Effect of frailty on short- and mid-term outcomes in vascular surgical patients.

BACKGROUND: Frailty is a multidimensional vulnerability resulting from age-associated decline. The impact of frailty on outcomes was assessed in a cohort of vascular surgical patients. METHODS: The study included patients aged over 65 years with length of hospital stay (LOS) greater than 2 days, who were admitted to a tertiary vascular unit over a single calendar year. Demographics, mode of admission, diagnosis, mortality, LOS and discharge destination were recorded, as well as a variety of frailty-specific characteristics. The impact of frailty on LOS, discharge destination, survival and readmission rate was assessed using multivariable regression techniques. The ability of the models to predict these outcomes was also assessed. RESULTS: In total, 413 patients of median age 77 years were followed for a median of 18 (range 12-24) months. The in-hospital, 3- and 12-month mortality rates were 3·6, 8·5 and 13·8 per cent respectively. Receiver operating characteristic (ROC) curve analysis revealed that frailty-based regression models were excellent predictors of 12-month mortality (area under the ROC curve (AUC) = 0·81), prolonged LOS (AUC = 0·79) and discharge to a care institution (AUC = 0·84). A simple additive frailty score using six key features retained strong predictive power for 12-month mortality (AUC = 0·83), discharge to a care institution (AUC = 0·78) and prolonged LOS (AUC = 0·74). This frailty score was also strongly associated with readmission rates (P < 0·001). CONCLUSION: Frailty in vascular surgery patients predicts a multiplicity of poorer outcomes. Optimal management should include identification of at-risk patients and treatment of modifiable risk factors.

Physicochemical effects of aldehydes on the human erythrocyte.

The effects of formaldehyde, acetaldehyde, and glutaraldehyde on human red blood cells were investigated. It was found that (a) The surface negative charge of the erythrocytes at pH 7 was increased 10% by glutaraldehyde, but not by the other two aldehydes. (b) The effect of incomplete fixation of the red blood cells was demonstrated by hemoglobin leakage studies The leakage of hemoglobin subsequent to formaldehyde treatment was especially pronounced Acetaldehyde-fixed cells showed some leakage of hemoglobin after an hour of exposure to the fixative, whereas glutaraldehyde-fixed cells showed no hemoglobin leakage. (c) All three aldehydes caused K(+) leakage during fixation. The concentrations of K(+) in the fixing solutions all reached the same level, but whereas the leakage with glutaraldehyde was immediate, that with formaldehyde was more gradual and that with acetaldehyde reached a steady state only after 24 hr. (d) The effects of the aldehydes on red cell deformability and swelling revealed that glutaraldehyde hardened the cells within 15 min, formaldehyde within 5 hr, while acetaldehyde required at least 24 hr to produce appreciable fixation. (e) The hematocrit changes accompanying the fixation process depended upon cell volume changes and loss of deformability.

D(--)-lactic acid and d(--)-lactate dehydrohgenase in octopus spermatozoa.

The spermatozoa of Octopus dofleini martini produce anaerobically D(-)-lactic acid and possess a very active D(-)-lactate dehydrogenase. In this respect, while resembling certain microorganisms, they differ strikingly from mammalian spermatozoa which produce L(+)-lactic acid and contain L(+)-lactate dehydrogenase.

International Equine Ophthalmology Consortium (IEOC) Symposium.

This first IEOC symposium met its goals of gathering a group of leading equine ophthalmology clinicians and researchers to identify the challenges of the field. To facilitate collaboration, notes from round-table discussions, including the ideas and plans that were discussed are being complied and will be distributed to the attendees. Development of an IEOC membership organisation and website was discussed and supported by the group in an effort further to advance the science of equine ophthalmology. To present results from the collaborations made at this first IEOC meeting, an IEOC mini-symposium will be held at the American College of Veterinary Ophthalmologists Annual Meeting in Chicago Illinois, on 6th November 2009. The second annual IEOC symposium will be held in Vienna, Austria on 4th and 5th June 2010.

Equine ulcerative keratitis with furrow: A review of the outcomes of 72 cases from 1987 to 2015.

BACKGROUND: Ulcerative keratitis with peripheral furrow formation is a poorly-described condition which has been associated with a grave prognosis due to rapid necrosis of the cornea. OBJECTIVE: To describe the infectious aetiologies associated with furrow-forming ulcerative keratitis, its overall clinical course and the efficacy of medical and surgical intervention in horses. STUDY DESIGN: Retrospective clinical case series. METHODS: Medical records of 72 horses which presented with furrow-forming ulcerative keratitis at the University of Florida between 1987 and 2015 were reviewed. RESULTS: Seventy-two horses (72 eyes) with furrow-forming ulcerative keratitis were treated at the University of Florida between 1987 and 2015. Of these, a definitive aetiologic diagnosis was available for 37 eyes. Ten of 37 eyes (27%) were diagnosed with fungal keratitis based on cytology of corneal scraping, culture, histopathology and/or fungal PCR. Fourteen of 37 eyes (38%) were diagnosed with a mixed fungal and bacterial keratitis. Thirteen of 37 eyes (35%) were diagnosed with bacterial keratitis. Overall, 26 of 72 total eyes were treated with medical therapy alone (36%). Forty-six of 72 eyes were treated medically and surgically (64%). Of the 26 eyes which received medical therapy, 20 healed with a positive visual outcome (77%) and 6 eyes were non-visual (23%). Of the 46 eyes which received surgical intervention, 40 healed with a positive visual outcome (87%), while six eyes were non-visual (13%). Altogether, 60 of 72 eyes healed with a positive visual outcome following medical or surgical treatment of furrow-forming ulcerative keratitis (83%). Twelve of 72 eyes failed treatment (17%), with six eyes requiring enucleation and six globes becoming phthisical after treatment. MAIN LIMITATIONS: Inconsistencies in available medical record data due to the large span of time (1987-2015) are inherent in this retrospective study, along with gradual evolution of corneal surgical techniques and medical therapies over the decades. CONCLUSIONS: Furrow-forming ulcerative keratitis was associated with a positive visual outcome in 83% of horses treated at the University of Florida between the years 1987 and 2015. Furrow formation may be associated with either fungal or bacterial infection.

Conjugation to hyperbranched polyglycerols improves RGD-mediated inhibition of platelet function in vitro.

RGD (arginine-glycine-aspartic acid) is a known peptide sequence that binds platelet integrin GPIIbIIIa and disrupts platelet-fibrinogen binding and platelet cross-linking during thrombosis. RGD peptides are unsuitable for clinical applications due to their high 50% inhibitory concentration (IC50) and low in vivo residence times. We addressed these issues by conjugating RGD peptides to biocompatible macromolecular carriers: hyperbranched polyglycerols (HPG) via divinyl sulfone. The GPIIbIIIa binding activity of RGD was maintained after conjugation and the effectiveness of the HPG-RGD conjugate was dependent upon molecular weight and the number of RGD peptides attached to each HPG molecule. These polyvalent inhibitors of platelet aggregation decreased the IC50 of RGD in an inverse linear manner based on the number of RGD peptides per HPG. Since HPG-RGD conjugates do not cause platelet activation by degranulation and certain substitution ratios do not increase fibrinogen binding to resting platelets, HPG-RGD may serve as a model for a novel class of antithrombotics.

Deep lamellar endothelial keratoplasty in 10 horses.

OBJECTIVE: To describe and evaluate a surgical technique utilized for the therapy of deep corneal stromal abscesses (DSA) in horses. The DSA is excised and replaced with a partial thickness corneal lamellar allograft. METHODS: A retrospective clinical study describing the indications for the surgical technique utilized and the outcomes of this procedure in 10 eyes of 10 horses. RESULTS: Each affected eye had a discrete DSA within the posterior stroma. An initial partial thickness semicircular corneal incision was made at the limbus, followed by anterior stromal lamellar dissection over the lesion. After excision of the DSA and replacement with a larger diameter split-thickness donor button, the anterior stroma was replaced into its original position and the initial corneal incision was repaired. All of the animals that underwent deep lamellar endothelial keratoplasty (DLEK) procedure healed appropriately and with subjectively less postoperative scarring and complications than previously described surgical approaches to DSA. CONCLUSIONS: This procedure is an effective technique for surgical removal of DSA in horses and, in most cases, results in a visual and cosmetically acceptable globe. The advantages of this technique compared to other surgical approaches to DSA are the peripheral location of the incision, shortened anesthesia times, the resultant minimal scarring and shorter healing times associated with DLEK.

Intranictitans tacking for replacement of prolapsed gland of the third eyelid in dogs.

OBJECTIVE: To evaluate a new procedure for fixation of prolapsed nictitans glands to the cartilage of the nictitans that will not interfere with the mobility of the nictitating membrane. METHODS: A prospective clinical trial utilizing a nonabsorbable suture to anchor the prolapsed gland to the cartilage of the third eyelid was undertaken. Fifteen eyes of 10 dogs were included in the study. A 4-0 nylon suture was passed from the anterior surface of the third eyelid through the base of the cartilage to the posterior aspect and then tunneled circumferentially beneath the conjunctiva over and around the prolapsed gland. The suture was then passed through the cartilage again to the anterior face of the third eyelid. The gland was replaced into its normal position as the suture was slowly tightened and then tied on the anterior aspect of the nictitans. RESULTS: Over a period of several weeks, the glands reduced in size and took on a normal appearance. All glands but one remained in place for the length of follow-up, which ranged from 2 weeks to 33 months. CONCLUSIONS: This procedure results in acceptable cosmetic effects with the return of the gland to its normal position posterior to the nictitating membrane. The advantage of this technique over traditional tacking to the orbital rim is that the third eyelid retains its normal mobility and, thus, its protective functions. The procedure once mastered is very quick and can be performed in less time than many of the traditional replacement techniques.

Corneal transplantation for inflammatory keratopathies in the horse: visual outcome in 206 cases (1993-2007).

OBJECTIVE: To evaluate the visual outcome of three techniques of corneal transplantation surgery in treating severe inflammatory keratopathies in the horse. DESIGN: Retrospective medical records study. ANIMALS STUDIED: Medical records of 206 horses that received corneal transplantation surgery at the University of Florida Veterinary Medical Center from 1993 to 2007 were reviewed. PROCEDURE: Data collected from the medical records included signalment, types of ocular lesions, type of transplant surgery performed, length of follow-up, complications, and visual outcomes. RESULTS: Full thickness penetrating keratoplasty (PK) was performed in 86 horses for melting ulcers, iris prolapse/descemetoceles, and medically nonresponsive full thickness stromal abscesses (SA). Posterior lamellar keratoplasty (PLK) and deep lamellar endothelial keratoplasty (DLEK) are split thickness penetrating keratoplasties that were utilized for medically nonresponsive deep stromal abscesses (DSA) in 54 and 66 eyes, respectively. The most common postoperative surgical complication was graft rejection and varying degrees of graft opacification. Wound dehiscence and aqueous humor leakage was also a common postoperative problem. A positive visual outcome was achieved for PK, PLK, and DLEK in 77.9%, 98.1%, and 89.4%, respectively. CONCLUSIONS: Corneal transplantation is a tectonically viable surgery in the horse with an overall success rate of 88.5% in maintaining vision when treating vascularized and infected corneal disease in the horse.

Effectiveness of commercially-available antibiotic-impregnated implants.

The aim of this study was to determine the effectiveness of antibiotic-impregnated implants in the prevention of bone infection. We used a model of contaminated fracture in goats to evaluate four treatment groups: no treatment, hand-made tobramycin-impregnated polymethylmethacrylate beads, commercially-available tobramycin-impregnated calcium sulphate pellets and commercially-available tobramycin-impregnated polymethylmethacrylate beads. Three weeks after intraosseous inoculation with streptomycin-resistant Staphylococcus aureus tissue cultures showed no evidence of infection in any of the antibiotic-treated groups. All of the cultures were positive in the untreated group. These results show that effective local antibiotic delivery can be obtained with both commercially-available products and with hand-made polymethylmethacrylate beads. The calcium sulphate pellets have the advantage of being bioabsorbable, thereby obviating the need for a second procedure to remove them.

Progesterone binding nano-carriers based on hydrophobically modified hyperbranched polyglycerols.

Progesterone (Pro) is a potent neurosteroid and promotes recovery from moderate Traumatic Brain Injury but its clinical application is severely impeded by its poor water solubility. Here we demonstrate that reversibly binding Pro within hydrophobically modified hyperbranched polyglycerol (HPG-Cn-MPEG) enhances its solubility, stability and bioavailability. Synthesis, characterization and Pro loading into HPG-Cn-MPEG is described. The release kinetics are correlated with structural properties and the results of Differential Scanning Calorimetry studies of a family of HPG-Cn-MPEGs of varying molecular weight and alkylation. While the maximum amount of Pro bound correlates well with the amount of alkyl carbon per molecule contributing to its hydrophobicity, the dominant first order rate constant for Pro release correlates strongly with the amount of structured or bound water in the dendritic domain of the polymer. The results provide evidence to justify more detailed studies of interactions with biological systems, both single cells and in animal models.

Characterization of a 22 kDa protein with widespread tissue distribution but which is uniquely present in secretions of the testis and epididymis and on the surface of spermatozoa.

The purification is reported of a 22 kDa protein which was first identified as one of the major components of the luminal secretion of the rat testis and epididymis. Antibodies against the 22 kDa protein cross-reacted with a protein of the same molecular weight in cytosolic extracts of other tissues from both male and female rats. However, since the protein could not be detected in blood, peritoneal fluid, saliva, milk, uterine fluid, seminal vesicle secretion, coagulating gland secretion or prostatic secretion, it would appear that the testis and epididymis may be unique in containing the protein in a soluble form within their luminal secretions. Proteins with slightly lower molecular weight were detected by the antibodies in cytosolic extracts of tissues from other animals (mice, rabbits, sheep, pigs, cattle), indicating that the protein may be conserved in a variety of species. However, in contrast to the rat, the protein was apparently not present in the testicular and epididymal secretions of these species. In addition to the occurrence of the 22 kDa protein as a soluble moiety in rat testicular and epididymal fluids, the protein was also located on sperm plasma membranes where its distribution was restricted to the surface of the flagellum. Amongst sperm surface proteins, the 22 kDa protein was the major protein containing sulphydryl groups and one of the major entities containing disulphide bonds. These properties may be of importance in the maintenance of sperm viability.

Enhanced concanavalin A agglutination of trypsinised erythrocytes is due to a specific class of aggregation.

Using the output of a rotational viscometer as a continuous index of aggregation, we have shown previously that the concanavalin A agglutination of native human erythrocytes can be resolved into three distinct classes of aggregation, static, type I and type II. Static aggregation occurs in the absence of shear forces while both type I and II aggregations are shear-induced. We now report that the increased concanavalin A agglutination of trypsinised erythrocytes is attributable to a specific enhancement in the development of type II aggregation. While type II formation in native cell suspensions requires high concanavalin A concentrations and continual shearing, an indistinguishable type of aggregation develops in suspensions of trypsinised red cells at considerably lower lectin concentrations and in the absence of applied shear forces.

Partition of proteins in aqueous polymer two-phase systems and the effect of molecular weight of the polymer.

The partition of substances in aqueous polymer two-phase systems is influenced by the molecular weight of the phase-forming polymers. We investigate how the effect of the molecular weight of the polymers depends on the molecular weight of the partitioned protein. We show that the magnitude of change of the partition is very small for proteins of molecular weights around 10,000, but increases almost linearly up to molecular weights of 250,000.

Composition of gangliosides from ovine testis and spermatozoa.

Gangliosides were extracted and purified from ovine testis and ejaculated spermatozoa which contained, respectively, 57 and 9 nmol lipid-bound sialic acid per gram wet weight. Fourteen gangliosides were resolved by thin-layer chromatography of testicular gangliosides, of which eleven were purified in sufficient quantity to enable a complete compositional analysis of the carbohydrate residues to be performed. None of the gangliosides contained fucose, but several contained N-glycolylneuraminic acid as a component of the sialic acid species. Relative migration on thin-layer chromatograms relative to known standards, compositional analysis, and selective degradation by specific enzymes were used as the basis for identification. Testis contained members of the ganglio series (GM1, GD1a, GD1b, GT1b, GQ1b), hematoside series (GM3, GD3), and sialosylparagloboside in the molar ratio of 54:40:6, respectively. Testicular GM3, GM1, GD3, GD1a, GD1b and GT1b ran as double bands on thin-layer chromatography which could be accounted for by observed differences in the fatty acid moiety. In addition, the slower migrating band of each pair contained some or all of its sialic acid residues as N-glycolylneuraminic acid, whereas the faster migrating band contained exclusively N-acetylneuraminic acid, except for GM3 where N-acetylneuraminic acid was the sole species in both bands. Thin-layer chromatography of sperm gangliosides revealed seven bands comigrating with equivalent testicular gangliosides. These coincided with the slower migrating bands of testicular GM3, GM1, GD3, GD1a, both bands of GD1b, and possibly both bands of GT1b. Sperm contained only trace amounts of sialosylparagloboside but, in addition, two unidentified bands which were absent from testis were also observed. The molar ratio of the ganglio series to the hematoside series in sperm was 42:58 with GM3 accounting for 42% of total gangliosides.

C1q binding to liposomes is surface charge dependent and is inhibited by peptides consisting of residues 14-26 of the human C1qA chain in a sequence independent manner.

Complement activation by anionic liposomes proceeds by antibody-independent, C1q-initiated activation of the classical pathway. Purified C1q bound to anionic liposomes in an acidic lipid concentration-dependent manner. Saturation binding, but not the apparent association constant, was enhanced by increasing the cardiolipin content of the liposomes or decreasing either the pH or ionic strength of the reaction mixture. These observations indicate the involvement of electrostatic factors in the binding. A highly cationic region in the collagen-like domain of C1q comprised of residues 14-26 of the C1qA polypeptide chain was assessed for involvement in liposome binding. This region has previously been shown to mediate C1q binding to other immunoglobulin-independent activators of the classical pathway of complement. Peptides containing residues 14-26 of C1qA, denoted C1qA14-26, inhibited C1q binding to and complement activation by anionic liposomes. The inhibitory capacity of these cationic peptides had no sequence or conformation specificity. Rather, the amount of positive charge on the peptides was the determining factor. When present in excess, peptides with five cationic residues inhibited C1q binding and complement activation; however, C1q peptides with only two cationic residues did not. In addition to the C1qA14-26 region, other parts of C1q that contain cationic residues may also be involved in C1q binding to anionic liposomes.

Can cytoplasm exist without undergoing phase separation?

Studies on such systems as the lens of the eye and theoretical considerations suggest that phase separation may well occur in cytoplasm. In this chapter, several issues relevant to this question are raised. It is suggested that while the interaction between water and the macromolecules in a mixture is proving crucial to their phase separation behavior, the abnormal water that is widely observed in cytoplasm and concentrated protein solutions is unlikely to constitute a thermodynamic phase in the sense of phase separation studies. The role of fixed structures in the cytoplasm, the likelihood that the volume of separated phases would be small and subject to spreading over the fixed structures and the expectation that much of the phase volume could be occupied and dominated by properties of the interface are also discussed. Finally, some experimental approaches to studying the existence of liquid-liquid phases in cytoplasm are proposed. While there is no proof that phase separation exists in cytoplasm, application of some of the techniques outlined might well provide more positive evidence for its presence.

Macromolecular crowding and its consequences.

Incompatible pairs of polymers separate into two phases in aqueous solution above a few percentage points total concentration. Protein pairs can also produce phase separation, but at somewhat higher concentrations. In this chapter, we explore the effect of high background concentrations of macromolecules on phase separation of pairs of species which would not be at sufficiently high concentration to separate in the absence of the uninvolved species. Effects produced by such high background concentrations are known as macromolecular crowding. Dramatic enhancements in various association reactions due to crowding have been predicted and observed but its effects on phase separation in biological mixtures typical of the cytoplasm have not been examined. Here, we describe a calculation based on the Flory-Huggins treatment of concentrated polymer solutions that sheds some light on this issue. We find that a background of 20 wt % of a high molecular weight species greatly reduces the concentrations needed to produce phase separation in a mixture of two incompatible macromolecules if one is more hydrophobic than the other. Given the high total concentration of macromolecules in cytoplasm, it is perhaps surprising that phases have not been observed. This issue is discussed and some explanations offered.

Hormonal regulation of rat androgen-binding protein (ABP) messenger ribonucleic acid and homology of human testosterone-estradiol-binding globulin and ABP complementary deoxyribonucleic acids.

Complementary DNA clones coding for rat androgen-binding protein (rABP) were isolated from a rat testis cDNA library constructed in the bacteriophage lambda gt11. The library was screened immunochemically, using two different antibodies against rABP. The identity of the isolated clones was confirmed by epitope selection and DNA sequence analysis. The mRNA encoding rABP could be detected in the testes of 20- and 46-day-old-rats, but not in the 10-day-old rats by hybridization with 32P-labeled rABP cDNA in a Northern blot of poly(A)+-RNA fractioned by agarose gel electrophoresis. No hybridization signal was seen with poly(A)+-RNA isolated from kidney and liver. The rABP mRNA appeared as a single species with a size of 1.65 kilobase, sufficient to encode a protein of 42,000 daltons. The concentration of rABP mRNA in the testes of 37-day-old hypophysectomized rats increased after treatment with testosterone and FSH, given alone or in combination. Sequence and hybridization analysis of cDNAs for rABP, human testosterone-estradiol-binding globulin, and human ABP demonstrates that the cDNAs for human testosterone-estradiol binding globulin and human ABP have greater sequence similarity with each other than either has with rABP.

Assessment of cocaine use in patients with chest pain syndromes.

BACKGROUND: Patients with myocardial ischemia may have different dispositions and/or pharmacologic interventions based on whether they have recently used cocaine. OBJECTIVE: To determine the prevalence of assessment of cocaine use in patients with acute chest pain syndromes. METHODS: In phase 1 of the study, we reviewed the medical records of all patients with chest pain who presented to the emergency department during February 1996 to assess historical documentation of the presence or absence of cocaine use. In phase 2, we evaluated whether cocaine questions were asked but not documented. After hospital admission, patients were interviewed to see if they were asked about cocaine use. In phase 3, we evaluated possible recall bias by using standardized questioning in the emergency department and used subsequent interviews to assess recall. Assessment of other cardiac risk factors served as the comparison group. RESULTS: In phase 1, 129 charts were reviewed, 13% of which revealed myocardial infarction. The presence or absence of cocaine use (13%) was less frequently documented than the presence or absence of hypercholesterolemia (58%), hypertension (82%), smoking (90%), diabetes (73%), or family history (77%) (chi 2, P < .05 for all comparisons). In phase 2, 27 (31%) of the 86 patients who were interviewed recalled cocaine questioning. There was chart documentation of the cocaine questioning in only 44% of the 27 cases. In phase 3, while in the emergency department, 20 patients were asked about cocaine use: 19 (95%) recalled being asked about cocaine use when interviewed the next day. CONCLUSIONS: Patients with chest pain often are not asked about recent cocaine use. When they are asked, their answers are poorly documented. These findings cannot be explained by poor recall. In cases of chest pain, efforts to improve questioning of patients about cocaine use are needed, since recent cocaine use may change treatment, disposition, and need for counseling.