Force-dependent detachment of kinesin-2 biases track switching at cytoskeletal filament intersections.
Intracellular trafficking of organelles often involves cytoskeletal track switching. Organelles such as melanosomes are transported by multiple motors including kinesin-2, dynein, and myosin-V, which drive switching between microtubules and actin filaments during dispersion and aggregation. Here, we used optical trapping to determine the unitary and ensemble forces of kinesin-2, and to reconstitute cargo switching at cytoskeletal intersections in a minimal system with kinesin-2 and myosin-V motors bound to beads. Single kinesin-2 motors exerted forces up to ∼5 pN, similar to kinesin-1. However, kinesin-2 motors were more likely to detach at submaximal forces, and the duration of force maintenance was short as compared to kinesin-1. In multimotor assays, force increased with kinesin-2 density but was not affected by the presence of myosin-V. In crossed filament assays, switching frequencies of motor-bound beads were dependent on the starting track. At equal average forces, beads tended to switch from microtubules onto overlying actin filaments consistent with the relatively faster detachment of kinesin-2 at near-maximal forces. Thus, in addition to relative force, switching probability at filament intersections is determined by the dynamics of motor-filament interaction, such as the quick detachment of kinesin-2 under load. This may enable fine-tuning of filament switching in the cell.


Thyroid Storm, a Mimicker of Guillain-Barre Syndrome: A Case Report.
A 60-year-old woman with diabetes mellitus presented to the emergency department (ER) with complaints of lower limb weakness, preceded by diarrhea. She complained of sweating and palpitations. There was no fever, cough, trauma, seizures, or headache. There was global weakness in all four limbs with absent reflexes and hypotonia. Examination of cranial nerves, the sensory system, and other systems was normal. Guillain-Barre syndrome (GBS) was suspected, but due to the patient's co-morbidities, treatment was withheld for 24 hours and the patient was kept under observation. Medical consultation was sought and thyroid function tests were ordered which showed thyroid-stimulating hormone (TSH) 0.019 uIU/ml (normal: 0.35-4.94 uIU/ml), free triiodothyronine (T3) 11.94 pg/ml (normal: 2.0-4.4 pg/ml), and free thyroxine (T4) >5 ng/dl (normal: 0.70-1.48 ng/dl). Thyroid storm was suspected and she was treated with hydrocortisone, propylthiouracil, Lugol iodine, and beta-blocker and her symptoms improved in 10 days with resolution of the weakness, confirming the diagnosis. Besides highlighting this association, this report demonstrates the importance of conducting thyroid function tests in patients presenting with axonal neuropathy. In patients having weakness in all four limbs and presenting with multiple comorbidities, we need to exclude medical reasons before starting treatment for GBS, such as in our case where it was thyrotoxicosis.


Separation and determination of anthraquinone derivatives in rhubarb and its preparations by micellar electrokinetic capillary chromatography.
A micellar electrokinetic capillary chromatographic method was set up for the quality control of rhubarb and its preparations. Anthraquinone derivatives were separated successfully within 10 min in the buffer solution of 50 mmol/L H3BO3-NaOH (pH 11) containing 25 mmol/L sodium deoxycholate. The established method, with a recovery of extraction of over 90%, has good linear relationship and reproducibility. The contents of anthraquinone derivatives in rhubarb and a tablet of Niu-huang-jie-du differed significantly, showing that the quality control of rhubarb and its preparations is necessary.


Developing a Prototype Knowledge-Based System for Diagnosis and Treatment of Diabetes Using Data Mining Techniques.
Diabetes is a disease that affects the body's ability to produce or use insulin. A total of 425 million people are suffering from diabetes in the world. Of this, more than 16 million people live in the Africa Region, which is estimated to be around 41 million by 2045. The main objective of this study was to design and develop a prototype knowledge-based system using data mining techniques for diagnosis and treatment of diabetes. For this study, experimental research design was employed, and the researchers used domain expert knowledge as a supplement of data mining techniques whereby three classification algorithms in WEKA; namely J48, PART and JRip were used, and finally the researchers decided to use the results of J48 classification algorithm. Ultimate Visual basic studio 2013 (Vb.net) was used to store knowledge and as front side of prototype. Common lisp prolog (Clisp) was used for obtained knowledge back end coding. Using a decision tree algorithm; namely J48, 2512 (95.1515%) of the instances were classified correctly, and 128 (4.8485 %) were classified incorrectly. The second most performing model was generated by JRip Classier. This model scored the 94.7348% accuracy on the general data to classify the status of diabetic patient datasets. It classified the 2501 instances of the records correctly. The J48 model was the best performing model with the best accuracy of results.


Dual-spot ratiometric microfluidic paper-based analytical device for accuracy and precision improvement.
Instrumental and environmental fluctuations are common sources of error in smartphone-based optical detection, significantly affecting the accuracy of analytical measurements. In this regard, spotting the sample and reference simultaneously and in close proximity compensates for the fluctuations. This "dual-spot" design is similar to the double-beam technique used in spectrophotometry, which reduces fluctuations in the results. The underlying hypothesis is that any instrumental and/or environmental factors influencing the color intensity in the detection zones will similarly impact the color intensity in the control zone under the same conditions. To test our design, a ratiometric microfluidic paper-based analytical device (μPAD), functionalized with a mixture of green-emissive carbon dots (CDs) and red-emissive ethidium bromide, was developed for the selective detection of ascorbic acid (AA). The green emission of the CDs is quenched by both AA and Fe3+; NaF was thus loaded onto the 3D connector as a masking agent to remove the interference effect of the Fe3+ ions. The color variations were monitored under a UV lamp, using a smartphone to capture the images, and the RGB intensities were processed using the Color Grab application. The proposed double-spot method greatly enhanced the analytical precision and accuracy of the device. A linear working range from 0 to 125 μM was obtained, and the limit of detection was 2.71 μM. The μPAD was successfully used for the detection of AA in human serum, with recoveries from 87.27 to 98.52 %.


[Automated cephalometric landmark identification and location based on convolutional neural network].
Objective: To develop an automated landmark location system applicable to the case of landmark missing. Methods: Four and eighty-one lateral cephalograms, which contained 240 males and 241 females, with an average age of (24.5±5.6) years, taken from January 2015 to January 2021 in the Department of Orthodontics, Capital Medical University School of Stomatology, and met the inclusion criteria were collected. Five postgraduate orthodontic students were the annotators to manually locate 61 possible landmarks in 481 lateral cephalograms. Two assistant professors in the department as reviewers performed calibration. Two professors as arbitrators, made final decision. Data sets were established (341 were used as training set, 40 as validation set, and 100 as test set). In this paper, an automatic landmarks identification and location model based on convolutional neural networks (CNN), CephaNET, was developed. The model was trained by feeding the original image into the feature extraction module and convolutional pose machine (CPM) module to locate landmarks with high accuracy using deep supervision. Training set was enhanced to 1 684 images by histogram equalization, cropping, and adjustment of brightness. The model was trained to compare the Gaussian heat maps output from the network with the set threshold to identify landmark missing cases. Test set of 100 lateral cephalograms was used to test the accuracy of the model. The evaluation criteria used were success detection rate of missing landmark, mean radial error (MRE) and success detection rate (SDR) in the range of 2.0, 2.5, 3.0, 3.5 and 4.0 mm. Results: The model identified and located 61 commonly used landmarks in 0.13 seconds on average. It had an average accuracy of 93.5% in identifying missing landmarks. The MRE of our testing set was (1.19±0.91) mm. SDR of 2.0, 2.5, 3.0, 3.5 and 4.0 mm were 85.4%, 90.2%, 93.5%, 95.4%, 97.0% respectively. Conclusions: The model proposed in this paper could adapt to the absence of landmark in lateral cephalograms and locate 61 commonly used landmarks with high accuracy to meet the requirements of different cephalometric analysis methods.


The Acute Hemolytic Anemias: The Importance of Emergency Diagnosis and Management.
Hemolytic anemias are defined by the premature destruction of red blood cells. These anemias have many causes that are mostly due to chronic diseases, but, occasionally, cases of acute life-threatening hemolysis can occur. The objectives of this article were to review the pathophysiology of hemolytic anemias, to discuss the general emergency department (ED) evaluation, and to discuss the assessment and treatment of important and "cannot miss" hemolytic diseases. Because hemolytic anemias are rarely seen, the emergency physician may consider a patient's anemia as due to blood loss rather than hemolysis, and the workup and treatment may not be appropriate. The primary goal for the emergency provider is to resuscitate, but he or she also must recognize that a hemolytic process is present. Appropriate laboratory work and specialist consultation should be obtained. While focused treatment is rarely necessary in the ED, the avoidance of certain treatments, such as early platelet transfusion in thrombotic thrombocytopenic purpura, may be necessary. Hemolytic anemias are rare, but should still be considered in the ED differential diagnosis of low hemoglobin. Emergency physicians should first resuscitate, but should also be able to identify the presence of hemolysis and obtain the appropriate laboratory tests. Occasionally, specific treatments are needed but should be discussed in conjunction with a specialist.


HLA-restricted cytotoxic T lymphocytes are an early immune response and important defense mechanism in cytomegalovirus infections.
Eighty-eight bone marrow transplant recipients were studied for development of cytomegalovirus (CMV) infections and associated cytotoxic lymphocyte responses. Sixty-one patients developed CMV infection that was diagnosed by virus isolation (18 patients), fourfold rises in serum antibodies (13), or both (30). Interstitial pneumonitis developed in 27 patients. Among patients tested during infection, HLA-restricted, CMV-specific cytotoxic T cell responses occurred in 25 patients, and non-restricted, non-T cell cytotoxic responses, or responses of undetermined cell type, occurred in 10 patients. Levels of CMV-specific cytotoxicity in infected and uninfected patients were compared in two ways. Eighty-nine percent of patients (32 of 36) with cytotoxicity greater than or equal to 15% lysis (50:1 effector-to-target-cell ratio) and 100% of patients (21 of 21) with sequential increases in cytotoxicity of greater than or equal to 15% lysis were infected. The median time of occurrence of CMV-specific cytotoxic responses preceded the median onset of viral shedding by 1.4 weeks, or rises in titers of serum antibody by 1.7 weeks, and of interstitial pneumonitis by 2.5 weeks. Immune competence, as demonstrated by survival from infection, was always associated with a CMV-specific cytotoxic response. The HLA-restricted cytotoxic T cell response appears to be important as an early host-defense mechanism and an early diagnostic evidence of infection.


Occurrence and temporal variations of TMDD in the river Rhine, Germany.
The chemical substance 2,4,7,9-tetramethyl-5-decyne-4,7-diol (TMDD) is a non-ionic surfactant used as an industrial defoaming agent and in various other applications. Its commercial name is Surynol 104 and the related ethoxylates are also available as Surfynol 420, 440, 465 and 485 which are characterized by different grades of ethoxylation of TMDD at both hydroxyl functional groups. TMDD and its ethoxylates offer several advantages in waterborne industrial applications in coatings, inks, adhesives as well as in paper industries. TMDD and its ethoxylates can be expected to reach the aquatic environment due its widespread use and its physico-chemical properties. TMDD has previously been detected in several rivers of Germany with concentrations up to 2.5 microg/L. In the United States, TMDD was also detected in drinking water. However, detailed studies about its presence and distribution in the aquatic environment have not been carried out so far. The aim of the present study was the analysis of the spatial and temporal concentration variations of TMDD in the river Rhine at the Rheingütestation Worms (443.3 km). Moreover, the transported load in the Rhine was investigated during two entire days and 7 weeks between November 2007 and January 2008. The sampling was carried out at three different sampling points across the river. Sampling point MWL1 is located in the left part of the river, MWL2 in the middle part, and MWL4 in the right part. One more sampling site (MWL3) was run by the monitoring station until the end of 2006, but was put out of service due to financial constrains. The water at the left side of the river Rhine (MWL1) is influenced by sewage from a big chemical plant in Ludwigshafen and by the sewage water from this city. The water at the right side of the river Rhine (MWL4) is largely composed of the water inflow from river Neckar, discharging into Rhine 14.9 km upstream from the sampling point and of communal and industrial wastewater from the city Mannheim. The water from the middle of the river (MWL2) is largely composed of water from the upper Rhine. Water samples were collected in 1-L bottles by an automatic sampler. The water samples were concentrated by use of solid-phase extraction (SPE) using Bond Elut PPL cartridges and quantified by use of gas chromatography-mass spectrometry (GC-MS). The quantification was carried out with the internal standard method. Based on these results, concentration variations were determined for the day profiles and week profiles. The total number of analyzed samples was 219. The results of this study provide information on the temporal concentration variability of TMDD in river Rhine in a cross section at one particular sampling point (443.3 km). TMDD was detected in all analyzed water samples at high concentrations. The mean concentrations during the 2 days were 314 ng/L in MWL1, 246 ng/L in MWL2, and 286 ng/L in MWL4. The variation of concentrations was low in the day profiles. In the week profiles, a trend of increasing TMDD concentrations was detected particularly in January 2008, when TMDD concentrations reached values up to 1,330 ng/L in MWL1. The mean TMDD concentrations during the week profiles were 540 ng/L in MWL1, 484 ng/L in MWL2, and 576 ng/L in MWL4. The loads of TMDD were also determined and revealed to be comparable in all three sections of the river. The chemical plant located at the left side of the Rhine is not contributing additional TMDD to the river. The load of TMDD has been determined to be 62.8 kg/d on average during the entire period. By extrapolation of data obtained from seven week profiles the annual load was calculated to 23 t/a. The permanent high TMDD concentrations during the investigation period indicate an almost constant discharge of TMDD into the river. This observation argues for effluents of municipal wastewater treatment plants as the most likely source of TMDD in the river. Another possible source might be the degradation of ethoxylates of TMDD (Surfynol series 400), in the WWTPs under formation of TMDD followed by discharge into the river. TMDD has to be considered as a high-production-volume (HPV) chemical based on the high concentrations found in this study. In the United States, TMDD is already in the list of HPV chemicals from the Environmental Protection Agency (EPA). However, the amount of TMDD production in Europe is unknown so far and also the biodegradation rates of TMDD in WWTPs have not been investigated. TMDD was found in high concentrations during the entire sampling period in the Rhine river at the three sampling points. During the sampling period, TMDD concentrations remained constant in each part of the river. These results show that TMDD is uniformly distributed in the water collected at three sampling points located across the river. 'Waves' of exceptionally high concentrations of TMDD could not be detected during the sampling period. These results indicate that the effluents of WWTPs have to be considered as the most important sources of TMDD in river Rhine. Based also on the occurrence of TMDD in different surface waters of Germany with concentrations up to 2,500 ng/L and its presence in drinking water in the USA, more detailed investigations regarding its sources and distribution in the aquatic environment are required. Moreover, the knowledge with respect to its ecotoxicity and its biodegradation pathway is scarce and has to be gained in more detail. Further research is necessary to investigate the rate of elimination of TMDD in municipal and industrial wastewater treatment plants in order to clarify the degradation rate of TMDD and to determine to which extent effluents of WWTPs contribute to the input of TMDD into surface waters. Supplementary studies are needed to clarify whether the ethoxylates of TMDD (known as Surfynol 400 series) are hydrolyzed in the aquatic environment resulting in formation of TMDD similar to the well known cleavage of nonylphenol ethoxylates into nonylphenols. The stability of TMDD under anaerobic conditions in groundwater is also unknown and should be studied.


Stress influences on anoikis.
It long has been suspected that psychosocial factors affect cancer development and progression. Although the connections between stress and cancer causation are not strong, epidemiologic and clinical studies have provided strong links between cancer progression and several stress-related factors including chronic stress, depression, and social isolation. Recent molecular and biological studies have identified specific signaling pathways that influence cancer growth and metastasis. In particular, stress hormones can have a significant impact on protecting cancer cells from undergoing the anoikis form of programmed cell death, thus, providing a mechanistic advantage for metastasis. This review provides an overview of the relationship between psychosocial factors and the avoidance of anoikis by cancer cells.


Inhibition of local effects of Indian Daboia/Vipera russelli venom by the methanolic extract of grape (Vitis vinifera L.) seeds.
Although anti-venom therapy is available for the treatment of fatal bite by snakes, it offers less or no protection against the local effects such as dermo- and myonecrosis, edema, hemorrhage and inflammation at the bitten region. The viper species are known for their violent local effects and such effects have been commonly treated with plant extracts without any scientific validation in rural India. In this investigation, the methanolic extract of grapes (Vitis vinifera L.) seed was studied against the Indian Daboia/Vipera russelli venom-induced local effects. The extract abolished the proteolytic and hyaluronidase activities and also efficiently neutralized the hemorrhage, edema-inducing and myonecrotic properties of the venom. In addition, the extract also inhibited partially the pro-coagulant activity of the venom and abolished the degradation of Aalpha and Bbeta chains of human fibrinogen. Thus, the extract possesses potent anti-snake venom property, especially against the local effects of viper bites.


Myocardial perfusion in compensated and failing hypertrophied left ventricle.
This study examined blood flow in the hypertrophied left ventricle with and without failure. Left ventricular hypertrophy was produced in 20 dogs by banding the ascending aorta at 6-7 wk of age; studies were performed after animals reached adulthood. Sixteen dogs had compensated hypertrophy, while four dogs had cardiac failure manifested by left ventricular dilatation and end-diastolic pressures greater than 18 mmHg. The degree of hypertrophy, assessed by left ventricular-to-body weight ratio, was similar in animals with compensated hypertrophy (7.29 +/- 0.26 g/kg) and failure (8.45 +/- 0.15); both were greater than control (4.50 +/- 0.15, P less than 0.01). Left ventricular systolic pressure was similar in compensated hypertrophy (184 +/- 9 mmHg) and failure (226 +/- 29), as compared with control (130 +/- 4; P less than 0.01). Left ventricular blood flow measured with microspheres was 0.89 +/- 0.07 ml X min-1 X g-1 in control animals, was increased to 1.34 +/- 0.05 with compensated hypertrophy (P less than 0.001), and was further increased with failure to 1.86 +/- 0.40 (P less than 0.05). The left ventricular wall thickness-to-cavity diameter ratio was increased to 0.63 +/- 0.04 with compensated hypertrophy but was only 0.40 +/- 0.05 in dogs with failure (P less than 0.01), suggesting that wall stress was greater in hearts with failure. These data suggest that increased blood flow rates in dogs with failure resulted from increased myocardial O2 requirements due to increased systolic wall stress. Need for increased blood flow during resting conditions in dogs with failure would impair the ability for further coronary vasodilation during periods of cardiac stress.


Increased vascular wall thrombogenicity combined with reduced blood flow promotes occlusive thrombus formation in rabbit femoral artery.
Plaque disruption does not always result in complete thrombotic occlusion. The mechanism of arterial thrombus propagation remains unclear. We studied how vascular wall thrombogenicity and blood flow reduction affect thrombus propagation using a rabbit model of single and repeated balloon injury. After balloon injury of the normal femoral artery, the blood flow was reduced to 50%, 25%, or 10% (n=5). Small mural thrombi composed of aggregated platelets were produced, but no occlusive thrombi developed in any flow reduction. Three weeks after the first balloon injury, neointima with tissue factor expression and increased procoagulant activity was developed. Balloon injury of the neointima with the same blood flow reduction (n=5) induced fibrin-rich thrombus formation. Additionally, injury with flow reduced to 25% and 10% promoted thrombus propagation resulting in vessel occlusion within 160+/-18 and 71+/-17 seconds, respectively. An injection of anti-von Willebrand factor (vWF) monoclonal antibody (AJW200; 1.0 mg/kg) prevented occlusive thrombus formation. Increased vascular wall thrombogenicity together with a substantial blood flow reduction is crucial for occlusive thrombus formation, and vWF plays an important role in thrombus propagation. Reduced blood flow at plaque disruption sites might contribute to thrombus propagation leading to acute coronary syndromes.


Detection of the deep biosphere in metamorphic rocks from the Chinese continental scientific drilling.
It is generally accepted that there is a vast, well-populated biosphere in the subsurface, but the depth limit of the terrestrial biosphere has yet to be determined, largely because of the lack of access to the subsurface. Here as part of the Chinese Continental Scientific Drilling (CCSD) project in eastern China, we acquired continuous rock cores and endeavored to probe the depth limit of the biosphere and the depth-dependent distribution of microorganisms at a geologically unique site, that is, a convergent plate boundary. Microbiological analyses of ultra-high-pressure metamorphic rock cores taken from the ground surface to 5,158-meter reveal that microbial distribution was continuous up to a depth of ~4,850 m, where temperature was estimated to be ~137°C. The metabolic state of these organisms at such great depth remains to be determined. Microbial abundance, ranging from 103 to 108 cells/g, was also related to porosity, but not to the depth and rock composition. In addition, microbial diversity systematically decreased with depth. Our results support the notion that temperature is a key factor in determining the lower limit of the biosphere in the continental subsurface.


Alpha 2-macroglobulin and serum preferentially counteract the mitoinhibitory effect of transforming growth factor-beta 2 in rat hepatocytes.
Transforming growth factors-beta 1 and beta 2 (TGF-beta 1 and TGF-beta 2) are potent, multifunctional modifiers of cellular proliferation and differentiation in many cell types. To evaluate factors which may alter the activity of the TGF-beta s during hepatocyte proliferation, we examined the influences of bovine serum and purified bovine or human alpha 2-macroglobulin (alpha 2M) on the mitoinhibitory effects of the TGF-beta in primary cultures of rat hepatocytes. The inhibitory activity of TGF-beta was evaluated by autoradiographic labeling index at 48 hours in hepatocyte cultures exposed to [3H]thymidine between hours 24 and 48 in culture. In the absence of serum or alpha 2M, TGF-beta 1 and TGF-beta 2 were equivalently potent in inhibiting S-phase DNA synthesis in hepatocytes cultured with or without epidermal growth factor. However, bovine serum and purified bovine or human alpha 2M consistently counteracted the mitoinhibitory effects of TGF-beta 2. S-phase DNA synthesis increased five- to six-fold when bovine serum (15%) or alpha 2M (200 micrograms/ml) were included with TGF-beta 2 (0.1 ng/ml) and epidermal growth factor (20 ng/ml). The mitoinhibitory effect of TGF-beta 1 was not influenced by the addition of purified bovine alpha 2M. Human alpha 2M or bovine serum counteracted inhibition by TGF-beta 1 to a lesser extent. [125I]TGF-beta 1 and 125I]TGF-beta 2 formed complexes with purified bovine alpha 2M and serum proteins migrating at identical positions to purified alpha 2M during nondenaturing polyacrylamide gel electrophoresis. However, [125I]TGF-beta 1 associated preferentially with the "fast" migrating form of alpha 2M, whereas [125I]TGF-beta 2 associated with both the "slow" and "fast" migrating forms. Inhibitory activity of TGF-beta 1 and TGF-beta 2 coeluted with alpha 2M in the high molecular weight fractions from a Sephacryl S-200 column. These results support the hypothesis that purified alpha 2M and alpha 2M in bovine serum binds both TGF-betas but preferentially counteracts the mitoinhibitory effect of TGF-beta 2 on rat hepatocytes.


Measurement of cochlear power gain in the sensitive gerbil ear.
The extraordinary sensitivity of the mammalian ear is commonly attributed to the cochlear amplifier, a cellular process thought to locally boost responses of the cochlear partition to soft sounds. However, cochlear power gain has not been measured directly. Here we use a scanning laser interferometer to determine the volume displacement and volume velocity of the cochlear partition by measuring its transverse vibration along and across the partition. We show the transverse displacement at the peak-response location can be >1,000 times greater than the displacement of the stapes, whereas the volume displacement of an area centred at this location is approximately tenfold greater than that of the stapes. Using the volume velocity and cochlear-fluid impedance, we discover that power at the peak-response area is >100-fold greater than that at the stapes. These results demonstrate experimentally that the cochlea amplifies soft sounds, offering insight into the mechanism responsible for the cochlear sensitivity.


Correlation of cardiac troponin T levels with inotrope requirement, hypoxic-ischemic encephalopathy, and survival in asphyxiated neonates.
Cardiac involvement in neonates with perinatal asphyxia not only complicates perinatal management but also contributes to increased mortality. To assess cardiac troponin T (cTnT) levels in asphyxiated neonates and their correlation with echocardiography findings, inotrope requirement, hypoxic-ischemic encephalopathy (HIE) stages, and mortality. cTnT levels, echocardiographic findings, the requirement of inotropes, HIE stages, and outcome were studied in neonates of gestational age ≥ 34 wk with perinatal asphyxia. Among 57 neonates with perinatal asphyxia, male gender, cesarean section, forceps/vacuum-assisted vaginal delivery and late preterm included 33 (57.9%), 23 (40.4%), 3 (5.3%), and 12 (21.1%) respectively. The mean gestational age was 38.4 wk (1.6 wk). HIE stages I, II, and III were observed in 7 (12.3%), 37 (64.9%), and 9 (15.8%) neonates respectively. 26 (45.6%) neonates had echocardiographic changes and 19 (33.3%) required inotropes. cTnT levels were elevated in 41 (71.9%) neonates [median (IQR); 0.285 (0.211-0.422) ng/mL]. The Median cTnT level showed an increasing trend with increasing changes in echocardiography (P = 0.002). Two neonates with mitral regurgitation and global hypokinesia had the highest cTnT levels (1.99 and 0.651 ng/mL). Of 31 neonates with normal echocardiography, 18 (58.06%) showed elevated cTnT. cTnT levels were significantly higher in those who required inotropic support than those who did not (P = 0.007). Neonates with HIE stage III had significantly higher cTnT levels compared to those with HIE stage I/II (P = 0.013). Survivors had lower median cTnT levels [0.210 (0.122-0.316) ng/mL] than who succumbed [0.597 (0.356-1.146) ng/mL]. cTnT levels suggestive of cardiac involvement were observed in 71.9% of asphyxiated neonates. cTnT levels correlated with echocardiography findings, inotrope requirement, HIE stages, and mortality.


Inducible nitric oxide synthase (iNOS) is expressed similarly in multiple aberrant crypt foci and colorectal tumors from the same patients.
Aberrant crypt foci (ACF) are the earliest identified neoplastic lesions in the colon. Aberrant expression of inducible nitric oxide synthase (iNOS or NOS2) has been documented in colorectal tumors, but expression of iNOS has not been reported in human ACF or multiple neoplastic lesions from the same patient. Immunohistochemical expression of iNOS was evaluated in 42 ACF, 14 adenomas, and 25 carcinomas and their adjacent normal mucosa. iNOS was strongly expressed in the normal colonic epithelial cells of all patients; it was markedly reduced in 21 of 42 (50%) ACF and in 14 of 25 (56%) carcinomas. The expression of iNOS was remarkably similar in multiple lesions from the same patient (P < 0.0001). These results suggest that the reduced expression of iNOS is a very early event in the development of some human colorectal tumors, and that host factors control the expression of iNOS similarly in premalignant and malignant colonic epithelial cells.


Prospective investigation of social support, coping, and depressive symptoms: A model of adherence to endocrine therapy among women with breast cancer.
Nonadherence reduces the effectiveness of behavioral change regimens for promoting health and decreasing morbidities and mortality. Such is the case with endocrine therapies, which lower the likelihood of recurrence in the approximately 70% of women with hormone receptor-positive breast cancers. This investigation tests a model of contributors to objective adherence. Women with breast cancer receiving their 1st endocrine therapy prescription (N = 130) were recruited from a large community oncology breast clinic. Participants completed three interview and questionnaire sessions at prescription initiation, one month and four months later. Questionnaires addressed patients' experience with cancer and included measures of coping (COPE and Emotional Approach Coping), social support (Interpersonal Support Evaluation List), and depressive symptoms (Center for Epidemiologic Studies Depression Scale). At the initial appointment, participants were provided a Medication Event Monitoring System (MEMS) cap to assess adherence; the MEMS cap was collected at the 4-month follow-up appointment. Structural equation modeling offered partial support for a mediational model, χ2(68, N = 130) = 77.47, p = .202, root-mean-square error of approximation = .03, comparative fit index = .97, standardized root-mean-square residual = .06. Greater social support at prescription initiation was associated with lower depressive symptoms 1 month later, which in turn were associated with higher objective adherence (MEMS) four months after study entry, controlling for significant covariates (p < .05). Use of avoidance-oriented coping was not statistically significantly related to depressive symptoms (p > .05). This model informs theory regarding medication adherence as well as intervention development for women prescribed adjuvant endocrine therapy. (PsycINFO Database Record


Choledochocele: diagnosis by magnetic resonance imaging.
The value of magnetic resonance imaging (MRI) and magnetic resonance cholangiopancreatography (MRCP) is demonstrated in a case of a 22-year-old female with a small choledochocele clinically presenting with acute cholecystitis and pancreatitis. Previous abdominal sonography and computed tomography were not diagnostic. MRI and MRCP showed a cystic dilatation of the distal common bile duct, intramurally located in the second portion of the duodenum and protruding into the duodenal lumen. These findings allowed a confident diagnosis of a choledochocele.


Dynamic model of heat inactivation kinetics for bacterial adaptation.
The Weibullian-log logistic (WeLL) inactivation model was modified to account for heat adaptation by introducing a logistic adaptation factor, which rendered its "rate parameter" a function of both temperature and heating rate. The resulting model is consistent with the observation that adaptation is primarily noticeable in slow heat processes in which the cells are exposed to sublethal temperatures for a sufficiently long time. Dynamic survival patterns generated with the proposed model were in general agreement with those of Escherichia coli and Listeria monocytogenes as reported in the literature. Although the modified model's rate equation has a cumbersome appearance, especially for thermal processes having a variable heating rate, it can be solved numerically with commercial mathematical software. The dynamic model has five survival/adaptation parameters whose determination will require a large experimental database. However, with assumed or estimated parameter values, the model can simulate survival patterns of adapting pathogens in cooked foods that can be used in risk assessment and the establishment of safe preparation conditions.


Mutant USA strain of porcine circovirus type 2 (mPCV2) exhibits similar virulence to the classical PCV2a and PCV2b strains in caesarean-derived, colostrum-deprived pigs.
In 2012, a mutant porcine circovirus type 2 (mPCV2) strain was identified in cases of PCV-associated disease (PCVAD) in the USA. The mPCV2 had an additional amino acid, lysine (K), in the capsid at position 234. The objectives of this study were to compare the pathogenicity of mPCV2, PCV2a and PCV2b in pigs using biologically pure infectious virus stocks derived from respective infectious DNA clones, and to investigate the importance of genotype-specific ORF2 and the presence of lysine at position 234 of the capsid. A total of 47, 2-week-old, caesarean-derived, colostrum-deprived (CDCD) pigs were assigned to one of seven groups. At 3 weeks of age, the pigs were experimentally inoculated with saline, PCV2a, PCV2b, mPCV2, PCV2b-234-K (lysine addition in ORF2), chimeric PCV2b-ORF1/mPCV2-ORF2 or reciprocal chimeric mPCV2-ORF1/PCV2b-ORF2. All pigs were necropsied 21 days post-infection (p.i.). Gross lesions were limited to visible icterus and loss of body condition in a portion of the mPCV2 pigs. The amount of PCV2 DNA was significantly higher in pigs inoculated with mPCV2 compared with PCV2b in sera at 7 days p.i. and faecal swabs at 14 days p.i. Based on lymphoid lesions, a higher prevalence of PCVAD was seen in pigs infected with PCV2s containing the additional 234-K (64.3 %) compared with those infected with a PCV2 with the regular 233 bp ORF2 (40 %). Results indicated that all PCV2 isolates were capable of inducing severe lesions and disease in the CDCD pig model, and there was no significant difference in virulence.


Extending cBioPortal for Therapy Recommendation Documentation in Molecular Tumor Boards: Development and Usability Study.
In molecular tumor boards (MTBs), patients with rare or advanced cancers are discussed by a multidisciplinary team of health care professionals. Software support for MTBs is lacking; in particular, tools for preparing and documenting MTB therapy recommendations need to be developed. We aimed to implement an extension to cBioPortal to provide a tool for the documentation of therapy recommendations from MTB sessions in a secure and standardized manner. The developed extension should be embedded in the patient view of cBioPortal to enable easy documentation during MTB sessions. The resulting architecture for storing therapy recommendations should be integrable into various hospital information systems. On the basis of a requirements analysis and technology analysis for authentication techniques, a prototype was developed and iteratively refined through a user-centered development process. In conclusion, the tool was evaluated via a usability evaluation, including interviews, structured questionnaires, and the System Usability Scale. The patient view of cBioPortal was extended with a new tab that enables users to document MTB sessions and therapy recommendations. The role-based access control was expanded to allow for a finer distinction among the rights to view, edit, and delete data. The usability evaluation showed overall good usability and a System Usability Scale score of 83.57. This study demonstrates how cBioPortal can be extended to not only visualize MTB patient data but also be used as a documentation platform for therapy recommendations.


The ubiquitin ligase c-Cbl down-regulates FcgammaRIIa activation in human neutrophils.
Little is known about the mechanisms that arrest FcgammaRIIa signaling in human neutrophils once engaged by immune complexes or opsonized pathogens. In our previous studies, we observed a loss of immunoreactivity of Abs directed against FcgammaRIIa following its cross-linking. In this study, we report on the mechanisms involved in this event. A stimulated internalization of FcgammaRIIa leading to the down-regulation of its surface expression was observed by flow cytometry and confocal microscopy. Immunoprecipitation of the receptor showed that FcgammaRIIa is ubiquitinated after stimulation. MG132 and clasto-lactacystin beta-lactone inhibited the loss of immunoreactivity of FcgammaRIIa, suggesting that this receptor was down-regulated via the proteasomal pathway. The E3 ubiquitin ligase c-Cbl was found to translocate from the cytosol to the plasma membrane following receptor cross-linking. Furthermore, c-Cbl was recruited to the same subset of high-density, detergent-resistant membrane fractions as stimulated FcgammaRIIa itself. Silencing the expression of c-Cbl by small interfering RNA decreased FcgammaRIIa ubiquitination and prevented its degradation without affecting the internalisation process. It also prolonged the stimulation of the tyrosine phosphorylation response to the cross-linking of the receptor. We conclude that c-Cbl mediates the ubiquitination of stimulated FcgammaRIIa and thereby contributes to the termination of FcgammaRIIa signaling via its proteasomal degradation, thus leading to the down-regulation of neutrophil signalisation and function (phagocytosis) through this receptor.


The role of post-translational modifications in fine-tuning BLM helicase function during DNA repair.
RecQ-like helicases are a highly conserved family of proteins which are critical for preserving genome integrity. Genome instability is considered a hallmark of cancer and mutations within three of the five human RECQ genes cause hereditary syndromes that are associated with cancer predisposition. The human RecQ-like helicase BLM has a central role in DNA damage signaling, repair, replication, and telomere maintenance. BLM and its budding yeast orthologue Sgs1 unwind double-stranded DNA intermediates. Intriguingly, BLM functions in both a pro- and anti-recombinogenic manner upon replicative damage, acting on similar substrates. Thus, BLM activity must be intricately controlled to prevent illegitimate recombination events that could have detrimental effects on genome integrity. In recent years it has become evident that post-translational modifications (PTMs) of BLM allow a fine-tuning of its function. To date, BLM phosphorylation, ubiquitination, and SUMOylation have been identified, in turn regulating its subcellular localization, protein-protein interactions, and protein stability. In this review, we will discuss the cellular context of when and how these different modifications of BLM occur. We will reflect on the current model of how PTMs control BLM function during DNA damage repair and compare this to what is known about post-translational regulation of the budding yeast orthologue Sgs1. Finally, we will provide an outlook toward future research, in particular to dissect the cross-talk between the individual PTMs on BLM.


Differences in real and illusory shape perception revealed by backward masking.
Illusory contours (ICs) are thought to be a result of processes involved in the perceptual recovery of occluded surfaces. Here, we investigate the relationship between real and illusory contour perception using a shape discrimination task and backward masking paradigm. ICs can mask other ICs when times between mask onset and stimulus onset, or SOAs, are very long ( approximately 300 ms), but real contours (RCs) are not similarly effective. Masking is absent for RC masks at perceptually salient contrasts, as well as for those with contrast lowered to match the perceived brightness of the illusory surface. We also find that RCs are not masked at long SOAs, either by ICs or by other RCs. Finally, the masking seen between ICs can occur for different sizes of target and mask. The cross-size masking would not be expected if the masking were at a level sensitive to retinal contour location. The late masking therefore may be related to a higher level of processing of shape categories and surfaces, the level at which shapes defined by ICs and RCs are differentially represented.


The value of joint ultrasonography in predicting arthritis in seropositive patients with arthralgia: a prospective cohort study.
The value of joint ultrasonography (US) in the prediction of clinical arthritis in individuals at risk of developing rheumatoid arthritis (RA) is still a point of debate, due to varying scanning protocols and different populations. We investigated whether US abnormalities assessed with a standard joint protocol can predict development of arthritis in seropositive patients with arthralgia. Anti-citrullinated protein antibodies and/or rheumatoid factor positive patients with arthralgia, but without clinical arthritis were included. US was performed at baseline in 16 joints: bilateral metacarpophalangeal 2-3, proximal interphalangeal 2-3, wrist and metatarsophalangeal (MTP) joints 2-3 and 5. Images were scored semi-quantitatively for synovial thickening and for positive signs on power Doppler (PD). Association between US abnormalities and arthritis development at the joint and at the patient level was evaluated. Also, we investigated the added value of US over clinical parameters. Out of 163 patients who underwent US examination, 51 (31%) developed clinical arthritis after a median follow-up time of 12 (interquartile range 5-24) months, of which 44 (86%) satisfied the 2010 American College of Rheumatology/European League Against Rheumatism classification criteria for RA. US revealed synovial thickening and PD in at least one joint in 49 patients (30%) and 7 patients (4%), respectively. Synovial thickening was associated with both development and timing of clinical arthritis in any joint (patient level) when MTP joints were excluded from the US assessment (odds ratio 6.6, confidence interval (CI) 1.9-22), and hazard ratio 3.4, CI 1.6-6.8, respectively, with a mean time to arthritis of 23 versus 45 months when synovial thickening was present versus not present). There was no association between US and arthritis development at the joint level. Predictive capacity was highest in the groups with an intermediate and high risk of developing arthritis based on a prediction rule with clinical parameters. Synovial thickening on US predicted clinical arthritis development at the patient level in seropositive patients with arthralgia when MTPs were excluded from the US assessment. Positive PD signs were infrequently seen in these at-risk individuals and was not predictive. In patients at intermediate risk of RA, US may help to identify those at higher risk of developing arthritis.


Working in the dead of night: exploring the transition to after-hours duty.
Transitions, although often difficult, represent integral components of medical training. New postgraduate trainees (first-year residents) find themselves in an especially challenging transition as they are expected to fulfil both learning and service expectations concurrently. Workplace learning theory has been suggested as a lens through which to understand this unique educational, yet service-oriented, role. This tension may be further amplified overnight when residents are on-call with little to no support. The aims of this study were to explore the transition from medical student to resident with respect to the on-call experience, and to provide theory-based suggestions to enhance learning during this unique transition. We conducted an interpretivist qualitative study by interviewing eight medical students and 10 first-year residents from six different specialty training programmes across four academic sites. Each semi-structured interview was transcribed verbatim and anonymised. Resident interview transcripts were initially coded for major themes, after which medical student interview transcripts were coded for consistencies and discrepancies. Four interrelated themes were identified in students' and residents' descriptions of on-call experiences: (i) shift in responsibility; (ii) supervisory support; (iii) contextual conditions, and (iv) clarity of expectations. Generally, students were not able to anticipate the challenges they would face as residents on-call, and residents perceived the transition as sudden with little emphasis placed on learning. First-year residents face multiple challenges during on-call, which may prevent optimal learning in this setting. These challenges are amplified by the large gap between the respective roles of medical students and residents. We identified promoters of and barriers to effective learning in this environment and, by using workplace learning theory, provide recommendations for how we might be able to enhance medical students' preparation for and first-year residents' learning during experiences of being on-call.


Human tracheal allotransplant with greater omentum for revascularization.
Human tracheal allotransplant is an option for treating long-segment tracheal defects, but no reliable and satisfactory tracheal transplant procedure has been developed. The purpose of this study is to review the results of 2-stage tracheal reconstruction in human tracheal allotransplant for tracheal defects > 5 cm with the use of greater omentum for revascularization. There were 3 patients who had tracheal allograft. In stage 1, a segment of donor trachea (6 cm) was excised and wrapped in the recipient greater omentum. In stage 2, at 3 to 5 weeks after stage 1, the tracheal allograft and omental pedicle were orthotopically transplanted to the cervical part of the trachea. All patients received immunosuppression with tacrolimus, mycophenolate mofetil, and methylprednisolone. Fiberoptic bronchoscopy was performed monthly during the 6 months after stage 2. In all 3 patients, no major postoperative complication was observed. Transverse section of the donor trachea showed a mechanically stable chondral framework. Histologic sections stained with hematoxylin-eosin showed complete cartilage rings covered by intact respiratory epithelium. A 2-stage procedure for human tracheal transplant with greater omentum is a safe and effective technique for tracheal reconstruction.


Acute hormonal responses before and after 2 weeks of HIT in well trained junior triathletes.
The aim was to compare the acute hormonal response to a single HIT session at the beginning and end of a HIT shock microcycle. 13 male junior triathletes (15.8±1.8 yrs.) performed 16 HIT sessions within a 2 week period. Venous blood samples were collected before and after the first and last HIT session. Significant increases in cortisol (first session +89.7%; last session +70.3%) and hGH (first session +435.1%; last session +314.6%) concentrations were observed after both training sessions (P<0.05). The acute responses of cortisol, hGH, T3, and fT3 were not different between the first and last HIT sessions (P=1.00). Although no acute changes in testosterone were detected after the training sessions, testosterone concentrations were significantly higher at all time points (62.6-80.1%) during the last compared to first training session (P≤0.001). Findings from the present study reveal that 16 sessions of HIT led to significant increases in baseline concentrations of serum testosterone. This might indicate a heightened anabolic state even in junior triathletes. Based on the hormonal data, we conclude that at the end of this 2 week microcycle no familiarization effect was evident and that the training stimulus produced by HIT was still great enough to "stress" the athletes and induce positive training adaptations.


Inhibition of alloxan action in isolated pancreatic islets by superoxide dismutase, catalase, and a metal chelator.
The possible participation of superoxide anions, hydrogen peroxide, and hydroxyl radicals in the action of alloxan was investigated using isolated rat pancreatic islets. Exposure of islets for 5 min to alloxan (0.15 or 0.2 mg/ml) inhibited subsequent glucose-stimulated insulin release. The presence of superoxide dismutase (1000 U/ml), catalase (50 microgram/ml), or a metal chelator diethylenetriaminepentacetic acid (1 mM) markedly attenuated this effect of alloxan. Use of these agents afforded complete protection from the lower concentration of alloxan and partial protection from the higher concentration of the toxic compound. Inactivation of the enzymes or addition of excess iron to the chelating agent before its use with alloxan eliminated the protective action of these agents. The results are consistent with the proposal that hydroxyl radicals, generated via reactions that involve superoxide anions, hydrogen peroxide, and iron, mediate the deleterious effect of alloxan in pancreatic islets.


Rapid and accurate diagnosis of Chlamydia trachomatis in the urogenital tract by a dual-gene multiplex qPCR method.
Introduction. Chlamydia trachomatis (C. trachomatis, CT) is an obligatory intracellular bacterium that causes urogenital tract infections and leads to severe reproductive consequences. Therefore, a rapid and accurate detection method with high sensitivity and specificity is an urgent requirement for the routine diagnosis of C. trachomatis infections.Aim. In this study, we aimed to develop a multiplex quantitative real-time PCR (qPCR) assay based on two target regions for accurate detection of C. trachomatis in urogenital tract infections.Methodology. Primers and probes based on the conserved regions of the cryptic plasmid and 23S rRNA gene were designed. Then, two qPCR assays were established to screen for the optimal probe and primers for each of the two target regions. Subsequently, the multiplex qPCR method was developed and optimized. For the diagnostic efficiency evaluation, 1284 urogenital specimens were tested by the newly developed multiplex qPCR method, an immunological assay and a singleplex qPCR assay widely used in hospitals.Results. The multiplex qPCR method could amplify both target regions in the range of 1.0×102-1.0×108 copies ml-1 with a strong linear relationship, and lower limits of detection (LODs) for both targets reached 2 copies PCR-1. For the multiplex qPCR method, the diagnostic sensitivity and specificity was 100.0 % (134/134) and 99.3 % (1142/1150), respectively. For the singleplex qPCR assay, the diagnostic sensitivity and specificity was 88.8 % (119/134) and 100.0 % (1150/1150), respectively. For the immunological assay, the diagnostic sensitivity and specificity was 47.0 % (63/134) and 100.0 % (1150/1150), respectively.Conclusion. In this study, a multiplex qPCR assay with high sensitivity and specificity for rapid (≤2.0 h) and accurate diagnosis of C. trachomatis was developed. The qPCR assay has the potential to be used as a routine diagnostic method in clinical microbiology laboratories.


Laparoscopic repair of an iliac artery injury during radical cystoprostatectomy.
Laparoscopy has experienced significant growth in the last 5 years and became more popular among practicing urologists. Even though laparoscopy is considered a safe technique it is not free of complications, some of which can be devastating. Vascular injuries are the second most frequent complication during laparoscopic surgery with a reported incidence of 0.22% to 1.1%. With the standardization of surgical techniques and the increasing proficiency of the teams performing it, serious surgical complications may be corrected without the need for conversion. We report a case in which the right external iliac artery was injured while pelvic lymph node dissection was carried out before a planned laparoscopic radical cystoprostatectomy. The complication was successfully managed entirely by means of laparoscopy.


Intracranial aneurysms treated with Guglielmi detachable coils: long-term imaging follow-up with contrast-enhanced magnetic resonance angiography.
The aim of this study was to assess the long-term results of intracranial aneurysms treated with Guglielmi detachable coils (GDCs) with the aid of contrast-enhanced magnetic resonance (MR) angiography. Between January 1998 and August 2001, 92 patients with 92 aneurysms treated by endovascular coiling with GDCs underwent contrast-enhanced MR angiography. These patients underwent long-term follow-up (range 32-78 months, mean 42.1 +/- 11.9 months [standard deviation]) after endovascular treatment. All images were compared with digital subtraction angiograms and contrast-enhanced MR angiograms that had been obtained during the short-term follow-up (range 5-25 months, mean 13 +/- 5.1 months after treatment). The MR angiograms were analyzed independently by 2 senior radiologists. Findings were assigned to 1 of 3 categories: complete obliteration (Class 1), residual neck (Class 2), or residual aneurysm (Class 3). Of 92 contrast-enhanced MR angiograms obtained at the long-term follow-up, complete obliteration of the aneurysm was noted in 57 patients (Class 1), a residual neck was seen in 22 (Class 2), and a residual aneurysm was observed in 13 (Class 3). One patient experienced aneurysm rehemorrhaging during the follow-up period. The comparison of short- and long-term follow-up angiograms demonstrated a change in aneurysm classification in 7 patients (7.6%), including 4 that progressed from Class 1 to Class 2 and 3 from Class 2 to Class 3. However, 4 (14.2%) of the 28 long-term recurrences were not detected on the short-term control images. Long-term follow-up with contrast-enhanced MR angiography after selective embolization of intracranial aneurysms can identify late aneurysm recanalization that is undetected at short-term follow-up.


Development and evaluation of a quality control system based on transdermal electrical resistance for skin barrier function in vitro.
In vitro skin permeation experiments are highly relevant for pharmaceutical, cosmetic, agricultural developments, and regulatory evaluation. A key requirement is the skin barrier integrity, that is accompanied by an intact stratum corneum (SC) which implements high skin quality. A variety of integrity tests are currently available, for example, measurement of transepidermal water loss, monitoring the permeation of tritiated water and the measurement of transdermal electrical resistance (TER). We aimed for a non-destructive examination of barrier integrity as quality control system, based on TER. Therefore, the in-house developed instrument SkinTER measures electrical resistance on excised human skin samples in a non-invasive and easy-to-use pattern. In this proof of concept study, we compared three human in vitro skin models with focus on their TER and permeation properties. The skin integrity was impaired to mimic conditions of skin during age, lifestyle (eg, shaving) or diseases (eg, obesity, psoriasis, and atopic dermatitis). The OECD permeation marker caffeine was correlated to the corresponding TER value. A correlation between both was obtained by having a Pearson coefficient of -0.830. Hereby, a minimum TER value for intact skin samples of ~1.77 kΩ*cm2 was suggested. Intact samples are significantly different (α = ≤0.05) to their impaired counterparts in flux and TER values. The new SkinTER instrument gives a quick and non-invasive feedback on skin quality before a permeation experiment.


Plant holo-(acyl carrier protein) synthase.
1. An improved method was developed for the assay of plant holo-(acyl carrier protein) synthase activity, using Escherichia coli acyl-(acyl carrier protein) synthetase as a coupling enzyme. 2. Holo-(acyl carrier protein) synthase was partially purified from spinach (Spinacia oleracea) leaves by a combination of (NH4)2SO4 fractionation and anion-exchange and gel-permeation chromatography. 3. The partially purified enzyme had a pH optimum of 8.2 and Km values of 2 microM, 72 microM and 3 mM for apo-(acyl carrier protein), CoA and Mg2+ respectively. Synthase activity was inhibited in vitro by the reaction product 3',5'-ADP. 4. Results from the fractionation of spinach leaf and developing castor-oil-seed (Ricinus communis) endosperm cells were consistent with a cytosolic localization of holo-(acyl carrier protein) synthase activity in plant cells.


Back to the future: encephalitis lethargica as an autoimmune disorder?
More than 100 years after its emergence, the exact pathophysiological mechanisms underlying encephalitis lethargica (EL) are still elusive and awaiting convincing and complete elucidation. This article summarizes arguments proposed over time to support or refute the hypothesis of EL as an autoimmune neuropsychiatric disorder triggered by an infectious process. It also provides a critical evaluation of modern cases labeled as EL and a comprehensive differential diagnosis of autoimmune neurological conditions that could mimic EL. The evidence supporting the autoimmune nature of historical EL is sparse and not entirely convincing. It is possible that autoimmune mechanisms were involved in the pathogenesis of this disease as an idiosyncratic response to a yet unidentified infectious agent in genetically predisposed individuals. Although there has been an increase in the incidence of presumed autoimmune encephalomyelitis since the peak of EL pandemics, most evidence does not support an underlying autoimmune mechanism. There are significant differences between historical and recent EL cases in terms of clinical symptomatology, epidemiology, and neuropathological features, suggesting that they are different entities with only superficial similarity. The term "encephalitis lethargica," still frequently used in the medical literature, should not be used for cases occurring at present in the sporadic form. Historical EL should be kept apart from recent EL, as they differ in important aspects.


Inside-out raising mucosal-tympanomeatal flap approach for the repair of large marginal perforations.
This study evaluated the graft success rate and hearing outcomes of the inside-out raising mucosal-tympanomeatal flap technique for the repair of large marginal perforations. Prospective case series. The study enrolled patients with large marginal perforations who underwent endoscopic cartilage myringoplasty with the inside-out raising mucosal-tympanomeatal flap technique. The graft success rate, hearing outcomes, and complications were evaluated at 12 months postoperatively. In total, 48 patients with large marginal perforations were included. 81% of the population had large perforation, 14.6 had subtotal and total perforation was seen in 4.2%. The mean operation time was 38.6 ± 7.1 min. At 12 months postoperatively, the graft success rate was 89.6% (43/48). The mean air-bone gap was 25.6 ± 5.2 dB preoperatively and 16.5 ± 4.1 dB at 12 months postoperatively, with significant differences between these values (p = 0.001). The functional success rate was 85.4% (41/48). None of the patients experienced worsened sensorineural hearing loss or graft-related complications, such as graft lateralization, significant blunting, and graft medialization, during follow-up. Endoscopic cartilage-perichondrium myringoplasty for the repair of large marginal perforations using the inside-out raising mucosal-tympanomeatal flap technique was associated with satisfactory graft outcomes and minimal complications.


Molecular Identification of Free-Living Amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) Isolated from Un-improved Hot Springs, Guilan Province, Northern Iran.
This study was conducted to determine the presence and molecular identify of Acanthamoeba, Naegleria and Vermamoeba in unimproved hot springs. From Jul to Aug 2017, 54 water samples were collected from hot springs in different parts of the Guilan Province, North Iran. For the isolation of Acanthamoeba, Naegleria and Vermamoeba approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane with a pore size of 0.45 μm. The filter was transferred onto non-nutrient agar plates seeded with Gram-negative bacteria (Escherichia coli) as a food source. The morphological key of page was used to identify free-living amoebae (FLA) using an inverted microscope, PCR amplification targeting specific genes for each genus and sequencing determined frequent species and genotypes base on NCBI database. Fifteen of the 54 samples were positive by culture and/or PCR for Acanthamoeba and other FLA from unimproved hot springs. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba castellanii (12 case isolates belonged to T4 genotype), 4 cases of V. vermiformis, and 3 cases of N. australiensis, 2 cases of N. pagei and 1 cases of N. gruberi. Although FLA-mediated illnesses are not as high as in environmental distribution, but because of a poor prognosis, more investigations about FLA distribution in hot springs is critical. Hot spring may enhance exposure of the amoebae in individuals. Hence, more attention to unimproved hot springs is needed to prevent free-living amoebae mediated diseases.


Immunization uptake and its determinants among the internal migrant population living in nonnotified slums of Hyderabad city, India.
The aim of this study is to assess the childhood immunization uptake and its determinants among the internal migrant population living in nonnotified slums of Hyderabad City, India. This is a cross-sectional study of 768 rural-urban migrant mothers with a child under 2 years of age residing for period minimum of 30 days, but not more than 10 years. Data were collected for sociodemographic details, health-seeking behavior, antenatal, postnatal services, and reception of vaccines appropriate for age. Full immunization coverage among the children of migrants was same as the general population of the State of Telangana (66.7%). The likelihood of child's reception of full immunization decreases with age of the mother and rose with the attainment of higher education. The head of household of salaried class, expectant mothers utilizing antenatal services, and the visit of postnatal health worker for counseling of expectant mothers, were significantly associated with reception of full immunization. Immunization coverage rates of children of 12-23 months age is lower than the general population of Telangana. The frequency of visits by health worker is low. Immunization uptake among the migrants and vulnerable segments of the population can be increased by locating new settlements, improving utilization of services and capacity building of health staff.


A poly (saccharide-ester-urethane) scaffold for mammalian cell growth.
Chitosan and poly(3-hydroxybutyrate) are non-toxic, biodegradable, and biocompatible polymers extensively used in regenerative medicine. However, it is unknown whether the chemical combination of these polymers can produce a biomaterial that induces an appropriate cellular response in vitro in mammalian cells. This study aimed to test the ability of a novel salt-leached polyurethane scaffold of chitosan grafted with poly(3-hydroxybutyrate) to support the growth of three mammalian cell lines of different origin: a) HEK-293 cells, b) i28 mouse myoblasts, and c) human dermal fibroblasts. The viability of the cells was assessed by either evaluation of their capacity to maintain the expression of the green fluorescent protein by adenoviral transduction or by esterase activity and plasma membrane integrity. The results indicated that the three cell lines attached well to the scaffold; however, when i28 cells were induced to differentiate, they did not produce morphologically distinct myofibers, and cell growth ceased. In conclusion, the findings reveal that, altogether, these observations suggest that this foam scaffold supports cell growth and proliferation but may not apply to all cell types. Hence, one crucial question yet to be resolved is a poly (saccharide-ester-urethane) derivative with a nano-topography that elicits a similar cellular response for different biological environments.


Vibrotactile thresholds at the fingertip, volar forearm, large toe, and heel.
Thresholds for the perception of vibration vary with location on the body due to the organization of tactile channels in hairy and non-hairy skin, and variations in receptor density. This study determined vibration thresholds at four locations on the body with two different contactors so as to assist the identification of the tactile channel determining the threshold at each location. Vibrotactile thresholds at six frequencies from 8 to 250 Hz were measured on the distal phalanx of the index finger, the volar forearm, the large toe, and the heel with two contactors: (i) a 1-mm diameter circular probe with a 1-mm gap to a fixed circular surround (i.e., 7.1-mm(2) excitation area), and (ii) a 6-mm diameter circular probe with a 2-mm gap to a fixed circular surround (i.e., 79-mm(2) excitation area). At all frequencies and with both contactors, thresholds on the fingertip were lower than thresholds on the volar forearm, the large toe, and the heel, consistent with a greater density of mechanoreceptors at the fingertip. Thresholds with the larger contactor were lower than thresholds with the smaller contactor on the fingertip at high frequencies (63, 125, and 250 Hz), on the large toe (except at 250 Hz), on the heel (at all frequencies), and on the volar forearm at 250 Hz. It is concluded that at least two tactile channels (Pacinian from 63 to 250 Hz, and non-Pacinian from 8 to 31.5 Hz) determined vibrotactile thresholds at the fingertip, whereas non-Pacinian channels had a dominant influence on vibrotactile thresholds at the volar forearm. The role of Pacinian and non-Pacinian channels could not be confirmed at the large toe or the heel despite some evidence of spatial summation.


Effects of Nosema apis, N. ceranae, and coinfections on honey bee (Apis mellifera) learning and memory.
Western honey bees (Apis mellifera) face an increasing number of challenges that in recent years have led to significant economic effects on apiculture, with attendant consequences for agriculture. Nosemosis is a fungal infection of honey bees caused by either Nosema apis or N. ceranae. The putative greater virulence of N. ceranae has spurred interest in understanding how it differs from N. apis. Little is known of effects of N. apis or N. ceranae on honey bee learning and memory. Following a Pavlovian model that relies on the proboscis extension reflex, we compared acquisition learning and long-term memory recall of uninfected (control) honey bees versus those inoculated with N. apis, N. ceranae, or both. We also tested whether spore intensity was associated with variation in learning and memory. Neither learning nor memory differed among treatments. There was no evidence of a relationship between spore intensity and learning, and only limited evidence of a negative effect on memory; this occurred only in the co-inoculation treatment. Our results suggest that if Nosema spp. are contributing to unusually high colony losses in recent years, the mechanism by which they may affect honey bees is probably not related to effects on learning or memory, at least as assessed by the proboscis extension reflex.


Effect of continuum-of-care intervention package on improving contacts and quality of maternal and newborn healthcare in Ghana: a cluster randomised controlled trial.
To evaluate the effect of a continuum-of-care intervention package on adequate contacts of women and newborn with healthcare providers and their reception of high-quality care. Cluster randomised controlled trial. 32 subdistricts in 3 rural sites in Ghana. The baseline survey involved 1480 women who delivered before the trial, and the follow-up survey involved 1490 women who received maternal and newborn care during the trial. The intervention package included training healthcare providers, using an educational and recording tool named 'continuum-of-care card', providing the first postnatal care (PNC) by retaining women and newborns at healthcare facility or home visit by healthcare providers. Adequate contacts were defined as at least four contacts during pregnancy, delivery with assistance of skilled healthcare providers at a healthcare facility and three timely contacts within 6 weeks postpartum. High-quality care was defined as receiving 6 care items for antenatal care (ANC), 3 for peripartum care (PPC) and 14 for PNC. The difference-in-difference method was used to assess the effects of the intervention on the study outcome. The percentage of adequate contacts with high-quality care in the intervention group in the follow-up survey and the adjusted difference-in-difference estimators were 12.6% and 2.2 (p=0.61) at ANC, 31.5% and 1.9 (p=0.73) at PPC and 33.7% and 12.3 (p=0.13) at PNC in the intention-to-treat design, whereas 13.0% and 2.8 (p=0.54) at ANC, 34.2% and 2.7 (p=0.66) at PPC and 38.1% and 18.1 (p=0.02) at PNC in the per-protocol design that assigned the study sample by possession of the continuum-of-care card. The interventions improved contacts with healthcare providers and quality of care during PNC. However, having adequate contact did not guarantee high-quality care. Maternal and newborn care in Ghana needs to improve its continuity and quality. ISRCTN90618993. .


Parental Stress as a Child With Diabetes Transitions From Adolescence to Emerging Adulthood.
This study examined parental and caregiver distress among families caring for children with type 1 diabetes as the child transitions into Emerging Adulthood. More than 96 hours of semistructured interviews were conducted with 19 adult caregivers including parents, grandparents, and other adult family members of 10 children. Each research partner participated in multiple face-to-face, 1- to 1.5-hour long-evolving interviews over the course of 4.5 years. Paradoxically, caregivers were found to experience significant increase in distress as their child with diabetes entered the developmental stage of Emerging Adulthood, 18 to 25 years old, by which time they should be masters of self-care, and parental distress should begin to decline. This increase in familial distress was associated with the emerging adults leaving the home, being unable to maintain an acceptable level of self-care, and experiencing declining health, frequent visits to the emergency department, and repeated hospitalizations. These findings suggest that parental distress from caring for a child with diabetes continues as the child ages, matures, and transitions into adulthood and may be exacerbated when the emerging adult with type 1 diabetes leaves the home and the direct observation and care of the parent.


Synthesis of α,α-difluoromethylene alkynes by palladium-catalyzed gem-difluoropropargylation of aryl and alkenyl boron reagents.
gem-Difluoropropargyl bromides are versatile intermediates in organic synthesis, but have rarely been employed in transition-metal-catalyzed cross-coupling reactions. The first palladium-catalyzed gem-difluoropropargylation of organoboron reagents with gem-difluoropropargyl bromides is now reported. The reaction proceeds under mild reaction conditions with high regioselectivity; it features a broad substrate scope and excellent functional-group compatibility and thus provides an attractive approach for the synthesis of complex fluorinated molecules, in particular for drug discovery and development.


Phosphite Integrated in Late Blight Treatment Strategies in Starch Potato Does Not Cause Residues in the Starch Product.
Currently available fungicides against potato late blight are effective but there are concerns about the sustainability of frequent applications and the risks of fungicide resistance. Therefore, we investigated how potassium phosphite can be integrated into late blight control programs with reduced fungicides in field trials. Phosphite was somewhat less effective than the conventional fungicides at suppressing late blight in the foliage, and the tubers contained less starch. However, when we reduced the amount of phosphite and combined it with reduced amounts of conventional fungicides, we observed no differences in disease suppression, total yields, and tuber starch contents compared with the full treatments with conventional fungicides. The amount of phosphite detected in the harvested tubers was linearly associated with the amount of phosphite applied to the foliage. Our analyses indicate that phosphite could replace some fungicides without exceeding the current European Union standards for the maximum residue levels in potato tubers. No phosphite was detected in the starch from the tubers. In 1 of 2 years, early blight (caused by Alternaria solani) was less severe in the phosphite treatments than in the treatments without phosphite. The integration of phosphite into current treatment strategies would reduce the dependence on conventional fungicides.


Perspectives on transgenic, herbicide-resistant crops in the United States almost 20 years after introduction.
Herbicide-resistant crops have had a profound impact on weed management. Most of the impact has been by glyphosate-resistant maize, cotton, soybean and canola. Significant economic savings, yield increases and more efficacious and simplified weed management have resulted in widespread adoption of the technology. Initially, glyphosate-resistant crops enabled significantly reduced tillage and reduced the environmental impact of weed management. Continuous use of glyphosate with glyphosate-resistant crops over broad areas facilitated the evolution of glyphosate-resistant weeds, which have resulted in increases in the use of tillage and other herbicides with glyphosate, reducing some of the initial environmental benefits of glyphosate-resistant crops. Transgenic crops with resistance to auxinic herbicides, as well as to herbicides that inhibit acetolactate synthase, acetyl-CoA carboxylase and hydroxyphenylpyruvate dioxygenase, stacked with glyphosate and/or glufosinate resistance, will become available in the next few years. These technologies will provide additional weed management options for farmers, but will not have all of the positive effects (reduced cost, simplified weed management, lowered environmental impact and reduced tillage) that glyphosate-resistant crops had initially. In the more distant future, other herbicide-resistant crops (including non-transgenic ones), herbicides with new modes of action and technologies that are currently in their infancy (e.g. bioherbicides, sprayable herbicidal RNAi and/or robotic weeding) may affect the role of transgenic, herbicide-resistant crops in weed management. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.


The effects of a combined strength and aerobic exercise program on glucose control and insulin action in women with type 2 diabetes.
The purpose of the present study was to investigate the short- and long-term effects of a combined strength and aerobic training program on glycemic control, insulin action, exercise capacity and muscular strength in postmenopausal women with type 2 diabetes. Nine postmenopausal women, aged 55.2 (6.7) years, with type 2 diabetes participated in a supervised training program for 4 months consisting of two strength training sessions (3 sets of 12 repetitions at 60% one-repetition maximum strength) and two aerobic training sessions (60-70% of maximum heart rate at the beginning, and 70-80% of maximum heart rate after 2 months). Anthropometrical measurements, percentage glycated hemoglobin, a 2-h oral glucose tolerance test, exercise stress testing and maximum strength were measured at the beginning, and after 4 and 16 weeks of the exercise program. Significant reductions were observed in both the glucose (8.1% P<0.01) and insulin areas under the curve (20.7%, P<0.05) after 4 weeks of training. These adaptations were further improved after 16 weeks (glucose 12.5%, insulin 38%, P<0.001). Glycated hemoglobin was significantly decreased after 4 weeks [7.7 (1.7) vs 7.1 (1.3)%, P<0.05] and after 16 weeks [7.7 (1.7) vs 6.9 (1.0)%, P<0.01] of exercise training. Furthermore, exercise time and muscular strength were significantly improved after 4 weeks (P<0.01) as well as after 16 weeks (P<0.001) of training. Body mass and body-mass index, however, were not significantly altered throughout the study. The results indicated that a combined training program of strength and aerobic exercise could induce positive adaptations on glucose control, insulin action, muscular strength and exercise tolerance in women with type 2 diabetes.


Anticardiolipin antibodies in stroke patients in Sudan. Appraisal of their significance in a region of high burden of endemic infections.
To screen and evaluate the significance of anticardiolipin seroprevalence in patients with acute ischemic stroke, in patients with infectious disease, and in healthy subjects resident in Sudan, a tropical country endemic for several infectious diseases. We conducted the study in Khartoum Teaching Hospital in Khartoum, Sudan between July 2003 and January 2005. We included 89 stroke cases, 30 infectious disease patients, and 30 asymptomatic healthy subjects. We estimated IgG and IgM anticardiolipin antibody titers in serum samples from all subjects in the 3 study categories at the time of hospital admission. We found a significantly higher prevalence of anticardiolipin antibodies in the stroke and infection groups compared to the healthy subjects. However, there was no significant difference in anticardiolipin seroprevalence between patients with stroke and patients with infectious disease. Caution is necessary when interpreting the presence of antiphospholipid antibodies as a stroke risk in patients harboring infection or living in places with high endemicity of infectious diseases.


Modelling of filamentous phage-induced antibiotic tolerance of P. aeruginosa.
Filamentous molecules tend to spontaneously assemble into liquid crystalline droplets with a tactoid morphology in environments with high concentration on non-adsorbing molecules. Tactoids of filamentous Pf bacteriophage, such as those produced by Pseudomonas aeruginosa, have been linked to increased antibiotic tolerance. We modelled this system and show that tactoids composed of filamentous Pf virions can lead to antibiotic tolerance by acting as an adsorptive diffusion barrier. The continuum model, reminiscent of descriptions of reactive diffusion in porous media, has been solved numerically and good agreement was found with the analytical results, obtained using a homogenisation approach. We find that the formation of tactoids significantly increases antibiotic diffusion times which may lead to stronger antibiotic resistance.


A model of locomotor-respiratory coupling in quadrupeds.
Locomotion and respiration are not independent phenomena in running mammals because locomotion and respiration both rely on cyclic movements of the ribs, sternum, and associated musculature. Thus, constraints are imposed on locomotor and respiratory function by virtue of their linkage. Specifically, locomotion imposes mechanical constraints on breathing that require the respiratory cycle to be synchronized with gait. Thus, many mammals, including humans, synchronize respiration with the movement of the limbs during locomotion. For example, quadrupeds synchronize locomotor and respiratory cycles at a 1:1 ratio (stride/breath) over a wide range of speeds. Interestingly, quadrupeds maintain an almost constant stride frequency (and therefore respiratory frequency) at different speeds. To increase speed, quadrupeds lengthen their stride. Accordingly, to increase minute ventilation, quadrupeds must increase tidal volume since respiratory rate is coupled with stride frequency. We developed a simple, inexpensive, and easy to build model to demonstrate this concept. A model was chosen because models significantly enhance student understanding. Students are drawn into discussion by the power of learning that is associated with manipulating and thinking about objects. Building and using this model strengthen the concept that locomotor-respiratory coupling provides a basis for the appropriate matching of lung ventilation to running speed and metabolic power.


Human regulatory T cells with alloantigen specificity are more potent inhibitors of alloimmune skin graft damage than polyclonal regulatory T cells.
Graft rejection by the immune system is a major cause of transplant failure. Lifelong immunosuppression decreases the incidence of graft rejection; however, nonspecific immunosuppression results in increased susceptibly to infection and cancer. Regulatory T cells (T(regs)), which suppress the activation of the immune system and induce tolerance, are currently under evaluation for use in clinical transplantation. Ex vivo expanded polyclonal T(regs) that are introduced into transplant recipients alter the balance of T effector cells to T(regs); however, experimental data suggest that alloantigen-specific T(regs) would be more effective at preventing graft rejection. We have developed a method to enrich alloantigen-specific human T(regs) based on the coexpression of activation markers, CD69 and CD71. These T(regs) could be readily expanded in vitro and demonstrated potent antigen-specific suppression. In a humanized mouse model of alloimmune-mediated injury of human skin grafts, alloantigen-specific T(regs) resulted in a significant reduction in clinically relevant indicators of dermal tissue injury when compared with polyclonal T(regs), restoring a histology comparable to healthy skin. This method of human allospecific T(reg) selection should be scalable to the clinic. The improved in vivo efficacy of alloantigen-specific T(regs) over polyclonal T(regs) shown here suggests that generating "customized" T(regs) with defined anti-donor allospecificities may improve current practice in clinical immunotherapy.


Therapeutic effects of Salvia miltiorrhiza injection combined with telmisartan in patients with diabetic nephropathy by influencing collagen IV and fibronectin: A case-control study.
Involvement of collagen IV (ColIV) and fibronectin (FN) in the occurrence and development of diabetic nephropathy (DN) and the effects of telmisartan and Salvia miltiorrhiza injection in the treatment of the patients were investigated. Two hundred and fifty-eight patients with stage IV DN were selected as the case group, and another 110 normal healthy subjects were incorporated as the control group. Involved patients were subdivided into different groups according to different treatment therapies; patients in the telmisartan group (T group) were given oral telmisartan; patients in the Salvia miltiorrhiza injection + telmisartan (S + T group) were administered with Salvia miltiorrhiza injection combined with telmisartan treatment, and there was a group of patients who received no intervention as the placebo group. After intervention, levels of glycemic indexes and renal damage indexes indicated downwards trends both in the T group and the S + T group when compared to the placebo group; besides, levels in the S + T group were much lower than those in the T group (all P<0.05). Additionally, in comparison among the above three intervention groups, differences in the fasting blood glucose, 2 h post-prandial blood glucose, glycosylated hemoglobin, blood urea nitrogen, serum creatinine and urinary albumin excretion rate were significant after treatment (all P<0.05). Further, before intervention, both Co1IV and FN in the urine were increased in the case group compared to the control group (all P<0.05). After intervention, both levels were apparently decreased. There were remarkable differences of Co1IV and FN levels in the urine when compared among three different intervention groups after treatment (P<0.05). Increased ColIV and FN levels may be partially responsible for the development of DN. Salvia miltiorrhiza injection with telmisartan have beneficial synergistic effects for DN patients through attenuating the increase in ColIV and FN, reversing hyperglycemia state and postponing ultrastructure changes of glomerular basement membrane.


Neonatal poisoning: causes, manifestations, prevention, and management.
Neonates are exposed to a growing list of potentially toxic environmental and therapeutic agents. Such exposure may be through mishap, misjudgment in both drug dosage and route of administration, inadvertent misuse of medications, or improper baby care. In addition, the newborn infant may show signs of adverse reactions to a variety of drugs that are given to the mother and cross the placenta, thereby affecting the fetus before or during both labor and delivery. Because of the immaturity of the neonate's liver, lungs, kidneys, and central nervous system, the process of metabolizing and eliminating drugs and poisons is delayed. Moreover, the clinical signs and symptoms of poisoning are not always immediately recognized. Through reviewing various drugs and environmental pollutants, this article places emphasis on prevention rather than treatment.


Safety and Efficacy of Andecaliximab (GS-5745) Plus Gemcitabine and Nab-Paclitaxel in Patients with Advanced Pancreatic Adenocarcinoma: Results from a Phase I Study.
Matrix metalloproteinase 9 (MMP9) expression in the tumor microenvironment is implicated in multiple protumorigenic processes. Andecaliximab (GS-5745), a monoclonal antibody targeting MMP9 with high affinity and selectivity, was evaluated in combination with gemcitabine and nab-paclitaxel in patients with advanced pancreatic adenocarcinoma. This phase I study was completed in two parts: part A was a dose-finding, monotherapy phase that enrolled patients with advanced solid tumors, and part B examined andecaliximab in combination with chemotherapy in specific patient cohorts. In the cohort of patients with pancreatic adenocarcinoma (n = 36), andecaliximab 800 mg every 2 weeks was administered in combination with gemcitabine and nab-paclitaxel. Patients were treated until unacceptable toxicity, withdrawal of consent, disease progression, or death. Efficacy, safety, and biomarker assessments were performed. Andecaliximab combined with gemcitabine and nab-paclitaxel appeared to be well tolerated and did not demonstrate any unusual toxicities in patients with pancreatic adenocarcinoma. The most common treatment-emergent adverse events were fatigue (75.0%), alopecia (55.6%), peripheral edema (55.6%), and nausea (50.0%). Median progression-free survival was 7.8 months (90% confidence interval, 6.9-11.0) with an objective response rate of 44.4% and median duration of response of 7.6 months. Maximal andecaliximab target binding, defined as undetectable, andecaliximab-free MMP9 in plasma, was observed. Andecaliximab in combination with gemcitabine and nab-paclitaxel demonstrates a favorable safety profile and clinical activity in patients with advanced pancreatic adenocarcinoma. The combination of andecaliximab, a novel, first-in-class inhibitor of matrix metalloproteinase 9, with gemcitabine and nab-paclitaxel in patients with advanced pancreatic adenocarcinoma provided a median progression-free survival of 7.8 months and objective response rate of 44.4%. The majority of systemic biomarkers related to matrix metalloproteinase 9 activity and immune suppression increased at 2 months, whereas biomarkers related to tumor burden decreased. Although this study demonstrates promising results with andecaliximab plus chemotherapy in patients with advanced pancreatic adenocarcinoma, andecaliximab was not associated with a survival benefit in a phase III study in patients with advanced gastric/gastroesophageal junction carcinoma.


Duplex real-time PCR assay for detection of Streptococcus pneumoniae in clinical samples and determination of penicillin susceptibility.
We have developed a duplex real-time PCR for the rapid diagnosis of Streptococcus pneumoniae infection from culture-negative clinical samples with the simultaneous determination of penicillin susceptibility. The assay amplifies a lytA gene target and a penicillin binding protein 2b (pbp2b) gene target in penicillin-susceptible organisms. The assay was shown to be sensitive (detects 0.5 CFU per PCR) and specific for the detection of S. pneumoniae DNA. The assay was validated by comparing pbp2b PCR results with MIC data for 27 S. pneumoniae isolates. All 5 isolates with penicillin MICs of > 1.0 mg/liter were pbp2b real-time PCR negative, as were 9 of the 10 isolates with penicillin MICs of 0.12 to 1.0 mg/liter. One isolate with a penicillin MIC of 0.12 to 1.0 mg/liter gave an equivocal pbp2b real-time PCR result. Twelve isolates were penicillin susceptible (MICs of < or = 0.06 mg/liter) and pbp2b real-time PCR positive. These data were used to establish an algorithm for the interpretation of penicillin susceptibility from the duplex PCR result. pbp2b real-time PCR results were also compared to an established PCR-restriction fragment length polymorphism (RFLP) method previously applied to these 27 isolates and 46 culture-negative clinical samples (containing S. pneumoniae DNA by broad-range 16S rRNA gene PCR). Discordant results were seen for four isolates and six culture-negative clinical samples, as PCR-RFLP could not reliably detect penicillin MICs of 0.12 to 1.0 mg/liter. We report prospective application of the duplex PCR assay to the diagnosis of S. pneumoniae infection from 200 culture-negative clinical specimens sent to the laboratory for diagnostic broad-range 16S rRNA gene PCR. One hundred six were negative in the duplex PCR. Ninety-four were lytA PCR positive, and 70 of these were also pbp2b PCR positive and interpreted as penicillin susceptible. Fourteen were pbp2b PCR negative and interpreted as having reduced susceptibility to penicillin. For the remaining 10 samples, susceptibility to penicillin was not determined.


A gene signature predicting for survival in suboptimally debulked patients with ovarian cancer.
Despite the existence of morphologically indistinguishable disease, patients with advanced ovarian tumors display a broad range of survival end points. We hypothesize that gene expression profiling can identify a prognostic signature accounting for these distinct clinical outcomes. To resolve survival-associated loci, gene expression profiling was completed for an extensive set of 185 (90 optimal/95 suboptimal) primary ovarian tumors using the Affymetrix human U133A microarray. Cox regression analysis identified probe sets associated with survival in optimally and suboptimally debulked tumor sets at a P value of <0.01. Leave-one-out cross-validation was applied to each tumor cohort and confirmed by a permutation test. External validation was conducted by applying the gene signature to a publicly available array database of expression profiles of advanced stage suboptimally debulked tumors. The prognostic signature successfully classified the tumors according to survival for suboptimally (P = 0.0179) but not optimally debulked (P = 0.144) patients. The suboptimal gene signature was validated using the independent set of tumors (odds ratio, 8.75; P = 0.0146). To elucidate signaling events amenable to therapeutic intervention in suboptimally debulked patients, pathway analysis was completed for the top 57 survival-associated probe sets. For suboptimally debulked patients, confirmation of the predictive gene signature supports the existence of a clinically relevant predictor, as well as the possibility of novel therapeutic opportunities. Ultimately, the prognostic classifier defined for suboptimally debulked tumors may aid in the classification and enhancement of patient outcome for this high-risk population.


Iron-independent phosphorylation of iron regulatory protein 2 regulates ferritin during the cell cycle.
Iron regulatory protein 2 (IRP2) is a key iron sensor that post-transcriptionally regulates mammalian iron homeostasis by binding to iron-responsive elements (IREs) in mRNAs that encode proteins involved in iron metabolism (e.g. ferritin and transferrin receptor 1). During iron deficiency, IRP2 binds IREs to regulate mRNA translation or stability, whereas during iron sufficiency IRP2 is degraded by the proteasome. Here, we identify an iron-independent IRP2 phosphorylation site that is regulated by the cell cycle. IRP2 Ser-157 is phosphorylated by Cdk1/cyclin B1 during G(2)/M and is dephosphorylated during mitotic exit by the phosphatase Cdc14A. Ser-157 phosphorylation during G(2)/M reduces IRP2 RNA-binding activity and increases ferritin synthesis, whereas Ser-157 dephosphorylation during mitotic exit restores IRP2 RNA-binding activity and represses ferritin synthesis. These data show that reversible phosphorylation of IRP2 during G(2)/M has a role in modulating the iron-independent expression of ferritin and other IRE-containing mRNAs during the cell cycle.


Critical dynamics in genetic regulatory networks: examples from four kingdoms.
The coordinated expression of the different genes in an organism is essential to sustain functionality under the random external perturbations to which the organism might be subjected. To cope with such external variability, the global dynamics of the genetic network must possess two central properties. (a) It must be robust enough as to guarantee stability under a broad range of external conditions, and (b) it must be flexible enough to recognize and integrate specific external signals that may help the organism to change and adapt to different environments. This compromise between robustness and adaptability has been observed in dynamical systems operating at the brink of a phase transition between order and chaos. Such systems are termed critical. Thus, criticality, a precise, measurable, and well characterized property of dynamical systems, makes it possible for robustness and adaptability to coexist in living organisms. In this work we investigate the dynamical properties of the gene transcription networks reported for S. cerevisiae, E. coli, and B. subtilis, as well as the network of segment polarity genes of D. melanogaster, and the network of flower development of A. thaliana. We use hundreds of microarray experiments to infer the nature of the regulatory interactions among genes, and implement these data into the Boolean models of the genetic networks. Our results show that, to the best of the current experimental data available, the five networks under study indeed operate close to criticality. The generality of this result suggests that criticality at the genetic level might constitute a fundamental evolutionary mechanism that generates the great diversity of dynamically robust living forms that we observe around us.


Detecting hyperbolic geometry in networks: Why triangles are not enough.
In the past decade, geometric network models have received vast attention in the literature. These models formalize the natural idea that similar vertices are likely to connect. Because of that, these models are able to adequately capture many common structural properties of real-world networks, such as scale invariance and high clustering. Indeed, many real-world networks can be accurately modeled by positioning vertices of a network graph in hyperbolic spaces. Nevertheless, if one observes only the network connections, the presence of geometry is not always evident. Currently, triangle counts and clustering coefficients are the standard statistics to signal the presence of geometry. In this paper we show that triangle counts or clustering coefficients are insufficient because they fail to detect geometry induced by hyperbolic spaces. We, therefore, introduce a differerent statistic, weighted triangles, which weighs triangles based on their evidence for geometry. We show analytically, as well as on synthetic and real-world data, that weighted triangles are a powerful statistic to detect hyperbolic geometry in networks.