Beneficial effect of zinc chloride and zinc ionophore pyrithione on attenuated cardioprotective potential of preconditioning phenomenon in STZ-induced diabetic rat heart.
Ischemic preconditioning (IPC) is well demonstrated to produce cardioprotection by phosphorylation and subsequent inactivation of glycogen synthase kinase-3β (GSk-3β) in the normal rat heart, but its effect is attenuated in the diabetic rat heart. This study was designed to investigate the effect of zinc chloride and zinc ionophore pyrithione (ZIP) on the attenuated cardioprotective potential of IPC in the diabetic rat heart. Diabetes mellitus (DM) was induced by a single intraperitoneal administration of streptozotocin (STZ) (50 mg/kg; i.p). The isolated perfused rat heart was subjected to 30 minutes of ischemia followed by 120 minutes of reperfusion. Myocardial infarct size was estimated by triphenyltetrazolium chloride (TTC) staining and cardiac injury was measured by estimating lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) in the coronary effluent. Also, GSK-3β was measured and neutrophil accumulation was measured by estimating myeloperoxidase (MPO) levels. IPC significantly decreased the myocardial infarct size, the release of LDH and CK-MB, the GSK-3β levels and the MPO levels in the normal rat heart. Pre- and post-ischemic treatment with zinc chloride and zinc ionophore pyrithione (ZIP) in the normal and diabetic rat hearts significantly decreased the myocardial infarct size, the level of CK-MB and LDH in the coronary effluent and GSK-3β and MPO levels. Our results suggest that pharmacological preconditioning with zinc chloride and ZIP significantly restored the attenuated cardioprotective potential of IPC in the diabetic rat heart.


Identification of perirenal or renal splenosis from undetermined masses: case report and review of the literature.
The incidental finding of tumor-llke perirenal or renal splenosis (PRS) creates a challenge to the renal physicians, because its benign nature has to be distinguished from a malignancy. This paper describes the case of a 40-year-old man referred from a local clinic for further evaluation of an incidental finding of left abdominal masses by ultrasonogram suspecting neoplasm, but was eventually confirmed as PRS by obtaining a history of splenectomy that pointed to splenosis and subsequently by a fusion image from single photon emission computed tomography using 99mTc-labelled heat-denatured erythrocytes and computed tomography (hybrid SPECT/CT). In addition, a review of 27 cases of PRS in a MEDLINE search including the present case revealed the following: all the masses were found incidentally and were associated with a history of previous splenectomy or splenic injury; the initial impressions were neoplastic tumor/PRS (n = 9), PRS (n = 10), and neoplastic tumor without consideration of splenosis (n = 8); surgical exploration was undertaken in all the 8 cases of suspected neoplastic tumor only, whereas non-invasive radiological or radionuclide imaging confirmed splenosis in the rest of the cases (n = 19). To avoid unnecessary tests and invasive surgery for undetermined perirenal or renal masses accompanying previous splenic injury, we stress the paramount importance of careful history-taking, physical examination, and a high index of suspicion for splenosis. Also, fusion imaging of hybrid SPECT/CT was reconfirmed as a useful diagnostic technique for accurately detecting and localizing splenic tissues by PRS.


[CD271 enrichment does not help isolating mesenchymal stromal cells from].
Reports on the isolation of mesencnymal stromal cells (MSCs) from granulocyte colony stimulating factor mobilized peripheral blood (G-CSF-mobilized PB) using regular culturing techniques are controversial. Enrichment techniques such as CD133 isolation have increased the success rates. CD271 is a well-known marker for enrichment of MSCs from bone marrow (BM). In the present study, we aimed to find out whether CD271 enrichment can help isolation of MSCs from G-CSF-mobilized PB. Five G-CSF-mobilized PB samples were collected from the remnant parts of the bags used for BM transplantation. Five BM samples were used as the control. Mononuclear cells (MNCs) from both resources were collected and underwent magnetic sorting for CD271-positive cells. The isolated cells were cultured, undergoing flowcytometry and differentiation assays to determine if they fulfill MSCs characteristics. CD271-positive portion of G-CSF-mobilized PB did not yield any cell outgrowth but the BM counterpart could successfully form MSC colonies. Although the percentage of CD271+ cells showed no difference between BM-MNCs and G-CSF-mobilized PB-MNCs, hematopoietic markers such as CD45, CD34 and CD133 composed a higher percentage of CD271-positive cells in the G-CSF-mobilized PB group. Results obtained indicated that CD271 enrichment does not help isolation of MSCs from G-CSF-mobilized PB. In this source, almost all of the CD271+ cells are from hematopoietic origin and the frequency of MSCs is so low that possibly during the process of cell isolation most of them are lost and the isolation fails.


Surveillance of Chigger Mite Vectors for Tsutsugamushi Disease in the Hwaseong Area, Gyeonggi-do, Republic of Korea, 2015.
Owing to global climate change, the global resurgence of vector-borne infectious diseases and their potential to inflict widespread casualties among human populations has emerged as a pivotal burden on public health systems. Tsutsugamushi disease (scrub typhus) in the Republic of Korea is steadily increasing and was designated as a legal communicable disease in 1994. The disease is a mite-borne acute febrile disease most commonly contracted from October to December. In this study, we tried to determine the prevalence of tsutsugamushi disease transmitted by chigger mites living on rodents and investigated their target vector diversity, abundance, and distribution to enable the mapping of hotspots for this disease in 2015. A total of 5 species belonging to 4 genera (109 mites): Leptotrombidium scutellare 60.6%, L. pallidum 28.4% Neotrombicula tamiyai 9.2%, Euschoengastia koreaensis/0.9%), and Neoschoengastia asakawa 0.9% were collected using chigger mite collecting traps mimicking human skin odor and sticky chigger traps from April to November 2015. Chigger mites causing tsutsugamushi disease in wild rodents were also collected in Hwaseong for the zoonotic surveillance of the vector. A total of 77 rodents belonging to 3 genera: Apodemus agrarius (93.5%), Crocidura lasiura (5.2%), and Micromys minutus (1.3%) were collected in April, October, and November 2015. The most common mite was L. pallidum (46.9%), followed by L. scutellare (18.6%), and L. orientale (18.0%). However, any of the chigger mite pools collected from rodent hosts was tested positive for Orientia tsutsugamushi, the pathogen of tsutsugamushi disease, in this survey.


Cell-mediated immunity in flavivirus infections. I. Induction of cytotoxic T lymphocytes in mice by an attenuated virus from the tick-borne encephalitis complex and its group-reactive character.
The lytic activity of splenocytes from C3H mice immunized with a highly attenuated line of Langat virus [tick-borne encephalitis (TBE) complex] was determined by the in vitro 51Cr release assay on TBE virus (western subtype)-infected target L929 cells. After the spleen cell suspensions were depleted of T cells with anti-mouse theta serum, or of B cells with anti-mouse immunoglobulin, the cytotoxic effect was found dependent on the presence of T lymphocytes. During immunization with Langat virus, cytotoxic T lymphocytes were generated, specific for both the foreign virus-specified antigen and the self component of the host system studied, coded by the K allele of the H-2 complex. The peak of T lymphocyte cytotoxic response was attained on the 6th day after administration of the live virus. T lymphocytes from mice, single-shot immunized with a flavivirus, displayed distinct cross-reactive lysis when studied on target cells infected with other flaviviruses but not on cells infected with an alphavirus.


MicroRNA Biomarkers as Promising Tools for Early Colorectal Cancer Screening-A Comprehensive Review.
Colorectal cancer (CRC) ranks as the third most prevalent cancer worldwide. Early detection of this neoplasia has proven to improve prognosis, resulting in a 90% increase in survival. However, available CRC screening methods have limitations, requiring the development of new tools. MicroRNA biomarkers have emerged as a powerful screening tool, as they are highly expressed in CRC patients and easily detectable in several biological samples. While microRNAs are extensively studied in blood samples, recent interest has now arisen in other samples, such as stool samples, where they can be combined with existing screening methods. Among the microRNAs described in the literature, microRNA-21-5p and microRNA-92a-3p and their cluster have demonstrated high potential for early CRC screening. Furthermore, the combination of multiple microRNAs has shown improved performance in CRC detection compared to individual microRNAs. This review aims to assess the available data in the literature on microRNAs as promising biomarkers for early CRC screening, explore their advantages and disadvantages, and discuss the optimal study characteristics for analyzing these biomarkers.


Comparison of buccal and intramuscular dexmedetomidine premedication for arthroscopic knee surgery.
To compare the sedative, anxiolytic, analgesic, hemodynamic, and respiratory effects of buccal dexmedetomidine with intramuscular (IM) dexmedetomidine for premedication in patients undergoing arthroscopic knee surgery during spinal anesthesia. Randomized, placebo-controlled trial. University medical center. 75 ASA physical status I and II patients undergoing arthroscopic knee surgery with spinal anesthesia. Patients were randomized to one of three groups for premedication: group B, buccal dexmedetomidine 2.5 microg kg(-1); group IM, IM dexmedetomidine 2.5 microg kg(-1); and group P, buccal 0.9% and NaCl 2 mL. Noninvasive blood pressure, heart rate, respiratory rate, and peripheral oxygen saturation were recorded. Sedation and anxiety levels were consecutively assessed with Ramsay sedation scores and Visual Analog Scale (VAS) scores of anxiety before premedication, before spinal anesthesia, during surgery, and at the end of surgery. Two, 4, and 8 hours after surgery, sedation levels, postoperative VAS pain scores, and consumption of analgesics (diclofenac sodium) were recorded. Before spinal anesthesia, during surgery, and at the end of surgery, sedation and anxiety scores of the patients receiving buccal or IM dexmedetomidine were, respectively, higher and lower than in group P. Patients receiving buccal dexmedetomidine (group B) had lower requirement of diclofenac sodium than group P and lower pain scores than groups P and IM. Mild hypotension and bradycardia were observed in the buccal and IM dexmedetomidine patients. Buccal dexmedetomidine for premedication in arthroscopic knee surgery provided equal levels of sedation and anxiolysis, and more evident analgesia compared with IM dexmedetomidine.


Resistance to extended-spectrum β-lactamases in Salmonella from a broiler supply Chain.
The prevalence of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae varies worldwide, however, the incidence of ESBL-producing environmental Salmonella isolates is increasing. Salmonella is still one of the most important pathogens that occur in the poultry supply chain. Therefore, this study analyzed the susceptibility of Salmonella isolates collected from a poultry supply chain to β-lactam antibiotics, and examined the phenotypes of the isolates based on enzyme-inducible AmpC β-lactamase analysis. All analysis of the putative positive isolates in the current study confirmed that 27.02% (77/285 analysis) of all ESBL tests realized with the isolates produced a profile of resistance consistent with β-lactamase production. All isolates of S. Minnesota serotype had ESBL phenotype. Aztreonam resistance was the least common amongst the Salmonella isolates, followed by ceftazidime. The presence of inducible chromosomal ESBL was detected in 14 different isolates of the 19 serotypes investigated. These results are very indicatives of the presence of ESBL genes in Salmonella isolates from a broiler supply chain, reaffirming the growing global problem of ESBL resistance.


Neck rigidity as a physical manifestation of higher brain hypo-function/dysfunction.
A total of 308 patients, who received inpatient rehabilitation in department of Neurology in Sapporo City General Hospital, were studied. The patients were divided into four subgroups according to the presence or absence of Parkinsonism and the location of organic neurological lesions: group A consisted of 47 patients with brain lesions who presented with Parkinsonism; group B consisted of 135 patients with manifestations deriving from brain disorders other than Parkinsonism; group C consisted of 68 patients with manifestations deriving from spinal cord, peripheral nerve, and muscle disorders; and group D consisted of 58 patients with non-organic functional disorders and patients with organic lesions not responsible for clinical manifestations. The patients were administered a battery of 5 tests for evaluating frontal lobe and/or higher brain function. In result, our study confirmed higher brain/frontal lobe dysfunction to affect the grade of neck rigidity unless there is an interruption, in the control of peripheral organs, by the brain. Consequently the association between higher brain/frontal lobe dysfunction and the grade of neck rigidity were confirmed. Although neck rigidity may be a minor clinical manifestation, it can provide important clues to brain function.


Breast Tumours Resembling the Tall Cell Variant of Thyroid Papillary Carcinoma: Are They Part of the Papillary Carcinoma Spectrum or a Distinct Entity?
Papillary tumours of the breast are diagnostically challenging lesions and represent a wide spectrum of diseases from papilloma to invasive papillary carcinoma. A rare subtype of breast papillary tumour resembling the tall cell variant of thyroid papillary carcinoma (BTRTPC) has been described. The nomenclature of this entity, its relationship to other papillary tumours, and its nature, whether in situ or invasive, remain unclear. Seventy-five papillary carcinomas (PCs) of the breast previously diagnosed in routine practice were reviewed and the presence of features (n = 10) characteristic of BTRTPC were assessed to determine whether BTRTPC comprises a distinct entity or is part of the spectrum of the previously defined PC variants. Nuclear overlapping and eosinophilic granular cytoplasm were seen in 81 and 75% of the cases, whereas nuclear grooves, nuclear clearing, and tall cells were noticed in 51, 42, and 38% of the cases, respectively; 27% of the cases showed macro- and micro-follicular architecture filled with colloid-like material. Five cases (7%) lacked oestrogen receptor (ER) expression. Co-existing invasive carcinoma was seen in 25 cases (33%). Two cases displayed several features characteristic of BTRTPC, and both were ER-negative. Features characteristic of BTRTPC overlap with other PCs of the breast. Molecular and immunohistochemical biomarkers are needed to provide objective diagnostic criteria for the characterisation of such lesions in routine practice.


Antiviral potential of interferon-omega on hepatitis B virus replication in human hepatoma cells.
Interferon-alpha (IFN-alpha) and other cytokines are able to interfere with hepatitis B virus (HBV) replication. However, a sustained antiviral effect is achieved only in 25% to 40% of the patients with chronic HBV infection and clearance of the virus rarely occurs, stressing the need for developing therapeutic alternatives. In this study the antiviral potential of a new recombinant interferon, IFN-omega was investigated. IFN-omega was assessed in comparison with IFN-alpha 2c, IFN-gamma, and TNF-alpha with respect to production of HBV proteins and DNA in HepG2.2.15 cells, a HBV-DNA transfected hepatoma cell line which produces infectious viral particles. Cells were seeded at different states of confluence (20%-90%) and treated with increasing concentrations of interferons (5 to 5,000 U/ml), TNF-alpha (5 to 500 ng/ml), or combinations of both for one to three days. IFN-omega reduced the production of HBsAg down to 59% of the untreated controls, which was comparable to the reduction obtained by treatment with IFN-alpha (60%), the standard interferon used for the treatment of chronic HBV infections. The strongest inhibition, however, was achieved by treatment with 500 ng/ml TNF-alpha (42%). Likewise, production of HBeAg and synthesis of HBV DNA were inhibited to similar degrees by the different interferons. In non-replicating high-density cultures only TNF-alpha was effective. IFN-omega is of similar antiviral potential as IFN-alpha in this in vitro experimental system.


LPS enhances TLR4 expression and IFN‑γ production via the TLR4/IRAK/NF‑κB signaling pathway in rat pulmonary arterial smooth muscle cells.
The aim of the present study was to investigate the role of the Toll‑like receptor (TLR)4 signaling pathway in cellular response to lipopolysaccharide (LPS) in rat pulmonary artery smooth muscle cells (PASMCs). Chronic obstructive pulmonary disease (COPD) rats were established with passive inhaling cigarette smoke plus injection of LPS. The TLR4 protein in lung tissues was determined with immunohistochemical staining and protein levels of the components of the TLR4 pathway in PASMCs were analyzed with western blotting. The production of interferon (IFN)‑γ upon LPS stimulation in PASMCs was measured with ELISA. TLR4 expression in lung tissue from COPD rats was increased obviously compared with that in normal group. LPS enhances TLR4 expression in rat PASMCs and induced production of IFN‑γ dramatically. LPS treatment resulted in increased phosphor‑interleukin‑1 receptor‑associated kinase (IRAK), IκB and IκB kinase, as well as the total protein of nuclear factor (NF)‑κB p65. TLR4 inhibitor TAK‑242, IRAK1/4 inhibitor and NF‑κB inhibitor Bay 117082 were capable of suppressing the effects of LPS. TLR4 signaling pathway is functional in PASMCs, and may be involved in the inflammatory response during the pathogenesis of COPD.


[Autologous blood donation].
Because of the risks and side reactions of homologous blood transfusion autologous blood donation/transfusion is always indicated when it is practicable. Appropriate performance has to be guaranteed. By adequate training of the responsible physicians drawing and storage of autologous blood as whole blood should be general possible. However, separation into buffy coat poor red cell concentrates and fresh frozen plasma (FFP) has clear advantages. Therefore the cooperation with a blood bank or a transfusion service should be intended to which production of blood components is restricted. There is an indication for autologous blood donation in all patients who plan to undergo an elective operation being cardially compensated without "hematogenous" infection and with at least 12 g/dl hemoglobin. The aptitude examination generally may be confined on history, physical examination including blood pressure and blood counting. In a preoperative interval of 2 to 28 days it is possible to reserve 1 to 4 units of whole blood or the same number of red cell concentrates and FFP when the patient has normal haematopoiesis. The greatest problems concern the organization. They are easily to be solved by adequate information of all persons involved and close cooperation between the various physicians sending the patient to the hospital, taking care of him in the clinic and drawing the autologous blood.


Quantitative analysis of the ankle strategy under translational platform disturbance.
The ankle strategy is one of the postural adjustment maneuvers humans utilize when the support platform is disturbed. This paper presents a quantitative analysis of the ankle strategy. A three-link sagittal biped model is considered. The first link represents the two legs locked together. The second link represents the two thighs locked together. The third link represents the hip, the torso, the upper limbs, the neck, and the head. The dynamics, control, and stability of the three-link biped, under platform translation, are considered. The disturbance of the platform is represented as an input and the effect of the muscular system is reduced to a set of torques applied to the joints and across the joints. Two digital computer simulations are presented to demonstrate the behavior of the biped under backward or forward platform disturbance. The simulations are compared with experimental measurements of humans subjected to postural disturbances. It is shown that the effect of a horizontal disturbance at the ankle appears to be about 40 times that of the effect of the disturbance at the knees and at least a few hundred times larger than the effect of a disturbance at the hip. This means that, under translational platform disturbance, the ankle angle is subjected to the largest excursion. The knee and the hip angle excursions are relatively minor. Consequently, the biped, as a whole, appears to move as a single inverted pendulum. Major postural corrections are initiated by the ankle excursion. Further, when the available ankle torque is limited or nonexistent, the stability requires resorting to the knee or hip strategies.


One-Pot Synthesis of Heteroatom-Bridged Cyclic Diaryliodonium Salts.
Two one-pot procedures for the construction of O- and N-bridged diaryliodonium triflates are described. An effective aryne-mediated arylation of o-iodophenols and -sulfonamides provides diarylether and diarylamine intermediates, which are subsequently oxidized and cyclized to the corresponding diaryliodaoxinium and -iodazinium salts. Different derivatizations were applied to demonstrate their capacity as useful building blocks and gain a deeper understanding toward the general reactivity of these underdeveloped but potentially highly useful compounds.


Bem1p, a scaffold signaling protein, mediates cyclin-dependent control of vacuolar homeostasis in Saccharomyces cerevisiae.
How proliferating cells maintain the copy number and overall size of their organelles is not clear. We had previously reported that in the budding yeast Saccharomyces cerevisiae the G1 cyclin Cln3p is required for vacuolar (lysosomal) homotypic fusion and loss of Cln3p leads to vacuolar fragmentation. The Cdc42p GTPase is also required for vacuole fusion. Here we show that the scaffold protein Bem1p, a critical regulator of Cdc42p activity, is a downstream effector of Cln3p and the cyclin-dependent kinase (Cdk) Cdc28p. Our results suggest that Bem1p is phosphorylated in a Cdk-dependent manner to promote vacuole fusion. Replacing Ser72 with Asp, to mimic phosphorylation at an optimal Cdk-consensus site located in the first SH3 domain of Bem1p, suppressed vacuolar fragmentation in cells lacking Cln3p. Using in vivo and in vitro assays, we found that Cln3p was unable to promote vacuole fusion in the absence of Bem1p or in the presence of a nonphosphorylatable Bem1p-Ser72Ala mutant. Furthermore, activation of Cdc42p also suppressed vacuolar fragmentation in the absence of Cln3p. Our results provide a mechanism that links cyclin-dependent kinase activity with vacuole fusion through Bem1p and the Cdc42p GTPase cycle.


Color stability of ceramic veneers as a function of resin cement curing mode and shade: 3-year follow-up.
The influence of curing mode and shade of resin cements on the color stability of minimum-thickness ceramic veneers after a three-year storage time in distilled water was evaluated in this study. Ninety-six 0.5-mm-thick feldspathic ceramic veneers (Mark II) were luted onto resin composite substrates (Filtek Z350 XT, shade A2E) with two light-cured (NX3 Light-cure and AllCem Veneer) and dual-cured resin cements (NX3 dual-cure and AllCem) in various shades. The specimens were stored in distilled water at 37°C. Color measurements were performed with a spectrophotometer at the following times: 1h and 24h; 7, 30, and 180 days; and 1, 2, and 3 years. Data for color difference (ΔEab) light-cured and dual-cured resin cements were analyzed by two-way ANOVA with repeated measures and Tukey's test (α = 0.05). For the light-cured cements, the ΔEab values were as follows: NX3-Yellow (2.37±1.35) = ACV-A1 (2.40±1.21) = ACV-Trans (2.52±1.46) = ACV-E-Bleach M (2.56±1.42) = NX3-White (2.69±1.49) = NX3-Clear (2.98±1.68). The lowest ΔE values were found for 1 h (0.61±0.36)a, followed by 24 h (1.15±0.55)b and 30 days (2.48±1.11)c. One year, 180 days, and 2 and 3 years presented higher ΔEab values (3.34±0.94, 3.52±1.04, 3.52±0.95 and 3.55±1.14, respectively)d. For the dual-cured cements, the ΔEab values varied as follows: NX3-Clear (2.32±1.24)a = NX3-Yellow (2.37±1.32)a = NX3-White (2.76±1.43)a < AC-Trans (3.77±1.91)b = AC-A3 (4.13±2.11)b < AC-A1 (5.38±2.92)c. Considering time, the lowest ΔEab values were found for 1 h (0.48±0.25)a, followed by 24 h (2.13 ±0.83)b, 30 days (3.54±1.31)c, and 180 days (3,70±1.73)c. The follow-up times of 1 (4.43±2.15)d, 2 (5.02±1.74)e, and 3 years (4.90±2.12)e presented higher ΔEab values. This study demonstrated that light-cured resin cements were less susceptible to color change than dual-cured cements. After 2 years of follow-up, all cements presented ΔEab values above the acceptability threshold.


Real-time visualization and characterization of tumor angiogenesis and vascular response to anticancer therapies.
In vivo angiogenesis assays provide more physiologically relevant information about tumor vascularization than in vitro studies because they take the complex interactions among cancer cells, endothelial cells, mural cells, and tumor stroma into consideration. Traditional microscopic assessment of vascular density conducted by immunostaining of tissue sections or by lectin angiogram visualization of tumor vessels is invasive and requires the sacrifice of tumor-bearing animals. Therefore, it prohibits longitudinal time-course observation in a single animal and requires a large number of animals at each time point to derive statistically-meaningful observations. Additionally, heterogenous behavior among different tumors will inevitably introduce individual biological variance that may obscure reliable interpretation of the results. While various artificial in vivo angiogenesis assays, such as the Matrigel implant assay, chick chorioallatoic membrane assay, and dorsal skin fold chamber assay have been developed and employed to more directly observe the progression of physiological angiogenesis, they can not appropriately assess tumor angiogenic progression or tumor vascular regression in response to therapeutic intervention. Here, we describe a noninvasive method and a detailed protocol that we have developed and optimized using the Olympus OV-100 in vivo imaging system for real-time high-resolution visualization and assessment of tumor angiogenesis and vascular response to anticancer therapies in live animals. We show that using this approach, tumor vessels can be monitored longitudinally through the whole vasculogenesis and angiogenesis process in the same mouse. Further, morphologic changes of the same vessel prior to and after drug treatments can be captured with microscopic high resolution. Moreover, the multichannel co-imaging capability of the OV-100 allows us to analyze and compare tumor vessel permeability before and after antiangiogenesis therapy by employing a near-infrared blood pool reagent, or by visualizing improved cytotoxic drug delivery upon tumor vessel normalization by using a fluorophore tagged drug. This noninvasive method can be readily applied to orthotopically transplanted breast cancer models as well as to subcutaneously-transplanted tumor models.


Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics.
Adverse reactions or lack of response to medications are important concerns for drug development programs. However, faithful predictions of drug metabolism and toxicity are difficult because animal models show only limited translatability to humans. Furthermore, current in vitro systems, such as hepatic cell lines or primary human hepatocyte (PHH) 2-dimensional (2D) monolayer cultures, can be used only for acute toxicity tests because of their immature phenotypes and inherent instability. Therefore, the migration to novel phenotypically stable models is of prime importance for the pharmaceutical industry. Novel 3-dimensional (3D) culture systems have been shown to accurately mimic in vivo hepatic phenotypes on transcriptomic and proteomic level, but information about their metabolic stability is lacking. Using a combination of targeted and untargeted high-resolution mass spectrometry, we found that PHHs in 3D spheroid cultures remained metabolically stable for multiple weeks, whereas metabolic patterns of PHHs from the same donors cultured as conventional 2D monolayers rapidly deteriorated. Furthermore, pharmacokinetic differences between donors were maintained in 3D spheroid cultures, enabling studies of interindividual variability in drug metabolism and toxicity. We conclude that the 3D spheroid system is metabolically stable and constitutes a suitable model for in vitro studies of long-term drug metabolism and pharmacokinetics.-Vorrink, S. U., Ullah, S., Schmid, S., Nandania, J., Velagapudi, V., Beck, O., Ingelman-Sundberg, M., Lauschke, V. M. Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics.


A new, non-destructive method for analysis of clinical samples with FT-IR microspectroscopy. Breast cancer tissue as an example.
A new method for infrared analysis of tissues and cells is presented. The method is based on Fourier transform infrared microspectroscopy coupled with attenuated total reflectance. The technique allows spectroscopic measurements on the same samples used by pathologists for histopathological evaluation, e.g. stained samples on plain glass slides. Since the same specimen can be used as for histopathology, the method does not require sample preparation or modification. Significantly, the sample is not damaged. Glass absorbs in the infrared and thus has not been used previously in infrared analysis of tissues and cells. Conventional infrared techniques utilize expensive substrates, such as BaF2 windows and gold coated slides which do not absorb infrared radiation. However, these measurements require special preparation and result in the destruction of the sample. Breast cancer tissues were examined to demonstrate the feasibility and reproducibility of the new method. Linear discriminant analysis was used to discriminate and classify three types of cells: benign, atypical hyperplasia and malignant. It was demonstrated that benign vs. malignant cells were discriminated with 100% accuracy, benign vs. atypical hyperplasia were discriminated with 100% accuracy and malignant vs. atypical hyperplasia were discriminated with an accuracy of 90% and higher.


Centrifugal Generation of Droplet-Based 3D Cell Cultures.
Quickly and easily producing uniform populations of microsphere-based 3D cell cultures using droplet-based templating methods has the potential to enable widespread use of such platforms in drug discovery or cancer research. Here, we advance the design of centrifuge-based droplet generation devices, describe the use of this platform for droplet generation with controlled cell occupancy, and demonstrate weeklong culture duration. Using simple-to-construct devices and easily implemented protocols, the initial concentration of encapsulated cells is adjustable up to hundreds of cells per microsphere. This work demonstrates the first instance of using centrifugal droplet-generating devices to produce large numbers of cell-encapsulating microspheres. Applications of this versatile methodology include the rapid formation of templated 3D cell culture populations suitable for suspension culture or large batch bioreactor studies that require uniform populations.


Spore germination in Dictyostelium discoideum. II. Effects of dimethyl sulfoxide on post-activation lag as evidence for the multistate model of activation.
Mutant spores of Dictyostelium discoideum, strain SG-10, differ from wild type spores in their ability to spontaneously germinate, to be activated with 5% Dimethyl Sulfoxide (DMSO), and to be deactivated with 0.2 M sucrose. Both heat-activated wild type and mutant spores began to swell after a lag of 60-75 min at ambient temperature. Suspension of heat activated spores in 5% DMSO resulted in blockage of spore swelling and a concomitant severe inhibition of respiration; removal of 5% DMSO allowed resumption of respiration and the spores began to swell after a lag of only 15 min. It was concluded that 5% DMSO allowed the early reactions (M) to proceed but blocked the later reactions (R) of post-activation lag. Treatment of one day old spores with 20% DMSO solution for 30-120 min quantitatively activated the population. The post-activation lag time was directly dependent on the time of 20% DMSO treatment. Spores activated with 20% DMSO treatment could be deactivated by incubation at 0 degrees C; the spores most quickly deactivated at 0 degrees C were those within 10 min of swelling. Mitochondrial transport inhibitors such as azide and cyanide caused deactivation in an analogous manner. It is hypothesized that spores proceed to the second portion of the lag phase called (R) before the environment determines if dormancy is reimposed or if germination will proceed. The sensitive strain (SG-10) showed a greater degree of "damage" than the wild type after supraoptimal treatment with 40% DMSO. The spores became more resistant with age to the "damaging" action of 40% DMSO. All the observed effects of DMSO treatment were compatible with our multistate model of activation which suggests that the early portion of the lag phase (M) may involve a relative uncoupling of oxidative phosphorylation while the later portion (R) may require tight coupling.


[Account of a macular hemorrhage in a 25-year-old Congolese man with homozygous sickle cell anemia].
The author reports a case of right macula hemorrhage that has brought a sudden impairment of the vision in a 25 years old African. Homozygous sickle-cell anemia has been diagnosed thanks to etiological investigations with a special mention of hemoglobinophoresis . Generally, the sickle lesions in the eye are delimited all around the retina. Macula localisation, like the case reported by the author, has rarely ever been described.


Spectral restoration and energy resolution improvement of electron energy-loss spectra by Pixon reconstruction: II. Application to practical ELNES analysis of low SNR.
We applied Pixon deconvolution as introduced in Part I to several practical, examples of low signal-to-noise ratio (SNR), electron energy-loss spectra with a goal toward restoring their fine spectral features and/or improving the energy resolution. We demonstrate that by directly fitting the two-dimensional spectral data recorded on the CCD; the method enables us to reveal fine spectral structures. Consequently, Pixon reconstruction extends the ability to probe electronic states in very spatially localized areas, a capability currently unique to our method.


Improved xenobiotic-degrading activity of Rhodococcus opacus strain 1cp after dormancy.
The goals of the present work were as follows: to obtain the dormant forms of R. opacus 1cp; to study the phenotypic variability during their germination; to compare phenotypic variants during the growth on selective and elective media; and to reveal changes in the ability of the strain to destruct xenobiotics that had not been degradable before dormancy. It was shown that Rhodococcus opacus 1cp (the strain degrading chlorinated phenols) became able to utilize a broader spectrum of xenobiotics after storage in the dormant state. Germination of the dormant forms of R. opacus 1cp on an agarized medium was followed by emergence and development of phenotypic variants that could grow on 4-chlorophenol and 2,4,6-trichlorophenol without adaptation. The cells of R. opacus 1cp phenotypic variants also utilized all of the tested chlorinated phenols: 2,3-, 2,5-, and 2,6-dichloro-, 2,3,4- and 2,4,5-trichloro-, pentachlorophenol, and 1,2,4,5-tetrachlorobenzene in concentrations up to 60 mg/L, though at the lower rates than 4-CP and 2,4,6-TCP. The improved degradation of chlorinated phenols by R. opacus strain 1cp exposed to the growth arrest conditions demonstrates the significance of dormancy for further manifestation of the adaptive potential of populations. A new principle of selection of variants with improved biodegradative properties was proposed. It embraces introduction of the dormancy stage into the cell life cycle with subsequent direct inoculation of morphologically different colonies into the media with different toxicants, including those previously not degraded by the strain.


Real-time in vivo optogenetic neuromodulation and multielectrode electrophysiologic recording with NeuroRighter.
Optogenetic channels have greatly expanded neuroscience's experimental capabilities, enabling precise genetic targeting and manipulation of neuron subpopulations in awake and behaving animals. However, many barriers to entry remain for this technology - including low-cost and effective hardware for combined optical stimulation and electrophysiologic recording. To address this, we adapted the open-source NeuroRighter multichannel electrophysiology platform for use in awake and behaving rodents in both open and closed-loop stimulation experiments. Here, we present these cost-effective adaptations, including commercially available LED light sources; custom-made optical ferrules; 3D printed ferrule hardware and software to calibrate and standardize output intensity; and modifications to commercially available electrode arrays enabling stimulation proximally and distally to the recording target. We then demonstrate the capabilities and versatility of these adaptations in several open and closed-loop experiments, demonstrate spectrographic methods of analyzing the results, as well as discuss artifacts of stimulation.


Exploring the Mediating Effect of Physical Activities on Built Environment and Obesity for Elderly People: Evidence From Shanghai, China.
There is still a scarcity of literature on the specific mechanisms of the linkage between the built environment and obesity. As a result, this study investigated whether and how physical activities mediate the associations between the objective built environment and the BMI of elderly people. To investigate the effect of the duration and intensity of physical activity on the effect of the built environment, the study made use of the bootstrap method. In general, we discovered that physical activity duration has a huge mediating effect on the elderly people in Shanghai, especially with respect to the density and accessibility of facilities (gyms, parks, fast-food restaurants) that can greatly stimulate physical activity in elderly people to reduce their BMI. There were both direct and indirect effects on their BMI, which means that the health benefits of green spaces for older people may be more complicated than first thought.


Identification of Five Immune-Related lncRNAs Predicting Survival and Tumor Microenvironment Characteristics in Breast Cancer.
A common cancer in females, breast cancer (BRCA) mortality has been recently reduced; however, the prognosis of BRCA patients remains poor. This study attempted to develop prognostic immune-related long noncoding RNAs (lncRNAs) for BRCA and identify the effects of these lncRNAs on the tumor microenvironment (TME). Gene expression data from The Cancer Genome Atlas (TCGA) database were collected in order to select differentially expressed lncRNAs. Immune-related lncRNAs were downloaded from the ImmLnc database, where 316 immune-related lncRNAs were identified, 12 of which were found to be significantly related to the prognosis of BRCA patients. Multivariate cox regression analysis was then applied to construct prognostic immune-related lncRNAs as the risk model, including C6orf99, LINC00987, SIAH2-AS1, LINC01010, and ELOVL2-AS1. High-risk and low-risk groups were distinguished according to the median of immune-related risk scores. Accordingly, the overall survival (OS) in the high-risk group was observed to be shorter than that in the low-risk group. qRT-PCR analysis demonstrated that lncRNA expression levels in BRCA cell lines were in basic agreement with predictions except for LINC00987. By validating numerous clinical samples, lncRNA C6orf99 was shown to be highly expressed in the advanced stage, while LINC01010 and SIAH2-AS1 decreased in the advanced T-stage and M-stage. Moreover, the expression of LINC0098 was found to be significantly decreased among the groups (>50 years old). Gene set enrichment analysis (GSEA) was applied to analyze the cancer hallmarks and immunological characteristics of the high-risk and low-risk groups. Importantly, the TIMER database demonstrated that this immune-related lncRNA risk model for breast cancer is related to the infiltration of immune cells. In conclusion, the results indicated that five immune-related lncRNAs could be used as a prognostic model and may even accelerate immunotherapy for BRCA patients.


Bilirubin production in healthy term infants as measured by carbon monoxide in breath.
To describe total bilirubin production in healthy term infants, we measured the end-tidal breath CO, corrected for ambient CO (ETCOc), with an automated sampler and electrochemical (EC) CO instrument. For infants of mothers with a negative Coombs' test, the ETCOc was 1.3 +/- 0.7 microL/L (n = 397) and the serum bilirubin on day 3 postpartum was 73 +/- 35 mg/L (n = 381). In contrast, the ETCOc for infants with ABO or Rh incompatibility, a positive direct Coombs' test, and bilirubin > 130 mg/L (n = 9) was significantly higher, 1.8 +/- 0.8 microL/L, than for those who had a positive Coombs' test result but whose bilirubin was < or = 130 mg/L (n = 12), 1.0 +/- 0.5 microL/L (P < 0.05). At 2 to 8 h postpartum seven term babies from mothers with insulin-dependent diabetes had ETCOc of 1.8 +/- 0.7 microL/L, significantly higher than that in the other term infants [1.3 +/- 0.7 microL/L (n = 390), P < 0.04]. Their bilirubin concentration at 72 +/- 12 h was also higher: 121 +/- 45 mg/L (n = 7) vs 73 +/- 34 mg/L (n = 374; P = 0.03). We conclude that ETCOc measurements may be helpful in understanding the mechanisms of jaundice in healthy term infants in a variety of conditions.


Nonsteroidal anti-inflammatory drugs misleading the diagnosis of periapical abscess.
Periapical abscesses are common but may rarely be a subclinical infection in dental clinics. If an infection progresses rapidly or slowly as a tooth ache of chronic periodontitis with clinical features misleading the diagnosis, the dentists are able to recognize the salient signs and identify the patients at risk. This article reviews a case of a rare innocuous periapical abscess presenting as angioedema of upper lip with history of consuming nonsteroidal anti-inflammatory drugs such as diclofenac sodium whenever needed for arthralgia on a short-term basis.


Assessment of impaired glucose tolerance prevalence with hemoglobin A₁c and oral glucose tolerance test in 252 Turkish women with polycystic ovary syndrome: a prospective, controlled study.
What is the prevalence of abnormalities in glucose metabolism in patients with polycystic ovary syndrome (PCOS) and controls in a Turkish population? The total prevalence of glucose abnormalities in PCOS patients was 16.3% [impaired glucose tolerance (IGT) 14.3%; type 2 diabetes mellitus (T2DM) 2%] and was higher than in healthy subjects (IGT 8.5%; T2DM 0%, respectively). One of the most common markers of chronic glycemia is hemoglobin Alc (HbA1c). However, little is known about whether the use of HbA1c results in diagnosis of more cases of glucose intolerance in the PCOS population than the oral glucose tolerance test (OGTT) alone. This was a prospective study, including 252 women with PCOS and 117 control women without PCOS. The study was carried out in the gynecological outpatient department of Namik Kemal University Hospital, Turkey, between 2010 and 2012. Women with PCOS (n = 252) were diagnosed according to Rotterdam criteria. The control group included 117 women (aged 17-45 years) who were selected randomly. BMI of participants ranged between 15.6 and 47.9 kg/m(2). Patients with PCOS were comparable to controls in terms of age (24.8 versus 25.9 years, respectively) and had higher BMI (26.1 versus 24.9 kg/m(2), respectively). Of 252 patients with PCOS, 41 had glucose intolerance (IGT 14.3%; T2DM 2%) when compared with 10 of the 117 control patients (IGT 8.5%; T2DM 0%; odds ratios = 2.08; P = 0.045) during the OGTT. When an HbA1c value ≥ 5.6% was used to divide the total population, the prevalence of abnormal glucose metabolism was 7.9% in the patients with PCOS, below the value detected in the control patients (8.5%), which showed that 20 of 41 patients with abnormal glucose tolerance would not have been diagnosed, if the HbA1c alone had been used. When compared with the OGTT results, HbA1c provided 52.4% sensitivity, 74.4% specificity, 67.1% positive and 60.9% negative predictive values with a threshold value of 5.6% in abnormal glucose tolerance. The receiver operating characteristic analysis suggested a threshold value of 5.35% in HbA1c (75.6% sensitivity and 52.6% specificity) for the prediction of abnormal glucose tolerance. This study did not involve weight-matched healthy subjects, which may cause a difference in prevalence of abnormal glucose metabolism between the groups, and the results are limited to an unselected population of patients who have the full PCOS phenotype. In addition, the incidence of T2DM among the first-degree relatives and 2-h insulin levels could not be reported in full. Further investigation of the efficacy of HbA1c for the prediction of abnormal glucose tolerance should be undertaken in long-term prospective studies and in different geographic populations. At present, the only way to reliably detect abnormal glucose metabolism in Turkish women with PCOS appears to be using the OGTT. No financial support. The authors have no competing interests to declare. Not applicable.


Giovanni Alfonso Borelli--the father of biomechanics.
Giovanni Alfonso Borelli is often described as the father of biomechanics. He was born in Naples in 1608. His De Motu Animalium, published in 1680, extended to biology the rigorous analytical methods developed by Galileo in the field of mechanics. Borelli calculated the forces required for equilibrium in various joints of the human body well before Newton published The Laws of Motion Borelli was the first to understand that the levers of the musculoskeletal system magnify motion rather than force, so that muscles must produce much larger forces than those resisting the motion. Borelli died in Rome on December 31, 1679, but his impressive body of original work helped inspire a great number of future scientists, microscopists, and inventors. The highest honor bestowed by the American Society of Biomechanics is the Giovanni Borelli Award.


Autocrine motility factor signaling induces tumor apoptotic resistance by regulations Apaf-1 and Caspase-9 apoptosome expression.
Autocrine motility factor (AMF) is a cytokine that regulates locomotion and metastasis of tumor cells. It is well known that expression levels of AMF secretion and its receptor (AMF R) are closely related to tumor malignancy and rheumatoid arthritis. We have established that AMF signaling induced anti-apoptotic activity and that human fibrosarcoma HT-1080 line that secreted high levels of AMF were resistant to drug-induced apoptosis. These cells did not express the apoptotic protease activating factor-1 (Apaf-1) and Caspase-9 genes that encode for the proteins that form the "apoptosome" complex. The disappearance of the Apaf-1 and Caspase-9 gene was recovered by a cellular signaling inhibitor of protein kinase C, phosphatidylinositol 3-phosphate kinase and mitogen-activated protein kinase of the in vitro cultured human fibrosarcoma HT-1080 line. Treatment with these inhibitors favored apoptotic cell death induced by anti-cancer drugs of the murine ascites Ehrlich line. Apoptotic resistance of tumor cells allows them to escape death from cancer chemotherapy, so an understanding of malignant anti-apoptotic activities is important. Antibodies against AMF induced Ehrlich ascites apoptosis in vitro, and effectively aided in vivo apoptosis induced by anti-cancer drugs. The results might indicate a novel route by which tumor cells protect themselves with products, such as AMF, and proliferate despite various stresses and chemical insults; AMF regulates expression of Apaf-1 and caspase-9 genes via a complex signaling pathway and indirectly regulates formation of the apoptosome.


Cascaded walks in protein sequence space: use of artificial sequences in remote homology detection between natural proteins.
Over the past two decades, many ingenious efforts have been made in protein remote homology detection. Because homologous proteins often diversify extensively in sequence, it is challenging to demonstrate such relatedness through entirely sequence-driven searches. Here, we describe a computational method for the generation of 'protein-like' sequences that serves to bridge gaps in protein sequence space. Sequence profile information, as embodied in a position-specific scoring matrix of multiply aligned sequences of bona fide family members, serves as the starting point in this algorithm. The observed amino acid propensity and the selection of a random number dictate the selection of a residue for each position in the sequence. In a systematic manner, and by applying a 'roulette-wheel' selection approach at each position, we generate parent family-like sequences and thus facilitate an enlargement of sequence space around the family. When generated for a large number of families, we demonstrate that they expand the utility of natural intermediately related sequences in linking distant proteins. In 91% of the assessed examples, inclusion of designed sequences improved fold coverage by 5-10% over searches made in their absence. Furthermore, with several examples from proteins adopting folds such as TIM, globin, lipocalin and others, we demonstrate that the success of including designed sequences in a database positively sensitized methods such as PSI-BLAST and Cascade PSI-BLAST and is a promising opportunity for enormously improved remote homology recognition using sequence information alone.


Vocal fold restoration after scarring: biocompatibility and efficacy of an MSC-based bioequivalent.
There is growing interest to application of regenerative medicine approaches in otorhinolaryngological practice, especially in the framework of the therapy of vocal fold (VF) scar lesions. The used conservative and surgical methods, despite the achieved positive outcomes, are frequently unpredictable and do not result in the restoration of the VF's lamina propria's structure, which provides the mechanical properties necessary for vibration. In this connection, the aim of this study was to ascertain the safety and efficacy of a bioequivalent in the treatment of VF scars using a rabbit model of chronic damage. The bioequivalent consisted of a hydrogel system based on a PEG-fibrin conjugate and human bone marrow-derived MSC. It was characterized and implanted heterotopically into rats and orthotopically into rabbits after VF scar excision. We showed that the fabricated bioequivalent consisted of viable cells retaining their metabolic and proliferative activity. While being implanted heterotopically, it had induced the low inflammatory reaction in 7 days and was well tolerated. The orthotopic implantation showed that the gel application was characterized by a lower hemorrhage intensity (p = 0.03945). The intensity of stridor and respiratory rate between the groups in total and between separate groups had no statistically significant difference (p = 0.96 and p = 1; p = 0.9593 and p = 0.97…1, respectively). In 3 days post-implantation, MSC were detected only in the tissues closely surrounding the VF defect. The bioequivalent injection caused that the scar collagen fibers were packed looser and more frequently mutually parallel that is inherent in the native tissue (p = 0.018). In all experimental groups, the fibrous tissue's ingrowth in the adjacent exterior muscle tissue was observed; however, in Group 4 (PEG-Fibrin + MSC), it was much less pronounced than it was in Group 1 (normal saline) (p = 0.008). The difference between the thicknesses of the lamina propria in the control group and in Group 4 was not revealed to be statistically significant (p = 0.995). The Young's modulus of the VF after the bioequivalent implantation (1.15 ± 0.25 kPa) did not statistically significantly differ from the intact VF modulus (1.17 ± 0.45 kPa); therefore, the tissue properties in this group more closely resembled the intact VF. The developed bioequivalent showed to be biocompatible and highly efficient in the restoration of VF's tissue.


Association of PYGO2 and EGFR in esophageal squamous cell carcinoma.
Wnt signaling is an important evolutionary conserved pathway that is not only involved in determination of cellular development, self-renewal, and fate, but also has significant roles in tumor development and progression. Deregulation of the Wnt/β-catenin signaling pathway and aberrant expression of its components is commonly observed in solid tumors. Such aberrant regulation of Wnt signaling is commonly related to either malfunction of its components or crosstalk with other cellular processes such as the epidermal growth factor receptor (EGFR) signaling cascade. Therefore, identification of the roles of major involved components may be useful to identify new therapeutic targets for cancer treatment. In this study, we assessed EGFR and PYGO2 mRNA expression in tumors and margin normal tissues from 55 esophageal squamous cell carcinoma (ESCC) patients using real-time qRT-PCR, and evaluated clinicopathology relative to the two genes' expression levels. Significant PYGO2 and EGFR overexpression was observed in 30.9 % (P = 0.017) and 38.2 % (P = 0.006) of tumors, respectively. PYGO2 and EGFR expression were significantly associated not only with each other (P < 0.001), but also with tumor staging and depth (P < 0.001). Furthermore, PYGO2 expression was significantly correlated with the tumor grade (P = 0.043) and size (P = 0.023). We identify PYGO2 as a new molecular marker of invasive tumors, introducing its probable oncogenic role in ESCC progression and aggressiveness. In line with other reports, we also illustrate the oncogenic function of EGFR in the development of ESCC through advance stages. We also observed a significant correlation between PYGO2 and EGFR in ESCC tumors, which reveals a mutual convergent influence of these factors in tumor progression and development. Considering aberrant expression, mutual positive feedback, and the significant clinical relevance of these genes in ESCC, we introduce them as appropriate therapeutic targets in adjuvant therapy of ESCC.


Melatonin Differentially Modulates NF-кB Expression in Breast and Liver Cancer Cells.
NF-kB (nuclear factor kappa B) is a transcription factor composed of two subunits, p50 and p65, which plays a key role in the inflammatory process. Melatonin has oncostatic, antiangiogenic and antimetastatic properties, and some recent studies have indicated an inhibitory effect of melatonin on NF-kB in some types of cancer. This work aims to investigate the effects of melatonin treatment on the expression of NFkB in breast and liver cancer models. The breast cancer xenographic model was performed using female Balb/c nude athymic mice injected with MDA-MB-231 cells. The animals were treated with 40 mg/Kg of melatonin for 21 days. Volume of the tumors was measured with a digital caliper. Hepatocarcinoma model was developed by using the HepG2 cells in vitro, treated with 1 mM melatonin for 24 h. The expression of NF-kB protein was verified by immunohistochemistry and immunocytochemistry and quantified by optical densitometry, in vivo study and in vitro study, respectively. NF-kB gene expression was performed by quantitative RT-PCR. The breast cancer xenografts nude mice treated with melatonin showed reduced tumor size (P=0.0022). There was a decrease in NF-kB protein staining (P=0.0027) and gene expression (P=0.0185) in mice treated with melatonin. The opposite results were observed for the hepatocarcinoma model. HepG2 cells treated with melatonin showed an increase in the NF-kB immunostaining when compared to control cells (P=0.0042). Our results indicated that the treatment with melatonin was able to decrease both gene and protein expressions of NF-kB in breast cancer cells and, conversely, increase the transcription factor protein expression in hepatocarcinoma cells. These data highlighted a double role in the expression of NF-kB, depending on the cell type. Further studies are needed to better elucidate the action of melatonin in NF-kB, since this transcription factor acts on different signaling pathways that are fundamental for carcinogenesis.


Highly dispersed pd catalyst locked in knitting aryl network polymers for Suzuki-Miyaura coupling reactions of aryl chlorides in aqueous media.
Highly dispersed palladium chloride catalysts locked in triphenylphosphine-functionalized knitting aryl network polymers (KAPs) are developed and exhibit excellent activity under mild conditions in the Suzuki-Miyaura cross-coupling reactions of aryl chlorides in aqueous media. This work highlights that the microporous polymers not only play the role of support materials, but also protect the Pd species from aggregation and precipitation, hence, positively effect the catalysis activity.


Incidence and risk factors of common viral infections among renal transplant recipients during the first year post-transplant in North-eastern Iran.
Despite major therapeutic advances, management of viral infections in renal transplant recipients is still a major challenge. Hence, it is urgently needed to establish protocols for appropriate control and the prevention of viral infection. We evaluated demographic/clinical characteristics, frequency, and risk factors of symptomatic viral infections in renal transplant recipients during the 1st year posttransplant, in northeastern Iran. We retrospectively reviewed medical files of 247 patients including 146 males and 101 females who had undergone renal transplantation at Montaserie organ transplantation hospital of Mashhad during 2012-2014. These patients were followed up for one year after transplantation for the detection of any symptomatic viral infection. Demographic and clinical characteristics of recipients were collected and analyzed using the Statistical Package for Social Sciences version 18 software; P < 0.05 was considered as statistically significant. Data were presented using descriptive statistics. Furthermore, logistic regression analysis was used to determine risk factors for infection. The mean age of the patients was 34.94 ± 13.89 years. During the 1st year posttransplant, 68 episodes of viral infections were detected in 64 patients (25.9%). Cytomegalovirus (CMV, 21.9%), Varicella Zoster virus (2.8%), herpes simplex virus (2.0%), and human polyomavirus BK virus (0.8%) were the most common symptomatic viral infections found. Age of the patients was the only significant risk factor for viral infections (odds ratio = 1.066; 95% confidence interval: 1.002-1.134; P = 0.042). The incidence of symptomatic viral infections, particularly CMV disease, is high in our center. Hence, it is recommended to use appropriate prophylaxis and monitor the patients during the first six months post-transplant.


Comparisons of reactivity of pulmonary vascular and airway smooth muscle of sheep and calf to tryptamine analogues, histamine, and antigen.
Serotonin (5-hydroxytryptamine) contracted the pulmonary artery, trachea, and bronchus in both cattle and sheep. Serotonin contracted the calf pulmonary vein, but it relaxed the pulmonary vein of sheep. Histamine constricted the 2 pulmonary blood vessels and trachea in calves and sheep. In contrast, histamine relaxed the ovine bronchus, but not the bronchus of the calf. Antigen (equine serum) constricted pulmonary veins from sensitized calves, but relaxed pulmonary veins from similarly sensitized sheep. Apparently, sheep are naturally more resistant than calves to both histamine and serotonin which contribute to the pulmonary inflammatory reaction.


Eye dominance and response latency in area V1 of the monkey.
We measured the latency of 35 cells from V1 in two rhesus monkeys, to dynamic random dot stimuli monocular and binocularly presented. Mean latencies after non-dominant eye stimulation (97.9 ms) were longer than those for dominant eye (78.2 ms) and binocular (70.7 ms) stimulation. Differences between latencies for dominant eye and binocular stimulation were not statistically significant. For dominant eye, there was a significant statistical correlation between dominance strength and latency (R = -0.36; p = 0.03). We failed to find significant statistical differences between latencies for cells with temporal and nasal dominant receptive-field. We conclude that, in V1, the response latency is largely determined by the dominant eye, whereas interocular interactions do not seem to play a relevant role regarding response latency.


Consonant categorization exhibits a graded influence of surrounding spectral context.
When spectral properties differ across successive sounds, this difference is perceptually magnified, resulting in spectral contrast effects (SCEs). Recently, Stilp, Anderson, and Winn [(2015) J. Acoust. Soc. Am. 137(6), 3466-3476] revealed that SCEs are graded: more prominent spectral peaks in preceding sounds produced larger SCEs (i.e., category boundary shifts) in categorization of subsequent vowels. Here, a similar relationship between spectral context and SCEs was replicated in categorization of voiced stop consonants. By generalizing this relationship across consonants and vowels, different spectral cues, and different frequency regions, acute and graded sensitivity to spectral context appears to be pervasive in speech perception.


Re-Setting the Circadian Clock Using Exercise against Sarcopenia.
Sarcopenia is defined as the involuntary loss of skeletal muscle mass and function with aging and is associated with several adverse health outcomes. Recently, the disruption of regular circadian rhythms, due to shift work or nocturnal lifestyle, is emerging as a novel deleterious factor for the development of sarcopenia. The underlying mechanisms responsible for circadian disruption-induced sarcopenia include molecular circadian clock and mitochondrial function associated with the regulation of circadian rhythms. Exercise is a potent modulator of skeletal muscle metabolism and is considered to be a crucial preventative and therapeutic intervention strategy for sarcopenia. Moreover, emerging evidence shows that exercise, acting as a zeitgeber (time cue) of the skeletal muscle clock, can be an efficacious tool for re-setting the clock in sarcopenia. In this review, we provide the evidence of the impact of circadian disruption on skeletal muscle loss resulting in sarcopenia. Furthermore, we highlight the importance of exercise timing (i.e., scheduled physical activity) as a novel therapeutic strategy to target circadian disruption in skeletal muscle.


Child mortality, hypothalamic-pituitary-adrenal axis activity and cellular aging in mothers.
Psychological challenges, including traumatic events, have been hypothesized to increase the age-related pace of biological aging. Here we test the hypothesis that psychological challenges can affect the pace of telomere attrition, a marker of cellular aging, using data from an ongoing longitudinal-cohort study of Kaqchikel Mayan women living in a population with a high frequency of child mortality, a traumatic life event. Specifically, we evaluate the associations between child mortality, maternal telomere length and the mothers' hypothalamic-pituitary-adrenal axis (HPAA), or stress axis, activity. Child mortality data were collected in 2000 and 2013. HPAA activity was assessed by quantifying cortisol levels in first morning urinary specimens collected every other day for seven weeks in 2013. Telomere length (TL) was quantified using qPCR in 55 women from buccal specimens collected in 2013. Shorter TL with increasing age was only observed in women who experienced child mortality (p = 0.015). Women with higher average basal cortisol (p = 0.007) and greater within-individual variation (standard deviation) in basal cortisol (p = 0.053) presented shorter TL. Non-parametric bootstrapping to estimate mediation effects suggests that HPAA activity mediates the effect of child mortality on TL. Our results are, thus, consistent with the hypothesis that traumatic events can influence cellular aging and that HPAA activity may play a mediatory role. Future large-scale longitudinal studies are necessary to confirm our results and further explore the role of the HPAA in cellular aging, as well as to advance our understanding of the underlying mechanisms involved.


Dendritic cell vaccination with a toll-like receptor agonist derived from mycobacteria enhances anti-tumor immunity.
Dendritic cell (DC)-based vaccines are considered useful in cancer immunotherapy, and the interaction of DC and adjuvants is important in the design of the next generation vaccines. In this study, whether DC combined with Rv2299c derived from mycobacteria could improve anti-tumor immune responses in a colon cancer mouse model was evaluated. MC38 cell lines were injected subcutaneously to establish colon-cancer-bearing mice and the following four groups were evaluated: PBS control, tumor antigen (TA) loaded-DC, Rv2299c, and a combination of TA-loaded-DC and Rv2299c. The combination treatment with TA-loaded-DC and Rv2299c exhibited greater inhibition of tumor growth compared to other groups. These effects were associated with the reduction of suppressor cells, such as myeloid-derived suppressor cells and regulatory T cells, and the induction of effector cells, such as CD4+ T cells and CD8+ T cells, in spleen, and with the activation of cytotoxic T Lymphocytes and NK cells. These results suggest that TA-loaded-DC vaccination with Rv2299c derived from mycobacteria enhanced anti-tumor immunity in a mouse colon cancer model by inhibiting the generation of immune-suppressive cells and recovering numbers of effector cells, and demonstrated superior polarization of the Th1/Th2 balance in favor of the Th1 immune response.


Chronic Kidney Disease After Liver Transplantation.
Chronic kidney disease among liver transplant recipients is common and associated with an increased mortality risk. Several risk factors and causes for the development of chronic kidney disease have been identified. They can be divided into perioperative factors, such as unresolved acute kidney injury; donor-related factors, such as the use of extended criteria liver allografts; and recipient-related factors, such as the use of calcineurin inhibitors and the presence of metabolic syndrome, diabetes, and obesity. There is a bimodal progression, more prominent during the initial post-transplant months, followed by a gradual but progressive decline over the subsequent years. Management strategies to prevent and treat chronic kidney disease in the general population can be reasonably applied to the liver transplant population and include addressing comorbidities such as hypertension and diabetes. Strategies to minimize or withdraw calcineurin inhibitors from the immunosuppressive regimen can slow progression of kidney dysfunction. Patients with advanced chronic kidney disease should be considered for kidney transplantation due to its survival advantage. Allocation policy in the United States confers safety-net allocation priority for liver transplant recipients who develop advanced chronic kidney disease within the first year of liver transplantation.


HorkaD, a chromosome instability-causing mutation in Drosophila, is a dominant-negative allele of Lodestar.
Correct segregation of chromosomes is particularly challenging during the rapid nuclear divisions of early embryogenesis. This process is disrupted by Horka(D), a dominant-negative mutation in Drosophila melanogaster that causes female sterility due to chromosome tangling and nondisjunction during oogenesis and early embryogenesis. Horka(D) also renders chromosomes unstable during spermatogenesis, which leads to the formation of diplo//haplo mosaics, including the gynandromorphs. Complete loss of gene function brings about maternal-effect lethality: embryos of the females without the Horka(D)-identified gene perish due to disrupted centrosome function, defective spindle assembly, formation of chromatin bridges, and abnormal chromosome segregation during the cleavage divisions. These defects are indicators of mitotic catastrophe and suggest that the gene product acts during the meiotic and the cleavage divisions, an idea that is supported by the observation that germ-line chimeras exhibit excessive germ-line and cleavage function. The gene affected by the Horka(D) mutation is lodestar, a member of the helicase-related genes. The Horka(D) mutation results in replacement of Ala777 with Thr, which we suggest causes chromosome instability by increasing the affinity of Lodestar for chromatin.


Pancreatic cancer proteome: the proteins that underlie invasion, metastasis, and immunologic escape.
Pancreatic cancer is a highly lethal disease that has seen little headway in diagnosis and treatment for the past few decades. The effective treatment of pancreatic cancer is critically relying on the diagnosis of the disease at an early stage, which still remains challenging. New experimental approaches, such as quantitative proteomics, have shown great potential for the study of cancer and have opened new opportunities to investigate crucial events underlying pancreatic tumorigenesis and to exploit this knowledge for early detection and better intervention. To systematically study protein expression in pancreatic cancer, we used isotope-coded affinity tag technology and tandem mass spectrometry to perform quantitative proteomic profiling of pancreatic cancer tissues and normal pancreas. A total of 656 proteins were identified and quantified in 2 pancreatic cancer samples, of which 151 were differentially expressed in cancer by at least 2-fold. This study revealed numerous proteins that are newly discovered to be associated with pancreatic cancer, providing candidates for future early diagnosis biomarkers and targets for therapy. Several differentially expressed proteins were further validated by tissue microarray immunohistochemistry. Many of the differentially expressed proteins identified are involved in protein-driven interactions between the ductal epithelium and the extracellular matrix that orchestrate tumor growth, migration, angiogenesis, invasion, metastasis, and immunologic escape. Our study is the first application of isotope-coded affinity tag technology for proteomic analysis of human cancer tissue and has shown the value of this technology in identifying differentially expressed proteins in cancer.


Therapeutic monoclonal antibodies and the need for targeted pharmacovigilance in India.
A growing number of innovative mAb therapeutics are on the global market, and biosimilar versions have now also been approved, including in India. Although efficacy and safety is demonstrated prior to approval, targeted pharmacovigilance is essential for the identification and assessment of risk for any mAb products. We analyzed the ADR data related to mAbs reported to the NCC-PvPI through the spontaneous reporting system Vigiflow during April 2011 to February 2014 to identify mAbs with the highest number of ADR including fatal/serious ADR. Only 0.72% reports were related to mAbs. Although 15 mAbs are approved in the country, only 6 mAbs were reported through Vigiflow. Rituximab was highly reported, and no fatal/serious ADR related to any mAbs were reported during the study period. Our study shows that PvPI is effective and robust system in the detection and assessment of risks associated with the use of mAbs.


Computational Analysis of Specific MicroRNA Biomarkers for Noninvasive Early Cancer Detection.
Cancer is a complex disease residing in various tissues of human body, accompanied with many abnormalities and mutations in genomes, transcriptome, and epigenome. Early detection plays a crucial role in extending survival time of all major cancer types. Recent advances in microarray and sequencing techniques have given more support to identifying effective biomarkers for early detection of cancer. MicroRNAs (miRNAs) are more and more frequently used as candidates for biomarkers in cancer related studies due to their regulation of target gene expression. In this paper, the comparative analysis is used to discover miRNA expression patterns in cancer versus normal samples on early stage of eight prevalent cancer types. Our work focuses on the specific miRNAs biomarkers identification and function analysis. Several identified miRNA biomarkers in this paper are matched well with those reported in existing researches, and most of them could serve as potential candidate indicators for clinical early diagnosis applications.


Detection of COX-2 in liquid biopsy in patients with breast cancer.
To determine the expression of the cyclooxygenase-2 (COX-2) gene in patients with breast cancer attended at the Centro Universitário Saúde ABC/Faculdade de Medicina do ABC (CUS-ABC/FMABC) outpatient clinic. Breast cancer is the most common cancer in women worldwide. More than two million new cases are reported annually. An overexpression of COX-2 has been observed in many cancers. COX-2 is related to parameters of cancer aggressiveness, including tumour size, positive nodal state and lower survival, and to angiogenesis and resistance to apoptosis. 15 mL of peripheral blood was obtained from 34 patients and 21 healthy women. The extracellular RNA of QIAamp RNA was submitted to an RNA sequestration kit for RNA reverse transcriptase. Quantitative real-time PCR was performed using COX-2-specific oligonucleotides and the endogenous Glyceraldehyde-3-Phosphate Dehydrogenase gene. The mean remission time was 53 years. The mean progression time was 33 months. The difference observed between the patient and control groups in median COX-2 expression (p<0.001) was significant. Patients with breast cancer showed a higher mean COX-2 expression in peripheral blood samples at diagnosis than the control group. Since this information could prove important in the diagnosis and prognosis of breast cancer, further research is required on larger patient samples.


Effects of Lycium barbarum Polysaccharides on Immunity and the Gut Microbiota in Cyclophosphamide-Induced Immunosuppressed Mice.
The mechanism of immunoregulation by Lycium barbarum polysaccharides (LBPs) was assessed by studying the effect of LBP on the immunity and the gut microbiota. LBP isolated and purified in this study was composed of nine monosaccharides, with an Mw 1,207 kDa. LBP showed immunomodulatory activity in cyclophosphamide (Cy)-treated mice by restoring the damaged immune organs and adjusting the T lymphocyte subsets. We also found that LBP increased the diversity of the gut microbiota and the relative abundances of bacteria, such as Rickenellaceae, Prevotellaceae, Bifidobacteriaceae, and so on, which were positively associated with immune traits. In addition, Caco2 cells model was used to explore the intestinal absorption of LBP. Results showed that LBP was hardly absorbed in the intestine, which suggesting that most LBP may interact with gut microbiota. These findings suggest that the immune response induced by LBP is associated with the regulation of the gut microbiota.


Growth kinetics of the photosynthetic bacterium Chlorobium thiosulfatophilum in a fed-batch reactor.
Hydrogen sulfide dissolved in water can be converted to elementary sulfur or sulfate by the photosynthetic bacterium Chlorobium thiosulfatophilum. Substrate inhibition occurred at sulfide concentrations above 5.7 mM. Light inhibition was found at average light intensities of 40,000 lux in a sulfide concentration of 5 mM, where no substrate inhibition occurred. Light intensity, the most important growth parameter, was attenuated through both scattering by sulfur particles and absorption by the cells. Average cell and sulfur particle sizes were 1.1 and 9.4 microm, respectively. Cells contributed 10 times as much to the turbidity as sulfur particles of the same weight concentration. The light attenuation factor was mathematically modeled, considering both the absorption and scattering effects based on the Beer-Lambert law and the Rayleigh theory, which were introduced to the cell growth model. Optimal operational conditions relating feed rate vs. light intensity were obtained to suppress the accumulation of sulfate and sulfide and save light energy for 2- and 4-L fed-batch reactors. Light intensity should be greater for the same performance (H(2)S removal rate/unit cell concentration) in larger reactors due to the scaleup effect on light transmission. Knowledge of appropriate growth kinetics in photosynthetic fed-batch reactors was essential to increase feed rate and light intensity and therefore cell growth. A mathematical model was developed that describes the cell growth by considering the light attenuation factor due to scattering and absorption and the crowding effect of the cells. This model was in good agreement with the experimental results.


Induced sputum in asthma.
This is a short review of the use of induced sputum in bronchial asthma. The clinical applications, the procedure of this method as well as the safety validity and reproducibility of the results are discussed in detail. It is shown that sputum induction is a practical and safe procedure and an important research tool of local inflammation of bronchial asthma.


[Molecular mechanism of SH2B1 in regulating JAK2/IRS2 during obesity development].
In order to investigate the effect of SH2B1 on leptin signal transduction JAK2/IRS2 and its biological function. Vitro kinase assay and Western blot were used to analyse tyrosine phosphorylatin of key molecule JAK2 and insulin receptor substrate-2 (IRS2). ELISA was used to measure the plasma leptin levels in mice. The postnatal growth of mice was monitored over 27 weeks. SH2B1 dramatically enhanced the leptin-stimulated tyrosine phosphorylation of JAK2 and IRS2 in HEK293 cells stably expressing LRb (HEK239(LRb)). Leptin-stimulated activation of hypothalamic JAK2 and phosphorylation of hyphothalamic IRS2 were significantly impaired in SH2B1(-/-) mice. The deletion of SH2B1 led to leptin resistance,and fasting and randomly fed plasma leptin levels were respectively 3.2 times and 5.1 times higher in SH2B1(-/-) males than wild-type littermates at 15 weeks of age. SH2B1(-/-) males gained body weight rapidly and exceeded wild-type littermates from 5th week. SH2B1(-/-) (at 21 weeks) was approximately twice heavier than wild-type littermates. SH2B1 is an endogenous enhancer of leptin sensitivity and required for maintaining normal bodyweight in mice via leptin JAK2/IRS2 pathway.


Participation of irradiated Anopheles arabiensis males in swarms following field release in Sudan.
The success of the sterile insect technique (SIT) depends the release of large numbers of sterile males, which are able to compete for mates with the wild male population within the target area. Unfortunately, the processes of colonisation, mass production and irradiation may reduce the competitiveness of sterile males through genetic selection, loss of natural traits and somatic damage. In this context, the capacity of released sterile Anopheles arabiensis males to survive, disperse and participate in swarms at occurring at varying distances from the release site was studied using mark-release-recapture (MRR) techniques. In order to assess their participation in swarms, irradiated and marked laboratory-reared male mosquitoes were released 50, 100 or 200 m from the known site of a large swarm on three consecutive nights. Males were collected from this large swarm on subsequent nights. Over the three days a total of 8,100 males were released. Mean distance travelled (MDT), daily probability of survival and estimated population size were calculated from the recapture data. An effect of male age at the time of release on these parameters was observed. Five per cent of the males released over three days were recaptured. In two-, three- and four-day-old males, MDT was 118, 178 and 170 m, and the daily survival probability 0.95, 0.90 and 0.75, respectively. From the recapture data on the first day following each release, the Lincoln index gives an estimation of 32,546 males in the natural population. Sterile An. arabiensis males released into the field were able to find and participate in existing swarms, and possibly even initiate swarms. The survival probability decreased with the age of male on release but the swarm participation and the distance travelled by older males seemed higher than for younger males. The inclusion of a pre-release period may thus be beneficial to male competitiveness and increase the attractiveness of adult sexing techniques, such as blood spiking.


The Utility of Hybrid SPECT/CT Lung Perfusion Scintigraphy in Pulmonary Embolism Diagnosis.
Pulmonary embolism (PE) is diagnosed either by ventilation/perfusion (V/Q) scintigraphy or pulmonary CT angiography. One of the imaging methods used in nuclear medicine is hybrid SPECT/CT scintigraphy. The aim of this study was to evaluate the utility of SPECT/CT(Q) scintigraphy in the diagnosis of PE and to compare SPECT/CT(Q) with planar(Q) and SPECT(Q) methods. The study group consisted of 109 consecutive patients suspected of having PE referred for performing lung scintigraphy. The inclusion criteria were: performance of perfusion planar, SPECT and SPECT/CT scans; availability of clinical data covering a 6-month follow-up period, and D-dimer level testing. The number of eligible patients was 84. PE was reported in patients with at least 1 segmental or 2 subsegmental perfusion defects without parenchymal abnormalities on CT scans. PE was excluded when there was a normal perfusion pattern or perfusion defects were caused by lung parenchymal abnormalities or were not arranged in accordance with the pulmonary vasculature. Twenty-six patients (31%) had a final diagnosis of PE. The sensitivity and specificity values of each method were as follows: planar(Q) 73 and 43%, SPECT(Q) 88 and 47% and SPECT/CT(Q) 100 and 83%. SPECT/CT(Q) yielded a significantly higher diagnostic accuracy than planar(Q) (p < 0.001) and SPECT(Q) (p < 0.001) scans. We conclude that hybrid SPECT/CT(Q) imaging has a high diagnostic efficacy in the diagnosis of PE. Lung perfusion scintigraphy performed with a hybrid SPECT/CT device has a significantly higher sensitivity and specificity than scanning performed with the planar or SPECT technique.


High throughput sequencing of small RNAs reveals dynamic microRNAs expression of lipid metabolism during Camellia oleifera and C. meiocarpa seed natural drying.
Camellia species are ancient oilseed plants with a history of cultivation over two thousand years. Prior to oil extraction, natural seed drying is often practiced, a process affecting fatty acid quality and quantity. MicroRNAs (miRNA) of lipid metabolism associated with camellia seed natural drying are unexplored. To obtain insight into the function of miRNAs in lipid metabolism during natural drying, Illumina sequencing of C. oleifera and C. meiocarpa small-RNA was conducted. A total of 274 candidate miRNAs were identified and 3733 target unigenes were annotated by performing a BLASTX. Through integrated GO and KEGG function annotation, 23 miRNA regulating 131 target genes were identified as lipid metabolism, regulating fatty acid biosynthesis, accumulation and catabolism. We observed one, two, and four miRNAs of lipid metabolism which were specially expressed in C. Meiocarpa, C. oleifera, and the two species collectively, respectively. At 30% moisture contents, C. meiocarpa and C. oleifer produced nine and eight significant differentially expressed miRNAs, respectively, with high fatty acid synthesis and accumulation activities. Across the two species, 12 significant differentially expressed miRNAs were identified at the 50% moisture content. Sequencing of small-RNA revealed the presence of 23 miRNAs regulating lipid metabolism in camellia seed during natural drying and permitted comparative miRNA profiles between C. Meiocarpa and C. oleifera. Furthermore, this study successfully identified the best drying environment at which the quantity and quality of lipid in camellia seed are at its maximum.


Spatial patterns of phylogenetic diversity.
Ecologists and conservation biologists have historically used species-area and distance-decay relationships as tools to predict the spatial distribution of biodiversity and the impact of habitat loss on biodiversity. These tools treat each species as evolutionarily equivalent, yet the importance of species' evolutionary history in their ecology and conservation is becoming increasingly evident. Here, we provide theoretical predictions for phylogenetic analogues of the species-area and distance-decay relationships. We use a random model of community assembly and a spatially explicit flora dataset collected in four Mediterranean-type regions to provide theoretical predictions for the increase in phylogenetic diversity - the total phylogenetic branch-length separating a set of species - with increasing area and the decay in phylogenetic similarity with geographic separation. These developments may ultimately provide insights into the evolution and assembly of biological communities, and guide the selection of protected areas.


Necrotizing Aspergillosis of large airways: CT findings in eight patients.
The aim of this study was to evaluate the CT findings of pathologically proven necrotizing aspergillosis of the large airways (necrotizing Aspergillus bronchitis). Medical records and imaging studies from two tertiary medical centers were reviewed for pathologically proven cases of necrotizing aspergillosis of the large airways. Fiberoptic bronchoscopic examination and CT scans of the chest were available in all cases. Two thoracic radiologists who were blinded to the clinical and pathologic data reviewed the thoracic CT scans retrospectively and reached a final decision. The CT images were evaluated for the presence, distribution, and extent of CT findings. The study included eight patients, seven men and one woman, ranging in age from 28 to 67 years (mean age 46 years). All patients had histopathologically proved necrotizing Aspergillus of the large airways and no other superimposed infections. Six patients had leukemia, one had chronic liver disease, and one had chronic obstructive lung disease. All patients had bronchial wall thickening and focal bronchial narrowing involving a lobar or segmental bronchus. The bronchial narrowing was irregular or nodular in seven patients and smooth in one. Atelectasis distal to a narrowed bronchus was present in five patients. The CT findings of necrotizing bronchial aspergillosis include bronchial wall thickening, which is often nodular, and narrowing of the bronchial lumen, which is often associated with distal atelectasis.


Investigation of the binding interactions mechanism between zein with chrysin by multispectroscopic techniques.
As a natural carrier protein, zein was intensively studied for the construction of a flavonoid delivery system. Chrysin has presented superior tumor-resistant, anti-inflammatory, and anti-oxidation potentials among the flavonoid candidates in clinical practice. However, due to inadequate research, the binding mechanism and structural affinity of zein to chrysin are still indeterminate. Therefore, multispectral methods were employed to explore the molecular interaction of zein and chrysin in this work. These techniques showed that chrysin reduced the intrinsic fluorescence of zein via a static process and that the interaction between zein and chrysin was mainly driven spontaneously by hydrophobic forces. Additionally, the experimental results revealed the changed microenvironment in the vicinity of tyrosine and affected secondary structure in the presence of chrysin, indicating zein's conformation were altered by chrysin. This work provided comprehensive insight into the combination of plant-derived protein (zein) and flavonoids (chrysin) and helped rationalize the protection, transportation, and release of chrysin through a zein-based delivery system.


Scattering of electromagnetic waves from 3D multilayer random rough surfaces based on the second-order small perturbation method: energy conservation, reflectivity, and emissivity.
A theoretical investigation of energy conservation, reflectivity, and emissivity in the scattering of electromagnetic waves from 3D multilayer media with random rough interfaces using the second-order small perturbation method (SPM2) is presented. The approach is based on the extinction theorem and develops integral equations for surface fields in the spectral domain. Using the SPM2, we calculate the scattered and transmitted coherent fields and incoherent fields. Reflected and transmitted powers are then found in the form of 2D integrations over wavenumber in the spectral domain. In the integrand, there is a summation over the spectral densities of each of the rough interfaces with each weighted by a corresponding kernel function. We show in this paper that there exists a "strong" condition of energy conservation in that the kernel functions multiplying the spectral density of each interface obey energy conservation exactly. This means that energy is conserved independent of the roughness spectral densities of the rough surfaces. Results of this strong condition are illustrated numerically for up to 50 rough interfaces without requiring specification of surface roughness properties. Two examples are illustrated. One is a multilayer configuration having weak contrasts between adjacent layers, random layer thicknesses, and randomly generated permittivity profiles. The second example is a photonic crystal of periodically alternating permittivities of larger dielectric contrast. The methodology is applied to study the effect of roughness on the brightness temperatures of the Antarctic ice sheet, which is characterized by layers of ice with permittivity fluctuations in addition to random rough interfaces. The results show that the influence of roughness can significantly increase horizontally polarized thermal emission while leaving vertically polarized emissions relatively unaffected.


Genetic risk factor for abdominal aortic aneurysm: HLA-DR2(15), a Japanese study.
Autoimmunity has been proposed to play a role in the pathogenesis of the abdominal aortic aneurysm (AAA). Several autoimmune diseases are associated with specific HLA DR alleles. These experiments were carried out to determine whether the same HLA DR types that have been reported to be associated with AAA in a mixed North American population are similarly associated with AAA in a more homogeneous group of patients in Japan. HLA DR typing was performed by a serologic method on samples of peripheral blood of patients with nonspecific infrarenal AAA in Nagasaki University Hospital in Japan. The frequencies of HLA DR antigens were compared with those of volunteers approximately matched for age and sex from the same referral area. HLA DR haplotypes were determined in 46 Japanese patients with AAA and in 50 patients in a control group. The HLA-DR2(15) antigen was observed in 27 (58.7%) patients (29 alleles 31.5%) with AAA and in 14 (28%) subjects (16 alleles 26.0%) in the control group (p < 0.005). The data suggest that HLA-DR2(15) has an important role as a genetic risk factor for AAA in Japanese patients, as previously reported in a mixed North American population.


Large Salt Dust Storms Follow a 30-Year Rainfall Cycle in the Mar Chiquita Lake (Córdoba, Argentina).
Starting in 2006, a new source of intense salt dust storms developed in Mar Chiquita (Córdoba, Argentina), the largest saline lake in South America. Storms originate from vast mudflats left by a 30-year expansion-retreat cycle of the lake due to changes in the regional rainfall regime. The annual frequency of salt dust storms correlated with the size of the salt mudflats. Events were restricted to the coldest months, and reached up to 800 km from the source. Occurrence of dust storms was associated with specific surface colors and textures easily identifiable in satellite images. High-emission surfaces were characterized by the presence of sodium sulfate hydrous/anhydrous crystals (mirabilite and thenardite), and a superficial and variable water table, which may result in the periodic development of a characteristic "fluffy" surface derived from salt precipitation-dissolution processes. HYSPLIT model simulation estimates a deposition maximum near the sources (of about 2.5 kg/ha/yr), and a decreasing trend from the emission area outwards, except for the relative secondary maximum modeled over the mountain ranges in southern Bolivia and northern Argentina due to an orographic effect. The 2009 total deposition of salt dust generated in Mar Chiquita was estimated at 6.5 million tons.


The in vitro interaction of dexmedetomidine with human liver microsomal cytochrome P4502D6 (CYP2D6).
The effect of dexmedetomidine DEX on cytochrome P4502D6 (CYP2D6)-dependent dextromethorphan O-demethylase (DEXTROase) activity was studied using native human liver microsomes. DEX (0.01-4.0 microM inhibited DEXTROase activity (IC50 = 1.8 +/- 0.25 microM; mean +/- SD; N = 5 livers) and was less potent than quinidine (QND), prototypical and clinically relevant CYP2D6 inhibitor (IC50 = 0.22 +/- 0.02 microM; mean Ki = 0.07 microM). Similar results were obtained with human B-lymphoblast microsomes containing cDNA-expressed CYP2D6 (DEX, IC50 = 2.2 microM; QND, IC50 0.15 microM). Formal kinetic analyses indicated that DEX was a reversible mixed (competitive/noncompetitive) inhibitor of DEXTROase activity in human liver microsomes, where Kies > Ki and alpha > 1 (Ki = 0.4 +/- 0.2 microM; Kies = 2.3 +/- 0.9 microM; alpha = 8.1 +/- 6.8; N = 3 livers). In addition, DEX elicited a Type IIb difference spectrum (lambda max approximately 436 nm; lambda min approximately 414 nm) when added to cDNA-expressed CYP2D6 under aerobic (oxidized) conditions. These data indicated that DEX was able to bind reversibly to the heme (ferric) iron of CYP2D6. It is postulated that binding occurs via the 4(5)-substituted imidazole moiety. In this instance, binding was characterized by a spectral dissociation constant (Ks) of 0.4 microM that was identical to the Ki obtained with native human liver microsomes.


Bifunctional charge transfer operated fluorescent probes with acceptor and donor receptors. 2. Bifunctional cation coordination behavior of biphenyl-type sensor molecules incorporating 2,2':6',2' '-terpyridine acceptors.
Based on donor (D)-acceptor (A) biphenyl (b) type molecules, a family of fluorescent reporters with integrated acceptor receptors and noncoordinating and coordinating donor substituents of varying strength has been designed for ratiometric emission sensing and multimodal signaling of metal ions and protons. In part 2 of this series on such charge transfer (CT) operated mono- and bifunctional fluorescent devices, the cation coordination behavior of the sensor molecules bpb-R equipped with a proton- and cation-responsive 2,2':6',2' '-terpyridine (bp) acceptor and either amino-type donor receptors (R = DMA, A15C5 = monoaza-15-crown-5) or nonbinding substituents (R = CF(3), H, OMe) is investigated employing the representative metal ions Na(I), Ca(II), Zn(II), Hg(II), and Cu(II) and steady-state and time-resolved fluorometry. The bpb-R molecules, the spectroscopic behavior and protonation behavior of which have been detailed in part 1 of this series, present rare examples for CT-operated bifunctional fluorescent probes that can undergo consecutive and/or simultaneous analyte recognition. The analyte-mediated change of the probes' intramolecular CT processes yields complexation site- and analyte-specific outputs, i.e., absorption and fluorescence modulations in energy, intensity, and lifetime. As revealed by the photophysical studies of the cation complexes of these fluoroionophores and the comparison to other neutral and charged D-A biphenyls, the spectroscopic properties of the acceptor chelates of bpb-R and A- and D-coordinated bpb-R are governed by CT control of an excited-state barrier toward formation of a forbidden charge transfer state, by the switching between analytically favorable anti-energy and common energy gap law type behavior, and by the electronic nature of the ligated metal ion. This accounts for the astonishingly high fluorescence quantum yields of the acceptor chelates of bpb-R equipped with weak or medium-sized donors and the red emission of D- and A-coordinated bpb-R observed for nonquenching metal ions.


Plasticity of representational maps in somatosensory cortex observed by in vivo voltage-sensitive dye imaging.
We investigated the effect of selective whisker trimming on the development of the cortical representation of a whisker deflection in layer 2/3 of rat somatosensory cortex using in vivo voltage-sensitive dye (vsd) imaging. Responses to deflection of D-row whiskers were recorded after trimming of A-row, B-row, and C-row whiskers, referred to as DE pairing, during postnatal development. Animals DE paired from postnatal day (p) 7 to p17 had a significant bias in the spread of the vsd signal, favoring spread toward the concomitantly nondeprived E-row columns. This resulted primarily from a strong decrease in signal spreading into the deprived C-row columns. In contrast, signal spread in control littermates was approximately symmetrical. DE pairing failed to elicit significant changes when begun after p14, thus defining a critical period for this phenomenon. The results suggest that sensory deprivation in this model results in lower connectivity being established between nondeprived columns and adjacent deprived ones.