-0.82; P<0.001) and older groups (HR, 0.62; 95% CI, 0.41-0.83; P<0.001) compared with controls. Anti-programmed death-1 (PD-1) agents appeared to be more efficient in older melanoma patients (HR, 0.34; 95% CI, 0.14-0.53) versus younger patients (HR, 0.52; 95% CI, 0.26-0.78). CONCLUSION: ICIs significantly prolonged the OS for melanoma patients in both younger and older groups than controls. Anti-PD-1 agents were more efficient in older melanoma patients versus younger patients. ICIs could be used for older melanoma patients.

Outbreak of a cutaneous Mycobacterium marinum infection in Jiangsu Haian, China.

Mycobacterium marinum is a slow-growing mycobacterium. In November 2008, we diagnosed a patient with M. marinum infection who worked at a fish farm in Jiangsu Haian, China. We conducted an investigation and found 18 patients with the same infection. In suspected cases, complete data were collected including medical history, clinical manifestations, laboratory features, and responses to treatment. Therapeutic regimens, including clarithromycin monotherapy or combined treatment with clarithromycin, rifampicin, and ethambutol, were prescribed. A total of 18 patients with M. marinum infection were found. All patients showed only skin lesions. Biopsies were performed and 16 patients showed infective granulomas. Acid-fast bacilli stain (Ziehl-Neelson stain) for cutaneous samples were positive in 7 patients. Ten patients were positive in purified protein derivative tests (tubercles were ≥10 mm in diameter). In 16 patients, colonies grew after tissue samples were incubated on Löwenstein-Jensen medium at 32 °C. All the isolates were identified as M. marinum by polymerase chain reaction-restriction fragment length polymorphism analysis, by direct gene sequencing, and by genotyping using mycobacterial interspersed repetitive units. Fifteen of the 18 patients were cured using clarithromycin-containing antibiotic regimens. The history of contact with fish and aquaria plays an important role in diagnosis. Clarithromycin-containing regimens were successful in most patients with M. marinum infections limited to the skin.