Open-label, phase II, multicenter study of lasofoxifene plus abemaciclib for treating women with metastatic ER+/HER2- breast cancer and an ESR1 mutation after disease progression on prior therapies: ELAINE 2.

BACKGROUND: Acquired ESR1 mutations in estrogen receptor-positive (ER+) metastatic breast cancer (mBC) drive treatment resistance and tumor progression; new treatment strategies are needed. Lasofoxifene, a next-generation, oral, endocrine therapy and tissue-specific ER antagonist, provided preclinical antitumor activity, alone or combined with a cyclin-dependent kinase 4/6 inhibitor (CDK4/6i) in ESR1-mutated mBC. PATIENTS AND METHODS: In the open-label, phase II, ELAINE 2 trial (NCT04432454), women with ESR1-mutated, ER+/human epidermal growth factor receptor 2-negative (HER2-) mBC who progressed on prior therapies (including CDK4/6i) received lasofoxifene 5 mg/day and abemaciclib 150 mg b.i.d until disease progression/toxicity. The primary endpoint was safety/tolerability. Secondary endpoints included progression-free survival (PFS), clinical benefit rate (CBR), and objective response rate (ORR). RESULTS: Twenty-nine women (median age 60 years) participated; all but one were previously treated with a CDK4/6i (median duration 2 years). The lasofoxifene-abemaciclib combination was well tolerated with primarily grade 1/2 treatment-emergent adverse events (TEAEs), most commonly diarrhea, nausea, fatigue, and vomiting. One patient (with no prior CDK4/6i) discontinued treatment due to grade 2 diarrhea. No deaths occurred during the study. Median PFS was 56.0 weeks [95% confidence interval (CI) 31.9 weeks-not estimable; ∼13 months]; PFS rates at 6, 12, and 18 months were 76.1%, 56.1%, and 38.8%, respectively. CBR at 24 weeks was 65.5% (95% CI 47.3% to 80.1%). In 18 patients with measurable lesions, ORR was 55.6% (95% CI 33.7% to 75.4%). ESR1-mutant circulating tumor DNA (ctDNA) allele fraction decreased from baseline to week 4 in 21/26 (80.8%) patients. CONCLUSIONS: Lasofoxifene plus abemaciclib had an acceptable safety profile, was well tolerated, and exhibited meaningful antitumor activity in women with ESR1-mutated, ER+/HER2- mBC after disease progression on prior CDK4/6i. Observed decreases in ESR1-mutant ctDNA with lasofoxifene concordant with clinical response suggest target engagement. If the ELAINE 2 findings are confirmed in the initiated, phase III, ELAINE 3 trial, these data could be practice-changing and help address a critical unmet need.

The effects of diet and beauty products on the uptake and storage of 14C in hair and nails: ramifications for the application of bomb pulse dating to forensic anthropological casework.

Radiocarbon dating is a useful tool in the examination of unknown human remains. Recent studies have shown that the analysis of hair and nail samples can provide a highly accurate estimation of the year of death (YOD). However, little research has examined factors that may influence the uptake and storage of 14C in these tissues, such as diet, or the use of beauty products. This study measured the level of 14C in human hair and nail samples collected from living individuals to determine whether diet, and the use of hair dye or nail polish, has a significant impact on the estimation of YOD. The results of this study showed that diet did not appear to impact the radiocarbon content in human hair and nail, and thus should not be considered a limitation when analysing samples obtained from unidentified human remains. The use of nail polish, and in the majority of cases, hair dye, did not significantly impact the 14C concentration in nails and hair. While the results of this study are preliminary, they suggest that in most cases, both hair and nail can be successfully analysed using radiocarbon dating to estimate an individual's YOD. However, best practice should involve the analysis of multiple tissue types, to minimise any error that may be introduced as a result of the decedent's use of beauty products.

Re-examining so-called 'secondary identifiers' in Disaster Victim Identification (DVI): Why and how are they used?

Disaster victim identification (DVI) refers to the identification of multiple deceased persons following an event that has a catastrophic effect on human lives and living conditions. Identification methods in DVI are typically described as either being primary, which include nuclear genetic markers (DNA), dental radiograph comparisons, and fingerprint comparisons, or secondary, which are all other identifiers and are ordinarily considered insufficient as a sole means of identification. The aim of this paper is to review the concept and definition of so-called 'secondary identifiers" and draw on personal experiences to provide practical recommendations for improved consideration and use. Initially, the concept of secondary identifiers is defined and examples of publications where such identifiers have been used in human rights violation cases and humanitarian emergencies are reviewed. While typically not investigated under a strict DVI framework, the review highlights the idea that non-primary identifiers have proven useful on their own for identifying individuals killed as a result of political, religious, and/or ethnic violence. The use of non-primary identifiers in DVI operations in the published literature is then reviewed. Because there is a plethora of different ways in which secondary identifiers are referenced it was not possible to identify useful search terms. Consequently, a broad literature search (rather than a systematic review) was undertaken. The reviews highlight the potential value of so-called secondary identifiers but more importantly show the need to scrutinise the implied inferior value of non-primary methods which is suggested by the terms "primary" and "secondary". The investigative and evaluative phases of the identification process are examined, and the concept of "uniqueness" is critiqued. The authors suggest that non-primary identifiers may play an important role in providing leads to formulating an identification hypothesis and, using the Bayesian approach of evidence interpretation, may assist in establishing the value of the evidence in guiding the identification effort. A summary of contributions non-primary identifiers may make to DVI efforts is provided. In conclusion, the authors argue that all lines of evidence should be considered because the value of an identifier will depend on the context and the victim population. A series of recommendations are provided for consideration for the use of non-primary identifiers in DVI scenarios.

The importance of bone sample selection when using radiocarbon analysis in cases of unidentified human remains.

The radiocarbon (14C) analysis of skeletonised human remains can provide vital information regarding the time since death. This study analysed the 14C content in both cortical and trabecular bone from the femur and rib of 18 human remains donated to the Australian Facility for Taphonomic Experimental Research. To determine the most appropriate bone to sample for 14C analysis when estimating the time since death, differences in turnover time between the bone types, and the difference between environmental 14C during the year of death (YOD) and the level of 14C found within the bone (i.e., the lag time) were examined. Overall, the average lag time for all donors was 25 years, with the cortical bone from femur samples providing the largest lag time (39 years) and the trabecular bone from rib samples containing the lowest level of 14C (most modern), with an average lag time of 13 years. The results of this study suggest that in cases of unidentified human remains, the analysis of trabecular bone from a rib may be preferred sample, as this bone contains the most recent 14C, and can provide an estimate of both the time since death, and a broad approximation of YOD.

Examining the use of different sample types following decomposition to estimate year of death using bomb pulse dating.

When human remains are discovered, confirming the identification of the decedent is the first part of the forensic medical investigation. In cases where the remains are skeletonised or badly decomposed, differential preservation often increases the difficulty of this task. Bomb pulse dating, which directly compares levels of 14 C within human tissues to atmospheric levels, can provide an estimate of the year of death, which may assist in the identification process. This study measured the 14 C content in samples of hair, nail and puparia collected from donors at the Australian Facility for Taphonomic Experimental Research (AFTER). The radiocarbon results demonstrated that the nail samples provided the most accurate year of death estimation, with 91% correctly predicting YOD, closely followed by hair, with a 79% correct prediction rate, with both hair and nails having a lag time of 0-1 years. This is consistent with the time taken for atmospheric CO 2 to enter the food chain, and be taken in by humans. Puparia was found to have the highest levels of 14 C, and was the least consistent with the actual YOD (46% correct). However, predicted YOD ranges were still within 4 years of the actual YOD. Based on the results of this study, hair, nail and puparia should be considered as useful samples to obtain accurate estimates for YOD using bomb pulse dating.

COVID-19 vaccination and breakthrough infections in patients with cancer.

BACKGROUND: Vaccination is an important preventive health measure to protect against symptomatic and severe COVID-19. Impaired immunity secondary to an underlying malignancy or recent receipt of antineoplastic systemic therapies can result in less robust antibody titers following vaccination and possible risk of breakthrough infection. As clinical trials evaluating COVID-19 vaccines largely excluded patients with a history of cancer and those on active immunosuppression (including chemotherapy), limited evidence is available to inform the clinical efficacy of COVID-19 vaccination across the spectrum of patients with cancer. PATIENTS AND METHODS: We describe the clinical features of patients with cancer who developed symptomatic COVID-19 following vaccination and compare weighted outcomes with those of contemporary unvaccinated patients, after adjustment for confounders, using data from the multi-institutional COVID-19 and Cancer Consortium (CCC19). RESULTS: Patients with cancer who develop COVID-19 following vaccination have substantial comorbidities and can present with severe and even lethal infection. Patients harboring hematologic malignancies are over-represented among vaccinated patients with cancer who develop symptomatic COVID-19. CONCLUSIONS: Vaccination against COVID-19 remains an essential strategy in protecting vulnerable populations, including patients with cancer. Patients with cancer who develop breakthrough infection despite full vaccination, however, remain at risk of severe outcomes. A multilayered public health mitigation approach that includes vaccination of close contacts, boosters, social distancing, and mask-wearing should be continued for the foreseeable future.

Quantifying center of pressure variability in chondrodystrophoid dogs.

The center of pressure (COP) position reflects a combination of proprioceptive, motor and mechanical function. As such, it can be used to quantify and characterize neurologic dysfunction. The aim of this study was to describe and quantify the movement of COP and its variability in healthy chondrodystrophoid dogs while walking to provide a baseline for comparison to dogs with spinal cord injury due to acute intervertebral disc herniations. Fifteen healthy adult chondrodystrophoid dogs were walked on an instrumented treadmill that recorded the location of each dog's COP as it walked. Center of pressure (COP) was referenced from an anatomical marker on the dogs' back. The root mean squared (RMS) values of changes in COP location in the sagittal (y) and horizontal (x) directions were calculated to determine the range of COP variability. Three dogs would not walk on the treadmill. One dog was too small to collect interpretable data. From the remaining 11 dogs, 206 trials were analyzed. Mean RMS for change in COPx per trial was 0.0138 (standard deviation, SD 0.0047) and for COPy was 0.0185 (SD 0.0071). Walking speed but not limb length had a significant effect on COP RMS. Repeat measurements in six dogs had high test retest consistency in the x and fair consistency in the y direction. In conclusion, COP variability can be measured consistently in dogs, and a range of COP variability for normal chondrodystrophoid dogs has been determined to provide a baseline for future studies on dogs with spinal cord injury.

Mechanical and Thermal Sensory Testing in Normal Chondrodystrophoid Dogs and Dogs with Spinal Cord Injury caused by Thoracolumbar Intervertebral Disc Herniations.

BACKGROUND: Intervertebral disc herniation is a common cause of spinal cord injury (SCI) causing paralysis and sensory loss. Little quantitative information is available on the loss and recovery of sensation in dogs with SCI. OBJECTIVES: To determine whether quantitative sensory testing (QST) can be used to establish thermal and mechanical sensory thresholds in chrondrodystrophoid dogs and compare thresholds among normal dogs and dogs with different grades of SCI. ANIMALS: Thirty-three client-owned chondrodystrophoid dogs: 15 normal and 18 SCI dogs. METHODS: Thermal testing was performed by placing a hot (49°C) and cold (5°C) probe on the dorsal metatarsus and mechanical thresholds were tested using calibrated forceps to apply force to the lateral digit. Stimuli were applied until acknowledged, and response rate, latency, and force applied to response were recorded. Test-retest repeatability was determined by calculating intraclass correlation coefficients. Response rates were compared using logistic regression and thresholds were compared using Kaplan-Meier Survival curves. RESULTS: Testing was feasible with moderate repeatability. Thresholds and response rates were significantly different between normal and SCI dogs for all modalities (P < .001). When dogs were grouped by their clinical grade, each grade was significantly different from normal dogs, and cold stimuli differentiated among all grades. CONCLUSION AND CLINICAL IMPORTANCE: Sensory thresholds can be measured reliably in chondrodystrophoid dogs and are altered by SCI. The differences in sensation among neurologic grades indicate that these techniques can be used to further characterize recovery of SCI dogs.

A comparison of toxicity and health care resource use between eribulin, capecitabine, gemcitabine, and vinorelbine in patients with metastatic breast cancer treated in a community oncology setting.

BACKGROUND: Capecitabine (C), gemcitabine (G), and vinorelbine (V) are commonly used as single agents in patients with metastatic breast cancer. Eribulin (E) is one of the most recent cytotoxic agents to gain regulatory approval for metastatic breast cancer in the United States as a single agent. EMBRACE - a large randomized trial demonstrated the safety and overall survival benefit of eribulin in heavily pretreated metastatic breast cancer patients compared to treatment of physician's choice. In this analysis, toxicity and the associated health care resource use were compared between the four agents in a sample of metastatic breast cancer patients treated in a US community oncology setting. METHODS: This study identified 411 patients (C=144, G=81, V=96, and E=90) who were treated in 19 community oncology clinics over the preceding two-year period. Data collection included baseline patient and disease characteristics, duration of therapy, use of supportive care drugs, type of dose limiting toxicities, and their impact on overall health care resource use. RESULTS: The median lines of therapy for C, G, V, and E were second, third, third, and fourth, respectively. Patients were comparable with respect to baseline comorbidities, performance status, serum creatinine, hemoglobin, neutrophil, and platelet counts. The proportion reporting at least one adverse event (any grade) with C, G, V, and E was 45%, 65%, 75%, and 63%. The most commonly reported toxicities (regardless of grade) for C, G, and V were diarrhea (19.4%), anemia (34.6%), and neutropenia (50.0%), respectively. The most common toxicity for E was neutropenia (32.2%). Overall, 5.6%, 19.8%, 22.9%, and 22.2% of patients receiving C, G, V, and E required at least one medical intervention to manage a toxic event. Toxicity was the cause of treatment discontinuation in 25.7%, 8.6%, 11.5%, and 8.9% of C, G, V, and E patients, respectively. The primary cause for treatment discontinuation in all four cohorts was disease progression. CONCLUSIONS: Eribulin demonstrated a comparable patient safety profile to gemcitabine and vinorelbine, even when administered after three lines of prior therapies. Capecitabine was generally used in earlier lines, had less neutropenia and anemia, but more treatment discontinuations due to toxicity.

The contributions of anthropology and mitochondrial DNA analysis to the identification of the human skeletal remains of the Australian outlaw Edward 'Ned' Kelly.

This paper details the anthropological and genetic analyses that contributed to the identification of the notorious Australian outlaw ('bushranger') Edward ('Ned') Kelly. In 1880 at the age of 25, Kelly was hanged and buried at the former Melbourne Gaol in Victoria, Australia. In 1929, the remains of executed prisoners (including those of Kelly) were haphazardly disinterred following the demolition of parts of the Melbourne Gaol and haphazardly reinterred in three distinct "pits" at the Pentridge Prison. In 1999 the Pentridge Prison was sold for commercial development and subsequently in 2008 and 2009 the human remains of prisoners were recovered. A total of 41 cases of unidentified human skeletal remains from Pentridge were examined using traditional anthropological techniques. At least one representative sample from each of the remains (mostly clavicles) from all three pits was selected for DNA analysis. Comparative ante-mortem reference samples were also located. Given the antiquity and condition of remains recovered from Pentridge, and the 130 years that had passed since Kelly's execution, mitochondrial DNA analysis was chosen as a suitable DNA analysis tool to examine the Pentridge cases to assist in the inclusion or exclusion of remains as being those of Ned Kelly. Only one of the Pentridge cases (Pen14) matched the HV1/HV2 mitochondrial DNA haplotype of the reference sample. Additional anthropological analyses indicated a number of pathological features that provided support that the remains of Pen14 are those of Edward ("Ned") Kelly.

Adsorption of cationized bovine serum albumin onto epithelial crypt fractions of the rat colon.

The purpose of the study was to characterize mucosal attachment of a cationized model protein, bovine serum albumin (BSA), onto the various fractions of colonic crypts epithelium in the rat. BSA was labeled with fluorescein isothiocyanate (FITC) and its surface net electric charge was modified from negative to positive. Attachment of the cationized protein (CF-BSA) onto rat colonic epithelium was performed by incubation of colonic everted sacs in medium containing cationized or non-cationized FITC-labeled BSA. Using a nonenzymatic isolation procedure, colonocytes were harvested from five horizontal fractions of the colonic crypts. BSA adhesion to the isolated colonocytes was quantified spectrofluorometrically. In addition, the effect of increasing concentrations of Mg(2+) on the adsorption of the cationized BSA onto the surface of colonic epithelium was evaluated by measuring its ability to displace the adhered BSA from its binding sites. BSA cationization facilitated protein adherence to the colon epithelium in a crypt depth-dependent manner. The largest extent of adherence was observed in the outer layer (first fraction) of the colon. Binding persisted to approximately half the depth of the crypts. The relation between CF-BSA concentration in the incubation medium and the amount of CF-BSA adsorbed onto the colonic epithelium was exponential in nature. The addition of electrolyte (Mg(2+)) caused a detachment of the CF-BSA. The adsorption process was characterized by Langmuir's adsorption isotherm. It is concluded that cationized BSA could be useful as a targetable drug platform in cases where the target site is the gastrointestinal epithelium.

Drug targeting by surface cationization.

Cationization of drug products and carriers involves a direct modification or attachment of conveying or accompanying components, either of which cause a charge modification. Cationization of macromolecules such as proteins and nucleotides and particulate drug carriers generally enhances their cellular uptake by endocytosis. The most common use of cationization today is in gene delivery. This is undertaken by either employing cationic polymers or entraping nucleotides in cationic carriers such as cationic liposomes. Cationized delivery systems are also used to overcome biological barriers and are suggested for drug targeting, in a nonspecific manner, to a variety of body organs, including brain, eyes, nose, and inflamed intestinal epithelium. Protein cationization is also suggested both for tumor immunotherapy and as a diagnostic tool in cancer therapy. Cationization has proven itself to be a straightforward tool for targeting to cells, tissues, and selected organs. This article reviews the extensive range of applications of cationization for improving drug and gene delivery and summarizes major technologies employed for that purpose.

The effect of local attachment of cationized antioxidant enzymes on experimental colitis in the rat.

PURPOSE: To investigate the possibility of local treatment of colitis with the adhesive antioxidant enzymes catalase and superoxide dismutase (SOD). METHODS: The net electric charge of the enzymes' surfaces was modified from negative to positive, to cause their adherence to the colon epithelium. The effects of this local administration were assessed in inflamed rat colon. Inflammation severity (colitis) was assessed by measuring colonic tissue myeloperoxidase (MPO) activity, amounts of tumour necrosis factor alpha (TNFalpha) and concentrations of reduced glutathione (GSH). The measurements were carried out in two types of protocols: preventive (pre-colitis induction) and treatment (post-colitis induction). In addition, the efficacy of treatment with the cationized enzymes was compared to 5-aminosalicylic acid (5-ASA) and betamethasone with similar administration routes. RESULTS: The two cationized antioxidant enzymes were found to be efficient in both prevention and treatment of experimental colitis. The two cationized enzymes caused a significant reduction in MPO activity. A reduction in TNFalpha concentration was noted only after the treatment protocol. No correlation was found between inflammation severity and tissue levels of GSH. In most cases the cationized enzymes were more effective than 5-ASA and betamethasone. CONCLUSION: Cationized catalase and cationized SOD have the potential to be efficient therapeutic tools in the local treatment of colitis.

Relation between colonic inflammation severity and total low-molecular-weight antioxidant profiles in experimental colitis.

Tissue antioxidant status is altered as a response to oxidative stress. This oxidative stress, caused by reactive oxygen species, is associated with inflammatory bowel disease (IBD). Our aim was to examine the relationship between total tissue low-molecular-weight antioxidant (LMWA) profile and inflammation severity in dinitrobenzene sulfonic acid (DNBS) experimental colitis in the rat. Rats were treated with three doses of DNBS: 1, 10, and 20 mg. Inflammation severity was assessed by tissue colonic wet weight, macroscopic evaluation, and tissue myeloperoxidase (MPO) activity. The capacity of water-soluble LMWA was assessed by measuring the reducing power of the tissues with cyclic voltammetry (CV) and by measuring tissue levels of reduced glutathione. While typical markers of inflammation (MPO, macroscopic examination, and colonic wet weight) indicated DNBS dose dependency, such dependency could not be demonstrated for the tissue LMWA as measured by reduced glutathione levels and by the tissues' reducing power. Mild colonic inflammation (induced by ethanol or by 1 mg of DNBS) caused an increase in the overall capacity of water-soluble LMWA. However, severe inflammation (induced by 20 mg of DNBS) caused a reduction in the tissue LMWA capacity. An intermediate dose of DNBS (10 mg) caused moderate inflammation, but did not cause a significant change in the tissue LMWA compared with a saline control treatment. In conclusion, LMWA changed in a biphasic pattern reflective of the severity of mucosal colonic inflammation. It is suggested that: low dose of DNBS (1 mg) and topical alcohol (25% v/v) caused an adaptation effect to the mild oxidative stress associated with mild inflammation. This resulted in an increase in the LMWA. A higher dose of DNBS (20 mg) caused more severe inflammation with an overall reduction in LMWA. The increased efflux of reactive oxygen species, associated with severe inflammation, led to an overall consumption of the tissue LMWA, which masked the increase in LMWA caused by the mild oxidative stress.

Differences in the reducing power along the rat GI tract: lower antioxidant capacity of the colon.

The ability of the gastrointestinal (GI) tract, as well as other tissues, to cope with reactive oxygen species (ROS) efflux in pathological events is determined partly by epithelial antioxidant levels. These levels are comprised of tissue antioxidant enzymes and low molecular weight antioxidants (LMWA). While glutathione levels and the activity of enzymatic antioxidants along the GI tract have been studied, the contribution of the overall LMWA to the total antioxidant capacity has not yet been determined. In this study the overall antioxidant activity in the mucosa/submucosa and muscularis/serosa of various sections along the small intestine and colon of the rat was evaluated by determining the reducing power, which reflects the total antioxidant activity derived from LMWA, using cyclic voltammetry. The activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase was also measured. The reducing power (total antioxidant activity) was higher in the mucosa/submucosa of the small intestine as compared to the mucosa/submucosa of the colon. Similarly, catalase and SOD activity in the mucosa/submucosa of the small intestine was significantly higher than in the mucosa/submucosa of the colon. Differences were also observed in the reducing power and SOD activity in the muscularis/serosa of the rat small intestine as compared to the colon. The low antioxidant capacity in the colon may facilitate reactive oxygen species (ROS)-mediated injury and lead to inflammatory diseases such as ulcerative colitis, specifically in the colon.

ARPP-21: murine gene structure and promoter identification of a neuronal phosphoprotein enriched in the limbic striatum.

ARPP-21 (cAMP-regulated phosphoprotein, Mr = 21,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis) is a phosphoprotein highly enriched in concentration in the neurons of the limbic striatum. It is likely a third messenger in the intracellular cascade of events following neuronal stimulation by first-messenger activators of the adenylate cyclase system, including dopamine via the D1 receptor. ARPP-21 expression is restricted to telencephalic post-mitotic, post-migrational neurons, and its precise pattern of temporal and spatial expression makes it an attractive candidate for the study of transcriptional regulation of neuronal maturation. To define genomic regions likely to contain functional promoter elements, we isolated the murine ARPP-21 gene. Primer extension and T2 RNase protection analyses identified multiple transcription start sites, but 1.3 kb of 5'-flanking DNA revealed few consensus transcription factor binding sequences. A series of transient transfection assays in clonal cell lines which do not express ARPP-21 identified a basal promoter active in both neuronal and non-neuronal lines. Expression in all lines was decreased by the inclusion of regions further upstream, and extinguished by the inclusion of the first intron. Further analyses are likely to reveal cell specific regulatory sequences.

Vasodilator provocation of occult urinary tract hemorrhage.

This case demonstrates the use of vasodilators to reactivate an intermittent urinary tract hemorrhage. The site of bleeding was demonstrated and treated with subselective embolization.

DARPP-32 promoter directs transgene expression to renal thick ascending limb of loop of Henle.

DARPP-32, a dopamine- and adenosine 3',5'-cyclic monophosphate (cAMP)-regulated inhibitor of protein phosphatase-1, is highly colocalized with neuronal and nonneuronal D1-type receptors. DARPP-32 concentration is enriched in the renal outer medulla and in the medium-size spiny neurons of the brain. In the ascending limb of the loop of Henle, DARPP-32 is phosphorylated following stimulation by dopamine and other first messengers, and in this form inhibits the activity of the Na(+)-K(+)-adenosinetriphosphatase pump. For functional analysis of the DARPP-32 promoter in the kidney, we characterized the murine gene. There are two groups of transcription start sites utilized in the brain, but the proximal set appears to be preferentially used in the kidney. In four of four lines of mice carrying a DARPP-32/lacZ transgene with 2.1 kb of 5'-flanking DNA, adult kidney lacZ transgene expression mimicked that of endogenous DARPP-32. There was no ectopic expression in peripheral organs. We conclude that the sequences necessary for direction of DARPP-32 expression to the medullary thick ascending limb are contained within this 2.1-kb fragment.

Bioprosthetic cardiac valve degeneration: role of inflammatory and immune reactions.

Inflammatory and immune reactions are thought to mediate both calcification and biodegradation of bioprosthetic cardiac valve implants. To investigate the mechanisms of implant degeneration, we evaluated the role of inflammatory and immune reactions and the effects of tissue preservative treatment in three series of experiments. In the first experiment, three kinds of implants, i.e. glutaraldehyde-treated autograft Sprague-Dawley (SD) rat skin, xenograft Swiss-Webster (SW) mouse skin, and saline-treated autograft (control) were subcutaneously implanted in ten weanling SD rats, and retrieved after 70 days. There was no significant difference in the level of calcification in the autograft (113.13 +/- 27.09 micrograms/mg dry weight) and xenograft (78.27 +/- 31.53 micrograms/mg dry weight) (p > 0.05), but both differed significantly from the control specimens (1.55 +/- 0.87 micrograms/mg dry weight). In the second experiment, the immunological response to glutaraldehyde-treated bovine pericardium (glut tBP) and glycerol treated bovine pericardium (glyc tBP) implants were tested in vivo and in vitro. A Gore-Tex implant was used as a control. Sections of these materials were implanted to the abdominal muscle wall of Lewis rats, with each group composed of twelve animals. Lymphocytes and sera from the animals were isolated, and histological examination was performed at two or four weeks post-implantation. Collagen type 1 (calf skin) was used as antigen. Tritiated thymidine incorporation was used to measure lymphocyte response to antigen collagen type 1 (calf skin), and an Enzyme Linked Immunosorbent Assay (ELISA) was used to test antibodies. The results showed that lymphocytes from both the glut tBP and the glyc tBP groups responded to collagen type 1. The ELISA results showed that the glyc tBP group produced more antibodies than did the glut tBP group, with the difference being significant at a level of p < 0.02. Histology revealed that the glyc tBP had greater inflammatory changes and collagen degeneration than did the glut tBP. In the third experiment, sections of glut tBP and glyc tBP were implanted subcutaneously in two groups of ten weanling SD rats, and retrieved after 70 days. The results showed that glut tBP caused more calcification (197.04 +/- 83.56 micrograms/mg dry weight) than did the glyc tBP (6.74 +/- 0.55 microgram/mg dry weight), with the difference being significant at a level of p < 0.05. From these investigations it is concluded that tissue treatment prior to implantation was very important in determining the tendency of tissue to calcify, and that there was no obvious relationship between bioprosthetic calcification and immunogenicity.

Use of biodegradable (PGA) fabric for repair of solid organ injury: a combined institution experience.

A fabric constructed of biodegradable mesh was used in the operative repair of injured intra-abdominal organs in 60 patients at two Level I Trauma Centres. Splenorrhaphy was performed in 44 patients, hepatorrhaphy in eight, renorrhaphy in five and one combined repair of spleen and liver and one kidney and liver. The age range for the patients was 5 to 61 years. Multiple-organ injury occurred in 21 patients. Mean emergency room systolic BP for the patient series was 120 +/- 24 mmHg (SD), Glasgow Coma Scale 14.3 +/- 1.9, haematocrit 37.2 +/- 6.4 per cent, Injury Severity Score (ISS) 28.1 +/- 16.3, Abdominal Trauma Index (ATI) 15.5 +/- 7.5. Postoperative complications occurred in 36.7 per cent of patients. Time for the operation averaged 165.1 + 72.1 min and preoperative and operative transfusion volume averaged 2248 ml. There were three deaths (5.4 per cent). The mesh organ repair technique is an alternative to conventional surgical procedures used to control bleeding from injured organ surfaces and to close organ parenchymal defects.