Immune cell populations residing in mesenteric adipose depots and mesenteric lymph nodes of lean dairy cows.

Inconsistent evidence of inflammatory immune cell infiltrates in adipose tissues with extensive triglyceride mobilization raises the possibility that regulatory or anti-inflammatory immune cell populations reside within the mesenteric adipose tissue (MAT) and mesenteric lymph nodes (MLN). These resident immune cell populations may be involved in attenuating the inflammatory response. We explored the immune cell population of MAT and MLN collected from lean, lactating Holstein cows without apparent disease in an abattoir (n = 42). Lean cows had a body condition score of 2.6 ± 0.1 (mean ± SD) with a greater frequency of adipocyte area occurring in small rather than large adipocytes. Cells were labeled with monoclonal antibodies specific to bovine leukocyte antigens for enumeration by flow cytometry. Within both lymph node and adipose tissues, relatively large subpopulations of cells expressed the ß2 integrins CD11b and CD11c, class II major histocompatibility antigens (MHCII), and the SIIRP-1α receptor (CD172a) typical of dendritic cells and macrophages. Macrophage/dendritic cell heterogeneity was marked by ß2 integrin expression alone or in conjunction with CD172a or MHCII across subpopulations from both tissues; CD209, the DC-SIGN c-type lectin receptor of dendritic cells, was not detected by fluorescence-activated cell sorting in either tissue. Lymphocytes comprised 74.1 ± 3.7% and 13.7 ± 3.7% of the MLN and MAT cell populations, respectively, and CD3+CD4+ lymphocytes accounted for 49.8 ± 9.9% of the MLN and 6.13 ± 1.23% of the MAT cells. Fox P3+ regulatory lymphocytes comprised 15.3 ± 1.1% and 6.73 ± 0.52% of the MLN and MAT cells, whereas γδ+ lymphocytes accounted for 6.65 ± 0.74% and 3.91 ± 0.43% of the MLN and MAT cells, respectively. Subpopulations of CD3+CD8+ cytotoxic T cells and CD3+CD11c+ innate lymphocytes were present in MLN but not MAT. These results show that subpopulations of resident tissue macrophages, dendritic cells, T helper lymphocytes, regulatory T lymphocytes (Tregs), and γδ lymphocytes reside in mesenteric lymph nodes and adipose tissues. Balance in the innate and adaptive immune functions embedded in these tissues could support metabolic health.

Pre-operative neutrophil count and neutrophil-lymphocyte count ratio (NLCR) in predicting the histological grade of paediatric brain tumours: a preliminary study.

INTRODUCTION: The neutrophil-lymphocyte count ratio (NLCR) is an established prognostic marker for renal, lung and colorectal carcinomas and has been suggested to be predictive of histological grade and outcome in adult intracranial tumours. The purpose of this study was to determine whether a correlation of the pre-operative neutrophil count (NC) and NLCR with the final histological grade exists in paediatric intracranial tumours. METHODS: A retrospective analysis was undertaken at a single centre. Patients less than 18 years old at the time of surgery who underwent tumour-related procedures from 2006 to 2015 were included. Patients with recurrent tumours, previous bone marrow transplant and metastases were excluded. Pre-operative full blood counts (FBC), collected before the diagnosis of intracranial pathology and before administration of steroids, were matched with histological diagnosis for each patient. Post-operative FBC was also recorded, together with survival data where applicable. RESULTS: A total of 116 patients (74 male, 42 female; mean age, 8 ± 0.9 years) with a diagnosis of primary intracranial tumours had pre-operative FBC that could be matched to final histological grade. Pre-operative NC and NLCR were higher with increasing grade of tumour: grade 1 (NC 4.29 109/l, NLCR 2.26), grade 2 (NC 4.59 109/l, NLCR 2.38), grade 3 (NC 5.67 109/l, NLCR 2.72) and grade 4 (NC 6.59 109/l, NLCR 3.31). Patients with WHO grade 1 and 2 tumours pooled together had a lower NC (4.37 95% CI ± 0.67 109/l) compared to WHO grade 3 and 4 patients (6.41 95% CI ± 0.99 109/l, p = 0.0013). The NLCR was lower in grade 1 and 2 tumours (2.29 ± 0.59) (compared to grade 3 and 4 tumours; 3.20 ± 0.76) but this did not reach significance (p = 0.069). The subgroup of patients with pilocytic astrocytoma had a significantly lower NC when compared to patients with high-grade tumours (p = 0.005). Medulloblastoma and supratentorial PNET subgroups had significantly higher NC compared to the low-grade group (p = 0.033, p = 0.002). Post-operative NC was significantly higher in the high-grade tumours (p = 0.034), but no difference was observed for NLCR (p = 0.28). CONCLUSIONS: No evidence exists to support the correlation of pre-operative NC or NLCR to histological diagnosis in paediatric intracranial tumours. Our results indicate that a higher pre-operative NC/NLCR correlates with a higher histological grade of tumour. This suggests that immunological mechanisms may be involved in the pathogenesis of paediatric brain tumours, and a further prospective study is required to substantiate and expand these findings.

High-harmonic fast-wave power flow along magnetic field lines in the scrape-off layer of NSTX.

A significant fraction of high-harmonic fast-wave (HHFW) power applied to NSTX can be lost to the scrape-off layer (SOL) and deposited in bright and hot spirals on the divertor rather than in the core plasma. We show that the HHFW power flows to these spirals along magnetic field lines passing through the SOL in front of the antenna, implying that the HHFW power couples across the entire width of the SOL rather than mostly at the antenna face. This result will help guide future efforts to understand and minimize these edge losses in order to maximize fast-wave heating and current drive.

Management strategies to optimize clinical outcomes after acute traumatic spinal cord injury: integration of medical and surgical approaches.

Although spinal cord injury (SCI) is approximately one-tenth as common as traumatic brain injury, its effects, at both a personal and societal level, are particularly devastating. At present, there is no single therapy that has demonstrated a uniform ability to improve neurological outcomes for SCI patients at long-term follow-up. In spite of this, the last 30 years have borne witness to numerous incremental advances within the field of spinal trauma including the incorporation of standardized neurological assessment tools, the completion of several large therapeutic efficacy trials and the development of modern day surgical classification systems. In this article we review the current evidence surrounding the medical and surgical management of SCI, as well as identify areas where future research is needed.

Early versus late surgery for traumatic spinal cord injury: the results of a prospective Canadian cohort study.

STUDY DESIGN: A multicenter Canadian cohort study. OBJECTIVES: The objective of this study is to evaluate the impact of early versus late surgical decompression on motor neurological recovery after traumatic spinal cord injury (SCI). SETTING: Canadian acute care and SCI rehabilitation facilities. METHODS: A prospective cohort study of patients within the Ontario Spinal Cord Injury Registry program was performed. We considered SCI patients with an admission American Spinal Injury Association (ASIA) Impairment Scale (AIS) grade of A through D, with magnetic resonance imaging-confirmed spinal cord compression. Grouped analysis was performed comparing the cohort of patients who received early surgery (<24 h after SCI) to those receiving delayed surgery (<24 h after SCI). The primary outcome was the change in ASIA motor score (AMS) occurring between hospital admission and rehabilitation discharge. RESULTS: A total of 35 (41.7%) patients underwent early surgery and 49 (58.3%) underwent late surgery. At admission, there was a greater proportion of patients within the early surgery group with more severe AIS grade A injuries. Of the 55 patients with neurological exam available at rehabilitation discharge, a greater proportion had at least a two-grade AIS improvement in the early-surgery group (P=0.01). The mean improvement in AMS at rehabilitation discharge was 20 points amongst early-surgery patients and 15 points amongst late-surgery patients (P=0.46). In the analysis investigating AMS improvement, adjusted for preoperative status and neurological level, there was a positive effect estimate for early surgical therapy that was statistically significant (P=0.01). CONCLUSION: The results here add weight to the growing body of literature, which supports the principle of early intervention in the setting of spinal trauma and SCI.

A novel HER2-positive breast cancer phenotype arising from germline TP53 mutations.

INTRODUCTION: The Li-Fraumeni Syndrome is caused by a germline TP53 mutation and is associated with a high risk of breast cancer at young ages. Basal (triple negative) breast cancers are now well recognised to be a typical sub-type of breast cancer developing in a large proportion of BRCA1 gene carriers. We considered whether a similar narrow sub-type of breast cancer was found in TP53 gene mutation carriers. OBJECTIVE: A hypothesis generating study to investigate whether there are specific breast tumour characteristics associated with germline TP53 mutations. METHODS: Pathological characteristics in 12 breast cancers arising in nine patients carrying pathogenic TP53 mutations were compared to a reference panel of 231 young onset breast tumours included in the POSH study. RESULTS: Patients carrying a TP53 mutation showed a significantly higher likelihood of developing a breast cancer with Human Epidermal growth factor Receptor (HER2) amplification (83%) when compared to the cohort of young onset breast cancer cases (16%); ER and PR status were equivalent between groups. CONCLUSION: These findings suggest that breast cancer developing on a background of an inherited TP53 mutation is highly likely to present with amplification of HER2.

Propagule pressure helps overcome adverse environmental conditions during population establishment.

The establishment success of a population is a function of abiotic and biotic factors and introduction dynamics. Understanding how these factors interact has direct consequences for understanding and managing biological invasions and for applied ecology more generally. Here we use a mesocosm approach to explore how the size of founding populations and the number of introduction events interact with environmental conditions (temperature) to determine the establishment success of laboratory-reared Drosophila melanogaster. We found that temperature played the biggest role in establishment success, eclipsing the role of the other experimental factors when viewed overall. Under optimal temperature conditions propagule pressure was of negligible importance to establishment success.  At adverse temperatures, however, establishment success increased with the total founding population size. This effect was considerably stronger at the cold than at the hot extreme. Whether the population was introduced all at once or by increments (changing the number of introduction events) had a negligible global effect. However, once again, a stronger effect of increasing number of introduction events was seen at adverse temperatures, with hot and cold extremes revealing opposite effects: adding flies incrementally decreased their establishment success at the hot extreme, but increased it at the cold extreme. These differing effects at hot and cold thermal extremes implies that different establishment mechanisms are at play at either extreme. These results suggest that the effort required to prevent (or conversely, to facilitate) the establishment of populations varies with the environment in ways that can be complicated but predictable.

alpha1-Antitrypsin: the presence of excess mannose in the Z variant isolated from liver.

The Z variant of alpha1-antitrypsin was isolated by a new technique from the liver of a patient homozygous for the Z allele of the protease inhibitor locus. The material was homogenous and antigenically competent but had no protease inhibiting capacity. An interesting correlation was found between the subcellular localization and the carbohydrate composition of the Z variant from liver. Carbohydate analysis of this glycoprotein showed an absence of galactose and sialic acid, an appreciable decrease in N-acetylglucosamine, and an almost twofold increase in mannose residues. These data indicate a considerable slowdown in the processing of the oligosaccharides of liver Z variant. In spite of the absence of sialyl residues, the liver Z varant was microheterogeneous by analytical isoelectric focusing. The isoproteins of liver Z variant coincided with those of asialo M variant in the focusing field.

Near identity of cognitive structure in two ethnic groups.

As part of a large-scale family study in Hawaii, Americans of either Japanese or European ancestry were administered a battery of 15 cognitive tests. Principal component analyses (varimax rotations) yielded the same four major cognitive factors for each of the two ethnic groups, and these factors are defined by strikingly similar factor loadings.

Commercially available gas-permeable cell culture bags may not prevent anoxia in cultured or shipped islets.

Prolonged anoxia has deleterious effects on islets. Gas-permeable cell culture devices can be used to minimize anoxia during islet culture and especially during shipment when elimination of gas-liquid interfaces is required to prevent the formation of damaging gas bubbles. Gas-permeable bags may have several drawbacks, such as propensity for puncture and contamination, difficult islet retrieval, and significantly lower oxygen permeability than silicone rubber membranes (SRM). We hypothesized that oxygen permeability of bags may be insufficient for islet oxygenation. We measured oxygen transmission rates through the membrane walls of three different types of commercially available bags and through SRM currently used for islet shipment. We found that the bag membranes have oxygen transmission rates per unit area about 100-fold lower than SRM. We solved the oxygen diffusion-reaction equation for 150-microm diameter islets seeded at 3000 islet equivalents per cm2, a density adequate to culture and ship an entire human or porcine islet preparation in a single gas-permeable device, predicting that about 40% of the islet volume would be anoxic at 22 degrees C and about 70% would be anoxic at 37 degrees C. Islets of larger size or islets accumulated during shipment would be even more anoxic. The model predicted no anoxia in islets similarly seeded in devices with SRM bottoms. We concluded that commercially available bags may not prevent anoxia during islet culture or shipment; devices with SRM bottoms are more suitable alternatives.

Devices and methods for maintenance of temperature and pressure during islet shipment.

UNLABELLED: Exposure to extreme temperatures and pressure fluctuations during shipment by air may have a detrimental impact on islet quality. In this study, we sought to assess the ability of methods and devices to provide better control of the internal environment of islet shipping containers in terms of temperature and pressure. METHODS: Experimental islet shipping containers were packed with 21 panels of commercially available TCP Phase 22 Phase Change Material (TCP). The containers were then exposed for at least 15 hours to three constant external temperature conditions, namely, -20 degrees C, 4 degrees C, and 40 degrees C, and then evaluated for their ability to maintain an internal temperature close to the desired value of 22 degrees C. Custom-designed pressure regulated gyroscopic shipping containers (PRGSC) placed in a vacuum chamber were exposed to an absolute pressure of 250 mm Hg (substantially lower than that experienced during shipment by air) for 25 minutes to assess their ability to control internal pressure under vacuum. Electronic data loggers were used to monitor internal and external temperatures and pressures under all conditions. RESULTS: Twenty-one TCP panels placed in a single islet shipping container were able to maintain the internal temperature between 17 degrees C and 24 degrees C for a minimum of 15 hours at all three external temperatures. The PRGSC tested were able to maintain a constant internal pressure of 760 mm Hg when exposed to vacuum. CONCLUSIONS: Our results demonstrated that the use of containers equipped with TCP and PRGSC exert excellent environmental control over islet shipments by minimizing temperature and eliminating pressure fluctuations.

Electrophysiological patterns of diabetic polyneuropathy.

OBJECTIVE: This study evaluates the pattern of electrodiagnostic (EDX) abnormalities in diabetic sensorimotor polyneuropathies. METHODS: EDX data from 112 consecutive patients with type 2 diabetes mellitus with distal, predominantly sensory, polyneuropathies were reviewed. Motor conduction velocities (CV), distal motor latencies (DML), compound muscle action potential (CMAP) amplitudes, distal to proximal amplitude ratios (PID), and F-wave latencies (FWL) were analyzed. Data were normalized based on normative reference values, and the proportion of nerves with abnormal values in the lower and upper limbs were evaluated. These data were also analyzed in relation to whether there was possible demyelinating versus axonal injury. Statistical analyses included comparison of the proportions of abnormal nerves in upper versus lower limbs as well as using Generalized Estimating Equations (GEE) to account for correlated observations for each patient between lower and upper limbs and adjusting for patient age effect. RESULTS: CVs were significantly more abnormal in the legs than the arms (p < 0.0006) and decreased CMAP amplitudes meeting criteria for axonal injury were also more frequent in the legs (p < 0.0001). Using the GEE model, axonal injury was more common in the legs while demyelinating injury was more common in the arms based on FWLs, especially in younger persons (e.g., 40 years old vs. 50 years old). These differences are not readily explained by the duration of the diabetes. CONCLUSION: Since in diabetics "axonal" type injury may be more common in the legs while "demyelinating" injury more frequent in the arms, this study emphasizes the limitation of this type of classification, and supports the idea that the pattern of EDX abnormalities in different types of neuropathies may be more helpful.

Use of an Ultrasonic Bone Curet for the Extraction of a Cervical Artificial Disc: A Novel Application: A Case Report.

CASE: Eighteen months after implantation of a C6-C7 cervical artificial disc (ProDisc-C; DePuy Synthes), a 44-year-old woman presented with right-sided C7 radiculopathy. Imaging revealed posterior subsidence of the prosthesis, with C7 foraminal stenosis. An ultrasonic curet was used to safely and effectively extract the prosthesis without neurologic complication; safe access for the insertion of an intervertebral cage was facilitated. CONCLUSION: The ultrasonic curet was a safe and effective adjunct for revision artificial cervical disc replacement surgery; this technique may help to reduce potential injury to neural structures in a procedure with a traditionally high complication rate.

The changing practice of transurethral resection of the prostate.

Introduction Transurethral resection of the prostate (TURP) is considered the gold standard surgical treatment for lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia. The number of TURPs performed has declined significantly over the last three decades owing to pharmaceutical therapy. TURP data from a single institution for the years 1990, 2000 and 2010 were compared to assess the difference in performance. Methods A retrospective analysis was undertaken of all patients who underwent TURP between January and December 2010. These findings were compared with historical data for the years 1990 and 2000: 100 sets of case notes were selected randomly from each of these years. Results The number of TURPs performed fell from 326 in 1990 to 113 in 2010. The mean age of patients increased from 70.6 years to 74.0 years. There was also a significant increase in the mean ASA grade from 1.9 to 2.3. The most common indication for TURP shifted from LUTS to acute urinary retention. No significant change in operating time was observed. The mean resection weight remained constant (22.95g in 1990, 22.55g in 2000, 20.76g in 2010). A reduction in transfusion rates was observed but there were higher rates of secondary haematuria and bladder neck stenosis. There was an increase from 2% to 11.5% of patients with long-term failure to void following TURP. Conclusions The number of TURPs performed continues to decline, which could lead to potential training issues. Urinary retention is still by far the most common indication. However, there has been a significant rise in the percentage of men presenting for TURP with high pressure chronic retention. The number of patients with bladder dysfunction who either have persistent storage LUTS or eventually require long-term catheterisation or intermittent self-catheterisation has increased markedly, which raises the question of what the long-term real life impact of medical therapy is on men with LUTS secondary to benign prostatic hyperplasia who eventually require surgery.

Factors affecting spinal cord stimulation outcome in chronic benign pain with suggestions to improve success rate.

For patient selection, psychological factors like fear avoidance, depression, secondary gain or refusal to be weaned off narcotics should be avoided. Trial Stimulation is an important tool to reduce the rate of failed permanent implants, and to improve cost-effectiveness. The etiology of pain has a strong influence on the success rate. The success rate is inversely proportional to the time interval from the initial onset of symptoms to the time of implantation. Multi-polar and multi-channel systems improve the long-term reliability and success rate and have proven to reduce the incidence of open surgery in case of electrode displacement. Third party coverage like the Worker's Compensation negatively affects the long term success. Reducing the complication rate directly benefits long term success rates. The electrode fracture rate can be reduced by using the paramedian approach, the use of three wing silicone anchor placed immediately at the point of exit of the lead from the deep fascia and avoiding a hard plastic twist lock anchor. The displacements can be reduced by fixing the anchor to the deep fascia firmly, supplemented by the use of silicone glue, and by placing the implantable pulse generator (IPG) in the abdominal wall, instead of the gluteal region. The use of prophylactic antibiotics tends to reduce the infection rate.

The importance of pollinators and autonomous self-fertilisation in the early stages of plant invasions: Banksia and Hakea (Proteaceae) as case studies.

Reproduction is a crucial stage in the naturalisation of introduced plant species. Here, using breeding system experiments and observations of floral visitors, we investigate whether a lack of pollinators or an inability to autonomously self-fertilise limits naturalisation in five Australian Banksia species and the co-familial Hakea salicifolia in South Africa. Banksia species were heavily utilised by native insects and nectar-feeding birds. Although Banksia produced fruit when pollinators were excluded, pollinators significantly increased seed set in four of the five species. H. salicifolia flowers were visited by 11 insect species; honeybees (Apis mellifera) were the main visitors. Flowers in naturalised H. salicifolia populations received almost four times the number of visits as flowers in non-naturalised populations; the latter showed both pollen limitation (PLI 0.40) and partial self-incompatibility. This should not prevent invasion, since H. salicifolia produces fruits via autonomous selfing in the absence of pollinators. The results suggest a limited role of breeding systems in mediating naturalisation of introduced Proteaceae species. Other factors, such as features of the recipient environments, appear to be more important. Spatial variation in rates of reproduction might, however, explain variation in the extent and rate of naturalisation of different populations.

Distribution, purification and characterization of rat intestinal UDPgalactose: N-acetylglucosaminyl(beta 1----4)galactosyltransferase.

Rat intestinal UDPgalactose: N-acetylglucosaminyl(beta 1----4)galactosyltransferase activity was studied as to its intestinal and villus-to-crypt distribution, and then purified and characterized. Rapid UDPgalactose hydrolysis was noted in the duodenum and jejunum; little to no breakdown was detected in the distal ileum, cecum and proximal colon. Product analysis suggested that UDPgalactose hydrolysis was due to nucleotide-sugar pyrophosphatase and galactose-1-phosphate phosphatase activities; ileum appeared to have little of the first activity and none of the latter. An aboral gradient of galactosyltransferase activity was noted, activity being 3-4-fold higher in the ileum, cecum and proximal colon. Total homogenate exogenous acceptor galactosyltransferase activities showed no villus-crypt differences but activity measured with intact isolated cells demonstrated higher activity with crypt cells; this was particularly evident in the ileum. Galactosyltransferase activity was purified from ileal-colonic mucosa. An over 4000-fold purification with 75 percent yield was achieved. Only one band of approx. 70-75 kDa was noted on sodium dodecyl sulfate polyacrylamide electrophoresis. As with other eukaryotic galactosyltransferase activities, there was an absolute requirement for Mn2+; the concentration required for half maximal activity was only 2.5 microM and higher concentrations did not inhibit. The Km for UDPgalactose was 30 microM.

The distribution and partial characterization of UDPgalactose: N-acetylgalactosamine mucin galactosyltransferase activity from rat small intestine and its sensitivity to Zn2+.

UDPgalactose: N-acetylgalactosamine mucin galactosyltransferase activity of the rat intestine was studied and purified using asialo-ovine submaxillary mucin as the acceptor substrate and inhibitors to suppress UDPgalactose breakdown by pyrophosphatase activities particularly prevalent in the duodenal-jejunal regions. Despite adequate suppression of UDPgalactose breakdown, significant intestinal region differences of mucin galactosyltransferase activity were observed. Elevations of activity were observed in the duodenum and distal ileum of the small intestine and the cecum and proximal colon; these elevations in activity correspond to areas of increased mucin production. Similarly, mucin galactosyltransferase activity of duodenal cells isolated along a crypt-to-villus axis showed a moderate increase (67.7%) in activity associated with cells in the crypt region. Small intestine mucin galactosyltransferase activity was purified 800-fold using a series of ion exchange (DEAE-Sepharose), gel filtration (S-200 Sephacryl) and affinity chromatographic steps to isolate the mucin galactosyltransferase activity from a Triton X-100/Nonidet P-40 extract of homogenized cells obtained by scraping everted intestines. The partially purified enzyme showed two distinct protein bands of 81.5 and 50 kDa and a faint band at 53.3 kDa. Kinetic analysis gave an apparent Km of 152 microM for UDPgalactose. The enzyme showed optimal activity with Mn2+ (20 mM) and partial activities using a number of other divalent cations. Higher concentrations of Mn2+ were slightly inhibitory. Mucin galactosyltransferase activity was inhibited by more then 90% in the presence of Zn2+ (4 mM) and this inhibition could not be reversed by additional Mn2+. Addition of Zn2+ (4 mM) to assays containing Mn2+ (20 mM) did not cause appreciable UDPgalactose breakdown, as measured by high-voltage paper electrophoresis, suggesting that Zn2+ inhibition is not a result of pyrophosphatase activation. In addition, Zn2+ does not appear to activate a protease or glycosidase activity in the partially purified enzyme preparation which could hydrolyze the galactosylated product prior to isolation.