Design and application of water-in-oil emulsions for use in lipstick formulations.

The addition of water to lipsticks in the form of a water-in-oil emulsion is an attractive opportunity for cosmetics manufacturers to deliver hydrophilic molecules to the consumers, as well as improving the moisturizing properties. In this work, the effect of the emulsifier type and water content on the structural properties of the designed products was investigated. It has been shown that PGPR leads to smaller droplets than the other emulsifiers tested. This was attributed to the ability of PGPR to form elastic interfaces that slow the coalescence between droplets during the process. It was also observed that crystals of wax tend to form structures at the interface upon cooling that prevent coalescence during storage. These structures also prevent leakage of water into the continuous phase. No effect of the water content on the melting properties of the emulsions was observed. Upon addition of more than 10% water, softening of the material was measured, due to the overall decrease in solid content. Addition of crystalline material (hard paraffin) was successfully used to reinstate the material properties.

Discovery of entry inhibitors for HIV-1 via a new de novo protein design framework.

A new (to our knowledge) de novo design framework with a ranking metric based on approximate binding affinity calculations is introduced and applied to the discovery of what we believe are novel HIV-1 entry inhibitors. The framework consists of two stages: a sequence selection stage and a validation stage. The sequence selection stage produces a rank-ordered list of amino-acid sequences by solving an integer programming sequence selection model. The validation stage consists of fold specificity and approximate binding affinity calculations. The designed peptidic inhibitors are 12-amino-acids-long and target the hydrophobic core of gp41. A number of the best-predicted sequences were synthesized and their inhibition of HIV-1 was tested in cell culture. All peptides examined showed inhibitory activity when compared with no drug present, and the novel peptide sequences outperformed the native template sequence used for the design. The best sequence showed micromolar inhibition, which is a 3-15-fold improvement over the native sequence, depending on the donor. In addition, the best sequence equally inhibited wild-type and Enfuvirtide-resistant virus strains.

New compstatin variants through two de novo protein design frameworks.

Two de novo protein design frameworks are applied to the discovery of new compstatin variants. One is based on sequence selection and fold specificity, whereas the other approach is based on sequence selection and approximate binding affinity calculations. The proposed frameworks were applied to a complex of C3c with compstatin variant E1 and new variants with improved binding affinities are predicted and experimentally validated. The computational studies elucidated key positions in the sequence of compstatin that greatly affect the binding affinity. Positions 4 and 13 were found to favor Trp, whereas positions 1, 9, and 10 are dominated by Asn, and position 11 consists mainly of Gln. A structural analysis of the C3c-bound peptide analogs is presented.

Seasonal patterns of traumatic brain injury deaths due to traffic-related incidents in the Slovak Republic.

Objective: Traumatic Brain Injuries (TBIs) are an important type of injury in terms of both morbidity and mortality. Road Traffic Incidents are one of the most frequent causes of TBI. This analysis seeks to quantify the number of such injuries occurring in the Slovak Republic, and examine patterns of TBI according to mode of transport and seasonality.Methods: Data concerning total numbers of TBIs occurring from the years 1996-2015 were obtained from the Statistical Office of the Slovak Republic. The events caused by road incidents were examined separately according the external cause stated on death certification. Events were classified into seasons according to the month of death. Summary statistics were produced concerning numbers of deaths according to sex, mode of transport and season. Analyses were performed to examine trends in TBI by season and type of road user.Results: During a period of 20 years from 1996, there were 17,047 recorded deaths involving TBI in the Slovak Republic. Of these, 5,370 were caused by road traffic incidents (RTIs). Age standardized rates tended to decrease from 8.3/100,000/year (1996) to 2.5/100,000/year (2015). Males made up approximately 79% of road traffic-caused TBIs. Summer and autumn showed significantly more events than any other season, with motorcyclists and cyclists in particular being more frequently injured at this time of year.Conclusions: The results show that Slovakia, like many countries, suffers a considerable burden of TBI and that RTIs are a major contributor to this, especially among young adults. Rates of TBI vary by season in Slovakia, and users of different modes of transport appear more or less likely to suffer such injury during different seasons. Considerable variability in rates of injury exists between road users and times of year. Improved understanding of the timing and sufferers of injuries may allow better planning of response and care services. Further research into transport modes and policies aimed at safer driving should be explored.

Dietary calcium has little effect on mineral balance and bone mineral metabolism through twenty weeks of lactation in Holstein cows.

Calcium and P balance and mobilization from bone were evaluated through 20 wk of lactation to determine the timing and extent of net resorption of bone mineral and mineral balance in lactating dairy cows. Eighteen Holstein cows were blocked by parity and calving date and randomly assigned to 1 of 3 dietary treatments: high (1.03%, HI), medium (0.78%, MED), or low (0.52%, LOW) dietary Ca. Dietary P was 0.34% in all diets. Cows consumed treatment diets from calving to 140 DIM. Total collection of milk, urine, and feces was conducted 2 wk before expected calving and in wk 2, 5, 8, 11, and 20 of lactation. Blood samples were collected at 14 and 10 d before expected calving and 0, 1, 3, 5, 10, 14, 21, 28, 35, 56, 70, 84, 98, and 140 d after calving. Blood samples were analyzed for Ca, P, and parathyroid hormone concentration. Serum concentrations of osteocalcin (OC), a marker of bone formation, and deoxypyridinoline (DPD), a marker of bone resorption, were measured to assess bone mobilization. Rib bone biopsies were conducted within 10 d postcalving and during wk 11 and 20 of lactation. Dietary Ca concentration affected Ca balance, with cows consuming the HI Ca diet in positive Ca balance for all weeks with the exception of wk 11. Interestingly, all cows across all treatments had a negative Ca balance at wk 11, possibly the result of timed estrous synchronization that occurred during wk 11. At wk 20, Ca balances were 61.2, 29.9, and 8.1 g/d for the HI, MED, and LOW diets, respectively. Phosphorus balances across all treatments and weeks were negative. Bone Ca content on a fat-free ash weight basis was least in cows consuming the MED diet, but bone P was not different. Serum Ca and P were not affected by treatment. Dietary Ca concentration did not affect P balance in the weeks examined, but there was a clear effect of parity on balance, markers of bone metabolism, and bone P. Primiparous cows had greater serum OC and DPD concentrations than multiparous cows. Regardless of dietary treatment, serum OC concentration peaked around d 35 of lactation. Simultaneously, DPD concentration began to decrease, which may indicate a switch from net bone resorption to formation after d 35. However, this was not reflected in balance measures. This information may help refine dietary mineral recommendations for lactating dairy cows and suggests that dietary P requirements are independent of dietary Ca.

Development and evaluation of a real-time PCR assay for detection of Pneumocystis jirovecii DNA in bronchoalveolar lavage fluid of HIV-infected patients.

BACKGROUND: Pneumocystis pneumonia (PCP) is conventionally diagnosed by identifying Pneumocystis jirovecii in lower respiratory tract samples using cytochemical stains. Molecular diagnosis of PCP is potentially more sensitive. METHODS: A study was undertaken to use an extensively optimised real-time polymerase chain reaction (PCR) using primers designed to hybridise with the P. jirovecii heat shock protein 70 (HSP70) gene to quantify P. jirovecii DNA in bronchoalveolar lavage (BAL) fluid from HIV-infected patients with and without PCP, and to compare this assay with conventional PCR targeting the P. jirovecii mitochondrial large subunit rRNA gene sequence (mt LSU rRNA). RESULTS: Sixty-one patients had 62 episodes of PCP (defined by detection of P. jirovecii in BAL fluid by cytochemical stains and typical clinical presentation). Quantifiable HSP70 DNA was detected in 61/62 (range approximately 13-18,608 copies/reaction; median approximately 332) and was detectable but below the limit of quantification (approximately 5 copies/reaction) in 1/62. Seventy-one other patients had 74 episodes with alternative diagnoses. Quantifiable HSP70 DNA was detectable in 6/74 (8%) episodes (range approximately 6-590 copies/reaction; median approximately 14) and detectable but below the limit of quantification in 34/74 (46%). Receiver-operator curve analysis (cut-off >10 copies/reaction) showed a clinical sensitivity of 98% (95% 91% to 100%) and specificity of 96% (95% CI 87% to 99%) for diagnosis of PCP. By contrast, clinical sensitivity of mt LSU rRNA PCR was 97% (95% CI 89% to 99%) and specificity was 68% (95% CI 56% to 78%). CONCLUSION: The HSP70 real-time PCR assay detects P. jirovecii DNA in BAL fluid and may have a diagnostic application. Quantification of P. jirovecii DNA by real-time PCR may also discriminate between colonisation with P. jirovecii and infection.

Blood mineral, hormone, and osteocalcin responses of multiparous Jersey cows to an oral dose of 25-hydroxyvitamin D3 or vitamin D3 before parturition.

Twenty-seven multiparous Jersey cows were randomly assigned to receive an oral bolus containing corn starch (control, CON), corn starch plus 15 mg of 25-hydroxyvitamin D(3) (25-OH), or 15 mg of cholecalciferol (D(3)) at 6 d before expected parturition. Cows were maintained in individual box stalls from 20 d before expected parturition and fed a common diet. Jugular blood samples were collected at -14, -13, -5, -4, -3, -2, -1 d before expected calving, at calving, and at 1, 3, 5, 7, 9, 11, 13, 28, 56, and 84 d postcalving. After calving, cows were housed in 1 pen in a free-stall barn and consumed a common diet. Colorimetric assays were used to analyze Ca, P, and Mg concentrations in serum. Serum concentrations of osteocalcin (OC), an indicator of bone formation, serum 25-hydroxyvitamin D(3), and parathyroid hormone (PTH) were determined in samples obtained from d -5 through d 13. The 9 control multiparous cows and 5 untreated primiparous cows were used to evaluate the effect of parity on the variables that were measured. There was no effect of parity on Ca, PTH, or 25-OH concentration. Compared with second-lactation cows and older cows (>2 lactations), first-lactation cows had greater serum OC (22.3, 32.0, and 48.3 ng/mL, respectively), indicating that younger animals were forming more bone. Blood Ca, P, and Mg decreased near the time of calving and then increased over time. Serum 25-hydroxyvitamin D(3) was greater for cows dosed with 25-OH (119.0 ng/mL) compared with those dosed with D(3) (77.5 ng/mL) or CON (69.3 ng/mL). Cows dosed with 25-OH tended to have lower serum PTH concentration, but treatments did not affect serum Ca, P, or Mg. Serum OC was greater in second-lactation cows compared with cows entering their third or fourth lactation but OC was unaffected by treatment. Although results indicated a 60% increase in serum 25-hydroxyvitamin D(3) due to a single oral dose of 25-OH before calving, the amount administered in this study apparently was not sufficient for initiation of any improvement in Ca homeostasis at parturition.

Manure nutrient excretion by lactating cows fed exogenous phytase and cellulase.

The effect of an exogenous phytase and cellulase-containing enzyme formulation on nutrient digestibility and excretion was evaluated in 24 Holstein cows. Cows were fed corn silage- and alfalfa silage-based diets with or without a cellulase-phytase blend for 31 d in a continuous random design. Treatment groups were balanced for parity, days in milk, and mature-equivalent projected milk yield. Diets contained 37% forage, 18.3% crude protein, 35.4% neutral detergent fiber, 18% acid detergent fiber, and 0.42% P (no supplemental P). Cows were fed once daily in Calan doors and milked 2 times daily. Body weight and milk yield were recorded at each milking. Milk samples were collected on d 28 to 31 at 8 consecutive milkings. On d 28 to 31, fecal grab samples were collected every 8 h, with sampling times advanced by 2 h each day. Feces samples were pooled by cow. Feed and feces samples were analyzed for acid detergent lignin (used as an internal marker) and for N, P, neutral detergent fiber, and acid detergent fiber. Days in milk were similar between treatments, and body weight and milk yield were unaffected by treatment. Cows fed the enzyme formulation had reduced fecal dry matter, neutral detergent fiber, and acid detergent fiber excretion and reduced fecal excretion of N and P. Apparent digestibility of dry matter, acid detergent fiber, neutral detergent fiber, and N tended to increase with the enzyme formulation. Addition of an exogenous phytase and cellulase enzyme formulation to diets for lactating cows reduced fecal nutrient excretion.

Characterization and comparative analysis of the EGLN gene family.

Rat Sm-20 is a homologue of the Caenorhabditis elegans gene egl-9 and has been implicated in the regulation of growth, differentiation and apoptosis in muscle and nerve cells. Null mutants in egl-9 result in a complete tolerance to an otherwise lethal toxin produced by Pseudomonas aeruginosa. This study describes the conserved Egl-Nine (EGLN) gene family of which rat SM-20 and C. elegans Egl-9 are members and characterizes the mouse and human homologues. Each of the human genes (EGLN1, EGLN2 and EGLN3) are of a conserved genomic structure consisting of five coding exons. Phylogenetic analysis and domain organization show that EGLN1 represents the ancestral form of the gene family and that EGLN3 is the human orthologue of rat Sm-20. The previously observed mitochondrial targeting of rat SM-20 is unlikely to be a general feature of the protein family and may be a feature specific to rats. An EGLN gene is unexpectedly found in the genome of P. aeruginosa, a bacterium known to produce a toxin that acts through the Egl-9 protein. The pathogenic bacterium Vibrio cholerae is also shown to have an EGLN gene suggesting that it is an important pathogenicity factor. These results provide new insights into host-pathogen interactions and a basis for further functional characterization of the gene family and resolve discrepancies in annotation between gene family members.

Vascular reactivity following ischemia/reperfusion.

Data in the literature supports the hypothesis that reactive oxygen species generated in the vascular wall alter vascular regulation. At present the majority of the literature tends to suggest that oxidant induced damage on the smooth muscle cell impair vasoconstriction. However, direct action of oxidants on the smooth muscle cell impair vasoconstrictor function. Differences in studies in the literature are likely to be reconciled when the target sites of reactive oxygen species are considered. Future research in this area should lead to a more comprehensive understanding on the impact of these pathways on vasoregulation in postischemic tissue.

Synthesis and characterization of self-catalyzed poly(ortho ester).

Poly(ortho esters) are currently under investigation as a carrier system for an antiproliferative agent in glaucoma filtering surgery. The present investigation illustrates the development of a series of self-catalyzed poly(ortho ester). These polymers contain short dimer segments of alpha-hydroxy acids in their backbone and are prepared by the addition of different polyols to the diketene acetal 3,9-diethylidene-2,4,8,10-tetra-oxaspiro-[5.5]-undecane. The structures were confirmed by NMR- and FT-IR-spectroscopy. The polymers were characterized by determination of the molecular weight, the glass transition temperature and the rheological behavior. The amount of residual solvents was also analyzed. The characteristics of the polymer can be varied by the type of polyol incorporated in its backbone. Since poly(ortho ester) is susceptible to acid-catalyzed degradation, the polymer hydrolysis can be controlled by the amount of incorporated portion of alpha-hydroxy acid. Due to the high hydrophobicity of the polymer structure, the ester bonds are more susceptible to hydrolysis than the ortho ester bonds in the polymer backbone. The hydrolysis proceeds via initial protonation of the exocyclic alkoxy group to yield pentaerythritol dipropionate and the free diol. In a next step, the pentaerythritol dipropionate hydrolysis to pentaerythritol and propionic acid. The molecular weight decrease, weight loss and the pH profile of the polymer in aqueous medium were monitored during the degradation.

Assessing public health practice: application of ten core function measures of community health in six states.

Factor analysis, combined with an evaluation of item difficulty and discrimination, can provide useful insights in questionnaire development. In 1993, as part of a study to develop a questionnaire to assess performance of the core functions of public health at a community level, 370 local health departments (LHDs) in six states completed a 26-item questionnaire (94% response). This study describes factor analysis results after controlling for item difficulty and discrimination. Fifteen items had intermediate difficulty and fair-to-good discrimination. Factor analysis of these 15 items identified four factors. Three of these factors included items from more than one of the core functions. These findings pose an interesting question for future research: are the core functions of public health better conceived as three discrete, distinguishable factors or as three interlocking factors that form a single, seamless unit?

Effect of 4 wk of deep water run training on running performance.

The purpose of this study was to determine whether trained competitive runners could maintain on-land running performance using 4 wk of deep water run training instead of on-land training. Eleven well-trained competitive runners (10 males, 1 female; ages, 32.5 +/- 5.4 yr; height, 179.8 +/- 9.3 cm; weight, 70.4 +/- 6.7 kg (mean +/- SD)) trained exclusively using deep water run training for 4 wk. Subjects trained 5-6 d.wk-1 for a total of 20-24 sessions (mean +/- SD, 22 +/- 1.5 sessions). Instruction and practice sessions were conducted prior to the training period. Before and after the deep water run training, subjects completed a 5-km race on the treadmill using a computer based system, a submaximal run at the same absolute workload to assess running economy, and a combined lactate threshold and maximal oxygen consumption test. No significant differences were found for (mean +/- SEM): 5-km run time (pre, 1142.7 +/- 39.5 s; post, 1149.8 +/- 36.9 s; P = 0.28), submaximal oxygen consumption (pre 44.8 +/- 1.2 mL.kg-1.min-1; post, 45.3 +/- 1.5 mL.kg-1.min-1; P = 0.47), lactate threshold running velocity (pre, 249.1 +/- 0.9 m.min-1; post, 253.6 +/- 6.3 m.min-1; P = 0.44), or maximal oxygen consumption (pre, 63.4 +/- 1.3 mL.kg-1.min-1; post, 62.2 +/- 1.3 mL.kg-1.min-1; P = 0.11). Also no differences were found among Global Mood State pre-training, each week during training, and post-training. Competitive distance runners maintained running performance using 4 wk of deep water run training as a replacement for on-land training.

Examination of the additivity assumption using the spiral and standard Salmonella assays to evaluate binary combinations of mutagens.

Binary combinations of pure chemicals and complex mixtures were evaluated for their ability to produce additive mutagenicity responses in the spiral and standard Salmonella mutagenicity assays. Single chemicals were selected that were representative of the primary chemical class responsible for much of the mutagenic activity of each complex mixture. The following agents were evaluated in the absence of S9: 1-nitropyrene, diesel exhaust extract, and the chlorinated drinking water mutagen 3-chloro-4-dichloromethyl-5-hydroxy-2-[5H]-furanone (MX). In the presence of S9, the following agents were evaluated: 4-aminobiphenyl, benzo[a]pyrene, and an organic extract from the particulate emissions resulting from the combustion of polyethylene in a rotary kiln incinerator. Binary combinations of the agents within each S9 group were tested. The results were analyzed for additivity by determining whether the difference between the expected response of the binary mixture was significantly different from the observed response. In general, the combinations of mutagens produced additive responses at low doses in both assays. However, at high doses, the results generally departed from additivity, especially for combinations of indirect-acting mutagens. A requirement for exogenous metabolic activation imposes conditions, such as different S9 optima for each mutagen, that complicate the application of the additivity assumption to binary combinations of indirect-acting mutagens. Although the two bioassays generally gave similar results for each binary combination of mutagens, the spiral assay permitted a gradient of mutagen and/or S9 concentrations to be evaluated, and it allowed the mutagens to be evaluated in the absence of solvents, eliminating any potential solvent-mutagen interactions. These features of the spiral assay may be advantageous when studying chemical interactions.

The role of distraction in improving the space available in the lumbar stenotic canal and foramen.

STUDY DESIGN: The terminal event in disc space narrowing and facet subluxation of the lumbar spine can produce clinical symptoms of spinal stenosis. Reversal of this process via distraction was performed on ten cadaveric motion segments with documented stenosis. METHODS: Computerized tomography and caliperic methods for the measurement of canal and foraminal areas were calculated in each segment after 5 and 10 mm of symmetrical distraction. Measurements were done in a blinded manner verified with orthographic software and statistical analysis. RESULTS: Decompression of foraminal space was statistically significant in 7 of 10 cadaveric specimens after 5 mm of distraction and 9 of 10 specimens after 10 mm of distraction. Minimal yet insignificant improvement in stenotic canal area was evident with distraction. CONCLUSION: The presence of posterior vertebral osteophytes was associated uniformly with poor improvement of space available in both stenotic canal and foramen.

The effect of back belt use on isometric lifting force and fatigue of the lumbar paraspinal muscles.

STUDY DESIGN: A repeated measures multivariate design to assess the effect of back belts on isometric lifting force and lumbar paraspinal muscle fatigue. OBJECTIVE: To determine the efficacy of lumbar back belts in minimizing a loss in isometric force production and fatigue of lumbar paraspinal muscles. SUMMARY OF BACKGROUND DATA: Increased prevalence of back belts in industry and lack of scientific evidence supporting them suggested additional research was needed. There has been no conclusive research to substantiate manufacturer claims that use of belts reduces fatigue of lumbar paraspinals. METHODS: Twenty-four subjects, 13 men and 11 women, were tested twice: once with and once without a belt. All subjects performed a dynamic lifting task at a rate of 10 lifts per minute for 20 minutes. Surface electromyograms from the lumbar paraspinals and isometric force measurements were recorded during a static maximal isometric lift at 0, 10, and 20 minutes. Load during the dynamic task was determined as 20% of initial maximum isometric force. Electromyographic data were used to detect changes in median power spectral frequencies. Muscular fatigue was indicated by a reduction in median power spectral frequency values or a decrease in isometric force-generating capacity. RESULTS: Multivariate and univariate analyses suggest that there is no effect on lumbar paraspinal muscle fatigue or isometric lifting force production as a result of back belt use. CONCLUSIONS: These findings do not support the use of back belts for the purpose of minimizing either lumbar paraspinal muscle fatigue or a loss in isometric lifting force production.

Co-amplification of the amelogenin and HLA DQ alpha genes: optimization and validation.

An optimized PCR-based system allowing the co-amplification of the HLA DQ alpha and the amelogenin genes has been developed and validated, enabling simultaneous identity testing and sex determination. Sensitivities below 100 pg of human DNA were obtained, using a convenient, high resolution agarose gel system and ethidium bromide staining. Comparison of several co-amplification methods revealed that the best sensitivities and most consistent results were obtained using a hotstart technique employing an inactivating antibody to Taq polymerase. HLA DQ alpha typing results were reliably obtained using the co-amplification process. The sensitivity and ease of this system rendered it directly applicable to forensic analyses. The optimized techniques described here have been validated and successfully applied to forensic cases including People vs. Trujillo, in which the California Superior Court accepted these techniques as scientifically reliable and admissible. Work currently in progress has demonstrated that the described protocol may also be used to co-amplify the amelogenin gene with the AmpliType PM (polymarker) system, allowing identity testing at six loci in addition to sex determination.

On the power of a method for comparing survival of trauma patients to a standard survival curve.

This paper investigates the power of a statistical procedure for comparing a unit's survival experience to a standard survival curve. The power of the statistical procedure provides a measure of how sensitive the test is to departures from the nominal values provided by the standard survival curve. This procedure has been used recently in a number of evaluation of care studies. A power study for thirty-two adult blunt trauma patients, contributed by a participating hospital to a national data base collected at Washington Hospital Center, is detailed.

Lumbar epidural sympathectomy for frostbite injuries of the feet.

Fourteen patients with pain and paresthesias secondary to frostbite injury were treated with lumbar sympathetic blockade. The majority of patients had an excellent response to 48 hours of continuous epidural blockade with no recurrence of symptoms after blockade. Those patients who had a limited response to epidural blockade responded well to surgical lumbar sympathectomy. Lumbar sympathetic blockade is a safe and effective technique for treating the symptoms associated with frostbite injury.

Cold weather injuries during peacetime military training.

Demographic data on 220 cold weather injuries seen over a 52-month period at the 67th Evacuation Hospital in Wuerzburg, Germany, was reviewed. Data were collected at the time of presentation and all diagnoses were made by a general/vascular surgeon. Statistics on age, gender, race, rank, unit, prior injury, use of tobacco products, classes on prevention, and activity at the time of injury were reviewed. Previously identified risk factors were confirmed except for tobacco use. There appeared to be no risk associated with gender or rank. Most injuries were sustained by soldiers performing low-risk activities for which no clear predisposing event could be ascribed. Prevention and early detection appear critical since injuries were not necessarily associated with specific actions or events.