Identification and characterization of multiple Spidroin 1 genes encoding major ampullate silk proteins in Nephila clavipes.

Spider dragline silk is primarily composed of proteins called major ampullate spidroins (MaSps) that consist of a large repeat array flanked by nonrepetitive N- and C-terminal domains. Until recently, there has been little evidence for more than one gene encoding each of the two major spidroin silk proteins, MaSp1 and MaSp2. Here, we report the deduced N-terminal domain sequences for two distinct MaSp1 genes from Nephila clavipes (MaSp1A and MaSp1B) and for MaSp2. All three MaSp genes are co-expressed in the major ampullate gland. A search of the GenBank database also revealed two distinct MaSp1 C-terminal domain sequences. Sequencing confirmed that both MaSp1 genes are present in all seven Nephila clavipes spiders examined. The presence of nucleotide polymorphisms in these genes confirmed that MaSp1A and MaSp1B are distinct genetic loci and not merely alleles of the same gene. We experimentally determined the transcription start sites for all three MaSp genes and established preliminary pairing between the two MaSp1 N- and C-terminal domains. Phylogenetic analysis of these new sequences and other published MaSp N- and C-terminal domain sequences illustrated that duplications of MaSp genes may be widespread among spider species.

Design and startup of a membrane-biological-reactor system at a Ford-engine plant for treating oily wastewater.

A wastewater-treatment facility at Ford (Dearborn, Michigan) was recently upgraded from chemical de-emulsification to ultrafiltration (UF) followed by a membrane-biological reactor (MBR). This paper describes the design, startup, and initial operational performance of the facility. Primary findings are as follows: (1) the MBR proved resilient; (2) the MBR removed approximately 90% of chemical-oxygen demand (COD) after primary UF; (3) the removal of total Kjeldahl nitrogen by MBR appeared to be more sensitive to operating conditions than COD removal; (4) nitrification and denitrification were established in one month; (5) the MBR removed oil and grease and phenolics to below detection levels consistently, in contrast to widely fluctuating concentrations in the past; (6) permeate fluxes of the primary and MBR UF were adversely affected by inadvertent use of a silicone-based defoamer; and (7) zinc concentrations in the effluent increased, which might have been a result of ethylenediaminetetraacetic acid used in membrane washing solutions and/or might have been within typical concentration ranges.

Literature review--efficacy of various disinfectants against Legionella in water systems.

There have been reported outbreaks of Legionnaires' disease at hospitals and industrial facilities, which prompted the development of various preventive measures. For example, Ford has been developing and implementing such a measure at its facilities worldwide to provide technical guidance for controlling Legionella in water systems. One of the key issues for implementing the measure is the selection of a disinfectant(s) and optimum conditions for its use. Therefore, available publications on various disinfectants and disinfection processes used for the inactivation of Legionella bacteria were reviewed. Two disinfection methods were reviewed: chemical and thermal. For chemical methods, disinfectants used were metal ions (copper and silver), oxidizing agents (halogen containing compounds [chlorine, bromine, iodine, chlorine dioxide, chloramines, and halogenated hydantoins], ozone, and hydrogen peroxide), non-oxidizing agents (heterocyclic ketones, guanidines, thiocarbamates, aldehydes, amines, thiocyanates, organo-tin compounds, halogenated amides, and halogenated glycols), and UV light. In general, oxidizing disinfectants were found to be more effective than non-oxidizing ones. Among oxidizing agents, chlorine is known to be effective and widely used. Among non-oxidizing agents, 2,2-dibromo-3-nitropropionamide appears to be the most effective followed by glutaraldehyde. Isothiazolin (known as Kathon), polyhexamethylene biguanide, and 2-bromo-2-nitropropionamide (known as Bronopol) were found to be less effective than glutaraldehyde. Thermal disinfection is effective at > 60 degrees C (140 degrees F).

The influence of supporting the Oxylog instrument on estimated maximal aerobic capacity during a step test and heart rate in a lifting test.

This study investigated the influence of wearing, an Oxylog mask and heart rate monitor while the Oxylog instrument was supported in a stand versus that of wearing the Oxylog mask, heart rate monitor, and the Oxylog instrument, on oxygen consumption (VO2) and heart rate response during a graduated submaximal step test protocol. Also, the predicted maximal aerobic capacities ( VO2max) produced by the two graduated step tests were compared. In addition, differences in the working heart rates in a submaximal lifting test were analyzed in two lifting tests, one with the participant wearing the Oxylog mask, heart rate monitor, and Oxylog instrument and the other required the participant to only wear a heart monitor. Seventeen experienced male manual materials handlers participated in the study, and each treatment was seen by each participant in a randomized Latin Square design. Results from the two investigations indicate that there was no significant difference in the estimated maximal oxygen consumption (p = 0.1384) and no significant difference in the heart rate between the two lift tests. The analysis did show that the 4th stage (participants reaching their physiological limits) of the step tests indicated a statistically significant difference (p = 0.0259 for oxygen consumption, and p = 0.0465 for heart rate).

Factors affecting minimum push and pull forces of manual carts.

The minimum forces needed to manually push or pull a 4-wheel cart of differing weights with similar wheel sizes from a stationary state were measured on four floor materials under different conditions of wheel width, diameter, and orientation. Cart load was increased from 0 to 181.4 kg in increments of 36.3 kg. The floor materials were smooth concrete, tile, asphalt, and industrial carpet. Two wheel widths were tested: 25 and 38 mm. Wheel diameters were 51, 102, and 153 mm. Wheel orientation was tested at four levels: F0R0 (all four wheels aligned in the forward direction), F0R90 (the two front wheels, the wheels furthest from the cart handle, aligned in the forward direction and the two rear wheels, the wheels closest to the cart handle, aligned at 90 degrees to the forward direction), F90R0 (the two front wheels aligned at 90 degrees to the forward direction and the two rear wheels aligned in the forward direction), and F90R90 (all four wheels aligned at 90 degrees to the forward direction). Wheel width did not have a significant effect on the minimum push/pull forces. The minimum push/pull forces were linearly proportional to cart weight, and inversely proportional to wheel diameter. The coefficients of rolling friction were estimated as 2.2, 2.4, 3.3, and 4.5 mm for hard rubber wheels rolling on smooth concrete, tile, asphalt, and industrial carpet floors, respectively. The effect of wheel orientation was not consistent over the tested conditions, but, in general, the smallest minimum push/pull forces were measured with all four wheels aligned in the forward direction, whereas the largest minimum push/pull forces were measured when all four wheels were aligned at 90 degrees to the forward direction. There was no significant difference between the push and pull forces when all four wheels were aligned in the forward direction.

Effectiveness of Waddell's nonorganic signs in predicting a delayed return to regular work in patients experiencing acute occupational low back pain.

STUDY DESIGN: Consecutive case series. OBJECTIVE: To determine whether the presence of Waddell's nonorganic signs in occupational, acute low back pain patients predicts a longer time before return to unrestricted regular work. SUMMARY OF BACKGROUND DATA: Waddell's nonorganic signs identify patients with chronic low back pain with a poor prognosis; however, they have not been used as an outcome predictor in patients with occupational, acute low back pain. METHODS: Standardized histories and physical examinations, including Waddell's signs were recorded at the first clinic visit (n = 143) by a single physician who was not blinded to the study's goals or methods. Those with chronic low back pain or complicating medical conditions (n = 88) were excluded. Data from patients exhibiting the nonorganic signs were compared with those from patients without the signs for time to return to regular work without restrictions and medical resource use. RESULTS: Fifty-five patients with acute work-related low back pain were included. One or more of the nonorganic signs were seen in 14 patients (25.5%) at the first appointment. The most common signs were simulated axial loading (78.6%) and simulated rotation (71.4%). Those with any nonorganic sign required a median 58.5 days to return to regular work compared with 15.0 days for those without (P < 0.0001). Patients exhibiting any nonorganic sign compared with those without used more physical therapy (50.0% vs. 12.2%; P < 0.01) and lumbar computed axial tomography (21.4% vs. 0.0%; P = 0.01). CONCLUSIONS: Patients with acute, occupational low back pain exhibiting Waddell's nonorganic signs had a four times lengthier time for return to unrestricted, regular work and a greater use of physical therapy and lumbar computed tomographic scans.

Respiratory allergenicity of detergent enzymes in the guinea pig intratracheal test: association with sensitization of occupationally exposed individuals.

A guinea pig intratracheal test was used to set occupational operating guidelines for new enzyme proteins used in the detergent industry. In these studies, animals were intratracheally dosed with different levels of enzyme protein and sera from the animals were titered for allergic antibody to the enzyme. The amount of antibody produced to an enzyme was compared to the amount of antibody produced to the same protein doses of Alcalase, for which effective operating guidelines exist. These comparisons were used to determine if a new enzyme was more potent, less potent, or equivalent to Alcalase; operating guidelines were then established for the new enzyme. Termamyl was about 10-fold more potent than Alcalase and the protease subtilisin B was shown to be less potent. Another protease, Savinase, was shown to be equivalent in potency to Alcalase. The operating guidelines for Termamyl were adjusted lower, whereas the operating guidelines for the proteases were set the same as that of Alcalase. Under these conditions, we would predict that sensitizations to new enzymes would be comparable to or lower than the sensitizations to Alcalase. Prospective evaluation of skin prick test data of factory workers showed that sensitizations to Termamyl and Savinase were similar to sensitizations to Alcalase. The sensitizations to subtilisin B were lower than those to Alcalase. During this time period (7 years), only three respiratory incidents (rhinitis) were reported, demonstrating that employees with positive skin prick tests can continue to work. These comparisons indicate that the guinea pig intratracheal test is a good animal model for evaluating enzymes as respiratory allergens and that the data generated can be used to set operating guidelines for occupational allergens.

Characterization of skin prick testing responses for detecting sensitization to detergent enzymes at extreme dilutions: inability of the RAST to detect lightly sensitized individuals.

We observed that a group of detergent enzyme workers with known exposure to the subtilisin enzyme, Alcalase (Novo Industries, Bagsvaerde, Denmark), exhibited percutaneous sensitivity to Savinase (Novo Industries), a microbial protease, to which there was no previous occupational exposure. This was attributed to either cross-reactivity between these enzymes or to foreign enzyme contaminants contained in the Savinase antigen. The aims of this study were to determine the range of concentrations eliciting percutaneous responses to Alcalase and to another enzyme, Rapidase (an alpha-amylase) (Gist Brocades, Belgie, Netherlands); to compare the sensitivity of RAST and skin prick testing; and to characterize the relationship between wheal size and antigen concentration. Prick testing was conducted over six log10 antigen dilutions of Alcalase and Rapidase in 30 workers with previous exposure and skin reactivity to enzymes (group 1) and compared to nonexposed control groups, which included 60 atopic subjects (group 2) and 30 nonatopic subjects (group 3). The RAST was performed with Alcalase and Rapidase antigens. The percutaneous threshold concentrations in group 1 subjects varied widely from 10(3) to 10(-3) micrograms of protein per milliliter. Of 19 group 1 workers with skin test reactivity to Alcalase, 84% had positive RAST results; 83% of 24 workers who were reactive to Rapidase had positive RAST results. It was concluded that skin prick testing is preferred over in vitro methods for longitudinal monitoring of human sensitization to workplace allergens. In addition, the data predicted that based on a known Alcalase level of 0.07% in Savinase, 26% of Alcalase-sensitized subjects could react to Savinase. An excellent correlation (r > 0.97) was found between log concentration of antigen and wheal size parameters, with the log diameter and log area performing equally as well (r > 0.98). Analysis of variance revealed that more than 60% of intragroup variation represented human variability in wheal parameters at each concentration tested, whereas at least 95% of intergroup variation was due to regression. The excellent correlations of both wheal diameter and area with antigen concentrations were attributed to the very small changes observed between test concentrations.

Chronic type A aortic dissection: an unusual complication of cocaine inhalation.

Acute aortic pathology temporally related to cocaine inhalation may lead to frank rupture or acute aortic dissection. This is a report of an unusual case of a 43-year-old man who presented 9 weeks after experiencing a tearing sensation in his chest while smoking cocaine. The diagnosis was chronic type A aortic dissection with 4+ aortic insufficiency. The successful surgical management included resuspension of the aortic valve and placement of a Dacron tube graft in the ascending aorta such that flow was maintained distally in both the true and false lumens.

Management of an innominate artery aneurysm during an open heart operation.

A 76-year-old woman was found to have a 4 x 2.5-cm saccular aneurysm at the origin of the innominate artery at the time of a reoperative open heart operation. The operative procedure was modified to include repair of the aneurysm with a Dacron patch. During the period of innominate artery occlusion, the patient was cooled to 25 degrees C and the mean arterial pressure was maintained at 90 mm Hg to maximize cerebral protection.

Perioperative right heart failure: treatment.

When right ventricular failure prohibits separation from cardiopulmonary bypass, standard methods of increasing pulmonary blood flow should be employed, including correction of hypoxia and acidosis, volume loading, and inotropic support of the right ventricle. Infusion of pulmonary vasodilators--particularly low-dose nitroprusside--should be used if the pulmonary vascular resistance is elevated. If pulmonary blood flow remains unsatisfactory, systemic intra-aortic balloon counterpulsation should be employed. In refractory cases, right-atrial-to-pulmonary-artery bypass using the Bio-Medicus centrifugal pump is the recommended therapy for those centers that do not have the Pierce-Donachy pneumatic ventricular assist-pump available. For refractory right ventricular failure following the surgical repair of congenital cardiac defects in which the placement of right atrial and pulmonary artery cannulae is not technically feasible, use of high-frequency high-volume ventilation appears to be quite promising. Utilization of pulmonary artery counterpulsation or the creation of an atrial septal defect may also be lifesaving, but is not recommended as the therapy of choice.

Pulmonary circulatory support. A quantitative comparison of four methods.

Profound right ventricular failure was produced in 16 goats by inducing ventricular fibrillation after the systemic circulation had been supported with a left atrial-aortic bypass pump. In each animal, four methods of providing pulmonary blood flow were compared quantitatively: passive flow through the pulmonary artery due to a right atrial to left atrial pressure gradient; pulmonary artery pulsation via a 40 ml intra-aortic type balloon within a 20 mm Dacron graft anastomosed to the main pulmonary artery; pulmonary artery pulsation via a 65 ml single-port, valveless, sac type pulsatile assist device; and right atrial-pulmonary arterial bypass via a valved pneumatic pulsatile pump. Average cardiac index of the 16 animals for each method was 31.1 +/- 12.9, 44.4 +/- 13.6, 64.3 +/- 16.9, and 102.0 +/- 20.7 ml/min/kg, respectively. Passive pulmonary artery flow alone provided inadequate pulmonary circulatory support. Addition of pulmonary artery pulsation via the intra-aortic balloon within a conduit increased cardiac index 13.3 ml/min/kg (43%) above passive pulmonary artery flow (p less than 0.0005); however, the cardiac index remained inadequate. Increasing pulmonary artery pulsation volume with a 65 ml sac device provided a 32.2 ml/min/kg (106%) increase in cardiac index above passive flow (p less than 0.0005) to a level that was marginally adequate. The valved right atrial-pulmonary arterial bypass pump increased cardiac index 70.9 ml/min/kg (228%) above passive pulmonary artery flow (p less than 0.0005) to a satisfactory level and is the recommended method of pulmonary circulatory support in profound right ventricular failure.

Inhibition of ionic transport and ATPase activities by serotonin analogues in the isolated toad lens.

The effects of serotonin and five other indoles were tested on the electrical parameters and ionic transport in the isolated toad lens. Serotonin, tryptophan and 5-hydroxy-L-tryptophan did not affect the electrical parameters of the lens at concentrations as high as 1 mM. Tryptamine, 5-methyltryptamine and 5-methoxytryptamine had dual effects: 1 mM in the posterior bathing solution depressed the potential difference of the posterior face of the lens, which resulted in an increase in the translenticular potential difference and short-circuit current; 1 mM in the anterior solution (in contact with the lens epithelium) produced a quick and pronounced reduction of the potential difference of the anterior face. This resulted in a 90-100% decline of the translenticular short-circuit current. Serotonin and tryptamine were then tested for their effect on the ATPases of lens epithelium. Both amines inhibited the enzymes with tryptamine at 5 mM completely inhibiting all ATPase activity. Since tryptophan is transported from the aqueous humor into the lens and may be converted by lens enzymes to serotonin and tryptamine, these findings may have physiological implications in cataractogenesis.