Cross talk between beta(1) and alpha(V) integrins: beta(1) affects beta(3) mRNA stability.

There is increasing evidence that a fine-tuned integrin cross talk can generate a high degree of specificity in cell adhesion, suggesting that spatially and temporally coordinated expression and activation of integrins are more important for regulated cell adhesive functions than the intrinsic specificity of individual receptors. However, little is known concerning the molecular mechanisms of integrin cross talk. With the use of beta(1)-null GD25 cells ectopically expressing the beta(1)A integrin subunit, we provide evidence for the existence of a cross talk between beta(1) and alpha(V) integrins that affects the ratio of alpha(V)beta(3) and alpha(V)beta(5) integrin cell surface levels. In particular, we demonstrate that a down-regulation of alpha(V)beta(3) and an up-regulation of alpha(V)beta(5) occur as a consequence of beta(1)A expression. Moreover, with the use of GD25 cells expressing the integrin isoforms beta(1)B and beta(1)D, as well as two beta(1) cytoplasmic domain deletion mutants lacking either the entire cytoplasmic domain (beta(1)TR) or only its "variable" region (beta(1)COM), we show that the effects of beta(1) over alpha(V) integrins take place irrespective of the type of beta(1) isoform, but require the presence of the "common" region of the beta(1) cytoplasmic domain. In an attempt to establish the regulatory mechanism(s) whereby beta(1) integrins exert their trans-acting functions, we have found that the down-regulation of alpha(V)beta(3) is due to a decreased beta(3) subunit mRNA stability, whereas the up-regulation of alpha(V)beta(5) is mainly due to translational or posttranslational events. These findings provide the first evidence for an integrin cross talk based on the regulation of mRNA stability.

Percutaneous regional compared with local anaesthesia for facial lacerations: a randomised controlled trial.

OBJECTIVE: Facial lacerations are usually repaired after local infiltration of an anaesthetic agent. Regional nerve blocks of the face offer several theoretical advantages over local infiltration. This study compared the pain of injection and anaesthetic efficacy of percutaneous regional and local anaesthesia for facial lacerations. STUDY DESIGN: Randomised clinical trial. PARTICIPANTS: Convenience sample of emergency department patients with facial lacerations requiring suturing in anatomical areas innervated by a regional nerve (supraorbital, infraorbital, or mental). INTERVENTIONS: Facial lacerations treated using standard wound care. Lacerations were randomised to local or regional infiltration of lidocaine (lignocaine) 1% with adrenaline (epinephrine) 1:100 000 using a number 27 needle. OUTCOMES: Pain of injection on 100 mm visual analogue scale (VAS) and need for rescue anaesthetic infiltration before suturing. DATA ANALYSIS: Group comparisons were with Student's t test and chi(2) test. This study had 80% power to detect a 20 mm difference in pain of injection (two tailed, alpha = 0.05). RESULTS: 36 patients were randomised to local (18) and regional (18) anaesthesia. Mean (SD) age was 20 (14); 19% were female. Groups were similar in baseline characteristics. Patients in the regional anaesthesia group experienced more pain during infiltration than patients in the local anaesthesia group (42.4 mm v 24.8 mm, mean difference 17.6 mm (95% CI 0.3 to 35.6 mm) and were more likely to require additional infiltration of a local anaesthetic (28% v 0%, (95% CI 6% to 50%)) than patients in the local anaesthetic group. CONCLUSIONS: Local infiltration of anaesthetics for facial lacerations is less painful and results in more effective anaesthesia than percutaneous regional infiltration.

Incremental benefit of individual American College of Surgeons trauma triage criteria.

OBJECTIVE: To determine the incremental benefit of individual American College of Surgeons (ACS) trauma triage criteria for prediction of severe injuries after consideration of concurrent physiologic, anatomic, mechanism, or "other" criteria. METHODS: A prospective cross-sectional study of motor vehicle crash victims transported to any of the 12 hospitals in a suburban/rural county by local ambulance services was performed. Demographic and individual ACS criteria were collected using structured data instruments. EDs provided patient disposition within 24 hours of patient arrival. Medical records were reviewed. Major outcomes were admission, operative interventions (OR), major nonorthopedic operative interventions or death (Maj-OR), and injury severity score (ISS). To optimize sensitivity and specificity of out-of-hospital triage decision rules, receiver operating characteristic (ROC) curves were derived. RESULTS: Of 1,545 patients, 13% were admitted; 6% had OR; 1% had Maj-OR; and 3% had ISSs > or = 16. For all outcomes, the most useful criteria were physiologic and anatomic. Some additional criteria (crash speed > 20 mph, > or = 30-inch vehicle deformity, axle displacement) substantially worsened specificity, with minimal or no improvement in sensitivity. For example, the optimal ROC curve for Maj-OR was determined by a systolic blood pressure < 90 mm Hg, Glasgow Coma Scale (GCS) score < 13, respiratory rate (RR) < 10 or > 29, death of a same-car occupant, penetrating injury, and/or > or = 24-inch opposite-side compartment intrusion (sensitivity, 85%; specificity, 87%). An ISS > or = 16 was predicted by GCS score < 13, RR < 10 or > 29, penetrating injury, 2 proximal long bone fractures, flail chest, > or = 24-inch opposite-side compartment intrusion, patient ejection, rollover, and/or age < 5 or > 55 years (sensitivity, 86%; specificity, 70%). CONCLUSION: Physiologic and anatomic trauma triage criteria predicted increased hospital resource utilization and severe injury. On the other hand, when used concurrently with physiologic, anatomic, and "other" criteria, some mechanism criteria worsen specificity with negligible improvement in sensitivity. In particular, crash speed > 20 mph and > or = 30-inch vehicle deformity had little predictive value for all outcomes.

Cellular components and tunic architecture of the solitary ascidian Styela canopus (Stolidobranchiata, Styelidae).

Cell distribution and tunic morphology in the ascidian Styela canopus were examined by electron microscopy. The observations showed that the outer covering is composed of a thin sinuous cuticle with several protrusions and a deep layer of ground substance. The fibrous component and its arrangement in the tunic were demonstrated: elementary fibrils exhibit a 'microtubular' structure and an elliptical cross-sectional shape. Four types of cells were described: clear vesicular tunic granulocytes, tunic microgranulocytes, unilocular tunic granulocytes, and globular tunic granulocytes. Morphofunctional aspects of the tunic tissue and certain phylogenetic relationships are discussed.

Encapsulation response of Ciona intestinalis (Ascidiacea) to intratunical erythrocyte injection. I. The inner capsular architecture.

Previous studies on the ascidian Ciona intestinalis have shown that an encapsulation response is experimentally induced by inserting vertebrate erythrocytes into the tunic, which initiates a massive inflammatory cell infiltration to isolate the injured area. Several hemocytes contribute to capsule formation, destruction of the foreign cells, tunic regeneration, and wound healing. The fine features of some inflammatory cell types are described although the complete capsular structure is not yet reported. Accordingly, the present investigation further examines various aspects of this cellular reaction against erythrocytes and, for the first time, presents the involvement of extratunical circulating hemocytes and mantle epithelium in capsule formation.

Isoelectric focusing studies on human pancreatic secretion.

Pure bile, pancreatic and duodenal human juices have been analyzed by isoelectric focusing, either at rest or upon stimulation with caerulein. In rats, stimulation has also been performed by secretin. Twenty bands have been resolved and quantified in the pancreatic secretion. By developing zymograms, a number of isozymes have been identified: 6 iso-amylases [pl's 7.2, 7.1 and 6.6 (major) and pl's 7.4, 6.7 and 5.8 (minor)], 3 lipases [pl's 7.0 and 6.8 (major) and 6.4 (minor)], two major alkaline proteases (pl's 9.8 and 8.4) and one major acidic protease (pl 4.3) and one band of RNAase activity (pl 8.6). The stimulation kinetics follow a mechanism according to Palade, indicating uniform response to secretogogues, parallel intracellular transport and parallel discharge of pancreatic exocrine proteins.

Computer-assisted assessment of the variations of sperm kinetics after semen treatment with Pellet Swim-up (PSu), and with centrifugation on a discontinuous Percoll gradient at reduced volumes (mini-CDPG).

The semen of the male partners of 37 infertile couples was analyzed with the use of the HTM-S Motility Analyzer (HTM-S MA). For each sample it was thus possible to assess the Total Concentration (TC), the Motile Cell Population Concentration (MCPC), the Progressive Cell Population Concentration (PCPC), the Average Path Velocity (VAP), the Straight Line Velocity (VSL), the Curvilinear Velocity (VCL), the Straightness (STR), the Linearity (LIN), the Lateral Head Displacement (ALH) and the Head Size (HS). The same parameters were then measured on the same semen after treatment with the Pellet Swim-up (PSu) and following Centrifugation on reduced-volume Discontinuous Percoll gradient (mini-CDPG). There was a significant difference in the TC (51.63 M/ml +/- 43.99 in the untreated ejaculate, 11.48 M/ml +/- 9.66 after PSu, 7.94 M/ml +/- 7.3 after mini-CDPG; chi r2 = 50.392, p < 0.05), in the MCPC (20.95 M/ml +/- 26.29 in the untreated ejaculate, 3.79 M/ml +/- 4.26 after PSu, and 2.74 M/ml +/- 3.73 after mini-CDPG; chi r2 = 33.55, p < 0.05), and in the PCPC (7.8 M/ml +/- 12.87 in the untreated ejaculate, 1.81 M/ml +/- 2.36 after PSu, and 1.28 M/ml +/- 1.73 after min-CDPG; chi r2 = 6.38; p < 0.05). The overall comparison between the couples showed a significant difference in the MCPC after PSu and after mini-CDPG (z = -2.09, p < 0.05) whereas no significant difference was found in the comparison off the results of either the TC after PSu and after mini-CDPG (z = -1.9; NS), or of the PCPC after PSu and after mini-CDPG (z = -1.68; NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Level of training, wound care practices, and infection rates.

This prospective, nonrandomized descriptive study compares the traumatic wound infection rates in patients based on level of training of emergency department (ED) practitioners. Wounds were evaluated in 1,163 patients. A wound registry data sheet was prospectively completed on all patients sutured in the ED. All practitioners were assigned a unique identification number. Follow-up data was obtained at the time of the return visit. Patients failing to return were contacted by telephone. Data were analyzed for patient wound infection rates by practitioner level of training. Patient wound infection rates by practitioner level of training were: medical students, 0/60 (0%); all resident physicians, 17/547 (3.1%); physician assistants, 11/305 (3.6%); and attending physicians 14/251 (5.6%), P was not significant. Comparison of junior (medical students and interns) to senior practitioners (all other practitioners) found no difference in infection rates (8/262 [3.1%] v 34/901 [3.8%], P = .58). In conclusion, carefully selected patients sutured by closely supervised medical students and junior residents have infection rates as low as those sutured by more experienced practitioners.

Comparison of plain, warmed, and buffered lidocaine for anesthesia of traumatic wounds.

STUDY OBJECTIVE: To compare pain on infiltration, need for additional anesthesia, and pain on suturing in patients given plain, warm, and buffered lidocaine preparations before the suturing of traumatic wounds in the emergency department. DESIGN: Randomized, prospective, single-blinded convenience sample. SETTING: University hospital ED. PARTICIPANTS: Patients with traumatic lacerations. Patients allergic to lidocaine and patients with abnormal mental status or altered pain sensorium were excluded. INTERVENTIONS: All wounds were anesthetized by use of a standard injection technique. Wound margins were anesthetized with plain, buffered, or warm lidocaine in a randomized fashion. Pain of infiltration was recorded for each margin by means of a previously validated visual-analogue pain scale. RESULTS: The main outcome parameter was pain of infiltration. Need for additional anesthesia and pain on suturing were secondary outcome parameters. We evaluated 45 patients. Pain on injection varied by the type of lidocaine (mean pain scores: plain, 8.2; buffered, 4.7 [P < .05 versus plain]; warm, 4.9 [P < .05 versus plain]). There was no significant difference between the mean pain scores of the groups given warm and buffered lidocaine (P = NS). Need for additional anesthesia and pain on suturing did not vary by the type of anesthesia. The order of injection was not found to influence results. Mean pain scores were not different for margins 1 and 2 in any of the groups (P = NS). CONCLUSION: Both buffered and warmed lidocaine were as efficacious as plain lidocaine, and they had significantly less pain associated with infiltration than did plain lidocaine. In EDs with fluid warmers, warmed lidocaine may be the most convenient, well-tolerated, efficacious lidocaine preparation for anesthesia of traumatic wounds.

Prospective countywide evaluation of the effects of motor vehicle safety device use on hospital resource use and injury severity.

STUDY OBJECTIVE: We evaluated the efficacy of safety devices (seat belts and air bags) on hospital resource use and injury severity while controlling for variables known to affect outcome of motor vehicle crashes. METHODS: We conducted a prospective observational study of motor vehicle crash victims who were transported to the hospital in a suburban-rural county with a population of 1.4 million. Out-of-hospital providers recorded patient demographic characteristics, mechanism of injury (based on American College of Surgeons criteria), and use of seat belts or air bags. Hospital charts were reviewed for hospital admission, length of hospital stay, ICU stay, surgery, major nonorthopedic surgery or death, and severe injury (Injury Severity Score > 15). RESULTS: The study group comprised 1,446 motor vehicle crash victims. Median age was 30 years, 47% were male, and 69% used safety devices. The use of safety devices reduced the likelihood of hospital admission (adjusted odds ratio [OR], .67; 95% confidence interval [CI], .46 to .98) surgery (adjusted OR, .52; 95% CI, .27 to 1.01), major surgery or death (adjusted OR, .14; 95% CI, .04 to .52), and severe injuries (adjusted OR of injury Severity Score > 15, .39; 95% CI, .19 to .83). CONCLUSION: The use of safety devices is associated with decreased hospital resource use and less severe injuries for victims of motor vehicle crashes who are transported to the hospital by emergency medical services providers, even after adjustment for many other variables known to affect the outcome of motor vehicle crashes. Continued efforts to promote safety device use are warranted.

Evaluation of American College of Surgeons trauma triage criteria in a suburban and rural setting.

In suburban and rural counties, patient transport to specialized facilities such as trauma centers may result in prolonged transport times with the resultant loss of ambulance coverage in the primary service area. We evaluated the American College of Surgeons trauma triage criteria as modified by New York State to determine the ability of these criteria to predict the need for trauma center care in victims of blunt traumatic injury. Blunt trauma patients were retrospectively identified through review of patient care reports for the presence either of mechanism or of physiological criteria for transport to a trauma center. Controls were randomly selected from patients with blunt trauma not meeting any of the criteria. Main outcome parameters were the emergency department (ED) disposition, length of hospital stay, need for intensive care unit (ICU) care, and major nonorthopedic operative interventions. There were 857 patients enrolled. The presence either of mechanism or of physiological criteria increased the likelihood of hospital admission (control, 11%; mechanism, 35%; and physiological, 33%). Relative to patients without any criteria, the presence of mechanism criteria alone did not identify patients who required a prolonged length of stay (67% vs 71%), intensive care unit services (13% vs 19%) or major nonorthopedic operative interventions (0.2% vs 1.6%). The presence of physiological criteria increased the likelihood of requiring all of these services. These comparisons held true for victims of motor vehicle accidents, pedestrians struck by motor vehicles, and people who fell from heights above ground level. Patients with physiologic criteria may benefit from transport directly to a trauma center. Because of the low need for operative intervention and ICU services, patients with no criteria or mechanism criteria at long distances from a trauma center may be initially evaluated at the closest hospital and transferred to a trauma center if hospitalization or ICU care is necessary. Further study to determine the predictive value of certain individual mechanism criteria is warranted.

Activation of the acrosome reaction after treatment with human follicular fluid. A morphofunctional evaluation useful for in vitro fertilization.

Studies aided by transmission electron microscopy were made in order to evaluate the occurrence and efficiency of the human follicular fluid in the activation of the processes leading to the acrosome reaction; quantitative, qualitative and morphological data and relative evaluations are here presented. The effects of the human follicular fluid were compared with those determined by other types of treatment, semen untreated, Pellet-Swim-up, and Centrifugation on Discontinuous Percoll Gradient. The transmission electron microscope observations permitted to evaluate the percentage of sperm with an activation of the acrosome reaction in the different groups. The analysis of the data showed a statistically relevant difference (P < 0.001) among the first group (Control group) and the SU group, the MP group and the hFF group, regarding the sperm with AR activation. The sperm treatment with 50% diluted hFF represents an important option to the preparation protocols of the semen samples useful for ART.

PAH gene mutations in the Sicilian population: association with minihaplotypes and expression analysis.

The molecular basis of PAH deficiency in the Sicilian population is characterized by a marked heterogeneity, with 44 mutations at a single locus identified by a "gene-scanning" approach and accounting for a detection rate of 91%. The remaining 9% of PAH alleles does not bear mutations in any of the 13 exons and 24 exon/intron junctions. Three mutations IVS10nt-11 G > A, R261Q, and A300S accounted for 30.5%, whereas the remaining mutations were found at relative frequencies of less than 5% and 20 mutations were observed once only. Five mutations have been detected only in Sicilians so far. By studying the association of mutations with intragenic STR-VNTR haplotypes ("minihaplotypes"), "identity by descent" has been established for 24 mutations also detected in other populations. This finding supports the hypothesis of a multipolar origin for a large proportion of PAH mutant alleles currently detected in Sicilians. In order to improve our understanding of the clinical heterogeneity of PAH deficiency in this population, we have for the first time analyzed three missense mutations L41F, T92I, and P211T in vitro by the pCDNA3/COS-7 eukaryotic expression system and found an activity of 10, 76, and 72%, respectively, compared to normal PAH. In two HPA patients with mild PKU and mild hyperphenylalaninemia (MHP), harboring respectively L41F/R261Q and T92I/P281L genotypes, the predicted biochemical effect of these genotypes appeared to be consistent with the metabolic phenotypes. In contrast, discordant metabolic phenotypes (mild PKU and MHP) were observed in two unrelated patients bearing the same R261Q/P211T genotype, a finding which underscores the complex relationship linking genotype to phenotype in PAH deficiency. Hypotheses on the possible mechanisms responsible for the observed discordance are discussed. The spectrum of PAH gene mutations in Sicily reflects the complex demographic history of this island at the crossroad of prehistoric and historical migrations in the Mediterranean sea. The data presented in this study also add to the present knowledge on the relationship between PAH genotypes and HPA phenotype and are expected to improve PAH genotyping among individuals with hyperphenylalaninemia.

Encapsulation response of Ciona intestinalis (Ascidiacea) to intratunical erythrocyte injection.

Electron microscopic studies on the encapsulation induced by erythrocyte injection into the tunic of the ascidian Ciona intestinalis were carried out. The observations reported in the present paper complete the description previously given of capsule architecture and contribute to the characterization of the cells involved in the inflammatory reaction. The inflamed area is surrounded by an ample and peculiar "three-layered coat" respectively composed of flattened and packed extratunical hemocytes, the monolayered epithelium, and a layer of intratunical electron-dense particles. The latter are also clustered, variously arranged, and distributed in the tunic ground substance. The epithelial cells appear to be undergoing an active secretory phase; in several regions they also show discontinuities and organule and surface changes. The infiltrating hemocytes, mainly globular and unilocular granulocytes, appear frequently to be degranulating and in relation with electron-dense particles, net-like fibrous materials, and fine membranes. The involvement of morula-shaped granulocytes is discussed, as well as possible relationships with the melanization process, and finally analogies in the structural organization with the inflammatory reactions induced in other invertebrates. A schematic drawing, based on all available observations of the capsule architecture, is presented in order to reconstruct the entire inflamed area and illustrate the relative fine features.