Differential scanning fluorimetry (DSF) screen to identify inhibitors of Hsp60 protein-protein interactions.

There are relatively few methods available for discovering inhibitors of the protein-protein interactions (PPIs) that hold together homo-oligomers. We envisioned that Differential Scanning Fluorimetry (DSF) might be a versatile way to discover this type of inhibitor because oligomers are often more thermally stable than monomers. Using the homo-heptameric chaperonin, Hsp60, as a model, we screened ∼5000 diverse compounds in 384-well plates by DSF, revealing molecules that partially inhibited oligomerization. Because DSF does not require protein labeling or structural information, we propose that it could be a versatile way to uncover PPI inhibitors.

High-throughput screen for inhibitors of protein-protein interactions in a reconstituted heat shock protein 70 (Hsp70) complex.

Protein-protein interactions (PPIs) are an important category of putative drug targets. Improvements in high-throughput screening (HTS) have significantly accelerated the discovery of inhibitors for some categories of PPIs. However, methods suitable for screening multiprotein complexes (e.g. those composed of three or more different components) have been slower to emerge. Here, we explored an approach that uses reconstituted multiprotein complexes (RMPCs). As a model system, we chose heat shock protein 70 (Hsp70), which is an ATP-dependent molecular chaperone that interacts with co-chaperones, including DnaJA2 and BAG2. The PPIs between Hsp70 and its co-chaperones stimulate nucleotide cycling. Thus, to re-create this ternary protein system, we combined purified human Hsp70 with DnaJA2 and BAG2 and then screened 100,000 diverse compounds for those that inhibited co-chaperone-stimulated ATPase activity. This HTS campaign yielded two compounds with promising inhibitory activity. Interestingly, one inhibited the PPI between Hsp70 and DnaJA2, whereas the other seemed to inhibit the Hsp70-BAG2 complex. Using secondary assays, we found that both compounds inhibited the PPIs through binding to allosteric sites on Hsp70, but neither affected Hsp70's intrinsic ATPase activity. Our RMPC approach expands the toolbox of biochemical HTS methods available for studying difficult-to-target PPIs in multiprotein complexes. The results may also provide a starting point for new chemical probes of the Hsp70 system.

Older Age and Larger Prostate Volume Are Associated with Stress Urinary Incontinence after Plasmakinetic Enucleation of the Prostate.

Background. To investigate the factors associated with the occurrence of and recovery from stress urinary incontinence (SUI) after plasmakinetic enucleation of the prostate (PKEP). Materials and Methods. This retrospective study enrolled 1,288 patients with benign prostatic hyperplasia treated with plasmakinetic enucleation from January 2008 to January 2015, collecting demographics and clinical parameters. SUI was defined as a patient complaint of involuntary urine leak, including stress or mixed urinary incontinence. Logistic regression analysis was used to investigate the factors associated with the occurrence of SUI. Results. SUI after PKEP occurred in 80 of 1,288 patients (6.2%), 73 of whom (91.3%) recovered within 3 months and 78 of whom (97.5%) recovered within 6 months. In multivariate regression analysis of factors that were significant in univariate analysis, the factors that were significantly associated with postoperative SUI were age ≥ 70 years (odds ratio [OR] = 9.239; 95% confidence interval [CI] = 4.616-18.495; P < 0.001) and prostate volume on transrectal ultrasound ≥ 90 mL (OR = 15.390; 95% CI = 8.077-29.326; P < 0.001). Conclusions. SUI occurred in 6.2% patients after PKEP and was associated with older age and larger prostate volume. We suggest that age and prostate volume be considered in preoperative candidate selection before PKEP to reduce the occurrence of postoperative SUI.

Bipolar transurethral enucleation and resection of the prostate versus bipolar resection of the prostate for prostates larger than 60gr: A retrospective study at a single academic tertiary care center.

OBJECTIVE: To evaluate the efficacy and safety of bipolar transurethral enucleation and resection of the prostate (B-TUERP) versus bipolar transurethral resection of the prostate (B-TURP) in the treatment of prostates larger than 60g. MATERIAL AND METHODS: Clinical data for 270 BPH patients who underwent B-TUERP and 204 patients who underwent B-TURP for BPH from May 2007 to May 2013 at our center were retrospectively analyzed. Outcome measures included operative time, decreased hemoglobin level, total prostate specific antigen (TPSA), International Prostate Symptom Score (IPSS), maximal urinary flow rate (Qmax), quality of life (QoL) score, post void residual urine volume (RUV), bladder irrigation duration, hospital stay, and the weight of resected prostatic tissue. Other measures included perioperative complications including transurethral resection syndrome (TURS), hyponatremia, blood transfusion, bleeding requiring surgery, postoperative acute urinary retention, urine incontinence and urinary sepsis. Patients in both groups were followed for two years. RESULTS: Compared with the B-TURP group, the B-TUERP group had shorter operative time, postoperative bladder irrigation duration and hospital stay, a greater amount of resected prostatic tissue, less postoperative hemoglobin decrease, better postoperative IPSS and Qmax, as well as lower incidences of hyponatremia, urinary sepsis, blood transfusion requirement, urine incontinence and reoperation (P<0.05 for all). CONCLUSIONS: B-TUERP is superior to B-TURP in the management of large volume BPH in terms of efficacy and safety, but this finding needs to be validated in further prospective, randomized, controlled studies.

Bipolar transurethral enucleation and resection of the prostate versus bipolar resection of the prostate for prostates larger than 60gr: A retrospective study at a single academic tertiary care center

ABSTRACT Objective: To evaluate the efficacy and safety of bipolar transurethral enucleation and resection of the prostate (B-TUERP) versus bipolar transurethral resection of the prostate (B-TURP) in the treatment of prostates larger than 60g. Material and Methods: Clinical data for 270 BPH patients who underwent B-TUERP and 204 patients who underwent B-TURP for BPH from May 2007 to May 2013 at our center were retrospectively analyzed. Outcome measures included operative time, decreased hemoglobin level, total prostate specific antigen (TPSA), International Prostate Symptom Score (IPSS), maximal urinary flow rate (Qmax), quality of life (QoL) score, post void residual urine volume (RUV), bladder irrigation duration, hospital stay, and the weight of resected prostatic tissue. Other measures included perioperative complications including transurethral resection syndrome (TURS), hyponatremia, blood transfusion, bleeding requiring surgery, postoperative acute urinary retention, urine incontinence and urinary sepsis. Patients in both groups were followed for two years. Results: Compared with the B-TURP group, the B-TUERP group had shorter operative time, postoperative bladder irrigation duration and hospital stay, a greater amount of resected prostatic tissue, less postoperative hemoglobin decrease, better postoperative IPSS and Qmax, as well as lower incidences of hyponatremia, urinary sepsis, blood transfusion requirement, urine incontinence and reoperation (P<0.05 for all). Conclusions: B-TUERP is superior to B-TURP in the management of large volume BPH in terms of efficacy and safety, but this finding needs to be validated in further prospective, randomized, controlled studies.

Anti-inflammatory activity of Gentiana striata Maxim.

The anti-inflammatory activity and the mechanism of action of Gentiana striata Maxim. has been investigated. The most active phase, the ethyl acetate extract of Gentiana striata Maxim. (EGS), displayed potent inhibitory activity on feet oedema of rheumatoid arthritis (RA) inflicted rats. This anti-inflammatory activity might be partly based on the notable reduction of prostaglandin E2 (PGE2) and nitric oxide (NO) levels. Six further compounds isolated from EGS have previously been reported as having anti-inflammatory activity.

[The utility of trnH-psbA gene for phylogenetic analysis of Huperziaceae and plant barcoding].

OBJECTIVE: The purpose of study was to discover the phylogenetic relations and plant barcoding of 17 plants from Huperziaceae. METHODS: Phylogenetic tree of chloroplast trnH-psbA gene of 17 plants from Huperziaceae was constructed by software. RESULTS: It showed that Huperziaceae could be divided into two genera Huperzia and Phlegmariurus and bootstrap value reached 91%. CONCLUSIONS: Holub and Qing' taxonomy was supported and 17 species in Huperziaceae were monophyletic groups and it suggested that trnH-psbA could be used as a DNA barcode to identify plants.

[Study on pharmacokinetics of emodin in Rhizoma Polygontum Cuspidatum and its compound].

OBJECTIVE: To study the difference in the pharmacokinetics of emodin in Zhiganning capsules and Rhizoma Polygontum Cuspidatum by nonaqueous RP-HPLC. METHOD: The rats were orally administered with the extraction of Rhizoma Polygontum Cuspidatum and Zhiganning capsules. After hydrolysis and extraction, the content of emodin in the plasma is determined by Nonaqueous RP-HPLC. RESULT: The concentration-time profiles of emodin fit two-compartment model. The pharmacokinetics parameters including, t1/2alpha, AUC(0-infinity), CL(s) and C(max) of emodin in the group of Rhizoma Polygontum Cuspidatum were significantly different from these in the group of its compounds. CONCLUSION: There is a significant difference in pharmacokinetics of emodin between zhiganning capsules and the extraction of Rhizoma Polygontum Cuspidatum.

Study on pharmacokinetics of emodin in Rhizoma Polygontum Cuspidatum and its compound / 中国中药杂志

<p><b>OBJECTIVE</b>To study the difference in the pharmacokinetics of emodin in Zhiganning capsules and Rhizoma Polygontum Cuspidatum by nonaqueous RP-HPLC.</p><p><b>METHOD</b>The rats were orally administered with the extraction of Rhizoma Polygontum Cuspidatum and Zhiganning capsules. After hydrolysis and extraction, the content of emodin in the plasma is determined by Nonaqueous RP-HPLC.</p><p><b>RESULT</b>The concentration-time profiles of emodin fit two-compartment model. The pharmacokinetics parameters including, t1/2alpha, AUC(0-infinity), CL(s) and C(max) of emodin in the group of Rhizoma Polygontum Cuspidatum were significantly different from these in the group of its compounds.</p><p><b>CONCLUSION</b>There is a significant difference in pharmacokinetics of emodin between zhiganning capsules and the extraction of Rhizoma Polygontum Cuspidatum.</p>