Double-lumen tracheal tubes and bougies: a bench study to investigate factors that influence the risk of shearing.

Tracheal intubation with a double-lumen tube can be more challenging than with a single-lumen tube. A bougie can be used to facilitate intubation. Case reports have described fragment shearing from bougies when they are removed from the tube after intubation. These fragments have the potential to cause harm. It has even been suggested that bougies and double-lumen tubes should not be used together. We conducted a benchtop trial to investigate factors that influence the risk of shearing. We investigated three brands of double-lumen tube (each in three sizes and both lateralities) and four brands of bougie. We simulated one intubation and 29 further insertions/removals of bougie with every bougie-double-lumen tube combination. We inspected the inside of the tube for evidence of shearing after first, tenth and thirtieth removals. We found that brand of bougie, brand of double-lumen tube and size of double-lumen tube (but not its laterality) all influenced the degree of shearing. Certain bougie-double-lumen tube combinations produced a particularly high degree of shearing, so these should be avoided.

Selection of baseline blood pressure to guide management of hypotension during spinal anaesthesia for caesarean section.

INTRODUCTION: Recommendations on vasopressor management during caesarean section under spinal anaesthesia suggest maintaining systolic arterial pressure ≥90% of an accurately measured baseline value. The baseline is often taken as the first reading in the operating room. We hypothesise that this reading may not reflect an accurate baseline value. METHODS: A retrospective case note review of 300 non-hypertensive women undergoing caesarean section with neuraxial anaesthesia, including spinal anaesthesia for elective delivery (n=100), and spinal (n=100) and epidural top-up anaesthesia (n=100) for emergency delivery. Systolic arterial pressure values recorded at various time points between the last antenatal visit and the first blood pressure value recorded in the operating room were compared. RESULTS: There was a stepwise and significant increase in systolic arterial pressure over three time points (last antenatal clinic, morning of surgery, operating room) before elective caesarean section (all P <0.001). In women having emergency caesarean under spinal anaesthesia, a stepwise increase over four time points (last antenatal clinic, first reading in labour, final reading in labour, operating room) was observed. A similar trend was seen over these time points for women having emergency caesarean under epidural top-up, although the systolic blood pressure did not rise during labour. CONCLUSIONS: Using the initial blood pressure reading in the operating room as the baseline value may lead to unnecessary vasopressor use and hypertension. Prospective research is required to clarify which reading represents the most accurate baseline to maintain homeostasis and reduce the hypotensive sequelae of neuraxial anaesthesia for both the mother and fetus.

Zonal winds in the central equatorial pacific and el nino.

Easterly trade winds from near-equatorial islands in the central Pacific weakened before each El Niño between 1950 and 1978, except for the 1963 El Niño. The weakening of the easterlies and their later collapse did not occur uniformly over several months, but rather through a series of strong westerly wind bursts lasting 1 to 3 weeks. The bursts may force equatorial Kelvin waves in the ocean that can both initiate and sustain the sea surface warming characteristics of El Niño events.

Effects of N-acetylcysteine and tirilazad mesylate on intestinal functional capillary density, leukocyte adherence, mesenteric plasma extravasation and cytokine levels in experimental endotoxemia in rats.

INTRODUCTION: The study's objective was to determine the effects of the administration of N-acetylcysteine (NAC) and of tirilazad mesylate (TM) on intestinal functional capillary density, mesenteric plasma extravasation, leukocyte adherence and on cytokine release during experimental endotoxemia in rats. METHODS: In a prospective, randomized, controlled animal study, 80 male Wistar rats were examined in 2 test series. Both series were divided into 4 groups. Group 1 served as control group (CON group). Group 2 (LPS group), group 3 (NAC group) and group 4 (TM group) received endotoxin infusions (10 mg/kg over 2 h). In NAC group 150 mg/kg body weight NAC was administered after the first 30 minutes of endotoxemia intravenously. In TM group, 10 mg/kg body weight TM was administered after the first 30 minutes of endotoxemia intravenously. Animals of the series 1 underwent studies of leukocyte adherence on submucosal venular endothelium of the small bowel wall and intestinal functional capillary density (FCD) in the intestinal mucosa and the circular as well as the longitudinal muscle layer by intravital fluorescence microscopy (IVM). Plasma levels of interleukin 1beta (IL-1beta), interferone gamma (IFN-gamma) and soluble intercellular adhesion molecule1 (s-ICAM 1) as well as white blood cell count (WBC) were estimated. In the animals of the series 2 mesenteric plasma extravasation was determined by IVM and plasma levels of tumor necrosis factor alpha (TNF-alpha), IL-4, IL-6, IL-10 and malondialdehyde (MDA) were estimated. RESULTS: After LPS administration, FCD in the villi intestinales was unchanged and in the longitudinal muscularis layer it was increased. There was no effect of NAC or TM administration on FCD.Although the plasma extravasation was not significantly influenced by LPS administration, TM administration resulted in a lower plasma extravasation in the TM group compared to the other groups. After endotoxin challenge, the firmly adherence of leukocytes to vascular endothelium as a parameter of leukocyte activation in endotoxemia was increased but NAC or TM administration had no influence on leukocyte adherence. The plasma levels of IL-1beta, IL-6, IL-10, TNF-alpha, IFN-gamma and sICAM-1 were increased in the endotoxemic groups (LPS group, NAC group and TM group) and the WBC was decreased compared to controls. IL-4 levels were unchanged during observation period. Plasma MDA levels were not influenced by LPS administration compared to controls. The administration of NAC resulted in lower sICAM-1 and MDA levels compared to the LPS group. The IL-1beta, IL-6, IL-10, TNF-alpha and IFN-gamma plasma levels were not influenced by NAC or TM administration. CONCLUSIONS: In this posttreatment sepsis model in rats, NAC administration resulted in lower sICAM-1 and MDA levels compared to the LPS treated animals. TM administration reduced the plasma extravasation in this model.

Delivery approaches for CRISPR/Cas9 therapeutics in vivo: advances and challenges.

INTRODUCTION: Therapeutic gene editing is becoming a viable biomedical tool with the emergence of the CRISPR/Cas9 system. CRISPR-based technologies have promise as a therapeutic platform for many human genetic diseases previously considered untreatable, providing a flexible approach to high-fidelity gene editing. For many diseases, such as sickle-cell disease and beta thalassemia, curative therapy may already be on the horizon, with CRISPR-based clinical trials slated for the next few years. Translation of CRISPR-based therapy to in vivo application however, is no small feat, and major hurdles remain for efficacious use of the CRISPR/Cas9 system in clinical contexts. AREAS COVERED: In this topical review, we highlight recent advances to in vivo delivery of the CRISPR/Cas9 system using various packaging formats, including viral, mRNA, plasmid, and protein-based approaches. We also discuss some of the barriers which have yet to be overcome for successful translation of this technology. EXPERT OPINION: This review focuses on the challenges to efficacy for various delivery formats, with specific emphasis on overcoming these challenges through the development of carrier vehicles for transient approaches to CRISPR/Cas9 delivery in vivo.

Protein expression by Streptococcus uberis in co-culture with bovine mammary epithelial cells.

Three strains of Streptococcus uberis isolated from dairy cows with mastitis were co-cultured with a bovine mammary epithelial cell line (MAC-T) in Dulbecco's modified Eagle's medium without fetal bovine serum. Protein profiles from culture supernatants and bacterial pellets among different treatments were compared by electrophoresis. There were proteins induced or having increased expression in both supernatant and surface-associated samples from S. uberis co-cultured with MAC-T cells. Some of these proteins were recognized by antibodies in serum obtained from a cow infected by S. uberis. In supernatant samples, there were two distinct protein bands at 35 and 36.8 kDa for all three strains of S. uberis co-cultured with MAC-T cells. These two bands were absent when bacterial protein synthesis was inhibited by chloramphenicol. This study clearly indicates that bacterial protein expression was regulated in response to co-culture with mammary epithelial cells.

Induction of surface-associated proteins of Streptococcus uberis by cultivation with extracellular matrix components and bovine mammary epithelial cells.

Surface-associated protein expression by Streptococcus uberis was influenced by the presence of collagen, laminin and bovine mammary epithelial cells in the culture medium. After electrophoresis and silver staining, four proteins stained more intensively in samples from S. uberis cultivated with epithelial cells and extracellular matrix components than in samples from S. uberis cultivated alone. Induction of these proteins was more obvious after multiple bacterial passages. The correlation between the phenotype of S. uberis and its potential virulence status as illustrated by an immunoblotting study with sera obtained from infected cows revealed that these proteins are probably expressed in vivo during infection.

Incubation of Streptococcus uberis with extracellular matrix proteins enhances adherence to and internalization into bovine mammary epithelial cells.

Two strains of Streptococcus uberis (UT 888 and UT 366) isolated from cows with clinical mastitis were co-cultured with bovine mammary epithelial cells (MAC-T) with and without laminin, fibrinogen, fibronectin or collagen. Incubation of S. uberis with extracellular matrix proteins (ECMPs) increased adherence to and internalization into MAC-T cells. Both strains of S. uberis exhibited greater adherence when co-cultured in the presence of collagen than with any other ECMP. However, adherence was always higher when strains were co-cultured with ECMP than in medium alone. S. uberis UT 888 adhered better to MAC-T cells than S. uberis UT 366. The influence of ECMPs on bacterial internalization into MAC-T cells was similar to adherence, however, differences among ECMPs were less noticeable. S. uberis UT 888 had a higher internalization index than S. uberis UT 366. It is possible that ECMPs induce or up-regulate proteins that selectively adhere to ECMPs which could serve as a bridge between the eukaryotic cell and the bacterial pathogen that leads to internalization of the ECMP-bound pathogen into the mammary epithelial cell.

Binding of bovine lactoferrin to Streptococcus dysgalactiae subsp. dysgalactiae isolated from cows with mastitis.

Three strains of Streptococcus dysgalactiae subsp. dysgalactiae (UT516, UT519, ATCC 27957) were used to determine if bovine lactoferrin (Lf) binds to bacterial cells by biotin avidin binding assay (BABA), enzyme-linked immunosorbent assay (ELISA), and binding inhibition assay. Binding assays revealed that all strains of S. dysgalactiae subsp. dysgalactiae (S. dysgalactiae) evaluated in this study bound to Lf. However, differences in Lf binding capability among strains and between methods used were detected. Binding of Lf was not inhibited by transferrin (Tf) and Lf moiety molecules (mannose, galactose, and lactose) but by Lf. This study demonstrates that S. dysgalactiae bound to bovine Lf in a specific manner.

Relative susceptibility of beef and dairy calves to infection by bovine leukemia virus via tabanid (Diptera: Tabanidae) feeding.

Differences in susceptibility of beef (mixed breeds) and dairy (Holstein) calves to infection by bovine leukemia virus (BLV) were compared. Transmission was accomplished by interrupted feeding of horse flies, Tabanus fuscicostatus Hine, on a donor cow exhibiting persistent lymphocytosis. Flies were transferred individually from the donor cow to each of 11 beef and 10 dairy calves. Transmission of BLV was accomplished with groups of 50 and 250 flies for beef calves and 75 and 250 for dairy calves. These findings indicate that susceptibility of beef and dairy calves to transmission of BLV by tabanids is equivalent and that BLV prevalence differences previously observed among cattle breeds may be caused by management practices.

Natural bovine lentivirus type 1 infection in Holstein dairy cattle. II. Lymphoid tissue lesions.

Bovine immunodeficiency virus (BIV) in Holstein cows was associated with morphologic evidence of lymphoid organ deficiency. Cows were subjected to normal management practices including parturition and lactation without adverse environmental stresses. During the clinical disease process there was marked weight loss and wasting with frequent and severe concurrent infections. Lymphoid follicular hyperplasia and dysplasia in lymph nodes, and hypertrophy and hyperplasia in hemal lymph nodes were characteristics of the lymphoid tissues. Atrophy of lymphoid cell compartments with depletion of lymphocytes and a lymphocytic lymphoid folliculitis were components of the lymphoid system pathology. The nodal tissue lesions resembled those observed in feline, simian, and human lentiviral disease. A functional correlation with immune system deficiency was the development of multiple bacterial infections which failed to resolve after appropriate therapy. The BIV-associated disease syndrome in dairy cows may be useful as a model system for investigation of the pathogenesis of the lymphoid organ changes that occur in humans and animals with lentiviral infection.

Natural bovine lentiviral type 1 infection in Holstein dairy cattle. I. Clinical, serological, and pathological observations.

Clinical, serological, and pathological abnormalities observed in Holstein cows naturally infected with bovine lentivirus 1 bovine immunodeficiency virus (BIV) and other infections were progressive and most commonly associated with weight loss, lymphoid system deficiency, and behavioral changes. Clinical evidence of meningoencephalitis was dullness, stupor, and occasional head or nose pressing postures. The polymerase chain reactions associated the BIV provirus with the lesions in the central nervous system and lymphoid tissues. Multiple concurrent infections developed in retrovirally infected cows undergoing normal stresses associated with parturition and lactation. A major functional correlate of the lymphoreticular alterations was the development of multiple secondary infections which failed to resolve after appropriate antibacterial therapy. The chronic disease syndrome in dairy cows associated with BIV may be useful as a model system for investigation of the pathogenesis of the nervous system lesions and lymphoid organ changes that occur in humans with lentiviral infection.

Encephalitis, lymphoid tissue depletion and secondary diseases associated with bovine immunodeficiency virus in a dairy herd.

Encephalitis, lymphoid tissue depletion and secondary infections occurred over a 5-yr-period in Holstein cows infected with bovine immunodeficiency virus (BIV). There were 59 cattle studied, the majority during 1991, when a severe environmental stress occurred, each with one or more primary causes of death, natural or by euthanasia, and most with several secondary diseases. The encephalitis was characterized by meningeal, perivascular and parenchymal infiltration with lymphocytes, occasional plasma cells and macrophages with perivascular edema in some cows. Affected areas included the cerebrum, cerebellum, and spinal cord with no particular distribution pattern recognized. The lymphoid depletion was primarily an absence of follicular development in nodes draining regions with secondary infections such as chronic mastitis and chronic suppurative pododermatitis. Paucity of lymphocytes in thymic-dependent regions of lymph nodes and the spleen suggested a primary depletion of T cells. Secondary infections were often multiple with each cow having several minor conditions, usually considered short-term and treatable. These included mastitis and pododermatitis, with many cows having non-responding abscesses, cellulitis and myositis attributed to injection site infections. A large number of the cattle had parturition difficulties such as dystocia, obturator paralysis, and metritis. Pulmonary, cardiovascular, and intestinal disease were recognized as both primary and secondary disease conditions. There was a high level of infection with bovine leukemia virus with 4 of the 59 cattle having lymphosarcoma. Under practical conditions, the infection with BIV has a different effect on the host than has been observed under experimental conditions. The presence of BIV combined with the stresses associated with parturition and a modern dairy production system were considered causal for the development of untreatable secondary diseases in immunocompromised cattle. The peak incidence in 1991 was attributed to increased environmental stress during renovation of the barn facility. During this time the cattle were kept on open pasture, exposed to an extremely wet winter, and spring weather conditions. The effect of co-infection with bovine leukemia virus, the influence of immunocompromise on the chronicity of mastitis, the relationship with laminitis and pododermatitis, and several questions related to viral transmission, complementarism with bovine leukemia virus, viral reactivation and immunoprophylaxis all remain as viable avenues for future investigations.

Determination of antimicrobial susceptibility of Pseudomonas aeruginosa by disk diffusion and microdilution methods.

Disk diffusion susceptibility tests were performed on 180 recent clinical isolates of Pseudomonas aeruginosa. Minimal inhibitory concentration values were determined at the same time by a broth microdilution method. All isolates were sensitive to colistin (< 4 migrogram/ml), but resistant to ampicillin (greater than or equal to 16 microgram/ml), cephalothin (greater than or equal to 64 microgram/ml), and nitrofurantoin (> microgram/ml). More than 90% of the isolates were sensitive to gentamicin (median, less than or equal to 0.25 microgram/ml), tobramycin (median, less than or equal to 0.25 microgram/ml), and amikacin (median, less than or equal to 1.0 microgram/ml) and more than 70% were sensitive to carbenicillin (median, 64 microgram/ml). When the resistant and intermediate categories were combined, over 90% of the isolates were resistant to tetracycline (median 16 microgram/ml), chloramphenicol (median, > 32 microgram/ml), kanamycin (median, 16 microgram/ml), and trimethoprim-sulfonamide combiantion (median, 4 microgram/ml; 76 microgram/ml). Differences between the disk diffusion and microdilution methods in distinguishing resistant isolates of P aeruginosa from sensitive isolates were minor. Complete agreement between the two methods was obtained in 87.0% of the observations.

Fatty acid composition of equine plasma.

Fatty acid composition of plasma lipids of normal horses was determined. Four fatty acids (C16:0, C18:0, C18:1, and C18:2) comprised 86.73% of the total, with C18:2 comprising 44.04% of the total. Eight other fatty acids were found in small amounts. Unsaturated fatty acids constituted 66% of the total. Marked variation was demonstrated in fatty acid occurrence and distribution in the sterol ester, triglyceride, phospholipid, and free fatty acid fractions.

Comparisons of serotests with calf inoculations for detection of carriers in anaplasmosis- vaccinated cattle.

Fifteen months after discontinuing vaccination for anaplasmosis in a herd of 131 cattle, 126 of the animals were reactive to the complement-fixation or card-agglutination tests. Inoculation of splenectomized calves proved that 9 animals were carriers and 117 animals were free of anaplasmosis. Anaplasmosis vaccination induced titers detectable by the standard serotests that persisted for at least 15 months.

Fatty acid composition of equine erythrocytes.

Fatty acid composition of erythrocytes of healthy horses was determined. Three fatty acids (C16:0, C18:0, and C18:1) were found in approximately equal quantities and comprised 72.17% of the total. Nine other fatty acids were found in small amounts. Saturated fatty acids constituted 67.2% of the total. Marked variation was demonstrated in the occurrence and distribution of fatty acids in the sterol ester, triglyceride, phospholipid, and free fatty acid fractions.

Screening for neonatal isohemolytic anemia in calves.

Anti-erythrocytic immunoglobulins in serum and colostrum of 124 anaplasmosis-vaccinated cows were detected with a saline agglutination test. Positive results were correlated with the occurrence of neonatal isohemolytic anemia (NIA) in calves and were used to predict the occurrence of NIA. The disease was prevented by withholding colostrum from calves with a high potential for NIA.