Novel azapeptide inhibitors of cathepsins B and K. Structural background to increased specificity for cathepsin B

Abstract: We have designed and synthesized a new series of azapeptides which act as potential inhibitors of cathepsin B and/or cathepsin K. Their structures are based upon the inhibitory sites of natural cysteine protease inhibitors, cystatins. For the synthesized azapeptides, the equilibrium constants for dissociation of inhibitor–enzyme complex, K_i, were determined. Comparison of these values indicated that all of the azainhibitors act much stronger toward cathepsin B. Z‐Arg‐Leu‐His‐Agly‐Ile‐Val‐OMe ( 7 ) proved to be approximately 500 times more potent for cathepsin B than for cathepsin K. To be able to explain the obtained experimental values we used the molecular dynamics procedures to analyze the interactions between cathepsin B and compound 7 . We also determined the structure of the most potent and selective cathepsin B azainhibitor by means of NMR studies and theoretical calculations. In this report, we describe SAR studies of azapeptide inhibitors indicating the influence of the conformational flexibility of the examined compounds on inhibition of cathepsins B and K.     

Synthesis and anti-tuberculous activity of diesters based on isosteviol and dicarboxylic acids

Diesters based on isosteviol and dicarboxylic acids were synthesized and tested for antituberculous activity. Isosteviol and some of its derivatives exhibit appreciable tuberculostatic properties in vitro, the activity being dependent on the length of the polymethylene spacer connecting two ent-beyeran fragments. The mechanism of the antituberculous action of isosteviol derivatives are discussed. __________ Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 40, No. 9, pp. 12–13, September, 2006.     

Fluconazole dosing in continuous veno-venous haemofiltration (CVVHF): need for a high daily dose of 800 mg

To cover intermediate sensitive Candida glabrata in ICU patients,fluconazole plasma peak levels at least in the range of 16–32µg/ml appear necessary for treatment. Previous studiesdid not reach these fluconazole levels under continuous veno-venoushaemofiltration (CVVHF) with dosages of 200–600 mg fluconzoledaily. In the present study, nine patients simultaneously requiringCVVHF for treatment of acute oligoanuric renal failure and antimycotictherapy of Candida septicemia received fluconazole 800 mg/day.Fluconazole plasma levels were determined to evaluate whetherthis dosage is adequate to reach the advised fluconazole levels.Patients were dialysed on two consecutive days with an ultrafiltrationrate (UF) of 1000 ml/h or 2000 ml/h, respectively, in a randomizedorder. The predilution was 800 ml/h and 1800 ml/h, respectively.The treatment was tolerated without adverse effects. All patientsreached plasma fluconazole concentrations between 16 and 32µg/ml, remaining in this range for a minimum of 1 up to24 h with a mean of 9.6 h and a UF rate of 2000 ml/h, and 15.7h with a UF rate of 1000 ml/h. So far, there are no in vivodata on the fluconazole plasma concentrations required for effectivetreatment. However, our data demonstrate, that at least thefluconazole concentrations desirable on the basis of in vitrosusceptibility testing can be reached in critically ill patientson CVVHF in an ICU setting. However, in these patients, 800mg fluconazole/day are necessary to achieve fungicidal drugconcentrations.     

18F-FDG PET/CT for detecting nodal metastases in patients with oral cancer staged N0 by clinical examination and CT/MRI.

(18)F-FDG PET has a high accuracy in staging head and neck cancer, but its role in patients with clinically and radiographically negative necks (N0) is less clear. In particular, the value of combined PET/CT has not been determined in this group of patients. METHODS: In a prospective study, 31 patients with oral cancer and no evidence of lymph node metastases by clinical examination or CT/MRI underwent (18)F-FDG PET/CT before elective neck dissection. PET/CT findings were recorded by neck side (left or right) and lymph node level. PET/CT findings were compared with histopathology of dissected nodes, which was the standard of reference. RESULTS: Elective neck dissections (26 unilateral, 5 bilateral; a total of 36 neck sides), involving 142 nodal levels, were performed. Only 13 of 765 dissected lymph nodes harbored metastases. Histopathology revealed nodal metastases in 9 of 36 neck sides and 9 of 142 nodal levels. PET was TP in 6 nodal levels (6 neck sides), false-negative in 3 levels (3 neck sides), true-negative in 127 levels (23 neck sides), and false-positive in 6 levels (4 neck sides). The 3 false-negative findings occurred in metastases smaller than 3 mm or because of inability to distinguish between primary tumor and adjacent metastasis. TP and false-positive nodes exhibited similar standardized uptakes (4.8 +/- 1.1 vs. 4.2 +/- 1.0; P = not significant). Sensitivity and specificity were 67% and 85% on the basis of neck sides and 67% and 95% on the basis of number of nodal levels, respectively. If a decision regarding the need for neck dissection had been based solely on PET/CT, 3 false-negative necks would have been undertreated, and 4 false-positive necks would have been overtreated. CONCLUSION: (18)F-FDG PET/CT can identify lymph node metastases in a segment of patients with oral cancer and N0 neck. A negative test can exclude metastatic deposits with high specificity. Despite reasonably high overall accuracy, however, the clinical application of PET/CT in the N0 neck may be limited by the combination of limited sensitivity for small metastatic deposits and a relatively high number of false-positive findings. The surgical management of the N0 neck should therefore not be based on PET/CT findings alone.     

The role of Self-Defined Race/Ethnicity in Population Structure Control

Population-based association studies are powerful tools for the genetic mapping of complex diseases. However, this method is sensitive to potential confounding by population structure. While statistical methods that use genetic markers to detect and control for population structure have been the focus of current literature, the utility of self-defined race/ethnicity in controlling for population structure has been controversial. In this study of 1334 individuals, who self-identified as either African American, European American or Hispanic, we demonstrated that when the true underlying genetic structure and the self-defined racial/ethnic groups were roughly in agreement with each other, the self-defined race/ethnicity information was useful in the control of population structure.     

The criminal purchase of firearm ammunition.

OBJECTIVE: Laws that prohibit certain individuals from owning firearms also pertain to ammunition. Whereas retail sales of firearms to criminals are regularly disrupted by instant background checks, sales of ammunition are essentially unchecked and the rate at which criminals acquire ammunition is unknown. This research describes the ammunition market and estimates the rate at which criminals are acquiring ammunition. DESIGN: Criminal background checks conducted on individuals purchasing ammunition in the City of Los Angeles in April and May 2004. SETTING: Los Angeles, CA, USA. SUBJECTS: Ammunition purchasers. MAIN OUTCOME MEASURES: Criminal activity that prohibits one from owning, purchasing, or possessing ammunition. RESULTS: 2.6% (95% CI 1.9% to 3.2%) of ammunition purchasers had a prior felony conviction or another condition that prohibited them from possessing ammunition. During the study period prohibited possessors purchased 10,050 rounds of ammunition in Los Angeles. CONCLUSIONS: These estimates suggest that monitoring ammunition transactions may help reduce the supply of ammunition to criminals and the frequency of injuries from felonious gun assaults. Such a record can also provide information for generating leads on illegal firearm possession.     

Acromioclavicular joint dislocation: a comparative biomechanical study of the palmaris-longus tendon graft reconstruction with other augmentative methods in cadaveric models

Background  

Acromioclavicular injuries are common in sports medicine. Surgical intervention is generally advocated for chronic instability of Rockwood grade III and more severe injuries. Various methods of coracoclavicular ligament reconstruction and augmentation have been described. The objective of this study is to compare the biomechanical properties of a novel palmaris-longus tendon reconstruction with those of the native AC+CC ligaments, the modified Weaver-Dunn reconstruction, the ACJ capsuloligamentous complex repair, screw and clavicle hook plate augmentation.     

Biocidal effect of bleach on Mycobacterium tuberculosis: a safety measure.

BACKGROUND: Although treatment of sputum using bleach has shown increased sensitivity in smear microscopy, the safety aspect of sodium hypochlorite (NaOCl) has not been addressed. OBJECTIVE: To determine the biocidal effect of NaOCl on Mycobacterium tuberculosis when used as a safety measure in direct sputum smear microscopy for the diagnosis of tuberculosis (TB). METHOD: A total of 156 smear- and culture-positive pooled sputum specimens were assessed for the viability of M. tuberculosis after treatment with NaOCl. Each specimen was divided into 11 equal portions. One portion was directly cultured. Each of five portions was treated with 3.5% NaOCl and the other five with 5% NaOCl. Specimens were then cultured at intervals of 15 min and 1, 3, 15 and 24 h. RESULTS: A total of 18 (11.5%) specimens showed growth after treatment with NaOCl. Of these, 5 (3.2%) showed growth upon treatment with 3.5% NaOCl at 15 min and 15 and 24 h. Ten (6.4%) showed growth after treatment with 5% NaOCl between 15 min and 3 h, but none showed growth after 15 h. Three (1.9%) specimens showed growth in both concentrations. CONCLUSION: Our preliminary findings show that exposure of sputum to 5% NaOCl for at least 15 h may be a safety procedure in smear microscopy for the diagnosis of TB. However, further studies are warranted.     

Tolerability of anti-tuberculosis treatment and HIV serostatus.

SETTING: Tuberculosis (TB) is frequent in human immunodeficiency virus (HIV) infected patients, but its treatment is hampered by adverse events and paradoxical reactions. OBJECTIVE: To examine the impact of HIV infection and other factors on the risk and spectrum of adverse events related to anti-tuberculosis treatment in a prospective cohort study conducted between January 2003 and August 2004. RESULTS: Of 105 patients treated for TB, 30 were HIV-infected. The overall incidence of adverse events was 122.5 +/- 18.5 per 100 patient-years (py) and the incidence of severe adverse events was 45.2 +/- 11.3/100 py. Age >50 years (OR 2.2, 95%CI 1.01-4.8, P = 0.046) and HIV infection (OR 3.9, 95%CI 2.1-7.5, P < 0.001) were independently associated with a higher risk of adverse events. Hepatitis (30.5/100 py) and neuropathy (28.6/100 py) were the most frequent adverse events. Hepatitis C virus infection was associated with hepatitis (OR 4.2, 95%CI 1.2-15.0, P = 0.028) and neuropathy with HIV infection (OR 3.8, 95%CI 1.1-13.7, P = 0.040). CONCLUSION: Adverse reactions to anti-tuberculosis drugs are frequent. HIV infection and age >50 years are factors associated with such reactions, while hepatitis C virus infection is a risk factor for hepatitis.