Search

INTRODUCTION: Knee dislocations (KD) have high rates of multi-ligamentous injury (MLI). Collateral ligaments rupture in 50-60% of KDs. Traditionally, collateral ligaments have undergone primary repair, though microscopic healing is not optimal. Artelon is a degradable, polyurethane urea bio- scaffold thought to decrease mechanical forces and promote healing, motion, and strength. Currently, little evidence exists regarding its indications or outcomes. MATERIALS AND METHODS: Thirty-two patients with KD and MLI undergoing collateral ligament repair at a level-I trauma centre between 2015-2020 were included. Patients age 15° of knee flexion. Only one gross failure occurred, in the AG. No differences were noted in infection or re-operation. Lysholm Knee Scale and Tegner Activity Scale were not significantly different, although Tegner scores in both cohorts decreased from pre-injury scores. CONCLUSION: In summary, Artelon appears to be safe without increasing risk for hypersensitivity or infection when used for collateral ligament augmentation. Additionally, Artelon appeared to be non-inferior and statistically equivalent to primary repair in this setting and may have promise with use in certain types of knee dislocations.

The polariton-polariton interaction strength is an important parameter for all kinds of applications using the nonlinear properties of polaritons, such as optical switching and single-photon blockade devices. In this paper, we review and compare the results of a series of experiments on polariton-polariton interactions in GaAs/AlxGa1-xAs microcavity polariton structures and present an updated analysis of these experiments. We show that not just the energy shift of the spectral lines but also the results of measurements sensitive to the polariton scattering rate are important for the calibration of the interaction parameter at low excitation density. We find that when adjustments are made to correct for recent understanding of the experiments, the value of the interaction parameter at low density is lower than previous reported, but still significantly higher than theoretically predicted.

Background: The diagnosis of infectious flexor tenosynovitis (FTS) has historically been made based on physical exam using Kanavel's signs. The specificity of these findings has come into question. We looked to evaluate the use of contrast-enhanced computed tomography (CT) in increasing the successful diagnosis of FTS., Methods: Two adult cohorts were formed, one of patients with FTS confirmed in the operating room and the second of patients with ICD.10 identified finger cellulitis (FC), without concomitant FTS. Demographics, laboratory values, CT scans, and examination findings were evaluated. Axial CTs were evaluated in the coronal and sagittal planes and tendon sheath/tendon width were measured. The tendon sheath/tendon was recorded as a ratio in the coronal (CR) and sagittal (SR) planes. Continuous and dichotomous variables were analyzed and measures of sensitivity, specificity, and predictivity were calculated. Seventy patients were included, 35 in the FTS cohort and 35 with FC., Result: A higher number of Kanavel signs were present in the FTS group (2.9 vs. 0.5, P < .05), with CR and SR both being significantly larger in the FTS group ( P < .05). CR and SR cutoffs ≥ 1.3 provided high sensitivity, specificity, and positive predictive value (PPV) for FTS. Likelihood of FTS increased 5.9% and 5.5% for every 0.1 increase in CR and SR, respectively, with a 14% increase for every additional Kanavel sign., Conclusion: In conclusion, CT ratios are useful in identifying FTS; and when used on their own or in combination with Kanavel's signs, CR and SR objectively improve the diagnosis of FTS., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: BT receives payments from ZimmerBiomet, Atricure, and Temple University. In addition to these he also has a relationship with Orthobullets.com. None of these are relevant to this manuscript. The remaining authors do not have any disclosures of a conflict of interest.

The white sturgeon (Acipenser transmontanus) is the largest freshwater fish in North America. Because of the unique life history characteristics of sturgeon, including longevity, late maturation and long spawning intervals, their aquaculture can be a significant investment of resources. As a result of habitat loss and overharvesting, natural populations of white sturgeon are threatened and there is a growing effort to improve conservation aquaculture programs. Germ cell transplantation is an innovative technology previously demonstrated in a variety of fish species to be able to produce a surrogate broodstock. The technique relies upon optimal donor germ cell recovery and transplantation into a recipient fish. In this study, we developed and optimized the harvest of donor cells for germline transplantation and evaluated methods for ovary cryopreservation for the first time in the white sturgeon. We found that harvesting gonads from juveniles between the ages of 1.5 and 2.5-years resulted in reliably high proportions of pre-meiotic cells regardless of sex, a critical feature for using white sturgeon for transplantation studies since the species shows no distinguishing external sex characteristics. From the viable cells, we identified germline cells using immunolabeling with the antibody DDX4, a marker specific to the germline. For in vivo tracking of donor cells during transplantations, gonadal cells were stained with a long half-life non-toxic cell membrane dye, PKH26, and microinjected into the peritoneal cavity of newly hatched white sturgeon larvae. Larvae were reared until 3 months post-transplantation to monitor for colonization and proliferation of PKH26-labeled cells within the recipient larval gonads. Furthermore, viable cell detection, assessment of germline-specificity, and transplantation was determined for cells recovered from cryopreserved ovarian tissue from sexually immature females. Transplantations using cells cryopreserved with media supplemented with dimethyl sulfoxide (DMSO) rather than ethylene glycol (EG) demonstrated the highest number of PKH26-labeled cells distributed along the gonadal ridges of the larval recipient. Determining optimal methods of tissue cryopreservation, and germ cell recovery and transplantation are foundational to the future development of germ cell transplantation as a strategy to improve the aquaculture and conservation of this species. Our study demonstrates that conservation actions, such as surrogate breeding, could be utilized by hatcheries to retain or improve natural gamete production without genetic modification, and provide an encouraging approach to the management of threatened sturgeon species., (© 2023. Springer Nature Limited.)

Intramedullary nailing of femur fractures has become the standard of care, with high union rates. Few high-level studies have discussed the effect that early weight bearing has on the healing of these fractures, regardless of nail size or fracture pattern. The goal of this study was to determine the clinical and radiographic outcomes of femoral shaft fractures for patients allowed immediate weight bearing after intramedullary nailing. We performed a retrospective review of 341 femoral shaft fractures, with 131 allowed immediate weight bearing, 99 allowed partial weight bearing, and 111 kept non-weight bearing. Demographic, intraoperative, and postoperative variables were collected and analyzed. Increased fracture complexity was associated with higher likelihood of delayed weight bearing. No significant difference was found for nail size or rate of failure with different nail sizes. A total of 50 nonunions were noted (14.7%), with no difference in nonunion rates between weight bearing cohorts. The only significant predictor of nonunion was Orthopaedic Trauma Association (OTA) classification of OTA32B fractures ( P =.02), which were 2 times and 4 times as likely to occur compared with OTA32A and OTA32C fractures, respectively. Failure of interlocking screws occurred among 15 patients (4.4%) and was more common with older patients, osteoporotic bone, and larger diameter nails. In summary, unilateral intramedullary nailing of adult femoral shaft fractures does not show a difference in fracture union rates or implant failure with unrestricted, immediate weight bearing, regardless of nail characteristics. [ Orthopedics . 2022;45(3):e148-e153.].

Significance: Deranged metabolism and dysregulated growth factor signaling are closely associated with abnormal levels of proliferation, a recognized hallmark in tumorigenesis. Fluorescence lifetime imaging microscopy (FLIM) of endogenous nicotinamide adenine dinucleotide (NADH), a key metabolic coenzyme, offers a non-invasive, diagnostic indicator of disease progression, and treatment response. The model-independent phasor analysis approach leverages FLIM to rapidly evaluate cancer metabolism in response to targeted therapy., Aim: We combined lifetime and phasor FLIM analysis to evaluate the influence of human epidermal growth factor receptor 2 (HER2) inhibition, a prevalent cancer biomarker, on both nuclear and cytoplasmic NAD(P)H of two squamous cell carcinoma (SCC) cultures. While better established, the standard lifetime analysis approach is relatively slow and potentially subject to intrinsic fitting errors and model assumptions. Phasor FLIM analysis offers a rapid, model-independent alternative, but the sensitivity of the bound NAD(P)H fraction to growth factor signaling must also be firmly established., Approach: Two SCC cultures with low- and high-HER2 expression, were imaged using multiphoton-excited NAD(P)H FLIM, with and without treatment of the HER2 inhibitor AG825. Cells were challenged with mitochondrial inhibition and uncoupling to investigate AG825's impact on the overall metabolic capacity. Phasor FLIM and lifetime fitting analyses were compared within nuclear and cytoplasmic compartments to investigate epigenetic and metabolic impacts of HER2 inhibition., Results: NAD(P)H fluorescence lifetime and bound fraction consistently decreased following HER2 inhibition in both cell lines. High-HER2 SCC74B cells displayed a more significant response than low-HER2 SCC74A in both techniques. HER2 inhibition induced greater changes in nuclear than cytoplasmic compartments, leading to an increase in NAD(P)H intensity and concentration., Conclusions: The use of both, complementary FLIM analysis techniques together with quantitative fluorescence intensity revealed consistent, quantitative changes in NAD(P)H metabolism associated with inhibition of growth factor signaling in SCC cell lines. HER2 inhibition promoted increased reliance on oxidative phosphorylation in both cell lines.

Current practice allows early weight bearing of unstable ankle fractures after fixation. This study offers a unique comparison of early weight bearing (EWB) vs late weight bearing (LWB) in operatively stabilized trimalleolar ankle fractures. The goal of this study was to evaluate union rates, clinical outcomes, and complications for patients who were managed with EWB vs LWB. The authors performed a retrospective review of 185 patients who underwent surgical stabilization for trimalleolar ankle fracture. Fixation of the posterior malleolus and weight bearing status were determined by surgeon preference. For this study, EWB was defined as 3 weeks or less and LWB was defined as greater than 3 weeks. Patients were evaluated for fracture union and implant failure. Complications and clinical outcomes included ambulatory status, infection rate, and return to surgery. The EWB group included 47 (25.4%) patients, and the LWB group included 138 (74.6%) patients. Of the 7 nonunions, 1 (14.3%) occurred in the EWB group and 6 (85.7%) in the LWB group. A total of 72 (38.9%) posterior malleolar fractures were operatively stabilized, and stabilization did not affect union rates. Syndesmotic fixation was required for 12.5% of patients, despite posterior malleolar stabilization. Syndesmotic fixation increased the union rate 2.5 times. Deep infection and open fracture decreased union. No difference was seen between groups in implant failure, union rate, infection, or return to the operating room. No deleterious effect of EWB in operatively treated trimalleolar ankle fractures was found for union, implant failure, infection, or reoperation. Syndesmotic fixation may offer an advantage over posterior malleolar fixation, with improved union rates. [ Orthopedics . 2021;44(3):160-165.].

Proximal humerus fractures, although common, have high rates of failure after open reduction and internal fixation. The use of a fibular allograft has been explored as a means to decrease complications, particularly varus collapse and the need for revision surgery. The authors performed a retrospective review of 133 proximal humerus fractures managed surgically with locking plates (n=72) or locking plates with fibular allograft intramedullary struts (n=61). Demographic, intraoperative, and postoperative variables were collected and analyzed. The fibular allograft group was more likely to be older (P<.01), be female (P=.04), and have a history of osteoporosis (P=.01). No differences were noted in the proportions of 2-, 3-, or 4-part fractures between groups. Average follow-up was 28 weeks. Medial calcar length was longer in the locking plate only group (P=.04); however, this group demonstrated a decreased head shaft angle (P=.01) and a trend toward increased rates of varus collapse (P=.06). No significant differences were found regarding other radiographic complications, irrespective of fracture complexity. A notable decrease in fluoroscopy time was seen with strut use (P=.04), but operative time and blood loss were similar between groups. A significant decrease in revision surgery rate was found with use of an allograft strut (P=.05). Using a strut appears to preserve the radiographic head shaft angle and decrease the risk of fracture collapse in 2-, 3-, and 4-part fractures, without increasing surgical time or morbidity. Use of an intramedullary strut appears to reduce the need for revision surgery, particularly in 3- and 4-part fractures. [Orthopedics. 2020;43(5):262-268.]., (Copyright 2020, SLACK Incorporated.)

Ankle fractures are common orthopedic injuries. Although operative indications and subsequent stabilization of these fractures have not significantly changed, postoperative protocols remain highly variable. Effects of early weight bearing (EWB) on fracture characteristics in operatively stabilized bimalleolar and bimalleolar equivalent ankle fractures remain poorly publicized. This study seeks to clarify postoperative fracture union rates, rates of hardware loosening or failure, and radiographic medial clear space changes when comparing EWB to late weight bearing (LWB) following open reduction and internal fixation (ORIF). A total of 95 patients with either bimalleolar (66%) or bimalleolar equivalent (34%) fractures who underwent ORIF were retrospectively reviewed. Weight bearing was allowed at three weeks in the EWB group and when signs of radiographic union were noted in the LWB group. Postoperatively, patients were evaluated at regular intervals for fracture union, signs of implant failure, and evidence of medial clear space widening radiographically. There were 38 patients (40%) in the EWB group and 57 patients (60%) comprising the LWB cohort. There were no significant demographic differences between groups. The EWB group on average began to weight bear at 3.1 + 1.4 weeks postoperatively, whereas the LWB group began at 7.2 + 2.1 weeks postoperatively (p<0.01). Union rate (p=0.51), time to union (p=0.23), and implant failure (p>0.1 at all time intervals) were not notably different between groups. No differences in medial clear space were detected at any postoperative interval between groups (p>0.1 at all time intervals). This study suggests that EWB at three weeks postoperatively does not increase markers of radiographic failure compared to six weeks of non-weight bearing (NWB), which has been regarded as the gold standard of treatment to allow for healing; this may represent an improvement to rehabilitation protocols after bimalleolar ankle ORIF of unstable ankle fractures., Competing Interests: The authors have declared financial relationships, which are detailed in the next section., (Copyright © 2020, Passias et al.)

Background: Genome-wide association studies have implicated variants in SCN10A, which encodes Nav1.8, as modulators of cardiac conduction. Follow-up work has indicated the SCN10A sequence includes an intronic enhancer for SCN5A. Yet the role of the Nav1.8 protein in the myocardium itself is still unclear. To investigate this, we use homozygous knockout mice (Scn10a -/- ) generated by disruption of exons 4 and 5, leaving the Scn5a enhancer intact., Methods and Results: We previously reported that pharmacologic blockade of Nav1.8 in wild-type animals blunts action potential prolongation by ATX-II at slow drive rates (≤1 Hz). Here we present evidence of the same blunting in Scn10a -/- compared to wild-type ventricular myocytes, supporting the conclusion that Nav1.8 contributes to late sodium current at slow rates. In contrast to earlier studies, we found no differences in electrocardiographic parameters between genotypes. Low-dose ATX-II exposure in lightly anesthetized animals and Langendorff-perfused hearts prolonged QTc and generated arrhythmias to the same extent in wild-type and Scn10a -/- . RNA sequencing failed to identify full-length Scn10a transcripts in wild-type or knockout isolated ventricular myocytes. However, loss of late current in Scn10a -/- myocytes was replicated independently in a blinded set of experiments., Conclusions: While Scn10a transcripts are not detectible in ventricular cardiomyocytes, gene deletion results in reproducible loss of late sodium current under extreme experimental conditions. However, there are no identifiable consequences of this Scn10a deletion in the intact mouse heart at usual rates. These findings argue that common variants in SCN10A that affect ventricular conduction do so by modulating SCN5A., (© 2016 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.)

Aims: Female gender is a risk factor for long QT-related arrhythmias, but the underlying mechanisms remain uncertain. Here, we tested the hypothesis that gender-dependent function of the post-depolarization 'late' sodium current (I(Na-L)) contributes., Methods and Results: Studies were conducted in mice in which the canonical cardiac sodium channel Scn5a locus was disrupted, and expression of human wild-type SCN5A cDNA substituted. Baseline QT intervals were similar in male and female mice, but exposure to the sodium channel opener anemone toxin ATX-II elicited polymorphic ventricular tachycardia in 0/9 males vs. 6/9 females. Ventricular I(Na-L) and action potential durations were increased in myocytes isolated from female mice compared with those from males before and especially after treatment with ATX-II. Further, ATX-II elicited potentially arrhythmogenic early afterdepolarizations in myocytes from 0/5 male mice and 3/5 female mice., Conclusion: These data identify variable late I(Na) as a modulator of gender-dependent arrhythmia susceptibility.

Rationale: Although the sodium channel locus SCN10A has been implicated by genome-wide association studies as a modulator of cardiac electrophysiology, the role of its gene product Nav1.8 as a modulator of cardiac ion currents is unknown., Objective: We determined the electrophysiological and pharmacological properties of Nav1.8 in heterologous cell systems and assessed the antiarrhythmic effect of Nav1.8 block on isolated mouse and rabbit ventricular cardiomyocytes., Methods and Results: We first demonstrated that Scn10a transcripts are identified in mouse heart and that the blocker A-803467 is highly specific for Nav1.8 current over that of Nav1.5, the canonical cardiac sodium channel encoded by SCN5A. We then showed that low concentrations of A-803467 selectively block "late" sodium current and shorten action potentials in mouse and rabbit cardiomyocytes. Exaggerated late sodium current is known to mediate arrhythmogenic early afterdepolarizations in heart, and these were similarly suppressed by low concentrations of A-803467., Conclusions: Scn10a expression contributes to late sodium current in heart and represents a new target for antiarrhythmic intervention.

Background: The D1275N SCN5A mutation has been associated with a range of unusual phenotypes, including conduction disease and dilated cardiomyopathy, as well as atrial and ventricular tachyarrhythmias. However, when D1275N is studied in heterologous expression systems, most studies show near-normal sodium channel function. Thus, the relationship of the variant to the clinical phenotypes remains uncertain., Methods and Results: We identified D1275N in a patient with atrial flutter, atrial standstill, conduction disease, and sinus node dysfunction. There was no major difference in biophysical properties between wild-type and D1275N channels expressed in Chinese hamster ovary cells or tsA201 cells in the absence or presence of β1 subunits. To determine D1275N function in vivo, the Scn5a locus was modified to knock out the mouse gene, and the full-length wild-type (H) or D1275N (DN) human SCN5A cDNAs were then inserted at the modified locus by recombinase mediated cassette exchange. Mice carrying the DN allele displayed slow conduction, heart block, atrial fibrillation, ventricular tachycardia, and a dilated cardiomyopathy phenotype, with no significant fibrosis or myocyte disarray on histological examination. The DN allele conferred gene-dose-dependent increases in SCN5A mRNA abundance but reduced sodium channel protein abundance and peak sodium current amplitudes (H/H, 41.0±2.9 pA/pF at -30 mV; DN/H, 19.2±3.1 pA/pF, P<0.001 vs. H/H; DN/DN, 9.3±1.1 pA/pF, P<0.001 versus H/H)., Conclusions: Although D1275N produces near-normal currents in multiple heterologous expression experiments, our data establish this variant as a pathological mutation that generates conduction slowing, arrhythmias, and a dilated cardiomyopathy phenotype by reducing cardiac sodium current.

Rationale: Although multiple lines of evidence suggest that variable expression of the cardiac sodium channel gene SCN5A plays a role in susceptibility to arrhythmia, little is known about its transcriptional regulation., Objective: We used in silico and in vitro experiments to identify possible noncoding sequences important for transcriptional regulation of SCN5A. The results were extended to mice in which a putative regulatory region was deleted., Methods and Results: We identified 92 noncoding regions highly conserved (>70%) between human and mouse SCN5A orthologs. Three conserved noncoding sequences (CNS) showed significant (>5-fold) activity in luciferase assays. Further in vitro studies indicated one, CNS28 in intron 1, as a potential regulatory region. Using recombinase-mediated cassette exchange (RMCE), we generated mice in which a 435-base pair region encompassing CNS28 was removed. Animals homozygous for the deletion showed significant increases in SCN5A transcripts, Na(V)1.5 protein abundance, and sodium current measured in isolated ventricular myocytes. ECGs revealed a significantly shorter QRS (10.7±0.2 ms in controls versus 9.7±0.2 ms in knockouts), indicating more rapid ventricular conduction. In vitro analysis of CNS28 identified a short 3' segment within this region required for regulatory activity and including an E-box motif. Deletion of this segment reduced reporter activity to 3.6%±0.3% of baseline in CHO cells and 16%±3% in myocytes (both P<0.05), and mutation of individual sites in the E-box restored activity to 62%±4% and 57%±2% of baseline in CHO cells and myocytes, respectively (both P<0.05)., Conclusions: These findings establish that regulation of cardiac sodium channel expression modulates channel function in vivo, and identify a noncoding region underlying this regulation.

Rationale: Voltage-gated sodium channels initiate action potentials in excitable tissues. Mice in which Scn5A (the predominant sodium channel gene in heart) has been knocked out die early in development with cardiac malformations by mechanisms which have yet to be determined., Objective: Here we addressed this question by investigating the role of cardiac sodium channels in zebrafish heart development., Methods and Results: Transcripts of the functionally-conserved Scn5a homologs scn5Laa and scn5Lab were detected in the gastrulating zebrafish embryo and subsequently in the embryonic myocardium. Antisense knockdown of either channel resulted in marked cardiac chamber dysmorphogenesis and perturbed looping. These abnormalities were associated with decreased expression of the myocardial precursor genes nkx2.5, gata4, and hand2 in anterior lateral mesoderm and significant deficits in the production of cardiomyocyte progenitors. These early defects did not appear to result from altered membrane electrophysiology, as prolonged pharmacological blockade of sodium current failed to phenocopy channel knockdown. Moreover, embryos grown in calcium channel blocker-containing medium had hearts that did not beat but developed normally., Conclusions: These findings identify a novel and possibly nonelectrogenic role for cardiac sodium channels in heart development.

Social workers in diverse community practice settings recruit and work with volunteers from religious congregations. This article reports findings from two surveys: 7,405 congregants in 35 Protestant congregations, including 2,570 who were actively volunteering, and a follow-up survey of 946 volunteers. It compares characteristics of congregation volunteers and nonvolunteers.Volunteers tended to be married, older, more highly educated, longer-term congregational members, and to score higher on all measures of faith maturity and faith practice than did nonvolunteers.Volunteers perceived their involvement as meaningful, important, and challenging. A large majority of volunteers (80 percent) reported changes in faith, attitudes and values, and behavior as results of their volunteer work. Findings provide insights into how religious individuals begin and continue to volunteer in service settings and how congregations promote high levels of community service among their members. These findings have implications for effective social work practice with congregation volunteers.

Three groups of beef cow and calf pairs were studied to determine plasma vitamin E and blood selenium (Se) concentrations of calves at 1 month old. Group 1 was managed on irrigated pasture and calves received no Se/vitamin E injections at birth. Group 2 was managed on irrigated pasture, and the calves were injected with Se/vitamin E at birth. Group 3 was managed on dry foothill grasslands, and these cows were supplemented with 56.3 mg vitamin E and 3 mg Se daily, and the calves received a Se/vitamin E injection at birth. The plasma concentration of vitamin E in group 1 and 2 cows (9.5 +/- 1.24 and 8.43 +/- 1.0 microg/ml, respectively) was significantly higher than that of the group 3 cows (2.28 +/- 0.42 microg/ml; P < 0.05). The blood Se concentrations in group 3 cows (169 +/- 37 ng/ml) were significantly higher than those in group 1 and 2 cows (36.4 +/- 15.9 and 31.1 +/- 12.5 ng/ml, respectively; P < 0.05). Calf Se was highly correlated to cow Se (r = 0.965), and calf vitamin E was moderately correlated to cow vitamin E (r = 0.605). Calf vitamin E concentrations were consistently lower than cow vitamin E concentrations, and many values would be considered deficient.

Background: The atrioventricular (AV) node is essential for the sequential excitation and optimized contraction of the adult multichambered heart; however, relatively little is known about its formation from the embryonic AV canal. A recent study demonstrated that signaling by Alk3, the type 1a receptor for bone morphogenetic proteins, in the myocardium of the AV canal was required for the development of both the AV valves and annulus fibrosus. To test the hypothesis that bone morphogenetic protein signaling also plays a role in AV node formation, we investigated conduction system function and AV node morphology in adult mice with conditional deletion of Alk3 in the AV canal., Methods and Results: High-resolution optical mapping with correlative histological analysis of 28 mutant hearts revealed 4 basic phenotypic classes based on electrical activation patterns and volume-conducted ECGs. The frequency of AV node conduction and morphological abnormalities increased from no detectable anomalies (class I) to severe defects (class IV), which included the presence of bypass tracts, abnormal ventricular activation patterns, fibrosis of the AV node, and twin AV nodes., Conclusions: The present findings demonstrate that bone morphogenetic protein signaling is required in the myocardium of the AV canal for proper AV junction development, including the AV node.

A 65-year-old woman developed a watery diarrhea syndrome with collagenous colitis. Later, weight loss and hypoalbuminemia were documented. This prompted small bowel biopsies that showed pathological changes of collagenous sprue. An apparent treatment response to a gluten-free diet and prednisone resulted in reduced diarrhea, weight gain and normalization of serum albumin. Later repeated biopsies from multiple small and large bowel sites over a period of over three years, however, showed reversion to normal small intestinal mucosa but persistent collagenous colitis. These results indicate that collagenous inflammatory disease may be a far more extensive process in the gastrointestinal tract than is currently appreciated. Moreover, collagenous colitis may be a clinical signal that occult small intestinal disease is present. Finally, collagenous sprue may, in some instances, be a completely reversible small intestinal disorder.

This study assessed the effectiveness of a model for diffusing the Life Skills Training (LST) Program into middle schools, examined implementation fidelity, and explored factors associated with involvement in training and program implementation. A convenience sample of master trainers (n = 44) and teachers (n = 45) from 16 Kentucky counties participated. Teachers were observed for content and process fidelity, and trainers and teachers completed questionnaires to assess factors related to training and program implementation. More than one-fourth (27%) of master trainers conducted training sessions, and 60% of teachers taught the curriculum. While implementation fidelity was relatively high, teachers were less likely to use the more innovative elements of the program. Trainers and teachers who conducted training and/or taught the LST Program were more enthusiastic toward the program than those who did nothing beyond being trained.

Many integrin adhesion receptors bind ligands containing the Arg-Gly-Asp (RGD) peptide motif. Most integrins exhibit considerable specificity for particular ligands and can distinguish among the many conformations of RGD. In this study we identify the domain of the integrin beta subunit involved in determining ligand binding specificity. Chimeras of beta3 and beta5, the most homologous integrin beta subunits, were expressed with alphav on the surface of human 293 cells. The ligand binding phenotype of each chimera was assessed using the ligands Fab-9 and fibrinogen, both of which have a binding preference for alphavbeta3. The results of the study show that when exons C and D of the beta3 subunit (residues 95-233) are substituted into beta5, the chimera gained the ability to bind Fab-9 with an affinity close to that of wild-type alphavbeta3. This chimera was able to mediate cell adhesion to fibrinogen. Furthermore, the swap of only a 39-residue segment of this larger domain, beta3 residues 164-202, into the backbone of beta5 enabled the chimeric integrin to bind soluble Fab-9. This small domain is highly divergent among the integrin beta subunits, suggesting that it may play a role in determining ligand selection by all integrins.

A comparison was made with the microslide gel diffusion technique, the complement fixation test, cultural isolation, and clinical examination in detecting ram epididymitis caused by Brucella ovis. The results of the gel diffusion method are shown to be similar to those obtained by the complement fixation test and isolation of B. ovis from cultures of semen. The technique offers a reliable diagnostic method adaptable to field use in controlling ovine brucellosis and is more practical.

The correlation between the modified counterimmunoelectrophoresis test and the indirect fluorescent-antibody test for detecting rabies virus antibodies in human sera was studied, using as a reference the standard mouse neutralization test. Higher sensitivity was observed with the counterimmunoelectrophoresis technique than with the indirect fluorescent-antibody method. The counterimmunoelectrophoresis test provides a practical quantitative alternative to the indirect fluorescent-antibody method.

A modified counterimmunoelectrophoresis (CIE/SN) technique was developed as a diagnostic procedure for the detection of serum-neutralizing antibodies to rabies virus. CIE/SN test results with sera from individuals undergoing postexposure rabies immunization compared favorably with those obtained by the standard mouse neutralization test procedure. Fractionation of selected sera by Sephadex G-200 column chromatography demonstrated that serological activity involved in both procedures was present in the immunoglobulin G-containing fractions of the sera. The mean advantage of the CIE/SN over the standard mouse neutralization test consists in considerably reducing the time required for results, a factor which can be of clinical importance in evaluating antibody levels to rabies virus. In addition, the CIE/SN method is simple to perform and inexpensive.

The counterimmunoelectrophoresis (CIE) technique employing a soluble extract antigen was found useful in screening human sera for antileptospiral antibodies to multiple leptospiral serovars. The CIE test results with sera from patients with suspected cases of leptospirosis and sera from a control group compared favorably (97.9%) with those obtained by the microscopic agglutination test using a battery of live leptospiral antigens. CIE is easy to perform, is highly sensitive, uses an antigen which is very stable at room temperature, and permits the screening of many sera in a short period of time. The CIE technique appears to be ideally suited to a small laboratory where facilities may be too limited for examining sera by the microscopic agglutination test.

The in vitro effectiveness of furazolidone combined with neomycin for the control of contaminants in Leptospira cultures was investigated. Enhanced additive or synergistic bactericidal activity was apparent with the combination when it was compared with each agent used separately or when compared with 5-fluorouracil. Combined furazolidone and neomycin, at 5-mug/ml concentrations each in Fletcher medium or in implanted sensitivity disks of 50 mug and 10 mug, respectively, was effective in inhibiting contaminants and did not interfere with the growth of leptospira belonging to 66 leptospiral collection serotypes. A simple technique for decontaminating swine kidneys from an abattoir for cultural isolation of leptospiras is described. The method involves exposing renal tissue diluted in saline containing furazolidone and neomycin in combination (each in a concentration of 25 mug/ml) for 1 h prior to inoculating culture media. The method is suitable for routine use in the isolation of leptospiras from contaminated clinical or pathological specimens.

The microslide gel-diffusion and macro-tube agglutination techniques to detect Brucella canis antibodies in dogs were compared. Sera from dogs experimentally infected with B. canis and a random sample of dog sera with unknown histories of exposure to this organism were examined. The results of the gel-diffusion method employing specific rough Brucella saline-extract antigens of B. canis and Brucella ovis were comparable to those obtained by the tube agglutination test. The easily prepared, stable R antigen in the freeze-dried form offers a convenient, simple, and reliable diagnostic method for the serological detection of canine brucellosis by the gel-diffusion test.

Three pathogenic leptosipras and 12 saprophytic Leptospira biflexa strains were isolated from 72 apparently normal horse kidneys collected at an abattoir in Argentina. Cross-agglutination reaction patterns of the pathogens showed that they were antigenically homologous with members of the Hebdomadis group. When one of the strains was compared to Hebdomadis serotypes in reciprocal agglutination-absorption tests, it was found to be serologically homologous to serotype hardjo. This is the first known report of an isolation of this serotype from horses. Serological tests were also carried out on randomly collected abattoir sera from 245 horses to determine the prevalence of equine leptospirosis. Significant antibody titers (1:100 or greater) were found in 74.6% of the sera. Predominant reactions occurred with the antigens pomona, hebdomadis group, pyrogenes, tarassovi, and canicola. Agglutination tests performed with antigen prepared with one of the saprophytic biflexa isolates showed seropositive reactions in 99.1% of the equine sera, with agglutination titers ranging from 1:100 to 1:3,200. Absorption of selected horse sera with the saprophytic strain removed the agglutinins to Leptospira interrogans serotypes. This suggests the possibility that L. biflexa strains may act as an antigenic stimulus and account for some of the persistent multiple cross-reaction patterns of equine sera with pathogenic serotypes.

A saprophytic Leptospira biflexa strain of equine origin was found which cross-reacts with immune rabbit antisera to 14 pathogenic Leptospira interrogans serotypes. Sera from goats experimentally inoculated with the saprophyte showed multiple low-level cross-agglutination reactions against a battery of live L. interrogans serotypes. Sonically treated and saline-extracted suspensions of the L. biflexa strain and serotypes canicola, icterohaemorrhagiae, and pomona yielded a common precipitating protein antigen that was detected by immunodiffusion and immunoelectrophoresis with all of the antileptospiral sera examined. In cross-absorption and gel diffusion tests, the precipitinogen from each of the strains was shown to be identical. Formaldehyde treatments and heating at 100 degree C suggest that the cellular location of the common antigen is either somatic or subsurface, and Sephadex G-200 gel filtration enabled the isolation of the active fraction of the L. biflexa antigen. Monoprecipitin sera against the common antigen of L. biflexa were produced by immunizing rabbits with specific precipitates in agar. In gel diffusion and immunoelectrophoresis tests the antisera with each of the soluble antigens developed a single precipitin formation, and the antisera agglutinated formolized and heated whole-cell suspensions of serotypes canicola, icterohaemorrhagiae, and pomona at low dilutions. The soluble L. biflexa antigen was evaluated as an immunogen and in passive immunity tests for protection against death and kidney infection in hamsters. No cross-protection occurred when the hamsters were challenged with virulent leptospires. In contrast, the animals vaccinated or administered hamster immune serum before challenge died earlier than the control animals.

The counterimmunoelectrophoresis (CIE) technique was developed as a diagnostic procedure for ram epididymitis caused by Brucella ovis. CIE test results with sera from naturally and experimentally infected male sheep compared favorably with those obtained by gel diffusion and complement fixation employing the same B. ovis surface R antigen. The main advantage of CIE over gel diffusion consists in a significant reduction of the time required to detect precipitin formation, whereas both methods obviate several of the difficulties encountered with complement fixation tests for B. ovis.

Sonically treated and saline-extracted antigens of Brucella ovis, B. canis, B. abortus, and B. melitensis were compared in gel diffusion, complement fixation, and serum absorption tests. All the sonically extracted antigens showed cross-reactions with sera from animals infected or immunized with these species, whereas the saline-extracted antigens were specific for the surface of the rough or smooth colonial phase of the species or strain. The saline-extracted antigens of B. ovis and B. melitensis were both eluted as a single peak in the void volume by Sephadex G-200 column chromatography, in gel diffusion had staining characteristics of lipoproteins, but in immunoelectrophoresis showed distinct mobility patterns. Serological activity for both gel diffusion and complement fixation tests was demonstrated in the immunoglobulin G-containing fraction of sera taken from sheep 12 to 412 days after infection with B. ovis. The gel diffusion test with saline extract of B. ovis is as sensitive as the complement fixation test for the diagnosis of ram epididymitis and is more practical.

Incorporation of neomycin to the culture medium was found to be effective in inhibiting Escherichia coli contaminants without interfering with the growth of serotype L. autumnalis. The growth of 12 other Leptospira serotypes was unaffected by the addition of 300 mug of neomycin per ml to Ellinghausen medium or 5 mug/ml to Fletcher medium. Neomycin-containing medium was found to be of value in the isolation of leptospiras from cultures of blood from infected laboratory animals. A higher percentage of isolates was obtained in swine kidneys from an abattoir in medium containing neomycin than resulted from the same medium without antibiotic or with 5-fluorouracil. Contaminated leptospiral cultures growing in media with 5-fluorouracil were purified by subculturing into neomycin-containing media.

A simple gel-diffusion technique is described for the diagnosis of ram epididymitis caused by Brucella ovis. The results are shown to be very similar to those obtained by the complement-fixation test, which is currently the standard method of diagnosis. The method is suitable for use in the field and is expected to facilitate the control of ram epididymitis in areas where laboratory facilities are not available.

Field strains of Brucella abortus were resistant to mitomycin C, whereas strain 19 was sensitive; therefore, the antibiotic was used to distinguish B. abortus strain 19 from other strains.

A comparison was made of the survival of 28 leptospiral serotypes in Fletcher semisolid medium and in the same medium containing a basal layer of Fletcher medium plus 0.7% of agar and 0.5% of activated animal charcoal. A year after culture, more motile leptospires were observed by microscope examination in the biphasic medium. Two years after culture, 4 serotypes grown in the biphasic medium and 11 in Fletcher medium did not show motile cells. Nineteen of the serotypes maintained in Fletcher medium and 25 in the biphasic medium for 2 years grew on subculture into Fletcher medium. Subcultures from the biphasic medium showed the characteristic leptospiral ring growth earlier during the incubation period.

A simple technique is described for drying sera on washed, dry sand or ordinary sugar cubes for the sero-diagnosis of leptospirosis. The results are shown to be very similar to those obtained with fluid sera or sera dried on filter paper discs. Sera adsorbed on sand or absorbed in sugar eliminated some of the problems associated with sera dried on paper. This method is suitable for use in the field and is expected to be of value in epizootiological studies where contamination and chemical denaturation of fluid serum samples held without refrigeration is a problem.