Toward safer prescribing: evaluation of a prospective drug utilization review system on inappropriate prescriptions, prescribing patterns, and adverse drug events and related health expenditure in South Korea.

OBJECTIVES: This study aimed to evaluate the effect of the prospective drug utilization review (DUR) system introduced in Korea in December 2010 as a real-time method to improve patient safety, in terms of changes in prescribing practices, adverse drug events (ADEs), and ADE-related healthcare expenditure, using non-steroidal anti-inflammatory drugs (NSAIDs) and their common ADEs as a guide. STUDY DESIGN: We used an interrupted time-series study design using generalized estimating equations to evaluate changes in prescription rate and ADE-related healthcare expenditure. Cox regression analysis was used to evaluate the probability of NSAID-associated ADEs. METHODS: A total of 154,585 outpatients with musculoskeletal or connective tissue disorders, without pre-existing gastric bleeding or ulcers were included in this study. The primary outcome was the level and trend change in prescription rate, drug-drug interactions, coprescribed gastro-protective drugs, and defined daily dose (DDD) of NSAIDs. The secondary outcome was the probability of ADEs and changes in ADE-related healthcare expenditure. RESULTS: There was a significant trend change after introducing the DUR system in terms of drug-drug interactions (-3.6%) and coprescribed gastro-protective drugs (+0.6%). The mean DDD of NSAIDs increased by 0.2. The probability of ADEs decreased overall (-1.7%) and in the high-risk group (age ≥65 years; -9.6%); however, only the latter was significant. There was no significant trend or level change in ADE-related health expenditure. CONCLUSIONS: The introduction of the DUR system was associated with more efficient prescribing, including a reduction in drug-drug interactions and an increase in the use of gastro-protective drugs. The system had a positive effect on patient outcome but was not associated with reduced ADE-related costs. Further studies are needed to evaluate the long-term effects of the DUR system in Korea.

The association between perceived unmet medical need and mental health among the Republic of Korea Armed Forces.

INTRODUCTION: We investigated the effect of unmet medical need on the mental health of Republic of Korea (ROK) Armed Forces personnel, as most of the service members work in remote areas and often experience such unmet needs. METHODS: This study used secondary data from the 2014 Military Health Survey (MHS), conducted by the ROK School of Military Medicine and designed to collect military health determinants. Descriptive statistics showed the general characteristics of the study populations by variable. We specifically compared the population after stratifying participants by suicide ideation. An analysis of variance was also carried out to compare Kessler Psychological Distress Scale 10 Scores. Additionally, dependent spouses and children of both active-duty service members and retirees are included among those entitled to Military Health System healthcare. RESULTS: Among the 4967 military personnel, 681 (13.7%) individuals reported an experience of unmet medical need within the past 12 months and gave reasons of 'no time (5.15%)', 'long office wait (2.6%)', 'no money (0.22%)', 'long distance from base (1.19%)', 'illness but not very serious (1.65%)', 'mistrust in doctors (1.95%)' and 'pressure due to performance appraisal (0.95%)'. Regression analysis revealed that unmet medical need was significantly associated with negative mental health (ß=1.753, p<0.0001) and increased suicide ideation (OR=2.649, 95% CI 1.84 to 3.82). Also, soldiers reporting unmet medical need due to 'no money', 'no time' or 'pressure due to performance appraisal' were significantly more likely to experience similar negative mental health effects. CONCLUSIONS: Our study indicates that unmet medical need is significantly associated with soldiers' mental health decline and suicide ideation, highlighting the importance of providing military personnel with timely, affordable and sufficient medical care.

The influence of sexual harassment on mental health among female military personnel of the Republic of Korea Armed Forces.

INTRODUCTION: Reports of sexual harassment are becoming more frequent in Republic of Korea (ROK) Armed Forces. This study aimed to analyse the impact of sexual harassment on mental health among female military personnel of the ROK Armed Forces. METHODS: Data from the 2014 Military Health Survey were used. Instances of sexual harassment were recorded as 'yes' or 'no'. Analysis of variance (ANOVA) was carried out to compare Kessler Psychological Distress Scale 10 (K-10) scores. Multiple logistic regression analysis was performed to identify associations between sexual harassment and K-10 scores. RESULTS: Among 228 female military personnel, 13 (5.7%) individuals experienced sexual harassment. Multiple logistic regression analysis revealed that sexual harassment had a significantly negative impact on K-10 scores (3.486, p<0.04). Higher K-10 scores among individuals experiencing sexual harassment were identified in the unmarried (including never-married) group (6.761, p<0.04), the short-term military service group (12.014, p<0.03) and the group whose length of service was <2 years (11.067, p<0.02). CONCLUSIONS: Sexual harassment has a negative impact on mental health. Factors associated with worse mental health scores included service classification and length of service. The results provide helpful information with which to develop measures for minimising the negative psychological effects from sexual harassment and promoting sexual harassment prevention policy.

Feasibility study on development of Cerenkov fiber-optic dosimeter for radiotherapy application.

To obtain real-time dose information in photon-beam therapy using a clinical linear accelerator, we fabricated a novel Cerenkov fiber-optic dosimeter using two plastic optical fibers without employing a scintillator. In this study, the light intensity and spectrum of Cerenkov radiation induced by a high-energy photon beam were measured as functions of the irradiation angle and the length difference between the two plastic optical fibers in the dosimeter probe. Also, we obtained a percentage depth dose curve for a 6 MV photon beam with a field size of 10 × 10 cm(2) according to the depth of the solid water phantom. Based on the results of this study, it is anticipated that the proposed Cerenkov fiber-optic dosimeter can be developed as a useful dosimeter to accurately obtain dose information prior to conducting radiotherapy.

Development of a fibre-optic dosemeter to measure the skin dose and percentage depth dose in the build-up region of therapeutic photon beams.

In this study, a fibre-optic dosemeter (FOD) using an organic scintillator with a diameter of 0.5 mm for photon-beam therapy dosimetry was fabricated. The fabricated dosemeter has many advantages, including water equivalence, high spatial resolution, remote sensing and real-time measurement. The scintillating light generated from an organic-dosemeter probe embedded in a solid-water stack phantom is guided to a photomultiplier tube and an electrometer via 20 m of plastic optical fibre. Using this FOD, the skin dose and the percentage depth dose in the build-up region according to the depths of a solid-water stack phantom are measured with 6- and 15-MV photon-beam energies with field sizes of 10 × 10 and 20 × 20 cm(2), respectively. The results are compared with those measured using conventional dosimetry films. It is expected that the proposed FOD can be effectively used in radiotherapy dosimetry for accurate measurement of the skin dose and the depth dose distribution in the build-up region due to its high spatial resolution.

Lung fibrosis in Sprague-Dawley rats, induced by exposure to manual metal arc-stainless steel welding fumes.

To investigate the disease process of pneumoconiosis induced by welding-fume exposure, a lung fibrosis model was established by building a stainless steel arc welding fume generation system and exposing male Sprague-Dawley rats for 90 days. The rats were exposed to welding fumes with concentrations of 57-67 mg/m3 (low dose) and 105-118 mg/m3 (high dose) total suspended particulates for 2 h per day in an inhalation chamber for 90 days. The concentrations of the main metals, Fe, Mn, Cr, and Ni, were measured in the welding fumes, plus the gaseous compounds, including nitrous gases and ozone, were monitored. During the exposure period, the animals were sacrificed after the initial 2-h exposure and after 15, 30, 60, and 90 days. Histopathological examinations were conducted on the animals' upper respiratory tract, including the nasal pathway and conducting airway, plus the gas exchange region, including the alveolar ducts, alveolar sacs, and alveoli. When compared to the control group, the lung weights did not increase significantly in the low-dose group, yet in the high-dose group there was a significant increase from day 15 to day 90. The histopathological examination combined with fibrosis-specific staining (Masson's trichrome) indicated that the lungs in the low-dose group did not exhibit any progressive fibrotic changes. Whereas, the lungs in the high-dose group exhibited early delicate fibrosis from day 15, which progressed into the perivascular and peribronchiolar regions by day 30. Interstitial fibrosis appeared at day 60 and became prominent by day 90, along with the additional appearance of pleural fibrosis. Accordingly, it would appear that a significant dose of welding-fume exposure was required to induce lung fibrosis.

Single molecule DNA sequencing in submicrometer channels: state of the art and future prospects.

We demonstrate a new method for single molecule DNA sequencing which is based upon detection and identification of single fluorescently labeled mononucleotide molecules degraded from DNA-strands in a cone shaped microcapillary with an inner diameter of 0.5 microm. The DNA was attached at an optical fiber via streptavidin/biotin binding and placed approximately 50 microm in front of the detection area inside of the microcapillary. The 5'-biotinylated 218-mer model DNA sequence used in the experiments contained 6 fluorescently labeled cytosine and uridine residues, respectively, at well defined positions. The negatively charged mononucleotide molecules were released by addition of exonuclease I and moved towards the detection area by electrokinetic forces. Adsorption of mononucleotide molecules onto the capillary walls as well as the electroosmotic (EOF) flow was prevented by the use of a 3% polyvinyl pyrrolidone (PVP) matrix containing 0.1% Tween 20. For efficient excitation of the labeled mononucleotide molecules a short-pulse diode laser emitting at 638 nm with a repetition rate of 57 MHz was applied. We report on experiments where single-stranded model DNA molecules each containing 6 fluorescently labeled dCTP and dUTP residues were attached at the tip of a fiber, transferred into the microcapillary and degraded by addition of exonuclease I solution. In one experiment, the exonucleolytic cleavage of 5-6 model DNA molecules was observed. 86 photon bursts were detected (43 Cy5-dCMP and 43 MR121-dUMP) during 400 s and identified due to the characteristic fluorescence decay time of the labels of 1.43+/-0.19 ns (Cy5-dCMP), and 2.35+/-0.29 ns (MR121-dUMP). The cleavage rate of exonuclease I on single-stranded labeled DNA molecules was determined to 3-24 Hz under the applied experimental conditions. In addition, the observed burst count rate (signals/s) indicates nonprocessive behavior of exonuclease I on single-stranded labeled DNA.

Pattern of deposition of stainless steel welding fume particles inhaled into the respiratory systems of Sprague-Dawley rats exposed to a novel welding fume generating system.

In order to investigate occupational diseases related to welding fume exposure, such as nasal septum perforation, pneumoconiosis and manganese intoxication, we built a welding fume exposure system that included a welding fume generator, exposure chamber and fume collector. The fume concentrations in the exposure chamber were monitored every 15 min during a 2-h exposure. Fume (mg/m(3)) concentrations of major metals, including Fe, Mn, Cr, and Ni were found to be consistently maintained. An acute inhalation toxicity study was conducted by exposing male Sprague-Dawley rats to the welding fumes generated in this apparatus by stainless steel arc welding. The rats were exposed in the inhalation chamber to a welding fume with a concentration of 62 mg/m(3) total suspended particulates for 4 h. Animals were sacrificed at 4 h and at 1, 3, 7, 10, and 14 days after exposure. Histopathological examinations were conducted on the animals' upper respiratory tracts, including the nasal pathway and the conducting airway, and on the gas exchange region including the alveolar ducts, alveolar sacs, and alveoli. Diameters of fume particles varied from 0.02 to 0.81 microm and were distributed log normally, with a mean diameter of 0.1 microm and geometric standard deviation of 1.42. Rats exposed to the welding fume for 4 h did not show any significant respiratory system toxicity. The mean particle diameter of 0.1 microm resulted in little adsorption of the welding fume particles in the upper respiratory tract. Particle adsorption took place principally in the lower respiratory tracts, including bronchioles, alveolar ducts, alveolar sacs, and alveoli.

Epidemiology and time trends of gestational trophoblastic disease in Korea.

OBJECTIVES: For the purpose of determining the annual incidence and time trends of gestational trophoblastic disease (GTD), the medical records from 24 university hospitals, 13 private general hospitals and the Korean Research Institute of Gestational Trophoblastic Disease (KRI-TRD) were analyzed from 1971 to 1995. MATERIALS & RESULTS: From a total of 7198 GTD cases (H-mole=3831, Invasive mole=2163, Choriocarcinoma=1177, PSTr=27) among 838659 deliveries between 1971 and 1995, the hospital-based incidence of H-mole per 1000 deliveries declined from 40.2 during 1971-975, to 2.3 during 1991-995. The population-based incidence of H-mole, however, revealed an average of 2.05 per 1000 deliveries during 1991-995. Old age and gravidities as factors in GTD patients both decreased significantly during the study period. Time trends for the incidence of GTD in Korea revealed significant changes, not only a decrease in the incidence of GTD, but also an improvement in the annual remission rate. Korea's socio-eonomic improvement in recent decades also contributed to the decreased incidence of GTD and the increased survival rates.

Effects of multiagent chemotherapy and independent risk factors in the treatment of high-risk GTT--25 years experiences of KRI-TRD.

UNLABELLED: A retrospective and comparative study of high-risk gestational trophoblastic tumor (GTT) treated with different chemoregimen from 1971 to 1995 was performed and to find most effective chemotherapy regimen and independent risk factors. Three hundred seven patients in scoring over 8 points in WHO classification were categorized into high-risk group among 802 GTT cases received chemotherapy in the 2,418 GTD patients registered at KRI-TRD (Korean Research Institute for Gestational Trophoblastic Disease), Catholic University Medical College in Korea. Study groups of multiagent combination chemotherapy in 227 patients of the high-risk GTT were divided such as 49 cases of combination chemotherapy with MTX + folinic acid and Act-D, 40 cases of MAC regimen, 42 cases of CHAMOCA regimen, and 96 cases of EMA/CO. Initial tumor response according to hCG titer decrease was found in good response (log fall) 69.8%, of EMA /CO regimen group. On the other hand, good response was shown in only 24.5% of MTX + ACT-D, 32.5% of MAC regimen, and 52.4%, of CHAMOCA regimen respectively. Remission rate of EMA/CO regimen was 90.6% (87/96) and courses of chemotherapy until remission was 8.5+/-2.2. However, remission rate of other regimens of MTX + Act-D, MAC, and CHAMOCA were 63.3%, (31/49) 67.5% (27/40) and 76.2% (32/45) respectively, with 10.0+/-4.0, 10.7+/-4.3, 9.1+/-3.9 chemotherapy courses respectively until remission. Therefore, EMA/CO regimen groups were found to have low drug toxicity, early remission and a low failure rate. In the study of independent risk factors in the 165 cases of high-risk gestational trophoblastic tumor patients received EMA/CO regimen, stepwise Coxs proportional hazard's regression of prognostic factors using multivariate analysis revealed tumor age, number of metastatic organs, metastatic site and inadequate previous chemotherapy. According to the performance of fitted logistic regression model, the prediction rate of death and survival was 80.5%. CONCLUSIONS: The most effective chemotherapy to high-risk GTT was EMA/CO regimen than other regimens. The following factors showed poor prognosis; 1) Tumor age is over 12 month, 2) more than 2 organs had metastatic lesion, 3) inadequate previous therapy that includes unplanned operation and inadequate previous chemotherapy.

Effects of multiagent chemotherapy and independent risk factors in the treatment of high-risk GTT -25 years experiences of KRI-TRD.

A retrospective and comparative study of high-risk gestational trophoblastic tumor (GTT) treated with different chemoregimen from 1971 to 1995 was performed and to find most effective chemotherapy regimen and independent risk factors. Three hundred seven patients in scoring over 8 points in WHO classification were categorized into high-risk group among 802 GTT cases received chemotherapy in the 2,418 GTD patients registered at KRI-TRD (Korean Research Institute for Gestational Trophoblastic Disease), Catholic University Medical College in Korea. Study groups of multiagent combination chemotherapy in 227 patients of the high-risk GTT were divided such as 49 cases of combination chemotherapy with MTX + folinic acid and Act-D, 40 cases of MAC regimen, 42 cases of CHAMOCA regimen, and 96 cases of EMA/CO. Initial tumor response according to hCG titer decrease was found in good response (log fall) 69.8%, of EMA /CO regimen group. On the other hand, good response was shown in only 24.5% of MTX + ACT-D, 32.5% of MAC regimen, and 52.4%, of CHAMOCA regimen respectively. Remission rate of EMA/CO regimen was 90.6% (87/96) and courses of chemotherapy until remission was 8.5 ± 2.2. However, remission rate of other regimens of MTX + Act-D, MAC, and CHAMOCA were 63.3%, (31/49) 67.5% (27/40) and 76.2% (32/45) respectively, with 10.0 ± 4.0, 10.7 ± 4.3, 9.1 ± 3.9 chemotherapy courses respectively until remission. Therefore, EMA/CO regimen groups were found to have low drug toxicity, early remission and a low failure rate. In the study of independent risk factors in the 165 cases of high-risk gestational trophoblastic tumor patients received EMA/CO regimen, stepwise Coxs proportional hazard's regression of prognostic factors using multivariate analysis revealed tumor age, number of metastatic organs, metastatic site and inadequate previous chemotherapy. According to the performance of fitted logistic regression model, the prediction rate of death and survival was 80.5%. CONCLUSIONS: The most effective chemotherapy to high-risk GTT was EMA/CO regimen than other regimens. The following factors showed poor prognosis; 1) Tumor age is over 12 month, 2) more than 2 organs had metastatic lesion, 3) inadequate previous therapy that includes unplanned operation and inadequate previous chemotherapy.

Epidemiology and time trends of gestational trophoblastic disease in Korea.

OBJECTIVES: For the purpose of determining the annual incidence and time trends of gestational trophoblastic disease (GTD), the medical records from 24 university hospitals, 13 private general hospitals and the Korean Research Institute of Gestational Trophoblastic Disease (KRI-TRD) were analyzed from 1971 to 1995. MATERIALS & RESULTS: From a total of 7198 GTD cases (H-mole = 3831, Invasive mole = 2163, Choriocarcinoma = 1177, PSTT = 27) among 838659 deliveries between 1971 and 1995, the hospital-based incidence of H-mole per 1000 deliveries declined from 40.2 during 1971-975, to 2.3 during 1991-995. The population-based incidence of H-mole, hwoever, revealed an average of 2.05 per 1000 deliveries during 1991-995. Old age and gravidities as factors in GTD patients both decreased significantly during the study period. Time trends for the incidence of GTD in Korea revealed significant changes, not only a decrease in the incidence of GTD, but also an improvement in the annual remission rate. Korea's socio-eonomic improvement in recent decades also contributed to the decreased incidence of GTD and the increased survival rates.

Human papillomavirus infection and TP53 gene mutation in primary cervical carcinoma.

Tumor specimens obtained from 136 patients with primary carcinoma of the uterine cervix were analyzed for the presence of human papillomavirus (HPV) sequences and for mutation of the TP53 gene. Polymerase chain reaction (PCR) showed that 130 of 136 (96%) tumors contained an oncogenic HPV 16 or 18 sequence. HPV 16 was the predominant type in cervical squamous cell carcinomas and HPV 18 was significantly associated with cervical adenocarcinomas (p < 0.05). The more dedifferentiated the primary tumor, the more frequent the HPV 16 infection and the more differentiated, the more frequent the HPV 18 infection (p < 0.05). Two out of 136 (1.5%) tumors demonstrated single-strand conformation polymorphism (SSCP) band shifts. One (positive for HPV 18) had a nonsense mutation of codon 101 in exon 4 from AAA to TAA transversion. Another (positive for L1 consensus primer set) showed a point mutation involving codon 179 in exon 5 changing CAT to CGT transition. The three specimens negative for HPV did not contain TP53 gene mutations. Our data show that mutation of TP53 is infrequent in primary cervical carcinoma and there is no inverse correlation between HPV infection and TP53 gene mutation. Other mechanisms independent of TP53 inactivation may also be implicated in tumorigenesis of the uterine cervix.

Presence of oncogenic HPV DNAs in cervical carcinoma tissues and pelvic lymph nodes associating with proliferating cell nuclear antigen expression.

The presence of oncogenic HPV DNAs (HPV-16/18) in cervical carcinomas and their normal and metastatic pelvic lymph nodes and the expression patterns of proliferating cell nuclear antigen (PCNA) in cervical carcinomas were retrospectively studied to elucidate the possible roles of them in malignant transformation and progression of the disease. HPV-16/18 DNAs were detected by polymerase chain reaction using HPV E6 type-specific primers in 79 patients with cervical cancer. 31 patients who had pelvic lymph node metastasis (group I) and 48 patients without pelvic lymph node metastasis (group II) who were proven by pathologic examination of surgical specimens. HPV-16 or -18 DNAs were detectable in cervical carcinoma tissues in 60 patients from 79 cervical cancer patients (75.9%; HPV-16 was 67.1% and HPV-18 was 8.9%). HPV DNAs were amplified from metastatic pelvic lymph nodes in 13 patients of group I (42%) and from nonmetastatic lymph nodes in 7 group I patients (22.5%). Recurrence was identified in 9 group I patients (29.0%) in 3 years of follow-up. HPV DNAs were amplified from nonmetastatic lymph nodes in 11 group II patients (22.9%). Two group II patients, who had HPV-16 DNA by PCR in nonmetastatic nodes, were recurrent. PCNA was overexpressed in 66.7% of HPV-16- or -18-positive cervical cancers and 16.7% of HPV-16- or -18-negative cervical cancers. However, the expression levels of PCNA in cervical cancers were not influenced by the presence of oncogenic HPV DNA or pathologic metastasis in the pelvic lymph nodes. In conclusion, HPV DNA could be amplified from some metastatic and nonmetastatic pelvic lymph nodes and the detectability of oncogenic HPV DNA in pelvic lymph nodes may represent the poor outcome in the treatment of disease. The expression of PCNA protein which was associated with presence of oncogenic HPV DNAs in cervical cancers, suggesting activation of S phase of cell cycle, may contribute to the malignant progression by HPV-16 or -18.

Sialyl Lewis(x) oligosaccharide reduces ischemia-reperfusion injury in the rabbit ear.

Ischemia-reperfusion injury in the rabbit ear is neutrophil (PMN)-mediated, and is significantly reduced by anti-adhesion agents directed against beta 2 integrins, P-selectin, or L-selectin. We further examined selectin-mediated adherence in this setting following the administration of soluble sialyl Lewis(x) (SLe(x)), the principal carbohydrate ligand for P-, L-, and E-selectin, at various times following reperfusion. Under constant ambient temperature conditions, the rabbit ear vascular supply was isolated and occluded with an atraumatic vascular clamp for 6 h, then allowed to reperfuse. Animals receiving i.v. SLe(x) (25 mg/kg bolus + 50 mg/kg infusion over 10 h) 1) at the time of reperfusion, 2) 1 h after reperfusion, 3) 4 h after reperfusion, or 4) 12 h after reperfusion were compared with control animals receiving either saline or sialyl lactosamine, an oligosaccharide structurally similar to SLe(x) but not involved in selectin recognition. Tissue injury was assessed by serial measurement of ear edema and by visual determination of ear necrosis over 7 days. Tissue edema and necrosis were significantly reduced in animals treated with SLe(x) immediately upon reperfusion or after a 1-h delay, but not in animals for whom SLe(x) administration was delayed by 4 or 12 h. Furthermore, SLe(x) administration alone had no effect on circulating leukocyte or PMN counts, or PMN expression of CD18 or L-selectin. We conclude that interruption of selectin-mediated adherence with soluble SLe(x) oligosaccharide attenuates reperfusion in the rabbit ear. The observation that SLe(x) is efficacious only if administered in the first hour after reperfusion suggests that the more immediately available P- and L-selectin participate in this PMN adhesion/injury process, whereas E-selectin, with its delayed endothelial expression, does not.

New fluorescent dyes in the red region for biodiagnostics.

The increased sensitivity together with the advent of low-cost optical sources and detectors in the visible-near IR region has led us to current efforts to develop new efficient fluorescent labels for biodiagnostics with absorption and emission beyond 600 nm. In view of the general fluorescence decrease with increasing emission wavelength, we investigated the possibility to shift the absorption of rhodamine dyes toward the region 620-670 nm. The hydrophobic nature of all known long-wavelength dyes results in the tendency to form intra- and intermolecular aggregates in hydrophilic solvents, especially in aqueous environment. Due to the aggregation with biological materials, fluorescence quenching of the dyes is often observed. New strategies for prevention of these processes are considered.

Ischemia reperfusion injury in the rabbit ear is reduced by both immediate and delayed CD18 leukocyte adherence blockade.

mAb blockade of CD18-mediated neutrophil adherence has previously been shown to reduce tissue injury in the rabbit ear as a result of ischemia followed by reperfusion. Similar injury reduction has been demonstrated whether treatment is given before ischemia or at the time of reperfusion. We examined the effects of delayed treatment with blocking CD18 mAb (60.3) after reperfusion of ischemic rabbit ears. The central neurovascular bundle of rabbit ears was isolated by microsurgery, the remainder of the ear devascularized, and all nerves cut to render the ear anesthetic. Arterial blood flow was occluded with a microvascular clamp for 6 h at an ambient temperature of 23 to 24 degrees C. The clamp was then removed and the ear allowed to reperfuse. Rabbits were divided into five treatment groups: 1) i.v. saline at reperfusion, 2) i.v. mAb 60.3 (2 mg/kg) at reperfusion, 3) i.v. mAb 60.3 1 h after reperfusion, 4) i.v. mAb 60.3 4 h after reperfusion, and 5) i.v. mAb 60.3 12 h after reperfusion. Ear edema (measured by volume displacement) was determined daily for 7 days. Edema in the immediate, 1 h, and 4 h mAb-treated groups was significantly less than in saline-treated controls, although less pronounced in the 4-h treatment group. Tissue necrosis measured at 7 days was significantly reduced in the same three mAb-treated groups compared with controls. However, edema and tissue necrosis in the 12 h mAb-treated group were similar to controls. We conclude that mAb blockade of CD18 at 1 h after reperfusion is as effective as immediate treatment in reducing ischemia reperfusion injury in the rabbit ear. Delaying treatment for 4 h is also effective but less so, whereas delaying treatment for 12 h results in no beneficial effects.

New fluorescent labels for time-resolved detection of biomolecules.

New dyes with characteristic fluorescence lifetimes have been developed for bioanalytical applications. Based upon the concept of "multiplex dyes," we have designed rhodamine dyes with nearly identical absorption and emission spectral characteristics but different fluorescence lifetimes. Extending this principle to applications with laser diodes, new rhodamines with functional groups for covalent coupling of analytes have been developed. The new labels exhibit absortion and fluorescence beyond 600 nm and have a high quantum efficiency, even in aqueous buffer systems.