Gait Abnormalities and the Risk of Falls in CKD.
Older adults with CKD are at high risk of falls and disability. It is not known whether gait abnormalities contribute to this risk. Quantitative and clinical gait assessments were performed in 330 nondisabled community-dwelling adults aged ≥65 years. CKD was defined as an eGFR <60 ml/min per 1.73 m2. Cox proportional hazards models were created to examine fall risk. A total of 41% (n=134) of participants had CKD. In addition to slower gait speed, participants with CKD had gait cycle abnormalities including shorter stride length and greater time in the stance and double-support phases. Among people with CKD, lower eGFR was independently associated with the severity of gait cycle abnormalities (per 10 ml/min per 1.73 m2 lower eGFR: 3.6 cm [95% confidence interval (95% CI), 1.4 to 5.8] shorter stride length; 0.7% [95% CI, 0.3 to 1.0] less time in swing phase; 1.1% [95% CI, 0.5 to 1.7] greater time in double-support phase); these abnormalities mediated the association of lower eGFR with slower gait speed. On clinical gait exam, consistent with the quantitative abnormalities, short steps and marked swaying or loss of balance were more common among participants with CKD, yet most had no identifiable gait phenotype. A gait phenotype defined by any of these abnormal signs was associated with higher risk of falls among participants with CKD: compared with people without CKD and without the gait phenotype, the adjusted hazard ratio was 1.72 (95% CI, 1.06 to 2.81) for those with CKD and the phenotype; in comparison, the adjusted hazard ratio was 0.71 (95% CI, 0.40 to 1.25) for people with CKD but without the phenotype (P value for interaction of CKD status and gait phenotype =0.01). CKD in older adults is associated with quantitative gait abnormalities, which clinically manifest in a gait phenotype that is associated with fall risk.


Longitudinal network structure of depression symptoms and self-efficacy in low-income mothers.
Maternal depression was recently conceptualized as a network of interacting symptoms. Prior studies have shown that low self-efficacy, as an index of maternal functioning, is one important source of stress that worsens depression. We have limited information, however, on the specific relationships between depression symptoms and self-efficacy. In this study, we used regularized partial correlation networks to explore the multivariate relationships between maternal depression symptoms and self-efficacy over time. Depressed mothers (n = 306) completed the Center for Epidemiological Studies Depression (CES-D) scale at four time points, between four and eight weeks apart. We estimated (a) the network structure of the 20 CES-D depression symptoms and self-efficacy for each time point, (b) determined the centrality or structural importance of all variables, and (c) tested whether the network structure changed over time. In the resulting networks, self-efficacy was mostly negatively connected with depression symptoms. The strongest relationships among depression symptoms were 'lonely-sleep difficulties' and 'inability to get going-crying'. 'Feeling disliked' and 'concentration difficulty' were the two most central symptoms. In comparing the network structures, we found that the network structures were moderately stable over time. This is the first study to investigate the network structure and their temporal stability of maternal depression symptoms and self-efficacy in low-income depressed mothers. We discuss how these findings might help future research to identify clinically relevant symptom-to-symptom relationships that could drive maternal depression processes, and potentially inform tailored interventions. We share data and analytical code, making our results fully reproducible.


Elevated Serum GAD65 and GAD65-GADA Immune Complexes in Stiff Person Syndrome.
Glutamic acid decarboxylase 65 (GAD65) and autoantibodies specific for GAD65 (GADA) are associated with autoimmune diseases including Stiff Person Syndrome (SPS) and Type 1 diabetes (T1D). GADA is recognized as a biomarker of value for clinical diagnosis and prognostication in these diseases. Nonetheless, it remains medically interesting to develop sensitive and specific assays to detect GAD65 preceding GADA emergence, and to monitor GADA-GAD65 immune complexes in blood samples. In the present study, we developed a highly sensitive proximity ligation assay to measure serum GAD65. This novel assay allowed detection of as little as 0.65 pg/ml GAD65. We were also able to detect immune complexes involving GAD65 and GADA. Both free GAD65 and GAD65-GADA levels were significantly higher in serum samples from SPS patients compared to healthy controls. The proximity ligation assays applied for detection of GAD65 and its immune complexes may thus enable improved diagnosis and better understanding of SPS.


Bradykinin is a mediator of anaphylactoid reactions during hemodialysis with AN69 membranes.
Anaphylactoid reactions (AR) are the most feared complications of hemodialysis. Recently, a high incidence of AR has been reported during dialysis with AN69 membranes in patients treated with ACE inhibitors. Plasma levels of C3a, histamine and bradykinin were measured in 12 patients at the onset of AR during dialysis with AN69. We also investigated bradykinin generation in 10 symptom-free patients dialyzed with four different membranes. None of the 12 patients studied during AR displayed excessive complement activation or histamine release. In contrast, high bradykinin plasma levels (2392 +/- 53 fmol/ml; mean +/- SEM) were observed in all nine patients of whom bradykinin was measured. One patient developed two consecutive episodes of hypersensitivity on AN69 membranes even without taking ACE inhibitors. Bradykinin levels were high in both episodes (5280 and 10467.7 fmol/ml). Furthermore, this patient showed no symptoms and normal bradykinin levels (123.4 fmol/ml) when dialyzed with other membranes. The role of the membrane type in the AR is further substantiated by the observation that AN69 also provoked a significantly higher bradykinin generation (327.6 +/- 18 fmol/ml; mean +/- SEM) during symptom-free sessions compared to other membranes like CuprophanR (5.1 +/- 7.3), HemophanR (17.2 +/- 6.3) and PolysulfoneR (39.7 +/- 6.6). Our findings strongly suggest that bradykinin is the principal mediator of AR during hemodialysis with AN69 membranes. To our knowledge it is the first time that data support the hypothesis of a more general role of bradykinin in shock-like symptoms. Furthermore, bradykinin generation must be regarded as a new marker of biocompatibility of extracorporeal treatments.


Differentiation of troponin in cardiac and skeletal muscles in chicken embryos as studied by immunofluorescence microscopy.
The differentiation of troponin (TN) in cardiac and skeletal muscles of chicken embryos was studied by indirect immunofluorescence microscopy. Serial sections of embryos were stained with antibodies specific to TN components (TN-T, -I, and -C) from adult chicken cardiac and skeletal muscles. Cardiac muscle began to be stained with antibodies raised against cardiac TN components in embryos after stage 10 (Hamburger and Hamilton numbering, 1951, J. Morphol. 88:49-92). It reacted also with antiskeletal TN-I from stage 10 to hatching. Skeletal muscle was stained with antibodies raised against skeletal TN components after stage 14. It also reacted with anticardiac TN-T and C from stage C from stage 14 to hatching. It is concluded that, during embryonic development, cardiac muscle synthesizes TN-T and C that possess cardiac-type antigenicity and TN-I that has antigenic determinants similar to those present in cardiac as well as in skeletal muscles. Embryonic skeletal muscle synthesizes TN-I that possesses antigenicity for skeletal muscle and TN-T and C which share the antigenicities for both cardiac and skeletal muscles. Thus, in the development of cardiac and skeletal muscles, a process occurs in which the fiber changes its genomic programming: it ceases synthesis of the TN components that are immunologically indistinguishable from one another and synthesizes only tissue-type specific proteins after hatching.


Mechanical allodynia induced by optogenetic sensory nerve excitation activates dopamine signaling and metabolism in medial nucleus accumbens.
The mesolimbic dopaminergic signaling, such as that originating from the ventral tegmental area (VTA) neurons in the medial part of the nucleus accumbens (mNAc), plays a role in complex sensory and affective components of pain. To date, we have demonstrated that optogenetic sensory nerve stimulation rapidly alters the dopamine (DA) content within the mNAc. However, the physiological role and biochemical processes underlying such rapid and regional dynamics of DA remain unclear. In this study, using imaging mass spectrometry (IMS), we observed that sensitized pain stimulation by optogenetic sensory nerve activation increased DA and 3-Methoxytyramine (3-MT; a post-synaptic metabolite obtained following DA degradation) in the mNAc of the experimental mice. To delineate the mechanism associated with elevation of DA and 3-MT, the de novo synthesized DA in the VTA/substantia nigra terminal areas was evaluated using IMS by visualizing the metabolic conversion of stable isotope-labeled tyrosine (13C15N-Tyr) to DA. Our approach revealed that at steady state, the de novo synthesized DA occupied >10% of the non-labeled DA pool in the NAc within 1.5 h of isotope-labeled Tyr administration, despite no significant increase following pain stimulation. These results suggested that sensitized pain triggered an increase in the release and postsynaptic intake of DA in the mNAc, followed by its degradation, and likely delayed de novo DA synthesis. In conclusion, we demonstrated that short, peripheral nerve excitation with mechanical stimulation accelerates the mNAc-specific DA signaling and metabolism which might be associated with the development of mechanical allodynia.


Involvement of sialic acid on endothelial cells in organ-specific lymphocyte recirculation.
Mouse lymphocytes incubated on cryostat-cut sections of lymphoid organs (lymph nodes and Peyer's patches) specifically adhere to the endothelium of high endothelial venules (HEV), the specialized blood vessels to which recirculating lymphocytes attach as they migrate from the blood into the parenchyma of the lymphoid organs. Treatment of sections with sialidase eliminated the binding of lymphocytes to peripheral lymph node HEV, had no effect on binding to Peyer's patch HEV, and had an intermediate effect on mesenteric lymph node HEV. These results suggest that sialic acid on endothelial cells may be an organ-specific recognition determinant for lymphocyte attachment.


Hepatitis C virus (HCV) core protein-induced, monocyte-mediated mechanisms of reduced IFN-alpha and plasmacytoid dendritic cell loss in chronic HCV infection.
IFN-alpha production by plasmacytoid dendritic cells (PDCs) is critical in antiviral immunity. In the present study, we evaluated the IFN-alpha-producing capacity of PDCs of patients with chronic hepatitis C virus (HCV) infection in treatment-naive, sustained responder, and nonresponder patients. IFN-alpha production was tested in PBMCs or isolated PDCs after TLR9 stimulation. Treatment-naive patients with chronic HCV infection had reduced frequency of circulating PDCs due to increased apoptosis and showed diminished IFN-alpha production after stimulation with TLR9 ligands. These PDC defects correlated with the presence of HCV and were in contrast with normal PDC functions of sustained responders. HCV core protein, which was detectable in the plasma of infected patients, reduced TLR9-triggered IFN-alpha and increased TNF-alpha and IL-10 production in PBMCs but not in isolated PDCs, suggesting HCV core induced PDC defects. Indeed, addition of rTNF-alpha and IL-10 induced apoptosis and inhibited IFN-alpha production in PDCs. Neutralization of TNF-alpha and/or IL-10 prevented HCV core-induced inhibition of IFN-alpha production. We identified CD14+ monocytes as the source of TNF-alpha and IL-10 in the HCV core-induced inhibition of PDC IFN-alpha production. Anti-TLR2-, not anti-TLR4-, blocking Ab prevented the HCV core-induced inhibition of IFN-alpha production. In conclusion, our results suggest that HCV interferes with antiviral immunity through TLR2-mediated monocyte activation triggered by the HCV core protein to induce cytokines that in turn lead to PDC apoptosis and inhibit IFN-alpha production. These mechanisms are likely to contribute to HCV viral escape from immune responses.


A carboxyl-terminally truncated mutant and nonglycosylated A2a adenosine receptors retain ligand binding.
The amino acids that comprise the ligand binding sites of adenosine receptors have not been identified. Adenosine and its agonist analogues differ from ligands for the well studied biogenic amine receptors and rhodopsin in that the adenosine receptor agonists are larger, contain a ribose moiety, and are uncharged at physiological pH. Thus, the locations of the ligand binding pockets of the adenosine receptors could differ significantly from those of the biogenic amine receptors. This report describes the characterization of a purification-amenable truncated mutant of the canine A2a adenosine receptor and demonstrates that neither the long carboxyl-terminal tail nor the glycosidic moiety appears to be required for ligand binding. The dog thyroid A2a adenosine receptor cDNA (RDC8) was subcloned into the mammalian expression vector pCMV4. A mutant A2a construct, in which six histidines replaced residues 310-412 as the carboxyl terminus of the protein, also was prepared. When overexpressed transiently in COS M6 cells, the wild-type and mutant A2a receptors exhibited similar 2-[p-(2-[3H]carboxyethyl)phenylethylamino]-5'-N- ethylcarboxamidoadenosine saturation binding and competition curve profiles. The following biochemical techniques confirmed that the COS M6 cells were transcribing and translating A2a receptors of the expected molecular masses: (a) immunoblotting with an antipeptide antibody directed against the putative carboxyl-terminal side of the second extracellular loop (Tyr155-Val172) of the canine A2a adenosine receptor, (b) photoaffinity labeling with the A2a-selective agonist 125I-2-[4-[2-[2-[(4-azidophenyl)methylcarbonylamino] ethylaminocarbonyl]ethyl]phenyl]ethylamino-5'-N-ethylcarboxamidoad enosine (125I-azido-PAPA-APEC), and (c) partial purification of the hexahistidine-tagged receptor on Ni2+.nitrilotriacetic acid resin. A presumed A2a receptor (44 kDa) from rabbit striatal membranes also was detected with the antisera against amino acids Tyr155-Val172 of the RDC8 receptor. Not only could the mutant A2a receptor be photolabeled specifically with 125I-azido-PAPA-APEC but so too could unglycosylated A2a receptors (i.e., from tunicamycin-treated COS M6 cells), either full length or truncated. In all of these cases, photolabeling was attenuated by both agonist and antagonist competitors.


The Escherichia coli DNA polymerase III holoenzyme contains both products of the dnaX gene, tau and gamma, but only tau is essential.
The replicative polymerase of Escherichia coli, DNA polymerase III, consists of a three-subunit core polymerase plus seven accessory subunits. Of these seven, tau and gamma are products of one replication gene, dnaX. The shorter gamma is created from within the tau reading frame by a programmed ribosomal -1 frameshift over codons 428 and 429 followed by a stop codon in the new frame. Two temperature-sensitive mutations are available in dnaX. The 2016(Ts) mutation altered both tau and gamma by changing codon 118 from glycine to aspartate; the 36(Ts) mutation affected the activity only of tau because it altered codon 601 (from glutamate to lysine). Evidence which indicates that, of these two proteins, only the longer tau is essential includes the following. (i) The 36(Ts) mutation is a temperature-sensitive lethal allele, and overproduction of wild-type gamma cannot restore its growth. (ii) An allele which produced tau only could be substituted for the wild-type chromosomal gene, but a gamma-only allele could not substitute for the wild-type dnaX in the haploid state. Thus, the shorter subunit gamma is not essential, suggesting that tau can be substitute for the usual function(s) of gamma. Consistent with these results, we found that a functional polymerase was assembled from nine pure subunits in the absence of the gamma subunit. However, the possibility that, in cells growing without gamma, proteolysis of tau to form a gamma-like product in amounts below the Western blot (immunoblot) sensitivity level cannot be excluded.


Development of a primary microglia screening assay and its use to characterize inhibition of system xc- by erastin and its analogs.
The inflammatory response in the central nervous system involves activated microglia. Under normal conditions they remove damaged neurons by phagocytosis. On the other hand, neurodegenerative diseases are thought to involve chronic microglia activation resulting in release of excess glutamate, proinflammatory cytokines and reactive oxygen species, leading to neuronal death. System xC- cystine/glutamate antiporter (SXC), a sodium independent heterodimeric transporter found in microglia and astrocytes in the CNS, imports cystine into the cell and exports glutamate. SXC has been shown to be upregulated in neurodegenerative diseases including multiple sclerosis, ALS, neuroAIDS Parkinson's disease and Alzheimer's disease. Consequently, SXC inhibitors could be of use in the treatment of diseases characterized by neuroinflammation and glutamate excitotoxicity. We report on the optimization of a primary microglia-based assay to screen for SXC inhibitors. Rat primary microglia were activated using lipopolysaccharides (LPS) and glutamate release and cystine uptake were monitored by fluorescence and radioactivity respectively. LPS-induced glutamate release increased with increasing cell density, time of incubation and LPS concentration. Conditions to screen for SXC inhibitors were optimized in 96-well format and subsequently used to evaluate SXC inhibitors. Known SXC inhibitors sulfasalazine, S-4CPG and erastin blocked glutamate release and cystine uptake while R-4CPG, the inactive enantiomer of S-4CPG, failed to inhibit glutamate release or cystine transport. In addition, several erastin analogs were evaluated using primary microglia and found to have EC50 values in agreement with previous studies using established cell lines.


Soft versus hard resting hand splints in rheumatoid arthritis: pain relief, preference, and compliance.
This study compared soft versus hard resting hand splints on pain and hand function in 39 persons with rheumatoid arthritis. Splint preference was also evaluated to determine its effects on splint wear compliance. A repeated measures research design was used to compare the two experimental conditions, wearing a soft splint versus a hard splint on the dominant hand for 28 days at night only, and an unsplinted control period of 28 days. Arthritis pain was considerably less during both splinted periods when compared with the pretest. Subjects identified fewer joints as being painful during the soft splint condition than during the unsplinted condition. There were no significant differences among conditions on hand function measures. Splint preference was 57% for the soft splint, 33% for the hard splint, and 10% for no splint. Splint wear compliance was significantly better with the soft splint (82%) than with the hard splint (67%). The findings indicate that resting hand splints are effective for pain relief and that persons with rheumatoid arthritis are more likely to prefer and comply with soft splint use for this purpose. Individualized splint prescription that focuses on client comfort and preference may enhance splint wear compliance.


A Rare Pediatric Case of Posttraumatic Pseudoaneurysm: Case Report and Literature Review.
Posttraumatic pseudoaneurysms are extremely rare in pediatric populations. In many cases, pseudoaneurysms may be confused with abscesses, epidermoid cysts, arteriovenous fistula, foreign objects, and ganglion cysts, as well as tumors. They are associated with distinguishing findings of "pulsatile mass, a palpable thrill, and an audible to-and-fro murmur" (1), which can be confirmed by various imaging techniques. In this report, we describe the case of a 4-year-old boy who presented to the pediatric emergency department 3 weeks after falling and subsequently getting cut by glass. Upon clinical examination, the patient presented with pulsatile, swollen mass in the left wrist. A Doppler ultrasound of the left wrist demonstrated that the area of clinical concern in the left wrist showed a pseudoaneurysm, and prominent arterial blood flow was seen within the pseudoaneurysm. Because pseudoaneurysms, particularly posttraumatic pseudoaneurysms, are extremely rare in the pediatric population, it may be easy to miss these cases during clinical examination. Misdiagnosis of the pseudoaneurysm can cause delayed treatment, a longer recovery period, and complications such as infection, rupture, and hemorrhage. It is important for physicians to consider this entity when evaluating patients with symptoms of asymptomatic bulges to painful pulsatile masses after trauma.


Physical separation from the mate diminishes male's attentiveness towards other females: a study in monogamous prairie voles Microtus ochrogaster.
We tested whether continuous cohabitation in monogamous voles affects the mated male's attentiveness to his breeding partner versus another female. Each male was housed in a 3-chamber apparatus with a Focal female (FF) and a Control female (CF) for 13 days then placed in a T-maze to assess his attentiveness to and memory of those females. The Distal male remained physically separated from both females, but received their distal cues. The Separate male cohabited with the FF for 3 days then remained physically separated from both females. The Disrupt male's continuous cohabitation with the FF was disrupted by having him physically separated from her after 10 days and placed with the CF for the last 3 days. The Continuous male cohabited continuously with the FF for 13 days. With females in the T-maze, the Separate and Disrupt males spent more time near the FF's box and the Disrupt males spent more time manipulating the FF's box than the CF's box. The Separate males groomed themselves more when near the FF's box than the CF's box. The Distal and Continuous males' attentiveness to the two females did not differ. Results suggest that physical distance from the partner may reduce male's attentiveness toward other potential mates. Prairie voles might be similar to socially monogamous primates in using tactile cues as a signal for maintaining their social bonds.


Tile-based self-assembly of a triple-helical polysaccharide into cell wall-like mesoporous nanocapsules.
Tile-based self-assembly is a robust system in the construction of three-dimensional DNA nanostructures but it has been rarely applied to other helical biopolymers. β-Glucan is an immunoactive natural polymer which exists in a triple helical conformation. Herein, we report that β-glucan, after modification using two types of short chain acyl groups, can self-assemble into tiles with inactivated sticky ends at the interface of two solvents. These tiles consist of a single layer of helices laterally aligned, and the sticky ends can be activated when a few acyl groups at the ends are removed; these tiles can further pack into mesoporous nanocapsules, in a similar process as the sticky DNA tiles pack into complex polyhedral nano-objects. These nanocapsules were found to have targeted effects to antigen presenting cells in a RAW264.7 cell model. Our study suggests that tile-based self-assembly can be a general strategy for helical biopolymers, and on fully exploiting this strategy, various new functional nanostructures will become accessible in the future.


Bovine viral diarrhea virus NS4B protein is an integral membrane protein associated with Golgi markers and rearranged host membranes.
Very little is known about BVDV NS4B, a protein of approximately 38 kDa. However, a missense mutation in NS4B has been implicated in changing BVDV from a cytopathic to noncytopathic virus, suggesting that NS4B might play a role in BVDV pathogenesis. Though this is one possible function, it is also likely that NS4B plays a role in BVDV genome replication. For example, BVDV NS4B interacts with NS3 and NS5A, implying that NS4B is part of a complex, which contains BVDV replicase proteins. Other possible BVDV NS4B functions can be inferred by analogy to hepatitis C virus (HCV) NS4B protein. For instance, HCV NS4B remodels host membranes to form the so-called membranous web, the site for HCV genome replication. Finally, HCV NS4B is membrane-associated, implying that HCV NS4B may anchor the virus replication complex to the membranous web structure. Unlike its HCV counterpart, we know little about the subcellular distribution of BVDV NS4B protein. Further, it is not clear whether NS4B is localized to host membrane alterations associated with BVDV infection. We show first that release of infectious BVDV correlates with the kinetics of BVDV genome replication in infected cells. Secondly, we found that NS4B subcellular distribution changes over the course of BVDV infection. Further, BVDV NS4B is an integral membrane protein, which colocalizes mainly with the Golgi compartment when expressed alone or in the context of BVDV infection. Additionally, BVDV induces host membrane rearrangement and these membranes contain BVDV NS4B protein. Finally, NS4B colocalizes with replicase proteins NS5A and NS5B proteins, raising the possibility that NS4B is a component of the BVDV replication complex. Interestingly, NS4B was found to colocalize with mitochondria suggesting that this organelle might play a role in BVDV genome replication or cytopathogenicity. These results show that BVDV NS4B is an integral membrane protein associated with the Golgi apparatus and virus-induced membranes, the putative site for BVDV genome replication. On the basis of NS4B Colocalization with NS5A and NS5B, we conclude that NS4B protein is an integral component of the BVDV replication complex.


Targeted Assessment of Enlargement of the Perivascular Space in Alzheimer's Disease and Vascular Dementia Subtypes Implicates Astroglial Involvement Specific to Alzheimer's Disease.
Waste clearance from the brain parenchyma occurs along perivascular pathways. Enlargement of the perivascular space (ePVS) is associated with pathologic features of Alzheimer's disease (AD), although the mechanisms and implications of this dilation are unclear. Fluid exchange along the cerebral vasculature is dependent on the perivascular astrocytic water channel aquaporin-4 (AQP4) and loss of perivascular AQP4 localization is found in AD. We directly measured ePVS in postmortem samples of pathologically characterized tissue from participants who were cognitively intact or had AD or mixed dementia (vascular lesions with AD). We found that both AD and mixed dementia groups had significantly increased ePVS compared to cognitively intact subjects. In addition, we found increased global AQP4 expression of the AD group over both control and mixed dementia groups and a qualitative reduction in perivascular localization of AQP4 in the AD group. Among these cases, increasing ePVS burden was associated with the presence of tau and amyloid-β pathology. These findings are consistent with the existing evidence of ePVS in AD and provide novel information regarding differences in AD and vascular dementia and the potential role of astroglial pathology in ePVS.


Lipid remodelling in human melanoma cells in response to UVA exposure.
Extensive exposure to UVA is thought to increase the risk of malignancy and the progression of melanoma, the most serious type of skin cancer. It is well known that alterations in lipid metabolism represent an early event in carcinogenesis, but the impact of UVA exposure on the lipid composition of cancer cells is still largely unknown. In this study we aimed at investigating lipid remodeling in human melanoma cells in response to UVA exposure. After UVA irradiation, lipid extracts were either immediately collected from SK-MEL-28 cells or collected after a recovery period of 2 h or 24 h. The lipid profiles for each event were determined by liquid chromatography or gas chromatography coupled to mass spectrometry. UVA exposure led to major alterations in both fatty acids (FA) and phospholipid profiles. An increase of monounsaturated FA (MUFA) and FA18:0, as well as a decrease of FA16:0, were observed 24 h after irradiation. Moreover, phosphatidylcholine (PC) decreased and phosphatidylinositol (PI) increased after UVA exposure. Molecular alterations in the PC, lysoPC, PI, phosphatidylethanolamine (PE), ether-linked PE and phosphatidylglycerol (PG) profiles were also observed. The absence of cleaved caspase-3 after 2 h and 24 h of re-incubation is correlated with impairment of apoptosis. Overall, these data showed changes in membrane lipids, which may be associated with lipogenesis after UVA exposure which, in turn, is usually a determinant for cell survival.


WNT ligands control initiation and progression of human papillomavirus-driven squamous cell carcinoma.
Human papillomavirus (HPV)-driven cutaneous squamous cell carcinoma (cSCC) is the most common cancer in immunosuppressed patients. Despite indications suggesting that HPV promotes genomic instability during cSCC development, the molecular pathways underpinning HPV-driven cSCC development remain unknown. We compared the transcriptome of HPV-driven mouse cSCC with normal skin and observed higher amounts of transcripts for Porcupine and WNT ligands in cSCC, suggesting a role for WNT signaling in cSCC progression. We confirmed increased Porcupine expression in human cSCC samples. Blocking the secretion of WNT ligands by the Porcupine inhibitor LGK974 significantly diminished initiation and progression of HPV-driven cSCC. Administration of LGK974 to mice with established cSCC resulted in differentiation of cancer cells and significant reduction of the cancer stem cell compartment. Thus, WNT/β-catenin signaling is essential for HPV-driven cSCC initiation and progression as well as for maintaining the cancer stem cell niche. Interference with WNT secretion may thus represent a promising approach for therapeutic intervention.


Bifemelane in the treatment of central retinal artery or vein obstruction.
Patients with retinal artery or vein obstruction received 150 mg of bifemelane hydrochloride daily for 1 to 11 months and other treatment (urokinase, steroid hormone, hyperbaric oxygen, or laser photocoagulation) or the latter without bifemelane. After treatment with bifemelane, improvements in visual acuity were seen in 12 of 17 eyes with central retinal artery obstruction, in three of four eyes with central retinal vein obstruction, and in five of nine eyes with branch retinal vein obstruction. Among the patients not receiving bifemelane, improvements were seen in two of six patients with central retinal artery disease, in 14 of 27 eyes with central retinal vein obstruction, and in 13 of 28 eyes with branch retinal vein obstruction. It is concluded that bifemelane can be used to improve visual acuity in many patients with central retinal artery obstruction.


Clinical Effectiveness and Safety Outcomes of Endovascular Treatment for Acute Anterior Circulation Ischemic Stroke in China.
This study was aimed at investigating the outcomes and predictors for the poor functional outcome after endovascular treatment (EVT) in a large, mostly Asian population. Between January 2014 and June 2016, acute stroke patients with anterior circulation occlusion and EVT were retrospectively enrolled from 21 stroke centers in China. The main outcomes were modified Rankin Scale (0-2 as functional independence, 3-6 as poor) at 90 days, symptomatic intracranial hemorrhage (sICH) at 72 h, and death at 90 days. Logistic regression was used to identify predictors for poor functional outcome at 90 days. Of the 698 patients, 304 (43.6%) patients had functional independence at 90 days. The sICH rate was 15.5% (108/698) and mortality rate at 90 days was 25.4% (177/698). Age (OR 1.04, 95% CI 1.02-1.07), National Institutes of Health Stroke Scale score at admission (11-20 vs. ≤10, OR 2.38, 95% CI 1.23-4.59; ≥21 vs. ≤10, OR 3.66, 95% CI 1.72-7.80), baseline glucose level (OR 1.09, 95% CI 1.01-1.18), onset to groin puncture >6 h (OR 1.88, 95% CI 1.06-3.31), sICH (OR 15.49, 95% CI 5.16-46.43), and pneumonia (OR 3.15, 95% CI 1.86-5.32) were independent predictors of poor functional outcomes, while good recanalization (OR 0.26, 95% CI 0.13-0.54), preoperative Alberta Stroke Program Early CT Score 8-10 (OR 0.48, 95% CI 0.28-0.83), and good collateral flow (OR 0.50, 95% CI 0.32-0.79) were protective factors. This study provides evidence in real world to support the performance of EVT in acute anterior circulation stroke patients in Chinese population. Patients with small infarct core, successful recanalization, good collateral status, and short treatment delay without sICH or pneumonia may benefit from EVT.


An eligible biological allograft patch in tension-free herniorrhaphy of swine.
Current patches made from macromolecular compounds or composix for tension-free herniorrhaphy are still unsatisfactory in biocompatibility. The ideal patch should be a biological patch with good biocompatibility. Herein allograft patches modified by tissue engineering were used in tension-free herniorrhaphy of swines. Tough membrane tissues from swine were modified with patented tissue engineering techniques to develop allograft patches for tension-free herniorrhaphy. Histological, and physical tests of the allograft patch were performed subsequently, which revealed that the allograft patch was sufficient and satisfactory for tension-free herniorrhaphy. The allograft patches were next used in tension-free herniorrhaphy of abdominal external hernia models of swines and compared to polypropylene patches. Serous CD4+, CD8+ T cells, interleukin-1β (IL-1β), and tumor necrosis factor α (TNF-α) were determined preoperatively and postoperatively. Local pathological changes were recorded postoperatively in swines. In vivo application of the allograft patches revealed that there were no significant serous cellular immune responses in swines, and inflammation induced by allograft patches was significantly lower compared to polypropylene patches, the allograft patches gradually degenerated and new collagen fibers appeared. Abdominal external hernias were cured with allograft patches and without relapse. The modified allograft patch with satisfactory biocompatibility was eligible and sufficient in tension-free herniorrhaphy of swine. Clinical trials should be performed for further evaluation of the allograft patch.


Revisiting the white blood cell count: immature granulocytes count as a diagnostic marker to discriminate between SIRS and sepsis--a prospective, observational study.
Sepsis is a serious disease condition and a major cause of intensive care unit (ICU) admission. Its diagnosis in critically ill patients is complicated. To diagnose an infection rapidly, and to accurately differentiate systemic inflammatory response syndrome (SIRS) from sepsis, is challenging yet early diagnosis is vital for early induction of an appropriate therapy. The aim of this study was to evaluate whether the immature granulocyte (IG) count is a useful early diagnostic marker of sepsis compared to other markers. Therefore, a total of 70 consecutive surgical intensive care patients were assessed. IGs were measured from whole blood samples using an automated analyzer. C-reactive protein (CRP), lipopolysaccharide binding protein (LBP) and interleukin-6 (IL-6) concentrations were also determined. The observation period was a maximum of 21 days and ended with the patients' discharge from ICU or death. Receiver operating characteristic (ROC) analyses were conducted and area under the curve (AUC) was calculated to determine sensitivities and specificities for the parameters. We found that the IG count significantly discriminates between infected and non-infected patients (P < 0.0001) with a sensitivity of 89.2% and a specificity of 76.4%, particularly within the first 48 hours after SIRS onset. Regarding the discriminative power for infection, the IG count was more indicative than other clinical parameters such as CRP, LBP and IL-6, which had a sensitivity of less than 68%. Additionally, the highest diagnostic odds ratio (DOR) with 26.7 was calculated for the IG count within the first 48 hours. During the course of the disease ROC curve analyses showed a superior positive predictive value of the IG count compared to the other measured parameters during the first five days following the fulfillment of SIRS criteria. However, the number of IGs was not correlated with ICU mortality. The total number of IG in peripheral blood from ICU patients is a good marker to discriminate infected and non-infected patients very early during SIRS. However, the IG count is not suitable as a prognostic marker for mortality. Routine and serial measurement of IGs may provide new possibilities for rapid screening of SIRS patients on ICU with suspected infections.


Glucagon-like peptide I receptors in the subfornical organ and the area postrema are accessible to circulating glucagon-like peptide I.
The intestinal incretin hormone glucagon-like peptide I (GLP-I) inhibits gastric motility and secretion in normal, but not in vagotomized subjects, pointing to a centrally mediated effect. Therefore, our aim was to study the availability of rat brain GLP-I receptors to peripherally injected 125I-labeled GLP-I. The specificity of the binding was tested by co-injection of excess amounts of unlabeled GLP-I. Using light microscopical autoradiography of rat brain sections, we found specific 125I-GLP-I binding exclusively in the subfornical organ and the area postrema. This binding was abolished when an excess amount of unlabeled GLP-I was co-injected with the labeled GLP-I. We conclude that cells in the subfornical organ and the area postrema could be responsive to blood-borne GLP-I. The observed binding of peripherally administered GLP-I to the subfornical organ and the area postrema, which both have close neuroanatomical connections with hypothalamic areas involved in water and appetite homeostasis, is consistent with the potential roles of circulating GLP-I in the central regulation of appetite and autonomic functions.


Formation of toroids by self-assembly of an α-α corner mimetic: supramolecular cyclization.
An α-α corner mimetic self-assembles to form a rod-like supramolecular structure which bends and closes end-to-end like a cyclization reaction to form uniform toroids. Each peptide fragment containing l-leucine, α-aminoisobutyric acid (Aib) and l-tyrosine forms rigid 310 helical structures stabilized by multiple intramolecular N-HO hydrogen bonds. Two 310 helices are connected by the spacer 3-aminomethyl-benzylamine and maintain an angular distance of 120° and therefore mimic the α-α corner motif of a protein super secondary structure. The individual α-α corner subunits are themselves regularly interlinked through multiple water mediated intermolecular hydrogen-bonding interactions to form the rod-like supramolecular structure and toroids. The formation of the supramolecular structure has been proven with X-ray crystallography and other spectroscopic techniques. The cyclization of the supramolecular structure and toroid formation were studied by optical microscope, AFM and FE-SEM experiments. Despite other assignments such as exfoliation of graphene from graphite, the compound exhibits significant memory to finally produce the toroids.


Compound-specific stable isotope analysis of organic contaminants in natural environments: a critical review of the state of the art, prospects, and future challenges.
Compound-specific stable isotope analysis (CSIA) using gas chromatography-isotope ratio mass spectrometry (GC/IRMS) has developed into a mature analytical method in many application areas over the last decade. This is in particular true for carbon isotope analysis, whereas measurements of the other elements amenable to CSIA (hydrogen, nitrogen, oxygen) are much less routine. In environmental sciences, successful applications to date include (i) the allocation of contaminant sources on a local, regional, and global scale, (ii) the identification and quantification of (bio)transformation reactions on scales ranging from batch experiments to contaminated field sites, and (iii) the characterization of elementary reaction mechanisms that govern product formation. These three application areas are discussed in detail. The investigated spectrum of compounds comprises mainly n-alkanes, monoaromatics such as benzene and toluene, methyl tert-butyl ether (MTBE), polycyclic aromatic hydrocarbons (PAHs), and chlorinated hydrocarbons such as tetrachloromethane, trichloroethylene, and polychlorinated biphenyls (PCBs). Future research directions are primarily set by the state of the art in analytical instrumentation and method development. Approaches to utilize HPLC separation in CSIA, the enhancement of sensitivity of CSIA to allow field investigations in the microg L(-1) range, and the development of methods for CSIA of other elements are reviewed. Furthermore, an alternative scheme to evaluate isotope data is outlined that would enable estimates of position-specific kinetic isotope effects and, thus, allow one to extract mechanistic chemical and biochemical information.


Active compensation of extrinsic polarization errors using adaptive optics.
We present a scheme for active compensation of complex extrinsic polarization perturbations introduced into an optical system. Imaging polarimeter is used to measure the polarization state across a beam profile and a liquid crystal spatial light modulator controls the polarization of the input beam. A sequence of measurements permits determination of the birefringence properties of a perturbing specimen. The necessary correction is calculated and fed back to the polarization modulator to compensate for the polarization perturbation. The system capabilities are demonstrated on a range of birefringent specimens.


Surveillance of artemether-lumefantrine associated Plasmodium falciparum multidrug resistance protein-1 gene polymorphisms in Tanzania.
Resistance to anti-malarials is a major public health problem worldwide. After deployment of artemisinin-based combination therapy (ACT) there have been reports of reduced sensitivity to ACT by malaria parasites in South-East Asia. In Tanzania, artemether-lumefantrine (ALu) is the recommended first-line drug in treatment of uncomplicated malaria. This study surveyed the distribution of the Plasmodium falciparum multidrug resistance protein-1 single nucleotide polymorphisms (SNPs) associated with increased parasite tolerance to ALu, in Tanzania. A total of 687 Plasmodium falciparum positive dried blood spots on filter paper and rapid diagnostic test strips collected by finger pricks from patients attending health facilities in six regions of Tanzania mainland between June 2010 and August 2011 were used. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to detect Pfmdr1 SNPs N86Y, Y184F and D1246Y. There were variations in the distribution of Pfmdr1 polymorphisms among regions. Tanga region had exceptionally high prevalence of mutant alleles, while Mbeya had the highest prevalence of wild type alleles. The haplotype YFY was exclusively most prevalent in Tanga (29.6%) whereas the NYD haplotype was the most prevalent in all other regions. Excluding Tanga and Mbeya, four, most common Pfmdr1 haplotypes did not vary between the remaining four regions (χ² = 2.3, p = 0.512). The NFD haplotype was the second most prevalent haplotype in all regions, ranging from 17% - 26%. This is the first country-wide survey on Pfmdr1 mutations associated with ACT resistance. Distribution of individual Pfmdr1 mutations at codons 86, 184 and 1246 varies throughout Tanzanian regions. There is a general homogeneity in distribution of common Pfmdr1 haplotypes reflecting strict implementation of ALu policy in Tanzania with overall prevalence of NFD haplotype ranging from 17 to 26% among other haplotypes. With continuation of ALu as first-line drug this haplotype is expected to keep rising, thus there is need for continued pharmacovigilance studies to monitor any delayed parasite clearance by the drug.


Intercomparison of oil spill prediction models for accidental blowout scenarios with and without subsea chemical dispersant injection.
We compare oil spill model predictions for a prototype subsea blowout with and without subsea injection of chemical dispersants in deep and shallow water, for high and low gas-oil ratio, and in weak to strong crossflows. Model results are compared for initial oil droplet size distribution, the nearfield plume, and the farfield Lagrangian particle tracking stage of hydrocarbon transport. For the conditions tested (a blowout with oil flow rate of 20,000 bbl/d, about 1/3 of the Deepwater Horizon), the models predict the volume median droplet diameter at the source to range from 0.3 to 6mm without dispersant and 0.01 to 0.8 mm with dispersant. This reduced droplet size owing to reduced interfacial tension results in a one to two order of magnitude increase in the downstream displacement of the initial oil surfacing zone and may lead to a significant fraction of the spilled oil not reaching the sea surface.


Trigonella foenum-graecum (Fenugreek) protects against selenite-induced oxidative stress in experimental cataractogenesis.
Cataract is the opacification in eye lens and leads to 50% of blindness worldwide. The present study was undertaken to evaluate the anticataract potential of Trigonella foenum-graecum Linn seeds (fenugreek) in selenite-induced in vitro and in vivo cataract. In vitro enucleated rat lenses were maintained in organ culture containing Dulbecco's modified Eagles medium (DMEM) alone or in addition with 100 microM selenite and served as the normal and control groups, respectively. For the test group, the medium was supplemented with selenite and T. foenum-graecum aqueous extract. The lenses were incubated for 24 h at 37 degrees C. After incubation, the lenses were processed for the estimation of reduced glutathione (GSH), lipid peroxidation product (malondialdehyde), and the antioxidant enzymes. In vivo selenite cataract was induced in 9-day-old rats by subcutaneous injection of sodium selenite (25 micromol/kg body weight). Animals in the test group were injected with different doses of aqueous extract of T. foenum-graecum 4 h before the selenite challenge. A fall in GSH and a rise in malondialdehyde levels were observed in control as compared to normal lenses. T. foenum-graecum significantly (P < 0.01) restored glutathione and decreased malondialdehyde levels. A significant restoration in the activities of antioxidant enzymes such as superoxide dismutase (P < 0.01), catalase, (P < 0.01), glutathione peroxidase (P < 0.01), and glutathione-S-transferase (P < 0.01) was observed in the T. foenum-graecum supplemented group as compared to control. In vivo, none of the eyes was found with nuclear cataract in treated group as opposed to 72.5% in the control group. T. foenum-graecum protects against experimental cataract by virtue of its antioxidant properties. Further studies are warranted to explore its role in human cataract.


LKB1 modulates lung cancer differentiation and metastasis.
Germline mutation in serine/threonine kinase 11 (STK11, also called LKB1) results in Peutz-Jeghers syndrome, characterized by intestinal hamartomas and increased incidence of epithelial cancers. Although uncommon in most sporadic cancers, inactivating somatic mutations of LKB1 have been reported in primary human lung adenocarcinomas and derivative cell lines. Here we used a somatically activatable mutant Kras-driven model of mouse lung cancer to compare the role of Lkb1 to other tumour suppressors in lung cancer. Although Kras mutation cooperated with loss of p53 or Ink4a/Arf (also known as Cdkn2a) in this system, the strongest cooperation was seen with homozygous inactivation of Lkb1. Lkb1-deficient tumours demonstrated shorter latency, an expanded histological spectrum (adeno-, squamous and large-cell carcinoma) and more frequent metastasis compared to tumours lacking p53 or Ink4a/Arf. Pulmonary tumorigenesis was also accelerated by hemizygous inactivation of Lkb1. Consistent with these findings, inactivation of LKB1 was found in 34% and 19% of 144 analysed human lung adenocarcinomas and squamous cell carcinomas, respectively. Expression profiling in human lung cancer cell lines and mouse lung tumours identified a variety of metastasis-promoting genes, such as NEDD9, VEGFC and CD24, as targets of LKB1 repression in lung cancer. These studies establish LKB1 as a critical barrier to pulmonary tumorigenesis, controlling initiation, differentiation and metastasis.


GAML: genome assembly by maximum likelihood.
Resolution of repeats and scaffolding of shorter contigs are critical parts of genome assembly. Modern assemblers usually perform such steps by heuristics, often tailored to a particular technology for producing paired or long reads. We propose a new framework that allows systematic combination of diverse sequencing datasets into a single assembly. We achieve this by searching for an assembly with the maximum likelihood in a probabilistic model capturing error rate, insert lengths, and other characteristics of the sequencing technology used to produce each dataset. We have implemented a prototype genome assembler GAML that can use any combination of insert sizes with Illumina or 454 reads, as well as PacBio reads. Our experiments show that we can assemble short genomes with N50 sizes and error rates comparable to ALLPATHS-LG or Cerulean. While ALLPATHS-LG and Cerulean require each a specific combination of datasets, GAML works on any combination. We have introduced a new probabilistic approach to genome assembly and demonstrated that this approach can lead to superior results when used to combine diverse set of datasets from different sequencing technologies. Data and software is available at http://compbio.fmph.uniba.sk/gaml.


Correlation of DNA adducts in blood mononuclear cells with tobacco carcinogen-induced damage in human lung.
The formation of carcinogen-DNA adducts within the respiratory epithelium is thought to be a critical factor in the induction of lung cancer from tobacco smoke. A reliable surrogate measure of carcinogen damage to the lung would be of great value in molecular epidemiological studies of cancer risk. The validity of measurements of DNA adducts formed from hydrophobic aromatic hydrocarbons in peripheral blood mononuclear cells (MNCs) was investigated by comparing the levels of aromatic DNA adducts detected in lung tissue from 31 lung cancer patients with those detected in MNCs from the same individuals using the 32P-postlabeling assay. The associations of smoking history and intake of dietary antioxidants with adduct levels also were assessed. Tissue-specific, as well as common DNA adducts were detected in lung and blood; total MNC adduct levels were highly correlated with total lung adducts. After smoking cessation, adduct levels appeared to decay in both tissues at similar rates. Multivariate analyses (Poisson regression modeling) indicated that dietary antioxidant intake (carotenoids, vitamin A, and retinol) modified the levels of aromatic DNA adducts in both the lungs and blood. Of all models tested, the optimal one for predicting lung adduct levels included the measure of blood MNC adduct levels only. Therefore, blood MNCs are a valid surrogate tissue for estimating the burden of DNA adducts in respiratory tissue in molecular epidemiological studies.


Influence of protein and RNA synthesis inhibitors on methylcholanthrene-mediated induction of aryl hydrocarbon hydroxylase activity in intestinal mucosa.
The sensitivity of the 3-methylcholanthrene (MC)-mediated induction of aryl hydrocarbon hydroxylase (AHH) activity to pretreatment with inhibitors of protein and RNA synthesis was studied. Cordycepin significantly inhibited the induction at 12 h but not at 3 h post-treatment with MC. Both cycloheximide and actinomycin D significantly inhibited the MC-mediated induction at 3 and 12 h after MC treatment. An MC-mediated stimulation of 3H-leucine incorporation into microsomal protein in vivo was observed at 12 h following MC treatment. A cell-free protein synthesis system employing intestinal ribosomes was developed. The rate of protein synthesis (3H-leucine incorporation/mg rRNA/3 min) and the number of active ribosomes (3H-peptidyl-puromycin formed/mg rRNA) were determined. No effect of in vivo MC treatment was observed in subsequently isolated intestinal ribosomes at either 3, 9 or 18 h post-treatment. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed on intestinal microsomes prepared from control, corn-oil- and MC-treated rats 18 h after treatment. The microsomal proteins depicted only slight quantitative changes in microsomal proteins following MC treatment, and no unique protein peaks. These results may explain the lack of detectable stimulation of the activity of isolated ribosomes. In addition, the methods used may lack sufficient sensitivity to detect the small net change in protein necessary to account for the increase in AHH activity seen after MC induction.


Waves, bumps, and patterns in neural field theories.
Neural field models of firing rate activity have had a major impact in helping to develop an understanding of the dynamics seen in brain slice preparations. These models typically take the form of integro-differential equations. Their non-local nature has led to the development of a set of analytical and numerical tools for the study of waves, bumps and patterns, based around natural extensions of those used for local differential equation models. In this paper we present a review of such techniques and show how recent advances have opened the way for future studies of neural fields in both one and two dimensions that can incorporate realistic forms of axo-dendritic interactions and the slow intrinsic currents that underlie bursting behaviour in single neurons.


Male-by-female interactions influence fertilization success and mediate the benefits of polyandry in the sea urchin Heliocidaris erythrogramma.
Numerous studies have reported that females benefit from mating with multiple males (polyandry) by minimizing the probability of fertilization by genetically incompatible sperm. Few, however, have directly attributed variation in female reproductive success to the fertilizing capacity of sperm. In this study we report on two experiments that investigated the benefits of polyandry and the interacting effects of males and females at fertilization in the free-spawning Australian sea urchin Heliocidaris erythrogramma. In the first experiment we used a paired (split clutch) experimental design and compared fertilization rates within female egg clutches under polyandry (eggs exposed to the sperm from two males simultaneously) and monandry (eggs from the same female exposed to sperm from each of the same two males separately). Our analysis revealed a significant fertilization benefit of polyandry and strong interacting effects of males and females at fertilization. Further analysis of these data strongly suggested that the higher rates of fertilization in the polyandry treatment were due to an overrepresentation of fertilizations due to the most compatible male. To further explore the interacting effects of males and females at fertilization we performed a second factorial experiment in which four males were crossed with two females (in all eight combinations). In addition to confirming that fertilization success is influenced by male x female interactions, this latter experiment revealed that both sexes contributed significant variance to the observed patterns of fertilization. Taken together, these findings highlight the importance of male x female interactions at fertilization and suggest that polyandry will enable females to reduce the cost of fertilization by incompatible gametes.


Are clinicians being prepared to care for abused women? A survey of health professional education in Ontario, Canada.
The current project undertook a province-wide survey and environmental scan of educational opportunities available to future health care providers on the topic of intimate partner violence (IPV) against women. A team of experts identified university and college programs in Ontario, Canada as potential providers of IPV education to students in health care professions at the undergraduate and post-graduate levels. A telephone survey with contacts representing these programs was conducted between October 2005 and March 2006. The survey asked whether IPV-specific education was provided to learners, and if so, how and by whom. In total, 222 eligible programs in dentistry, medicine, nursing and other allied health professions were surveyed, and 95% (212/222) of programs responded. Of these, 57% reported offering some form of IPV-specific education, with undergraduate nursing (83%) and allied health (82%) programs having the highest rates. Fewer than half of undergraduate medical (43%) and dentistry (46%) programs offered IPV content. Postgraduate programs ranged from no IPV content provision (dentistry) to 41% offering content (nursing). Significant variability exists across program areas regarding the methods for IPV education, its delivery and evaluation. The results of this project highlight that expectations for an active and consistent response by health care professionals to women experiencing the effects of violence may not match the realities of professional preparation.


Multi-scale marine biodiversity patterns inferred efficiently from habitat image processing.
Cost-effective proxies of biodiversity and species abundance, applicable across a range of spatial scales, are needed for setting conservation priorities and planning action. We outline a rapid, efficient, and low-cost measure of spectral signal from digital habitat images that, being an effective proxy for habitat complexity, correlates with species diversity and requires little image processing or interpretation. We validated this method for coral reefs of the Great Barrier Reef (GBR), Australia, across a range of spatial scales (1 m to 10 km), using digital photographs of benthic communities at the transect scale and high-resolution Landsat satellite images at the reef scale. We calculated an index of image-derived spatial heterogeneity, the mean information gain (MIG), for each scale and related it to univariate (species richness and total abundance summed across species) and multivariate (species abundance matrix) measures of fish community structure, using two techniques that account for the hierarchical structure of the data: hierarchical (mixed-effect) linear models and distance-based partial redundancy analysis. Over the length and breadth of the GBR, MIG alone explained up to 29% of deviance in fish species richness, 33% in total fish abundance, and 25% in fish community structure at multiple scales, thus demonstrating the possibility of easily and rapidly exploiting spatial information contained in digital images to complement existing methods for inferring diversity and abundance patterns among fish communities. Thus, the spectral signal of unprocessed remotely sensed images provides an efficient and low-cost way to optimize the design of surveys used in conservation planning. In data-sparse situations, this simple approach also offers a viable method for rapid assessment of potential local biodiversity, particularly where there is little local capacity in terms of skills or resources for mounting in-depth biodiversity surveys.


Focused ultrasound delivers targeted immune cells to metastatic brain tumors.
Natural killer (NK) cells are cytotoxic lymphocytes involved in innate immunity. NK-92, a human NK cell line, may be targeted to tumor-associated antigens in solid malignancies where it exhibits antitumor efficacy, but its clinical utility for treating brain tumors is limited by an inability to cross the blood-brain barrier (BBB). We investigated the potential for focused ultrasound (FUS) to deliver targeted NK-92 cells to the brain using a model of metastatic breast cancer. HER2-expressing human breast tumor cells were implanted into the brain of nude rats. The NK-92-scFv(FRP5)-zeta cell line expressing a chimeric HER2 antigen receptor was transfected with superparamagnetic iron oxide nanoparticles before intravenous injection, before and following BBB disruption using focused ultrasound (551.5 kHz focused transducer, 0.33 MPa average peak rarefaction pressure) in the presence of a microbubble contrast agent. Baseline and posttreatment 1.5T and 7T MR imaging was done, and histology used to identify NK-92 cells post-mortem. Contrast-enhanced MRI showed reproducible and consistent BBB disruption. 7T MR images obtained at 16 hours posttreatment revealed a significant reduction in signal indicating the presence of iron-loaded NK-92 cells at the tumor site. The average ratio of NK-92 to tumor cells was 1:100 when NK cells were present in the vasculature at the time of sonication, versus 2:1,000 and 1:1,000 when delivered after sonication and without BBB disruption, respectively. Our results offer a preclinical proof-of-concept that FUS can improve the targeting of immune cell therapy of brain metastases.


Predictive markers of asthma exacerbation during stepwise dose reduction of inhaled corticosteroids.
To determine predictors for failed reduction of inhaled corticosteroids (ICS), in 50 subjects with well-controlled asthma (age 43.7 [18-69]; 22 males) taking a median dose of 1,000 microg ICS/d (100-3,600 microg/d), ICS were halved every 8 wk. Airway hyperresponsiveness (AHR) to a bronchial provocation test (BPT) with histamine was measured at baseline. AHR to BPT with mannitol, spirometry, exhaled nitric oxide (eNO), and, in 31 subjects, sputum inflammatory cells were measured at baseline and at monthly intervals. Thirty-nine subjects suffered an asthma exacerbation. Seven subjects were successfully weaned off ICS. Using a Kaplan- Meier survival analysis, the significant predictors of a failure of ICS reduction were being hyperresponsive to both histamine and mannitol at baseline (p = 0.039), and being hyperresponsive to mannitol during the dose-reduction phase of the study (p = 0.02). Subjects older than 40 yr of age tended to be at greater risk of ICS reduction failure (p = 0.059). Response to mannitol and percentage sputum eosinophils were significantly greater before a failed ICS reduction than before the last successful ICS reduction, whereas there were no significant differences in symptoms, spirometry, or eNO. These findings suggest that documentation of patient's AHR or sputum eosinophils may be useful in guiding the reduction of ICS doses.


Secondary Prevention of Nonvalvular Atrial Fibrillation: A Retrospective Cohort Study.
Secondary prevention of atrial fibrillation (AF) could be carried out by means of antiarrhythmic drugs; however this strategy has not received any endorsement because these drugs are burdened by a high risk of proarrhythmic events (flecainide, sotalol) or extracardiac effects (amiodarone). In our retrospective cohort study we have compared amiodarone 200 mg per day with the strategy implying the renunciation of any specific drug as well as with the approach using oral anticoagulant (rivaroxaban) or a combined approach including amiodarone plus rivaroxaban. A total of 255 patients with a history of AF (paroxysmal, persistent or long-lasting persistent) successfully treated with achievement of sinus rhythm have been gathered. Amiodarone has been the most effective option for AF secondary prevention, with regard to the recurrences of AF as well as rehospitalizations: P (Kruskal-Wallis test) < 0.05 for both, over a median follow-up of 24 months. Patients kept free from any specific drug therapy have been shown to experience more numerous AF relapses and related rehospitalizations. On the contrary, the amiodarone use has been associated with a decreased risk of AF recurrences and hospital admissions. Thus, amiodarone might be an efficacious tool for realizing a successful long-term AF secondary prevention.


Is reactive gliosis a property of a distinct subpopulation of astrocytes?
We have shown previously that the A2B5 monoclonal antibody distinguishes two types of glial fibrillary acidic protein-containing astrocytes in semithin frozen sections of adult rat optic nerve: A2B5- (type-1) astrocytes are found mainly at the periphery of the nerve, where they form the glial limiting membrane, while A2B5+ (type-2) astrocytes are found mainly in the interior of the nerve and constitute more than 65% of the astrocytes in the adult optic nerve. In the present study we show that although most astrocytes in semithin frozen sections of adult rat corpus callosum and optic nerve are A2B5+, the great majority of reactive astrocytes in similar sections of corpus callosum examined 20 weeks after a stab lesion, and in optic nerve examined 20 weeks after adult transection, are A2B5-. Although both A2B5+ and A2B5- astrocytes are stimulated to synthesize DNA in the first week after transection, adult optic nerves examined 20 weeks after transection contain only half as many astrocytes as do normal optic nerves: While A2B5+ astrocytes are reduced almost 10-fold, A2B5- astrocytes are increased by about 25%. We consider the simplest interpretation of these findings to be that type-1 astrocytes are largely responsible for forming glial scars in adult white matter following either a stab lesion or Wallerian degeneration and that in transected optic nerves, most type-2 astrocytes eventually die, possibly because they depend on axons for their long-term survival.


Nutrient reference value: non-communicable disease endpoints--a conference report.
Nutrition is complex-and seemingly getting more complicated. Most consumers are familiar with "essential nutrients," e.g., vitamins and minerals, and more recently protein and important amino acids. These essential nutrients have nutrient reference values, referred to as dietary reference intakes (DRIs) developed by consensus committees of scientific experts convened by the Institute of Medicine of the National Academy of Sciences, Engineering, and Medicine and carried out by the Food and Nutrition Board. The DRIs comprise a set of four nutrient-based reverence values, the estimated average requirements, the recommended dietary allowances (RDAs), the adequate intakes and the tolerable upper intake levels for micronutrient intakes and an acceptable macronutrient distribution range for macronutrient intakes. From the RDA, the US Food and Drug Administration (FDA) derives a labeling value called the daily value (DV), which appears on the nutrition label of all foods for sale in the US. The DRI reports do not make recommendations about whether the DV labeling values can be set only for what have been defined to date as "essential nutrients." For example, the FDA set a labeling value for "dietary fiber" without having the DV. Nutrient reference values-requirements are set by Codex Alimentarius for essential nutrients, and regulatory bodies in many countries use these Codex values in setting national policy for recommended dietary intakes. However, the focus of this conference is not on essential nutrients, but on the "nonessential nutrients," also termed dietary bioactive components. They can be defined as "Constituents in foods or dietary supplements, other than those needed to meet basic human nutritional needs, which are responsible for changes in health status (Office of Disease Prevention and Health Promotion, Office of Public Health and Science, Department of Health and Human Services in Fed Regist 69:55821-55822, 2004)." Substantial and often persuasive scientific evidence does exist to confirm a relationship between the intake of a specific bioactive constituent and enhanced health conditions or reduced risk of a chronic disease. Further, research on the putative mechanisms of action of various classes of bioactives is supported by national and pan-national government agencies, and academic institutions, as well as functional food and dietary supplement manufacturers. Consumers are becoming educated and are seeking to purchase products containing bioactives, yet there is no evaluative process in place to let the public know how strong the science is behind the benefits or the quantitative amounts needed to achieve these beneficial health effects or to avoid exceeding the upper level (UL). When one lacks an essential nutrient, overt deficiency with concomitant physiological determents and eventually death are expected. The absence of bioactive substances from the diet results in suboptimal health, e.g., poor cellular and/or physiological function, which is relative and not absolute. Regrettably at this time, there is no DRI process to evaluate bioactives, although a recent workshop convened by the National Institutes of Health (Options for Consideration of Chronic Disease Endpoints for Dietary Reference Intakes (DRIs); March 10-11, 2015; http://health.gov/dietaryguidelines/dri/ ) did explore the process to develop DVs for nutrients, the lack of which result in increased risk of chronic disease (non-communicable disease) endpoints. A final report is expected soon. This conference (CRN-International Scientific Symposium; "Nutrient Reference Value-Non-Communicable Disease (NRV-NCD) Endpoints," 20 November in Kronberg, Germany; http://www.crn-i.ch/2015symposium/ ) explores concepts related to the Codex NRV process, the public health opportunities in setting NRVs for bioactive constituents, and further research and details on the specific class of bioactives, n-3 long-chain polyunsaturated fatty acids (also termed omega-3 fatty acids) and their constituents, specifically docosahexaenoic acid and eicosapentaenoic acid.


Micromechanical String Resonators: Analytical Tool for Thermal Characterization of Polymers.
Resonant microstrings show promise as a new analytical tool for thermal characterization of polymers with only few nanograms of sample. The detection of the glass transition temperature (Tg) of an amorphous poly(d,l-lactide) (PDLLA) and a semicrystalline poly(l-lactide) (PLLA) is investigated. The polymers are spray coated on one side of the resonating microstrings. The resonance frequency and quality factor (Q) are measured simultaneously as a function of temperature. Change in the resonance frequency reflects a change in static tensile stress, which yields information about the Young's modulus of the polymer, and a change in Q reflects the change in damping of the polymer-coated string. The frequency response of the microstring is validated with an analytical model. From the frequency independent tensile stress change, static Tg values of 40.6 and 57.6 °C were measured for PDLLA and PLLA, respectively. The frequency-dependent damping from Q indicates higher Tg values of 62.6 and 88.8 °C for PDLLA and PLLA, respectively, at ∼105 Hz. Resonant microstrings facilitate thermal analysis of nanogram polymer samples measuring the static and a dynamic glass transition temperature simultaneously.


Relation between consumption of sugar-sweetened drinks and childhood obesity: a prospective, observational analysis.
The rising prevalence of obesity in children has been linked in part to the consumption of sugar-sweetened drinks. Our aim was to examine this relation. We enrolled 548 ethnically diverse schoolchildren (age 11.7 years, SD 0.8) from public schools in four Massachusetts communities, and studied them prospectively for 19 months from October, 1995, to May, 1997. We examined the association between baseline and change in consumption of sugar-sweetened drinks (the independent variables), and difference in measures of obesity, with linear and logistic regression analyses adjusted for potentially confounding variables and clustering of results within schools. For each additional serving of sugar-sweetened drink consumed, both body mass index (BMI) (mean 0.24 kg/m2; 95% CI 0.10-0.39; p=0.03) and frequency of obesity (odds ratio 1.60; 95% CI 1.14-2.24; p=0.02) increased after adjustment for anthropometric, demographic, dietary, and lifestyle variables. Baseline consumption of sugar-sweetened drinks was also independently associated with change in BMI (mean 0.18 kg/m2 for each daily serving; 95% CI 0.09-0.27; p=0.02). Consumption of sugar-sweetened drinks is associated with obesity in children.


Globally optimal grouping for symmetric closed boundaries by combining boundary and region information.
Many natural and man-made structures have a boundary that shows a certain level of bilateral symmetry, a property that plays an important role in both human and computer vision. In this paper, we present a new grouping method for detecting closed boundaries with symmetry. We first construct a new type of grouping token in the form of symmetric trapezoids by pairing line segments detected from the image. A closed boundary can then be achieved by connecting some trapezoids with a sequence of gap-filling quadrilaterals. For such a closed boundary, we define a unified grouping cost function in a ratio form: the numerator reflects the boundary information of proximity and symmetry and the denominator reflects the region information of the enclosed area. The introduction of the region-area information in the denominator is able to avoid a bias toward shorter boundaries. We then develop a new graph model to represent the grouping tokens. In this new graph model, the grouping cost function can be encoded by carefully designed edge weights and the desired optimal boundary corresponds to a special cycle with a minimum ratio-form cost. We finally show that such a cycle can be found in polynomial time using a previous graph algorithm. We implement this symmetry-grouping method and test it on a set of synthetic data and real images. The performance is compared to two previous grouping methods that do not consider symmetry in their grouping cost functions.


Survivin protein as predictor of pathologic response in patients with locally advanced cervical cancer treated with chemoradiation followed by radical surgery.
We investigated the correlation of pathologic response and immunohistochemically assessed expression of survivin protein in 71 patients with locally advanced cervical cancer treated with chemoradiation (CT/RT) followed by radical surgery. The prognostic role of survivin expression was also evaluated. Immunohistochemical analysis of survivin expression was carried out using the polyclonal rabbit antisurvivin antibody. Cytoplasmic survivin immunoreaction was observed in 69 (97.2%) of 71 cases and nuclear staining in 7 (9.8%) of 71 women. Median cytoplasmic survivin expression was 160 (range, 0-280), and higher levels were observed in patients with residual disease (≥3 mm) in the cervix (survivin level, 160 versus 120; P = .016) and in women with metastatic lymph nodes (survivin levels, 160 versus 150; P = .032). No differences were documented in the distribution of patients with positive nuclear staining, according to clinicopathological variables. In multivariate analysis, cytoplasmic survivin expression emerged as an independent predictor of residual cervical disease and lymph node status after CT/RT. During a follow-up period of 83 months (range, 8-175 months), recurrences occurred in 24 (33.8%) women, and all patients died of disease. Women with high cytoplasmic survivin experienced shorter disease-free survival compared with patients with low levels (5-year disease-free survival, 80.8% versus 55.3%; P = .033). Only a trend was observed for greater overall survival in patients with high expression (5-year overall survival, 81.0% versus 55.3%; P = .069). No survival differences were documented for nuclear survivin status. The immunohistochemically assessed survivin cytoplasmic levels at diagnosis represent a reliable and easily assessable tool to predict response to CT/RT in patients with locally advanced cervical cancer.


Green leaf volatiles and oxygenated metabolite emission bursts from mesquite branches following light-dark transitions.
Green leaf volatiles (GLVs) are a diverse group of fatty acid-derived compounds emitted by all plants and are involved in a wide variety of developmental and stress-related biological functions. Recently, GLV emission bursts from leaves were reported following light-dark transitions and hypothesized to be related to the stress response while acetaldehyde bursts were hypothesized to be due to the 'pyruvate overflow' mechanism. In this study, branch emissions of GLVs and a group of oxygenated metabolites (acetaldehyde, ethanol, acetic acid, and acetone) derived from the pyruvate dehydrogenase (PDH) bypass pathway were quantified from mesquite plants following light-dark transitions using a coupled GC-MS, PTR-MS, and photosynthesis system. Within the first minute after darkening following a light period, large emission bursts of both C(5) and C(6) GLVs dominated by (Z)-3-hexen-1-yl acetate together with the PDH bypass metabolites are reported for the first time. We found that branches exposed to CO(2)-free air lacked significant GLV and PDH bypass bursts while O(2)-free atmospheres eliminated the GLV burst but stimulated the PDH bypass burst. A positive relationship was observed between photosynthetic activity prior to darkening and the magnitude of the GLV and PDH bursts. Photosynthesis under (13)CO(2) resulted in bursts with extensive labeling of acetaldehyde, ethanol, and the acetate but not the C(6)-alcohol moiety of (Z)-3-hexen-1-yl acetate. Our observations are consistent with (1) the "pyruvate overflow" mechanism with a fast turnover time (<1 h) as part of the PDH bypass pathway, which may contribute to the acetyl-CoA used for the acetate moiety of (Z)-3-hexen-1-yl acetate, and (2) a pool of fatty acids with a slow turnover time (>3 h) responsible for the C(6) alcohol moiety of (Z)-3-hexen-1-yl acetate via the 13-lipoxygenase pathway. We conclude that our non-invasive method may provide a new valuable in vivo tool for studies of acetyl-CoA and fatty acid metabolism in plants at a variety of spatial scales.


Self-Targeting, Immune Transparent Plasma Protein Coated Nanocomplex for Noninvasive Photothermal Anticancer Therapy.
Cancer cells exhibit specific physiological differences compared to normal cells. Most surface membranes of cancer cells are characterized by high expression of given protein receptors, such as albumin, transferrin, and growth factors that are also present in the plasma of patients themselves, but are lacking on the surface of normal cells. These distinct features between cancer and normal cells can serve as a niche for developing specific treatment strategies. Near-infrared (NIR)-light-triggered therapy platforms are an interesting novel avenue for use in clinical nanomedicine. As a photothermal agent, conducting polymer nanoparticles, such as polypyrrole (PPy), of great NIR light photothermal effects and good biocompatibility, show promising applications in cancer treatments through the hyperthermia mechanism. Autologous plasma proteins coated PPy nanoparticles for hyperthermia therapy as a novel core technology platform to treat cancers through secreted protein acid and rich in cysteine targeting are developed here. This approach can provide unique features of specific targeting toward cancer cell surface markers and immune transparency to avoid recognition and attack by defense cells and achieve prolonged circulation half-life. This technology platform unveils new clinical options for treatment of cancer patients, supporting the emergence of innovative clinical products.


Fitting hearing aids to first-time users.
Clinical experience indicates that first-time hearing aid users prefer less gain and lower maximum output levels than experienced users. This hypothesis was tested on 20 subjects being fitted with their first aids. The study was double blinded by using a programmable hearing aid, set to either the standard setting according to the manufacturer's software or to reduced gain and maximum output. Half of the subjects started with one hearing aid and half with the other, changing to the other hearing aid after 3 days trial with each setting. At the end of the study, subjects stated preference in specified situations and overall. No significant differences in APHAB, sound quality, estimated communication ability or perceived loudness scores were seen for the two settings. Nine subjects preferred the standard setting, seven the reduced setting and four were undecided. No correlation could be found between preference and audiological variables.


Nanospectroscopy Captures Nanoscale Compositional Zonation in Barite Solid Solutions.
Scientists have long suspected that compositionally zoned particles can form under far-from equilibrium precipitation conditions, but their inferences have been based on bulk solid and solution measurements. We are the first to directly observe nanoscale trace element compositional zonation in <10 µm-sized particles using X-ray fluorescence nanospectroscopy at the Hard X-ray Nanoprobe (HXN) Beamline at National Synchrotron Light Source II (NSLS-II). Through high-resolution images, compositional zonation was observed in barite (BaSO4) particles precipitated from aqueous solution, in which Sr2+ cations as well as HAsO42- anions were co-precipitated into (Ba,Sr)SO4 or Ba(SO4,HAsO4) solid solutions. Under high salinity conditions (NaCl ≥ 1.0 M), bands contained ~3.5 to ~5 times more trace element compared to the center of the particle formed in early stages of particle growth. Quantitative analysis of Sr and As fractional substitution allowed us to determine that different crystallographic growth directions incorporated trace elements to different extents. These findings provide supporting evidence that barite solid solutions have great potential for trace element incorporation; this has significant implications for environmental and engineered systems that remove hazardous substances from water.


Multigenerational effects of the anticancer drug tamoxifen and its metabolite 4-hydroxy-tamoxifen on Daphnia pulex.
Tamoxifen and its metabolite 4-hydroxy-tamoxifen (4OHTam) are two potent molecules that have anticancer properties on breast cancers. Their medical use is expected to increase with the increasing global cancer rate. After consumption, patients excrete tamoxifen and the 4OHTam metabolite into wastewaters, and tamoxifen has been already detected in wastewaters and natural waters. The concentrations of 4OHTam in waters have never been reported. A single study reported 4OHTam effects on the microcrustacean Daphnia pulex. The effects of tamoxifen and 4OHTam over more than two generations are unknown in aquatic invertebrates. The main goal of this study was to assess the long-term sensitivity of the microcrustacean D. pulex over four generations, based on size, reproduction, viability and the intrinsic rate of natural increase (r). Additional experiments were carried out to observe whether the effects of tamoxifen and 4OHTam were reversible in the next generation after descendants were withdrawn from chemical stress (i.e., recovery experiment), and whether the lowest test concentration of each chemical induced toxic effects when both concentrations were combined (i.e., mixture experiments). Our results showed that tamoxifen and 4OHTam induced the adverse effects at environmentally relevant concentrations. Tamoxifen and 4OHTam impaired size, viability, reproduction and the r in four generations of treated D. pulex, but these effects were not clearly magnified over generations. Tamoxifen was more potent than 4OHTam on D. pulex. When used in a mixture, the combination of tamoxifen and 4OHTam induced effects in offspring, whereas no effects were observed when these chemicals were tested individually. In the recovery experiment, the reproduction and size were reduced in offspring withdrawn from chemical exposures. Our results suggested that tamoxifen and its metabolite may be a relevant pharmaceutical to consider in risk assessment.


The effect of intravenous magnesium sulfate infusion on reduction of pain after abdominal hysterectomy under general anesthesia: a double-blind, randomized clinical trial.
Post-surgical pain is a physiological response to tissue trauma that produces unpleasant physiological effects with manifestations on various organic systems. According to the effect of magnesium sulfate on the N-methyl-d-aspartate (NMDA) receptor, this study examined the effect of magnesium sulfate on the reduction of pain and the mean amount of narcotics consumed by patients after abdominal hysterectomies. This double-blind clinical trial study was performed on 60 patients who had undergone abdominal hysterectomies in Shahid Sadoughi Hospital in Yazd, Iran, from 2013 to 2015. The patients were divided randomly into two groups of 30 members each. All of the patients received 2 mg of Midazolam and 2 mcg/kg of Fentanyl as the induction of anesthesia with propofol (2-2.5 mg/kg) and Atracurium 0.5 mg/kg was conducted. All of the patients received 5 mg of intravenous morphine 30 min after induction of anesthesia. Afterwards, the study group received 50 mg/kg of magnesium sulfate in 500 cm(3) of Ringer's serum during the 20 minutes, and 500 cm(3) of Ringer's serum was administered to the members of the placebo group. Visual analogue scale VAS scores were evaluated to reach the minimum difference of 0.8 in mean pain score. The results of this study indicated that the mean pain scores immediately after surgery and at 1, 2, 6, and 12 hr after surgery were lower in the study group than in the placebo group. The mean value of narcotic consumption at all measured time points was higher in the placebo group. No significant differences were found between two groups concerning drug complications. The results of this study indicated that the intravenous injection of magnesium sulfate can reduce pain, reduce morphine consumption, and reduce the side effects of morphine in patients after surgery. This study was funded by Shahid Sadoughi University of Medical Sciences, Yazd, Iran. The trial was registered at the Thai Clinical Trials Registry (http://www.clinicaltrials.in.th) with the registration ID: TCTR20160308001.


Obstruction and perforation in colorectal adenocarcinoma: an analysis of prognosis and current trends.
In adenocarcinoma of the colon and rectum, obstruction and perforation may occur either alone or together at the site of the neoplasm or proximally. Both events carry a poor prognosis. This retrospective study sought to determine whether a correlation exists between perforation and obstruction, and between these conditions and different clinicopathological factors in colorectal adenocarcinoma. The medical records of 1950 patients with colorectal adenocarcinoma treated in our hospital during a 7-year period were retrospectively analyzed. One hundred patients (5%) were excluded from this study because of a loss of follow-up. Data on clinicopathological factors including age, sex, tumor location, surgical mortality, pathological type, stage, and long-time cancer-free rate were simultaneously analyzed. Patients were grouped as follows: Group 1, complete colonic obstruction without perforation (n = 120). Group 2, complete obstruction with perforation at the site of the cancer (n = 35); Group 3, complete obstruction with perforation proximal to the cancer (n = 13); and Group 4, nonobstructing, nonperforated cancers (n = 1682). When compared with Group 4, Group 1 had a more advanced Dukes' stage, older age, greater incidence of colonic versus rectal cancers, and a poorer cancer-free survival (P < or = .005). Groups 2 and 3 had a greater incidence of colonic versus rectal cancers (P < or = .004), and Group 3 had a greater operative mortality (P < .001). No significant differences were found between Groups 1, 2, and 3. Multivariate analysis revealed that the independent factors favorable to cancer-free survival (> 5-year survival) were female gender (P = .035), well-differentiated pathology (P < .001), uncomplicated cases (P = .004), colon versus rectal location (P < .001), and early stage (P < .001). The perioperative mortality rate for perforated colorectal cancer at the site of the cancer was 9%; for obstructive colorectal cancer, 5%. Perioperative mortality was much greater for perforations of the colon and rectum occurring proximal to the cancer (31%). Survival was worse (P < .001) for patients with obstruction (33%) or perforation proximal to the cancer (33%). The site of perforation did not appear to impact the 5-year survival, although the numbers are relatively small.


Impaired sweating responses to a passive whole body heat stress in individuals with multiple sclerosis.
Multiple sclerosis (MS) is an autoimmune disease that affects the central nervous system (CNS), disrupting autonomic function. The aim of this study was to test the hypothesis that individuals with MS have blunted control of thermoregulatory reflex increases in sweat rate (SR) and cutaneous vasodilation compared with controls during a passive whole body heat stress (WBH). Eighteen individuals with relapsing-remitting MS and 18 healthy controls (Con) participated in the study. Core temperature (Tcore), skin temperature, heart rate, arterial blood pressure (10-min intervals), skin blood flow (laser-Doppler flux, LDF), and SR were continuously measured during normothermic baseline (34°C water perfusing a tube-lined suit) and WBH (increased Tcore 0.8°C via 48°C water perfusing the suit). Following WBH, local heaters were warmed to 42°C, inducing peak cutaneous vasodilation at the site of LDF collection. Cutaneous vascular conductance (CVC) was calculated as the ratio of LDF to mean arterial pressure and expressed as a percentage of peak achieved during local heating. Individuals with MS had attenuated SR responses to WBH (ΔSR from baseline: Con, 0.65 ± 0.27; MS, 0.42 ± 0.17 mg·cm-2·min-1, P = 0.003), whereas Δ%CVC42C from baseline was similar between groups (Con, 42 ± 16%; MS, 38 ± 12%, P = 0.39). SR responses were blunted as a function of Tcore in MS (interaction: group × Tcore, P = 0.03), of which differences were evident at ΔTcore 0.7°C and 0.8°C (P < 0.05). No interaction was observed in Δ%CVC42C Taken together, the findings show MS blunts sweating responses, whereas control of the cutaneous vasculature is preserved, in response to WBH.NEW & NOTEWORTHY This study is the first to assess the reflex control of the thermoregulatory system in individuals living with multiple sclerosis (MS). The novel findings are twofold. First, attenuated increases in sweat rate in subjects with MS compared with healthy controls were observed in response to a moderate increase (0.8°C) in core temperature via passive whole body heat stress. Second, it appears the reflex control of the cutaneous vasculature is preserved in MS.


Taking directions: the role of microtubule-bound nucleation in the self-organization of the plant cortical array.
The highly aligned cortical microtubule array of interphase plant cells is a key regulator of anisotropic cell expansion. Recent computational and analytical work has shown that the non-equilibrium self-organization of this structure can be understood on the basis of experimentally observed collisional interactions between dynamic microtubules attached to the plasma membrane. Most of these approaches assumed that new microtubules are homogeneously and isotropically nucleated on the cortical surface. Experimental evidence, however, shows that nucleation mostly occurs from other microtubules and under specific relative angles. Here, we investigate the impact of directed microtubule-bound nucleations on the alignment process using computer simulations. The results show that microtubule-bound nucleations can increase the degree of alignment achieved, decrease the timescale of the ordering process and widen the regime of dynamic parameters for which the system can self-organize. We establish that the major determinant of this effect is the degree of co-alignment of the nucleations with the parent microtubule. The specific role of sideways branching nucleations appears to allow stronger alignment while maintaining a measure of overall spatial homogeneity. Finally, we investigate the suggestion that observed persistent rotation of microtubule domains can be explained through a handedness bias in microtubule-bound nucleations, showing that this is possible only for an extreme bias and over a limited range of parameters.


BioFlow: a non-invasive, image-based method to measure speed, pressure and forces inside living cells.
Cell motility is governed by a complex molecular machinery that converts physico-chemical cues into whole-cell movement. Understanding the underlying biophysical mechanisms requires the ability to measure physical quantities inside the cell in a simple, reproducible and preferably non-invasive manner. To this end, we developed BioFlow, a computational mechano-imaging method and associated software able to extract intracellular measurements including pressure, forces and velocity everywhere inside freely moving cells in two and three dimensions with high spatial resolution in a non-invasive manner. This is achieved by extracting the motion of intracellular material observed using fluorescence microscopy, while simultaneously inferring the parameters of a given theoretical model of the cell interior. We illustrate the power of BioFlow in the context of amoeboid cell migration, by modelling the intracellular actin bulk flow of the parasite Entamoeba histolytica using fluid dynamics, and report unique experimental measures that complement and extend both theoretical estimations and invasive experimental measures. Thanks to its flexibility, BioFlow is easily adaptable to other theoretical models of the cell, and alleviates the need for complex or invasive experimental conditions, thus constituting a powerful tool-kit for mechano-biology studies. BioFlow is open-source and freely available via the Icy software.


Joint modelling of time-to-event and multivariate longitudinal outcomes: recent developments and issues.
Available methods for the joint modelling of longitudinal and time-to-event outcomes have typically only allowed for a single longitudinal outcome and a solitary event time. In practice, clinical studies are likely to record multiple longitudinal outcomes. Incorporating all sources of data will improve the predictive capability of any model and lead to more informative inferences for the purpose of medical decision-making. We reviewed current methodologies of joint modelling for time-to-event data and multivariate longitudinal data including the distributional and modelling assumptions, the association structures, estimation approaches, software tools for implementation and clinical applications of the methodologies. We found that a large number of different models have recently been proposed. Most considered jointly modelling linear mixed models with proportional hazard models, with correlation between multiple longitudinal outcomes accounted for through multivariate normally distributed random effects. So-called current value and random effects parameterisations are commonly used to link the models. Despite developments, software is still lacking, which has translated into limited uptake by medical researchers. Although, in an era of personalized medicine, the value of multivariate joint modelling has been established, researchers are currently limited in their ability to fit these models routinely. We make a series of recommendations for future research needs.


Animal Trauma Triage Score, Modified Glasgow Coma Scale, age, and weight were associated with outcome in feline bite wounds (1,065 cases): a VetCOT registry study.
To identify associations between admission variables, Animal Trauma Triage (ATT) score, and Modified Glasgow Coma Scale (MGCS) score with need for transfusion or surgical interventions and survival to discharge in cats with bite wounds. 1,065 cats with bite wounds. Records of cats with bite wounds were obtained from the VetCOT registry from April 2017 to June 2021. Variables included point of care laboratory values, signalment, weight, illness severity scores, and surgical intervention. Associations between admission parameters, terciles of MGCS, quantiles of ATT scores, and death or euthanasia were assessed using univariable and multivariable logistic regression analysis. 872 cats (82%) survived to discharge, while 170 (88%) were euthanized and 23(12%) died. In the multivariable model, age, weight, surgical treatment, ATT and MGCS scores were associated with nonsurvival. For every 1 year of age, odds of nonsurvival increased by 7% (P = .003) and for every 1 kg of body weight, odds of nonsurvival decreased by 14% (P = .005). Odds of dying increased with lower MGCS and higher ATT scores (MGCS: 104% [95% CI, 116% to 267%; P < .001]; ATT: 351% [95% CI, 321% to 632%; P < .001). Odds of dying decreased by 84% (P < .001) in cats that underwent surgery versus those that did not. This multicenter study indicated association of higher ATT and lower MGCS with worse outcome. Older age increased the odds of nonsurvival, while each kilogram increase in bodyweight decreased odds of nonsurvival. To our knowledge, this study is the first to describe associations of age and weight with outcome in feline trauma patients.