Quantifying regeneration in patients following peripheral nerve injury.

Healthy nerve function provides humans with the control of movement; sensation (such as pain, touch and temperature) and the quality of skin, hair and nails. Injury to this complex system creates a deficit in function, which is slow to recover, and rarely, if ever, returns to what patients consider to be normal. Despite promising results in pre-clinical animal experimentation effective translation is challenged by a current inability to quantify nerve regeneration in human subjects and relate this to measurable and responsible clinical outcomes. In animal models, muscle and nerve tissue samples can be harvested following experimental intervention. This allows direct quantification of muscle mass and quality and quantity of regeneration of axons; such an approach is not applicable in human medicine as it would ensure a significant functional deficit. Nevertheless a greater understanding of this process would allow the relationship that exists between neural and neuromuscular regeneration and functional outcome to be more clearly understood. This article presents a combined commentary of current practice from a specialist clinical unit and research team with regard to laboratory and clinical quantification of nerve regeneration. We highlight how electrophysiological diagnostic methods (which are used with significant recognised limitations in the assessment of clinical medicine) can potentially be used with more validity to interpret and assess the processes of neural regeneration in the clinical context, thus throwing light on the factors at play in translating lab advances into the clinic.

Tetrazolium reduction allows assessment of biofilm formation by Campylobacter jejuni in a food matrix model.

AIMS: To develop a staining method for specific detection of metabolically active (viable) cells in biofilms of the foodborne pathogen Campylobacter jejuni. METHODS AND RESULTS: Conversion of 2,3,5 triphenyltetrazolium chloride (TTC) to insoluble, red 1,3,5-triphenylformazan (TPF) was dependent on metabolic activity of Camp. jejuni. When used with chicken juice, TTC staining allowed quantification of Camp. jejuni biofilm levels, whereas the commonly used dye, crystal violet, gave high levels of nonspecific staining of food matrix components (chicken juice). The assay was optimized to allow for monitoring of biofilm levels and adapted to monitor levels of Camp. jejuni in broth media. CONCLUSIONS: Staining with TTC allows for the quantification of metabolically active Camp. jejuni and thus allows for quantification of viable cells in biofilms and food matrices. The TTC staining method can be adapted to quantify bacterial cell concentration in a food matrix model, where the accepted method of A600 measurement is not suitable due to interference by components of the food matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: 2,3,5 Triphenyltetrazolium chloride (TTC) staining is a low-cost technique suitable for use in biofilm analysis, allowing rapid and simple imaging of metabolically active cells and increasing the methods available for biofilm assessment and quantification.

Cervical dilation as a predictor of pregnancy outcome following emergency cerclage.

OBJECTIVE: To determine whether cervical dilation at the time of physical examination indicated cerclage placement can predicts latency and gestational age at delivery. METHODS: A retrospective cohort study of all women who underwent physical examination indicated cerclage placement from 1996 to 2011 at Duke University Hospital (DUH) was performed. Physical examination indicated cerclage was defined as cerclage placement after 16 weeks in women with a cervical length of less than 2.5 cm and/or cervical dilation greater than or equal to 1 cm at time of procedure. Subjects were divided into two groups depending on cervical dilation at time of procedure (2 cm, <2 cm) for comparison. A multivariate linear regression model for the outcome gestational age of delivery was constructed, controlling for confounding variables. RESULTS: A total of 110 women with complete data were available for analysis. Median gestational age at cerclage placement was similar between the two groups (20.3 vs. 20.3 weeks, p = 0.8). Women with cervical dilatation ≥ 2 cm dilation delivered at an earlier median gestational age than women with cervical dilation <2 cm (27.0 vs. 35.6 weeks, p < 0.001). Cervical dilation at the time of cerclage placement independently predicted gestational age at delivery while controlling for use of intracervical Foley balloon catheter for membrane reduction, cerclage suture type, history of prior preterm birth, race, insurance status, and tobacco use. CONCLUSIONS: Women who receive a rescue cerclage are more likely to deliver at an earlier gestational age when cervical dilation is ≥ 2 cm at the time of procedure.

Climate change and air quality: the potential impact on health.

The objectives of the study were to: consider the potential health impacts in Australia and the region arising from changes in air quality occurring as a result of climate change, identify vulnerable groups and potential adaptation measures and discuss the implications for policy. The authors provide an overview of international and national information on the potential health impacts of air pollutants that would most likely be affected by climate change and a discussion of the policy implications. Climate change is likely to have an impact on levels of ozone and possibly particulates, both of which are associated with increased mortality and a range of respiratory and cardiovascular health effects. One of the implications is therefore a possible increase in adverse health effects due to air pollutants. Regional health impact assessments of climate change should address the issue of air quality, consider current coping capacity, and determine the need for adaptation, particularly for vulnerable groups. Implications for policy include the need for improved modeling and forecasting of air pollutant levels, increased efforts to reduce emissions of air pollutants, continued monitoring of air pollutant levels, and monitoring of the incidence of health effects associated with air pollutants in all countries in the region.

Cervicovaginal fistula presenting during miscarriage.

Cervicovaginal fistula is a recognized complication of induced mid-trimester termination of pregnancy, but more recently it has also been recognized as representing a complication of prior cervical cerclage. We report the ultrasound findings of prolapse of the amniotic sac through a cervicovaginal fistula in a woman with prior cervical cerclage. A woman with cervical incompetence and prior failed McDonald cerclage presented for prophylactic Shirodkar cerclage. Before the procedure, transvaginal ultrasonography revealed a live intrauterine pregnancy at 14 weeks' gestation. Upon further ultrasound examination, the amniotic sac appeared to protrude through the posterior wall of the cervix into the vaginal vault. Pelvic examination verified prolapse of the amniotic sac through a cervicovaginal fistula. The patient underwent an uncomplicated dilatation and evacuation. Women with a history of cervical cerclage are at risk for the development of cervicovaginal fistula, the detection of which is important to prevent potential morbidity.

Indoor air pollution from biomass combustion and respiratory symptoms of women and children in a Zimbabwean village.

UNLABELLED: Rural areas of developing countries are particularly reliant on biomass for cooking and heating. Women and children in these areas are often exposed to high levels of pollutants from biomass combustion that is associated with a range of respiratory symptoms. Domestic exposure to carbon monoxide (CO) and respirable particles (RSPs) in association with respiratory symptoms among women and children in Zimbabwe was investigated in 48 households. Health status and household characteristics were also recorded. In this study, indoor levels of CO and RSPs exceeded World Health Organization (WHO) air quality guidelines in over 95% of kitchens. The level of indoor air pollutants was associated with the area of kitchen windows and the length of cooking time combined with the level of fire combustion. Prevalence of respiratory symptoms was 94% for women and 77% for children. In addition, women reporting respiratory symptoms were exposed to higher levels of RSPs when compared with those reporting no respiratory symptoms. The study results indicated that levels of indoor air pollutants in rural Zimbabwe may contribute to respiratory symptoms in both women and children. PRACTICAL IMPLICATIONS: Levels of respirable particles and carbon monoxide in kitchens in rural Zimbabwe are unacceptably high and measures to reduce levels should be undertaken. Based on the study findings, recommendations for increasing the area of kitchen windows may be considered as a practical method of reducing indoor air pollutants in rural Zimbabwe.

CD4+CD45RBHi T cell transfer induced colitis in mice is accompanied by osteopenia which is treatable with recombinant human osteoprotegerin.

BACKGROUND AND AIMS: Transfer of CD4+CD45RBHi T cells into semi syngeneic immunodeficient mice represents a model of inflammatory bowel disease (IBD). As patients with IBD often suffer from osteopenia, we studied if this T cell transfer in mice results in osteopenia in addition to colitis, and if treatment with osteoprotegerin (OPG) has effects on the bone mineral density of T cell transferred mice. We also investigated whether osteopenia was due to malabsorption as a result of a dysregulated digestive tract or as a consequence of the inflammatory process. METHODS: CD4+CD45RBHi or CD4+CD45RBLo T cells (4 x 10(5)) were sorted from CB6F1 and transferred into C.B.17 scid/scid mice. Recipient mice were treated with human IgG1 Fc (control) or Fc-OPG three times per week in a prophylactic regimen as well as a therapeutic regimen (after 10% body weight loss) and were evaluated for osteopenia and colitis. RESULTS: Mice that received CD4+CD45RBHi T cells developed osteopenia (as indicated by decreased bone density accompanied by decreased osteoblasts and increased osteoclasts) and colitis (as indicated by histological changes in the large intestine). Mice that received CD4+CD45RBLo T cells developed neither osteopenia nor colitis. All animals consumed, on average, the same amount of food and water over the course of the study. Prophylactic treatment with Fc-OPG increased bone density in mice that received either CD4+CD45RBHi or CD4+CD45RBLo T cells but had no effects on the gastrointestinal tract. Fc-OPG treatment of osteopenic mice with established IBD caused the normalisation of bone density. Osteopenia in CD4+CD45RBHi T cell recipients was accompanied by hypoparathyroidism that was partially normalised by treatment with Fc-OPG. CD4+CD45RBHi T cell recipients also had a bone marrow inflammatory cell infiltrate expressing tumour necrosis factor alpha which was unaffected by treatment with Fc-OPG. CONCLUSIONS: CD4+CD45RBHi T cell transfer results in osteopenia in addition to colitis. Evidence suggests that this osteopenia was induced by inflammatory cell infiltration and not by malabsorption of calcium. Recombinant human osteoprotegerin effectively treated the osteopenia. OPG may be a useful therapeutic option for treating osteopenia in patients with IBD.

Outcome of infants delivered between 24 and 28 weeks' gestation in women with severe pre-eclampsia.

OBJECTIVE: To determine whether there are differences in neonatal outcome between infants born to mothers with severe pre-eclampsia and those born to normotensive mothers with preterm labor and intact membranes between 24 and 28 weeks' gestation. MATERIALS AND METHODS: Over a 4-year period between 1991 and 1995, neonates of women with severe pre-eclampsia delivering between 24 and 28 weeks were matched for maternal age, antenatally assigned gestational age and mode of delivery to normotensive women delivering during the same period. RESULTS: Fifty-eight women with severe pre-eclampsia were matched to 58 normotensive controls who delivered as a result of preterm labor. Antenatal steroids were used more often in pre-eclamptic women (75% vs. 47%, p < 0.01). The mean birth weight of pre-eclamptic neonates was significantly lower than that of controls, 767 g vs. 989 g, respectively. Other neonatal complications were similar for both groups. Neonates of pre-eclamptics required longer ventilator support (21 vs. 16 median days, p = 0.03). Neonatal survival was similar for both groups (72% and 79% for pre-eclamptics and normotensives, respectively). CONCLUSIONS: Neonates born to patients with severe pre-eclampsia have similar survival but a lower birth weight and require longer ventilator support than neonates born to women with preterm labor.

Evaluation of the Affirm Ambient Temperature Transport System for the detection and identification of Trichomonas vaginalis, Gardnerella vaginalis, and Candida species from vaginal fluid specimens.

The objective of this study was to measure the performance of the Affirm Ambient Temperature Transport System (ATTS) over time and to estimate the length of time the system can preserve a vaginal specimen containing the three common organisms causing vaginitis: Trichomonas vaginalis, Candida species, and Gardnerella vaginalis (one of the causative agents of bacterial vaginosis). Women with symptoms of vaginitis presenting to one of three clinical centers were evaluated over a 4- to 8-week period. Four simultaneously obtained swabs were collected and tested by the Affirm VPIII assay at time zero with and without a preservative reagent, at 24 h with reagent, and at either 48 or 72 h with reagent. For each of the three organisms, Trichomonas, Gardnerella, and Candida, positivity at each time point was evaluated and compared to that at reference time zero with and without the ATTS. A total of 940 specimens were obtained from the three clinical sites. Eight hundred three were positive for one or more of the three organisms. Gardnerella had the highest overall positive rate (62%), followed by Candida with 18% and Trichomonas at 9%. The percent sensitivity versus control for Trichomonas ranged from 100% at time zero with and without reagent to 91% by 72 h. Gardnerella and Candida sensitivity remained at 100% for each time period. The Affirm VPIII ATTS system performed within 10% of the control swab (no transport reagent) at all four time points (0, 24, 48, and 72 h) for Trichomonas, Gardnerella, and Candida.

Expression of protein kinase C isozymes in nonpregnant and pregnant human myometrium.

OBJECTIVE: The aim of this study was to compare the distributions of protein kinase C isozymes in human nonpregnant and pregnant myometrial tissues and primary cell cultures. STUDY DESIGN: Myometrial tissues were obtained at hysterectomy from nonpregnant women and at cesarean delivery from women both before and during early labor at term. Western immunoblot analysis was performed on homogenates of myometrial tissues and primary cell cultures with monoclonal antibodies specific for protein kinase C isozymes. Redistribution and translocation of protein kinase C were examined by means of immunocytochemical methods. RESULTS: Nonpregnant myometrial tissues contained protein kinase C isozymes alpha, gamma, delta, mu, iota, and zeta but not beta(1), beta(2), theta, or epsilon. Pregnant myometrial tissues both before and during early labor contained the same protein kinase C isozymes and also beta(1) and beta(2). The protein kinase C isozyme distribution in primary myometrial cell cultures was identical to that in the myometrial tissues. Protein kinase C redistribution and translocation were demonstrated in these cultured myometrial cells. CONCLUSION: Both human myometrial tissues and primary cell cultures expressed a broad range of protein kinase C isozymes. Protein kinase C isozymes beta(1) and beta(2) were absent in nonpregnant myometrium but were induced during pregnancy. Labor at term did not alter protein kinase C isozyme expression.

Quantitation of the five forms of plasma high molecular weight angiotensinogen in women with pregnancy-induced hypertension.

In the human pregnant state a high molecular weight form of angiotensinogen (HMrA) is present in significant quantities in addition to the usual low molecular weight angiotensinogen (LMrA). In a previous study involving a small number of white women, it was found that women who had developed pregnancy-induced hypertension (PIH) had significantly higher levels of plasma HMrA. It has been determined that there are five isoforms of HMrA. The objectives of this study were to expand the previous study with the inclusion of black women and to determine which isoform(s) of plasma HMrA are elevated in PIH. Plasma LMrA and HMrA were quantitated in 24 normotensive pregnant women and 65 women with PIH. The PIH group had higher levels of HMrA and somewhat lower levels of LMrA than the normotensive group. The HMrA/LMrA ratio was elevated in 47% of the PIH group. The five isoforms of HMrA were quantitated in plasma from 10 white women with PIH, 10 black women with PIH, and 6 normotensive pregnant white women. Half of both the white and black women with PIH had an elevated HMrA/LMrA ratio. The relative proportion of the HMrA isomers was similar in all groups. These studies show that half the women with PIH have a distinct abnormality in their renin angiotensin system. Both white and black women show this abnormality. In those women who have an elevated total HMrA, all five isoforms of HMrA are equally elevated.

Outcome of patients undergoing transabdominal cerclage: a descriptive study.

OBJECTIVE: Transabdominal cerclage is now evolving as an alternative to transvaginal cerclage in patients with cervical incompetence. The purpose of our study was to evaluate and describe our experience in patients selected to undergo transabdominal cerclage placement. METHODS: Outcome data for all patients who underwent transabdominal cerclage from January, 1990, through December, 1994, was collected. Indications for transabdominal cerclage included patients with prior failed vaginal cerclage, extremely shortened cervix, or anatomical defects judged unsuitable for transvaginal cerclage. RESULTS: Eleven patients underwent 12 transabdominal cerclage procedures. The mean gestational age at the time of cerclage placement was 12.5 weeks (range 10-14 weeks). The mean gestational age at delivery was 34 weeks. There were two neonatal deaths, which were related to extreme prematurity. The mean birthweight was 2,622 g. The fetal salvage rate was 83%. CONCLUSIONS: These findings suggest that in a highly select group of patients, the transabdominal cerclage can be a safe and effective procedure.

Effect of cocaine on intracellular calcium regulation in myometrium from pregnant women.

OBJECTIVE: To evaluate the effect of cocaine on intracellular free calcium ([Ca2+]i) regulation in human myometrial cells by determining the sources of Ca2+ it might mobilize, as well as assess the role cocaine might play in the catecholamine's effect on the cell's [Ca2+]i. METHODS: Primary culture of myometrial cells from pregnant women was used as an experimental model. [Ca2+]i relative changes in response to cocaine and norepinephrine were measured with fura-2 fluorometry and analyzed by means of one-way analysis of variance. RESULTS: Cocaine alone (10(-8) to 10(-3) mol/L) increased [Ca2+]i by up to 43 +/- 18% over basal level in a dose-dependent manner. Norepinephrine also elevated [Ca2+]i in a concentration-dependent manner (202 +/- 24% over basal level at 10(-4) mol/L). The norepinephrine-evoked increase was inhibited in Ca(2+)-free media by 48%, whereas the cocaine response was not affected. The Ca(2+)-channel antagonist nifedipine caused decrease in the [Ca2+]i response to 10(-5) mol/L of norepinephrine by 84%, whereas the [Ca2+]i rise to 10(-5) mol/L cocaine was not significantly changed. Inhibitor of the sarcoplasmic reticulum Ca2+ pump, thapsigargin, completely blocked cocaine-evoked increases in [Ca2+]i, whereas norepinephrine responses were greatly reduced. At the same time, cocaine (10(-8) to 10(-3) mol/L) did not potentiate norepinephrine-evoked Ca2+]i increases in the cells. CONCLUSION: These results indicate that cocaine increases [Ca2+]i in pregnant human myometrial cells, primarily by stimulating release of Ca2+ from intracellular stores rather than by direct stimulation of Ca2+ influx.

A randomized comparison of home uterine activity monitoring in the outpatient management of women treated for preterm labor.

OBJECTIVE: The aim of the study was to evaluate home uterine activity monitoring as an intervention in reducing the rate of preterm birth among women treated for preterm labor. STUDY DESIGN: A total of 186 women were treated in the hospital with magnesium sulfate for preterm labor and were prospectively randomly assigned to study groups; among these, 162 were ultimately eligible for comparison. Eighty-two of these women were assigned to the monitored group and 80 were assigned to an unmonitored control group. Other than monitoring, all women received identical prenatal follow-up, including daily perinatal telephone contact and oral terbutaline therapy. Outcome comparisons were primarily directed toward evaluation of preterm birth at <35 weeks' gestation. Readmissions for recurrent preterm labor and observations lasting <24 hours were evaluated in monitored and unmonitored groups. Compliance with monitoring was also evaluated in the monitored group. RESULTS: The monitored and control groups were demographically similar. According to a multivariate logistic regression model, women with cervical dilatation of >/=2 cm were 4 times more likely to be delivered at <35 weeks' gestation (P <.05). Gestational ages at delivery were similar in the monitored and control groups. There was no significant difference in the overall rate of preterm delivery at <35 weeks' gestation between the monitored group (10.9%) and the control group (15.0%). The overall rates of delivery at <37 weeks' gestation were high (48.8% and 60.0% for monitored and control groups, respectively), and the difference was not significant. The numbers of women with >/=1 instance of readmission and treatment for recurrent preterm labor were equal in the monitored and control groups. The numbers of women with >/=1 hospital observation lasting <24 hours were not different between the groups. Compliance with monitoring did not significantly differ for women who were delivered at <35 weeks' gestation, women with >/=2 cm cervical dilatation at enrollment, or for African American women. CONCLUSION: A reduction in the likelihood of preterm delivery at <35 weeks' gestation was not further enhanced by the addition of home uterine monitoring to the outpatient management regimens of women treated for preterm labor.

Chorioamnionitis: is continuation of antibiotic therapy necessary after cesarean section?

OBJECTIVE: Our purpose was to determine whether the continuation of antibiotics postoperatively after cesarean section in patients whose labors were complicated by chorioamnionitis would reduce the incidence of endometritis. STUDY DESIGN: Patients with a clinical diagnosis of chorioamnionitis treated with ampicillin during labor and who required cesarean delivery for obstetric indications received preoperative intravenous clindamycin and gentamicin and were randomized into 2 groups. Group 1 received no scheduled postoperative antibiotics and group 2 continued to receive clindamycin 900 mg every 8 hours and gentamicin 1.5 mg/kg every 8 hours until afebrile for a minimum of 24 hours (temperature

Localization of a baculovirus-induced chitinase in the insect cell endoplasmic reticulum.

Confocal immunofluorescence microscopy was used to demonstrate that the Autographa californica nucleopolyhedrovirus (AcMNPV) chitinase was localized within the endoplasmic reticulum (ER) of virus-infected insect cells. This was consistent with removal of the signal peptide from the chitinase and an ER localization motif (KDEL) at the carboxyl end of the protein. Chitinase release from cells, a prerequisite for liquefaction of virus-infected insect larvae, appears to be aided by synthesis of the p10 protein. Deletion of p10 from the AcMNPV genome delayed the appearance of chitinase activity in the medium of virus-infected cells by 24 h and also delayed liquefaction of virus-infected Trichoplusia ni larvae by the same period.

The association of aneuploidy and unexplained elevated maternal serum alpha-fetoprotein.

OBJECTIVE: The purpose of this study was to examine fetal chromosomal abnormalities in pregnancies complicated by unexplained elevated maternal serum alpha-fetoprotein (MSAFP). STUDY DESIGN: We reviewed, using a computerized database, 58,162 obstetrical ultrasounds that were performed for various indications. Fetuses with MSAFP multiples of the median (MOM) > or = 2.5 and normal extensive ultrasounds were identified. Maternal demographic data and fetal karyotype were obtained. RESULTS: Seven hundred eighty-nine patients received ultrasounds for evaluation of elevated MSAFP. Of the 595 patients with normal scans, 195 (32.8%) underwent amniocentesis and cytogenetic evaluation. Two chromosomal abnormalities were detected (1.0%), including an inversion and a balanced translocation. CONCLUSION: The two karyotypic abnormalities identified in our study consisted of structural rearrangements. Patients undergoing karyotype analysis for unexplained elevated MSAFP should be counseled that the types of aneuploidy detected under this circumstance differ from those associated with advanced age and specific fetal anomalies (trisomy and triploidy).

Anticoagulation.

Outpatient management of women requiring treatment and prophylaxis against thromboembolic conditions during pregnancy and the postpartum period requires a coordinated effort between the patient, her obstetrician and, in certain cases, a hematologic consultant. The anticoagulation regimen should be tailored to the clinical situation, with patient compliance and cost taken into consideration.

Methadone maintenance in pregnancy: a reappraisal.

OBJECTIVE: The purpose of this study is to evaluate women receiving methadone maintenance during pregnancy. STUDY DESIGN: Thirty-two pregnancies in women receiving methadone maintenance were matched by gestational age to women with a positive urine screen for cocaine at delivery and to drug-free controls. Pregnancy outcome variables were compared, including birth weight and neonatal morbidity. Analysis was by chi2 and t test with significance set at .05. RESULTS: Birth weight of methadone-exposed infants was 2748 g versus 2925 g for cocaine and 3032 g for controls, P = not significant. Birth weight comparison with a 50-mg maternal methadone cutoff dose was not different. A head circumference for methadone infants of 32.4 +/- 4.7 cm was significantly less than controls, 33.5 +/- 4.0 cm, P < .04, but not different from infants of cocaine users, 32.8 +/- 3.1 cm. Women using cocaine had a significantly higher incidence of meconium in labor compared with methadone and controls. Of women taking methadone 27 of 32 (84.3%) were positive for other drugs of abuse in the last screen before or at delivery. Cocaine 12 of 32 (37.5%), other opiates 13 of 32 (40.6%), and marijuana 14 of 32 (43.7%) were the most prevalent. Neonatal withdrawal occurred in 23 of 32 (72%) women taking methadone. The neonates of women using < 50 mg of methadone were as likely to withdraw as those women using > or = 50 mg, 61.5% versus 79.0%, P not significant. Three neonates in the methadone group (9.3%) had major congenital anomalies, with 2 of the 3 (66.6%) resulting in mortality. CONCLUSIONS: Birth outcome is not significantly different between methadone and cocaine users. Women receiving methadone maintenance are likely to abuse other illicit drugs.

Cytomegalovirus infection.

Cytomegalovirus (CMV) infection is the most common perinatal infection and may result in severe injury to the fetus. Forty percent to 50% of infants delivered to mothers with primary CMV will have congenital infections. Of these, 5% to 18% will be overtly symptomatic at birth. The mortality rate in these children is almost 30%; approximately 80% of the survivors have severe neurological morbidity. The majority of congenitally infected infants will be asymptomatic at birth; 10% to 15% of these children subsequently have sequelae such as visual and auditory defects. If recurrent or reactivated CMV infection develops during pregnancy, the risk of serious fetal injury is very low. Similarly, neonatal infection acquired during delivery or from breast feeding also poses minimal risk to the child. Because antimicrobial therapy and immunoprophylaxis for CMV infection are unsatisfactory, pregnant women must be educated about preventive measures.