Development of a sensitive enzyme immunoassay for measuring taipan venom in serum.

The detection and measurement of snake venom in blood is important for confirming snake identification, determining when sufficient antivenom has been given, detecting recurrence of envenoming, and in forensic investigation. Venom enzyme immunoassays (EIA) have had persistent problems with poor sensitivity and high background absorbance leading to false positive results. This is particularly problematic with Australasian snakes where small amounts of highly potent venom are injected, resulting in low concentrations being associated with severe clinical effects. We aimed to develop a venom EIA with a limit of detection (LoD) sufficient to accurately distinguish mild envenoming from background absorbance at picogram concentrations of venom in blood. Serum samples were obtained from patients with taipan bites (Oxyuranus spp.) before and after antivenom, and from rats given known venom doses. A sandwich EIA was developed using biotinylated rabbit anti-snake venom antibodies for detection. For low venom concentrations (i.e. <1 ng/mL) the assay was done before and after addition of antivenom to the sample (antivenom difference method). The LoD was 0.15 ng/mL for the standard assay and 0.1 ng/mL for the antivenom difference method. In 11 pre-antivenom samples the median venom concentration was 10 ng/mL (Range: 0.3-3212 ng/mL). In four patients with incomplete venom-induced consumption coagulopathy the median venom concentration was 2.4 ng/mL compared to 30 ng/mL in seven patients with complete venom-induced consumption coagulopathy. No venom was detected in any post-antivenom sample and the median antivenom dose prior to this first post-antivenom sample was 1.5 vials (1-3 vials), including 7 patients administered only 1 vial. In rats the assay distinguished a 3-fold difference in venom dose administered and there was small inter-individual variability. There was small but measurable cross-reactivity with black snake (Pseudechis), tiger snake (Notechis) and rough-scale snake (Tropidechis carinatus) venoms with the assay for low venom concentrations (<1 ng/mL). The use of biotinylation and the antivenom difference method in venom EIA produces a highly sensitive assay that will be useful for determining antivenom dose, forensic and clinical diagnosis.

Impact of hospital-wide infection rate, invasive procedures use and antimicrobial consumption on bacterial resistance inside an intensive care unit.

We performed a 30-month ecological study to determine the impact of hospital-wide antibiotic consumption, invasive procedure use and hospital-acquired infections (HAIs) on antibiotic resistance in an intensive care unit (ICU). Microbiological isolates from ICU patients with established diagnosis of hospital infection were monitored throughout the study. Overall hospital consumption per 100 patient-days of piperacillin-tazobactam, fluoroquinolones and cephalosporins increased from 1.9 to 2.3 defined daily doses (DDD) (P<0.01), from 4.7 to 10.3 DDD (P<0.01) and from 12.1 to 16.4 DDD (P<0.01), respectively. Bacterial multiresistance in ICU was identified in 31.3% (N=466) of isolates, with increasing resistance demonstrated for meropenem-resistant Klebsiella spp. (P=0.01) and meropenem-resistant Acinetobacter spp. (P=0.02). There was a positive correlation between multiresistance rate and DDD of cephalosporins (P<0.01) and fluoroquinolones (P=0.03). The rate of ceftazidime-resistant Klebsiella spp. correlated with DDD of fluoroquinolones and cephalosporins; the rate of ceftazidime-resistant Pseudomonas spp. correlated with consumption of cephalosporins, and rate of meticillin-resistant Staphylococcus aureus (MRSA) correlated with fluoroquinolone use. During the studied period, 36.9% (P<0.001) and 34.5% (P<0.01) of the changing multiresistance rate in ICU was associated with use of invasive procedures and overall HAI rate, respectively. Multiresistance rates in ICU are influenced by the variation in overall HAI rate, hospital-wide invasive procedures and antibiotic consumption outside the ICU.

Renal failure is a risk factor for cefepime-induced encephalopathy.

BACKGROUND: Cefepime is a widely used antibiotic. However, it can cause encephalopathy, which has been increasingly described in the literature, occurring mainly in patients with impaired renal function. The primary objective in this study was to measure the incidence of cefepime-induced encephalopathy and determine potential risk factors for its occurrence. METHODS: In the period from February 2005 to February 2006, a prospective cohort study was conducted, which followed 498 patients using cefepime. Other metabolic problems were ruled out for all patients with clinical suspicion of encephalopathy and, when cefepime was the probable cause, electroencephalographic (EEG) tests were performed to assist in the diagnosis, with the first performed during cefepime use and another performed at least 48 hours following drug discontinuation and/or clinical improvement. RESULTS: Among patients selected for this study (n=498), 5 were diagnosed with cefepime-induced encephalopathy, thus indicating a cumulative incidence of approximately 1% (0.01), 387 had glomerular filtration rate (GFR) >or=60 ml/min and 111 had GFR <60 ml/min. Among the latter, 5 patients developed cefepime-induced encephalopathy. Mean GFR value in patients with encephalopathy (n=5) was 17.20 ml/min (SD +/-10.75 ml/min) and, in patients without encephalopathy (n=106) it was 32.59 ml/min (SD +/-14.89 ml/min) (p=0.025). CONCLUSION: The development of cefepime-induced encephalopathy seems to be related to the severity of impairment in glomerular filtration.

Critical steps in fluoroquinolones and carbapenems prescriptions: results from a prospective clinical audit.

Antibiotic misuse is associated with emergence of resistance and high expenditures. Fluoroquinolones (FQ) and carbapenems (CP) are drugs with considerable potential of resistance development and its disseminated use is a concern. We undertook a prospective clinical audit to evaluate prescriptions of FQ and CP in a multistep process. Each prescription was unfolded in the following steps: indication for antimicrobial therapy; adequacy of initial prescription, dosage and route; previous cultures; and parenteral-oral transition. There was no antibiotics indication in 8.9% of FQ and 1.5% of CP group (p = 0.07). In CP 25.8% of initial schemes were inappropriate (21% in FQ). Lack of switch to oral therapy comprised 25% of monthly costs of FQ. Inadequacy in initial choice accounted for 13.6% of CP expenses. We concluded that, in spite of infection control restrictive policies, inappropriateness of antibiotic usage is worrisome. Clinical audit in a multistep approach may identify possible flaws in this process.

Bonding characteristics to dentin walls of class II cavities, in vitro.

OBJECTIVES: The objective of this study was to test the hypothesis that bond strengths to intact class II cavity wall surfaces are lower than those measured on corresponding flat surfaces isolated by cutting away the rest of the cavity walls. METHODS: Class II (MOD) cavities were prepared in extracted human third molars and then divided into two groups: Intact cavity bonding group or flat surface group. All prepared surfaces were acid-etched and bonded with Scotchbond Multi-Purpose Plus adhesive system and restored with Z100 resin composite. After storage in water for 2 days, the teeth were divided mesio-distally into two equal halves. One half was vertically serially sectioned, while the other half was horizontally serially sectioned to yield a series of 0.5mm thick slabs. Each slab was trimmed into an hour-glass outline with the narrowest cross-sectional area at the region of interest (i.e., axial resin-dentin interface). RESULTS: The mean bond strengths obtained in the cavity bonding group were significantly lower (p<0.05) than those of the flat bonding group. However, within either group, there were no significant consistent differences among the various regions. SIGNIFICANCE: The large flat surfaces used in most laboratory bonding studies may overestimate the bond strengths that can be achieved in complex cavities prepared and restored under clinically relevant conditions.

Treatment of traumatic injuries in the front teeth: restorative aspects in crown fractures.

Crown fractures are the most common form of traumatic dental injuries encountered in permanent dentition. Restorative treatment modalities incorporate adhesive materials to effectively maintain function and aesthetics. While uncomplicated injuries of the enamel and/or dentin can be treated solely with adhesive procedures, complicated trauma that involves pulp exposure requires the incorporation of a multidisciplinary treatment approach. Fragment reattachment is facilitated by the utilization of bonding agents that enhance retention and aesthetics. This article discusses the application of provisional and permanent restorative options for the treatment of complications following traumatic injuries.

Noninvasive mechanical body contouring: (Endermologie) a one-year clinical outcome study update.

LPG Endermologie is a machine-assisted massage system that allows positive pressure rolling, in conjunction with applied negative pressure to the skin and subcutaneous tissues (LPG Endermologie U.S. A. (800-222-3911). Endermologie was originally developed in the late 1970s in France to soften scars and standardize physical therapy; however, patients treated with the LPG machine also showed improvement in body contour and skin texture. Since then, Endermologie machines have been used in France, the United States, and many other nations as an alternative method to altering fat distribution in the subcutaneous plane. The authors have continued their study of determining the safety and efficacy of this machine. Since our last report in March 1997 (Ersek RA et al., Aesth. Plast. Surg. 21(2):61-67, 1997), we have compiled records of 85 additional patients. With this larger patient pool, we can expect more statistically accurate results. This study is composed of 85 women between the ages of 21 to 61. The study group exhibited a wide range of body habitus, initial weights, and final results. Out of 85 patients, 46 patients completed seven sessions of treatment and showed a mean index reduction in body circumference of 1.34 cm, while 39 patients who completed 14 sessions of treatments showed a mean index reduction in body circumference of 1.83 cm. A decrease in mean body circumference index was seen regardless of loss or gain in patients' weight in most cases.

Two protein-tyrosine phosphatases inactivate the osmotic stress response pathway in yeast by targeting the mitogen-activated protein kinase, Hog1.

Protein phosphatases inactivate mitogen-activated protein kinase (MAPK) signaling pathways by dephosphorylating components of the MAPK cascade. Two genes encoding protein-tyrosine phosphatases, PTP2, and a new phosphatase, PTP3, have been isolated in a genetic selection for negative regulators of an osmotic stress response pathway called HOG, for high osmolarity glycerol, in budding yeast. PTP2 and PTP3 were isolated as multicopy suppressors of a severe growth defect due to hyperactivation of the HOG pathway. Phosphatase activity is required for suppression since mutation of the catalytic Cys residue in Ptp2 and Ptp3, destroys suppressor function and biochemical activity. The substrate of these phosphatases is likely to be the MAPK, Hog1. Catalytically inactive Ptp2 and Ptp3 coprecipitate with Hog1 from yeast extracts. In addition, strains lacking PTP2 and PTP3 do not dephosphorylate Hog1-phosphotyrosine as well as wild type. The latter suggests that PTP2 and PTP3 play a role in adaptation. Consistent with this role, osmotic stress induces expression of PTP2 and PTP3 transcripts in a Hog1-dependent manner. Thus Ptp2 and Ptp3 likely act in a negative feedback loop to inactivate Hog1.