Comparison of green pit viper and Agkistrodon halys antivenom in inhibition of coagulopathy due to Trimeresurus albolabris venom: an in-vitro study using human plasma.

INTRODUCTION: There are two antivenoms that may be administered in Hong Kong following a bite by Trimeresurus albolabris: the green pit viper antivenom from the Thai Red Cross Society in Thailand and the Agkistrodon halys antivenom from the Shanghai Institute of Biological Products in China. Both are recommended by the Central Coordinating Committee of Accident and Emergency Services of the Hospital Authority for treating patients with a bite by Trimeresurus albolabris. The choice of which antivenom to use is based on physician preference. This study aimed to compare the relative efficacy of the two antivenoms. METHODS: This in-vitro experimental study was carried out by a wildlife conservation organisation and a regional hospital in Hong Kong. Human plasma from 40 adult health care worker volunteers was collected. The Trimeresurus albolabris venom was added to human plasma and the mixture was assayed after incubation with each antivenom (green pit viper and Agkistrodon halys) using saline as a control. Fibrinogen level and clotting time in both antivenom groups were studied. RESULTS: The mean fibrinogen level was elevated from 0 g/L to 2.86 g/L and 1.11 g/L after the addition of green pit viper antivenom and Agkistrodon halys antivenom, respectively. When mean clotting time was measured, the value was 6.70 minutes in the control, prolonged to more than 360 minutes by green pit viper antivenom and to 19.06 minutes by Agkistrodon halys antivenom. CONCLUSIONS: Green pit viper antivenom was superior to Agkistrodon halys antivenom in neutralisation of the thrombin-like and hypofibrinogenaemic activities of Trimeresurus albolabris venom.

Atmospheric deposition of phosphorus to land and freshwater.

We compiled published and newly-obtained data on the directly-measured atmospheric deposition of total phosphorus (TP), filtered total phosphorus (FTP), and inorganic phosphorus (PO4-P) to open land, lakes, and marine coasts. The resulting global data base includes data for c. 250 sites, covering the period 1954 to 2012. Most (82%) of the measurement locations are in Europe and North America, with 44 in Africa, Asia, Oceania, and South-Central America. The deposition rates are log-normally distributed, and for the whole data set the geometric mean deposition rates are 0.027, 0.019 and 0.14 g m(-2) a(-1) for TP, FTP and PO4-P respectively. At smaller scales there is little systematic spatial variation, except for high deposition rates at some sites in Germany, likely due to local agricultural sources. In cases for which PO4-P was determined as well as one of the other forms of P, strong parallels between logarithmic values were found. Based on the directly-measured deposition rates to land, and published estimates of P deposition to the oceans, we estimate a total annual transfer of P to and from the atmosphere of 3.7 Tg. However, much of the phosphorus in larger particles (principally primary biological aerosol particles) is probably redeposited near to its origin, so that long-range transport, important for tropical forests, large areas of peatland and the oceans, mainly involves fine dust from deserts and soils, as described by the simulations of Mahowald et al. (Global Biogeochemical Cycles 22, GB4026, 2008). We suggest that local release to the atmosphere and subsequent deposition bring about a pseudo-diffusive redistribution of P in the landscape, with P-poor ecosystems, for example ombrotrophic peatlands and oligotrophic lakes, gaining at the expense of P-rich ones. Simple calculations suggest that atmospheric transport could bring about significant local redistribution of P among terrestrial ecosystems. Although most atmospherically transported P is natural in origin, local transfers from fertilised farmland to P-poor ecosystems may be significant, and this requires further research.

Chemical fluxes in time through forest ecosystems in the UK - soil response to pollution recovery.

Long term trend analysis of bulk precipitation, throughfall and soil solution elemental fluxes from 12 years monitoring at 10 ICP Level II forest sites in the UK reveal coherent national chemical trends indicating recovery from sulphur deposition and acidification. Soil solution pH increased and sulphate and aluminium decreased at most sites. Trends in nitrogen were variable and dependant on its form. Dissolved organic nitrogen increased in bulk precipitation, throughfall and soil solution at most sites. Nitrate in soil solution declined at sites receiving high nitrogen deposition. Increase in soil dissolved organic carbon was detected - a response to pollution recovery, changes in soil temperature and/or increased microbial activity. An increase of sodium and chloride was evident - a possible result of more frequent storm events at exposed sites. The intensive and integrated nature of monitoring enables the relationships between climate/pollutant exposure and chemical/biological response in forestry to be explored.

Oral bacterial flora of the Chinese cobra (Naja atra) and bamboo pit viper (Trimeresurus albolabris) in Hong Kong SAR, China.

OBJECTIVE: To determine the oral bacterial flora associated with two common local venomous snakes in Hong Kong, namely the Chinese cobra (Naja atra) and the bamboo pit viper (Trimeresurus albolabris). DESIGN: Cross-sectional study. SETTING: A non-government organisation and a regional hospital in Hong Kong. SUBJECTS: Thirty-two Chinese cobras and seven bamboo pit vipers. MAIN OUTCOME MEASURES: Species identification of bacteria in the oral cavity of both snakes and their antibiotic susceptibilities. RESULTS: The oral cavity of Chinese cobra harbour a wide range of pathogenic bacteria, including: Gram-negative bacterial species like Morganella morganii, Aeromonas hydrophila and Proteus, and Gram-positive bacteria like Enterococcus faecalis, coagulase-negative Staphylococcus as well as anaerobic species (clostridia). The oral cavity of the Chinese cobra is more likely than that of the bamboo pit viper to harbour pathogenic bacteria associated with snakebite infection (P<0.001). The median number of pathogenic bacteria per snake was significantly higher in the Chinese cobra (P<0.001). All pathogenic Gram-negative bacteria isolated were susceptible to levofloxacin. Amoxicillin/clavulanate provided good coverage against pathogenic Gram-positive bacteria (Enterococcus faecalis) and anaerobes. CONCLUSION: 'Prophylactic' antibiotic treatment for Chinese cobra bites may be beneficial, owing to the multiple pathogenic bacteria in its oral cavity and the higher risk of ensuing necrosis. The regimen of levofloxacin plus amoxicillin/clavulanate appears promising for this purpose, but further study is required to confirm its clinical utility in patients.

The use of Raman spectroscopy to differentiate between different prostatic adenocarcinoma cell lines.

Raman spectroscopy (RS) is an optical technique that provides an objective method of pathological diagnosis based on the molecular composition of tissue. Studies have shown that the technique can accurately identify and grade prostatic adenocarcinoma (CaP) in vitro. This study aimed to determine whether RS was able to differentiate between CaP cell lines of varying degrees of biological aggressiveness. Raman spectra were measured from two well-differentiated, androgen-sensitive cell lines (LNCaP and PCa 2b) and two poorly differentiated, androgen-insensitive cell lines (DU145 and PC 3). Principal component analysis was used to study the molecular differences that exist between cell lines and, in conjunction with linear discriminant analysis, was applied to 200 spectra to construct a diagnostic algorithm capable of differentiating between the different cell lines. The algorithm was able to identify the cell line of each individual cell with an overall sensitivity of 98% and a specificity of 99%. The results further demonstrate the ability of RS to differentiate between CaP samples of varying biological aggressiveness. RS shows promise for application in the diagnosis and grading of CaP in clinical practise as well as providing molecular information on CaP samples in a research setting.

Assessment of fiberoptic near-infrared raman spectroscopy for diagnosis of bladder and prostate cancer.

OBJECTIVES: To determine whether a fiberoptic Raman system, suitable for in vivo use, is able to differentiate between benign and malignant bladder and prostate pathologic findings in vitro. Raman spectroscopy is an optical technique that provides a measure of the molecular composition of tissue by analyzing the way that tissue scatters laser light. Laboratory studies have shown that the technique can be used to identify and characterize transitional cell carcinoma and prostate adenocarcinoma in vitro. METHODS: A total of 220 Raman spectra were recorded from 29 snap-frozen bladder samples collected at cystoscopic procedures, and 197 Raman spectra were recorded from 38 snap-frozen prostate samples collected at transurethral resection of the prostate. The spectra were correlated with the histologic features and used to construct separate diagnostic algorithms for the bladder and prostate. These algorithms were tested as to their ability to determine the pathologic finding of a sample from its Raman spectrum. RESULTS: The bladder algorithm was able to differentiate benign samples (normal and cystitis) from malignant samples (transitional cell carcinoma), with an overall accuracy of 84%. The prostate algorithm was able to differentiate benign samples (benign prostatic hyperplasia and prostatitis) from malignant samples (prostate cancer), with an overall accuracy of 86%. CONCLUSIONS: The results of this study have demonstrated that the clinical Raman system can provide an accurate and objective method to diagnose prostate and bladder cancer in vitro. Because the Raman probe is suitable for use during endoscopic, laparoscopic, or open procedures, this work paves the way for in vivo studies.

The use of Raman spectroscopy to identify and characterize transitional cell carcinoma in vitro.

OBJECTIVE: To determine whether Raman spectroscopy can be used to differentiate between normal, inflammatory and malignant bladder pathologies in vitro, and secondly if it can used to grade and stage transitional cell carcinoma (TCC). MATERIALS AND METHODS: In all, 1525 Raman spectra were measured from 75 bladder samples comprising normal bladder, cystitis, carcinoma in situ (CIS), TCC and adenocarcinoma. Multivariate analysis was applied to the spectral dataset to construct diagnostic algorithms; these were then tested for their ability to determine the histological diagnosis of each sample from its Raman spectrum. RESULTS: The diagnostic algorithms could be used to accurately differentiate among the pathological groups, in particular, a three-group algorithm differentiated among normal bladder, cystitis and TCC/CIS with sensitivities and specificities of > 90%. Algorithms could also accurately characterize TCC in terms of splitting them into low (G1/G2) or high (G3) grade and superficial (pTa) or invasive (pT1/pT2) stage. CONCLUSION: Raman spectroscopy can be used to accurately identify and grade/stage TCC in vitro. The technique therefore shows promise for use as an objective method to assist the pathologist in assessing bladder pathologies. Raman spectroscopy also has potential to provide immediate pathological diagnoses during surgical procedures. Following the promising results of this in vitro study, in vivo cystoscopic studies are planned.

National and international variation in the registration of bladder cancer.

OBJECTIVE: To determine whether variations exist in the methods by which different cancer registries record information on bladder cancers. METHODS: The registration practices of the various cancer registries within the UK, Europe and the USA were investigated by consulting the available publications and by correspondence with registry staff. In addition, a telephone survey was carried out within the UK to determine whether the national guidelines on bladder cancer coding were being followed. RESULTS: There is variation in the registration of bladder cancers both among regions within the UK and between the UK and other regions. The telephone survey showed that only four of the 11 UK regional registries were correctly following the national bladder cancer coding guidelines. Bladder cancer registration also varies between the cancer registries within mainland Europe. When comparing registration practices in the UK and the USA the major difference is that cases of bladder carcinoma in situ and pTa transitional cell carcinoma are included in the North American cancer statistics but not in the British cancer statistics. CONCLUSION: Much needs to be done before it can be claimed that the registration of bladder cancers has been standardized either nationally or internationally. In particular, the differences in registration practices between the UK and the USA will tend to give a falsely low impression of British incidence and survival rates compared with the equivalent North American figures. This confounding factor must be considered if these incidence and survival values are to be compared.

The use of Raman spectroscopy to identify and grade prostatic adenocarcinoma in vitro.

Raman spectroscopy is an optical technique, which provides a measure of the molecular composition of tissue. Raman spectra were recorded in vitro from both benign and malignant prostate biopsies, and used to construct a diagnostic algorithm. The algorithm was able to correctly identify each pathological group studied with an overall accuracy of 89%. The technique shows promise as a method for objectively grading prostate cancer.

Comparison of POSSUM scoring and the Hardman Index in selection of patients for repair of ruptured abdominal aortic aneurysm.

BACKGROUND: The aim was to assess to what extent the POSSUM (Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity) and Hardman scoring systems were predictive of outcome after surgery for ruptured abdominal aortic aneurysm (RAAA). METHODS: From January 1990 to December 2001, 232 patients presented with RAAA. Forty-one were treated conservatively and all died; the remainder had emergency surgery. The case notes of all but three of these patients were reviewed retrospectively. POSSUM and Hardman scores were calculated and related to mortality. RESULTS: The mortality rate after emergency repair was 54 per cent (104 of 191). The physiology-only POSSUM score specific for RAAA and the Hardman Index score were both significantly associated with increased mortality after operation (P < 0.001). Most non-operated patients were in the highest risk bands. CONCLUSION: Both POSSUM and Hardman scoring systems predicted outcome after emergency surgery for RAAA. The Hardman Index was simpler to calculate, but POSSUM identified a higher number of patients at risk. Risk scoring may help identify patients with RAAA for whom surgery is futile.

The influence of histological diagnosis on the postoperative complication rate following trans-urethral resection of prostate (TURP).

BACKGROUND: A number of studies have shown that the early mortality following TURP is higher for patients with prostate cancer than those with benign disease. This study examines the effect of the histological diagnosis on the predischarge complication rate following TURP. METHODS: Information on the postoperative, predischarge complications of 3036 patients, who underwent TURP over the last decade at our institution, was collated from the urology department database (AuditBase for Windows). The information on this database is collected prospectively, at the point of care and validated at monthly audit meetings. Statistical analyses were performed using chi2 and difference of proportion where n > 60. Statistical significance was taken as P < 0.05. RESULTS: The postoperative, predischarge major complication rate for patients with benign disease was 2.1%. This was not statistically different from the 2.3% complication rate seen in patients with malignant disease. Patients suffering a postoperative complication stayed in hospital significantly longer than those who had a straightforward postoperative course (P < 0.001); however, patients with malignant histology suffering a postoperative complication did not stay statistically significantly longer than those with benign histology suffering a postoperative complication (P < 0.1). CONCLUSIONS: Patients undergoing TURP for prostate cancer do not suffer more postoperative, predischarge complications or stay in hospital longer than patients undergoing TURP for benign disease.

A single normal ultrasonographic scan at age 65 years rules out significant aneurysm disease for life in men.

BACKGROUND: Screening for abdominal aortic aneurysm (AAA) has been carried out in Gloucestershire since 1990. All men in the county are offered aortic ultrasonography in their 65th year. Men with an aortic diameter of less than 26 mm are considered 'normal' and no follow-up is arranged. The aim of this study was to ascertain if men with 'normal' aortic diameters at age 65 years ever develop a clinically significant aneurysm. METHODS: A cohort study was performed on 223 65-year-old men who had an aorta of less than 26 mm in diameter in 1988. These men had repeat ultrasonography in 1993 and 2000. The causes of death in men who died during this interval were investigated. RESULTS: Eight men were lost to follow-up. As far as it was possible to ascertain, none of the 86 men who died over the 12-year interval did so from ruptured AAA. There was no clinically significant increase in mean aortic diameter in the remaining 129 men who had three serial ultrasonographic scans over the 12-year interval. CONCLUSION: A single, 'normal' ultrasound scan at age 65 years effectively rules out the risk of clinically significant aneurysm disease for life in men.

Acquired autoantibodies to coagulation factors.

Acquired coagulation factor antibodies are either alloantibodies or autoantibodies. Alloantibodies are formed when the body reacts to an external antigen. Commonly, alloantibodies arise when a patient who has a congenital clotting factor deficiency is infused with a blood product. Alternately, patients exposed to a coagulation protein from a different species may develop alloantibodies to the animal protein that cross-reacts with their own protein. On the other hand, autoantibodies develop spontaneously in people without pre-existing factor deficiencies and without exposure to an external antigen. These antibodies may neutralize function of a clotting factor, promote rapid clearance of a clotting factor from the blood, or alter the clotting factor in such a way that the protein-antibody complex acquires a unique function. This review focuses on recent reports of autoantibodies directed against fibrinogen, prothrombin, factor V, factor VII, factor X, and von Willebrand factor, in which these various activities of autoantibodies are illustrated.

Purification and initial characterization of peptidyl-tRNA hydrolase from rabbit reticulocytes.

We have identified an activity in rabbit reticulocyte lysate as peptidyl-tRNA hydrolase, based upon its ability to hydrolyze native reticulocyte peptidyl-tRNA, isolated from polyribosomes, and N-acylaminoacyl-tRNA, and its inability to hydrolyze aminoacyl-tRNA, precisely the same substrate specificity previously reported for peptidyl-tRNA hydrolase from bacteria or yeast. The physiological role of the reticulocyte enzyme may be to hydrolyze and recycle peptidyl-tRNA that has dissociated prematurely from elongating ribosomes, as suggested for the bacterial and yeast enzymes, since reticulocyte peptidyl-tRNA hydrolase is completely incapable of hydrolyzing peptidyl-tRNA that is still bound to polyribosomes. We have purified reticulocyte peptidyl-tRNA hydrolase over 5,000-fold from the postribosomal supernatant with a yield of 14%. The purified product shows a 72-kDa band upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis that has co-purified with enzyme activity and comprises about 90% of the total stained protein, strongly suggesting that the 72-kDa protein is the enzyme. Sucrose density gradient analysis indicates an apparent molecular mass for the native enzyme of 65 kDa, implying that it is a single polypeptide chain. The enzyme is almost completely inactive in the absence of a divalent cation: Mg2+ (1-2 mM) promotes activity best, Mn2+ is partly effective, and Ca2+ and spermidine are ineffective. The hydrolase shows a Km of 0.60 microM and Vmax of 7.1 nmol/min/mg with reticulocyte peptidyl-tRNA, a Km of 60 nM and Vmax of 14 nmol/min/mg with Escherichia coli fMet-tRNA(fMet), and a Km of 100 nM and Vmax of 2.2 nmol/min/mg with yeast N-acetyl-Phe-tRNA(Phe). The enzyme has a pH optimum of 7.0-7.25, it is inactivated by heat (60 degrees C for 5 min), and its activity is almost completely inhibited by pretreatment with N-ethylmaleimide or incubation with 20 mM phosphate. The fact that the enzyme hydrolyzes E. coli but not yeast or reticulocyte fMet-tRNA(fMet) may be explained, at least in part, by structural similarities between prokaryotic tRNA(fMet) and eukaryotic elongator tRNA that are not shared by eukaryotic tRNA(fMet).

The site of hydrolysis by rabbit reticulocyte peptidyl-tRNA hydrolase is the 3'-AMP terminus of susceptible tRNA substrates.

The preceding paper (Gross, M., Starn, T.K., Rundquist, C., Crow, P., White, J., Olin, A., and Wagner, T. (1992) J. Biol. Chem. 267, 2073-2079) reported the purification and partial characterization of rabbit reticulocyte peptidyl-tRNA hydrolase. In this article we demonstrate that, unlike bacterial and yeast peptidyl-tRNA hydrolase which act by deacylation, the reticulocyte enzyme hydrolyzes N-acylaminoacyl-tRNA to N-acylaminoacyl-AMP. Reticulocyte lysate has a separate enzyme, that we have isolated and termed aminoacyl-AMP deacylase, which hydrolyzes N-acylaminoacyl-AMP and aminoacyl-AMP, recycling the amino acid and nucleotide components. The action of this enzyme is relatively specific for the N-acylaminoacyl-AMP generated by peptidyl-tRNA hydrolase, since it is much less active with N-acylaminoacyl-adenosine and inactive with N-acylaminoacyl-ACCAC, N-acylaminoacyl-tRNA, or aminoacyl-tRNA. The tRNA product of peptidyl-tRNA hydrolase action is tRNA missing only its 3'-AMP terminus (tRNA(c-c)), since reaminoacylation requires tRNA nucleotidyltransferase but not CTP. The 3' exonucleolytic action of reticulocyte peptidyl-tRNA hydrolase is specific to susceptible tRNA substrates, since it does not hydrolyze CACCA, CACCA-N-acylamino acid, polyuridylic acid, or the 3' polyadenylate tail of globin mRNA, and, since its ability to hydrolyze Escherichia coli f[3H]Met-tRNA(fMet) is not reduced by excess 5 S or 28 S ribosomal RNA and is reduced only slightly by excess tRNA(c-c). Reticulocyte peptidyl-tRNA hydrolase also hydrolyzes th 3'-AMP terminus of deacylated tRNA. This property may explain why the 3'-terminal AMP of tRNA undergoes turnover in reticulocytes and reticulocyte lysate, since we find that such turnover in gel-filtered reticulocyte lysate is increased under conditions where aminoacylation is reduced.