Molecular epidemiology of HTLV-I in the world.
The geographic distribution of human T-cell lymphotropic/leukemia virus type I (HTLV-I) was initially believed to be limited to southwestern Japan, the Caribbean basin, and Africa. However, extensive searches in recent years have discovered its existence in other areas of the world as well as in isolated, ethnic populations such as South Amerindians, Australo-Melanesian aborigines, religiously segregated Jews, and Pygmies. Previous genetic analyses indicated that HTLV-I can be phylogenetically classified into three major lineages: Melanesian, Central African, and Cosmopolitan groups. Recently, more detailed characterization using long terminal repeat sequences (the most variable genomic region) has revealed that the Cosmopolitan group consists of four subtypes: (A) Transcontinental, (B) Japanese, (C) West African, and (D) North African. Most HTLV-I isolates of the same ethnic group from distant locations and those from different groups inhabiting the same area showed phylogenetic similarities. These observations indicate the present distribution of this virus should be interpreted from the anthropological backgrounds of the virus-possessing populations as well as spatial contact among them. Thus, the molecular epidemiology of HTLV-I and its simian counterpart, STLV-I, provides us with important clues for understanding not only the origin and dissemination of this retrovirus but past human movements over the globe.


Glial fibrillary acidic protein in the fish optic nerve.
The intermediate filament glial fibrillary acidic protein (GFAP) is the predominant cytoskeletal protein of mature glial cells in the mammalian nervous system. The nervous systems of lower vertebrates, such as fish, have been examined for the presence of GFAP and several investigators have shown that goldfish (Carassius auratus) brain contains GFAP-positive astrocytes. The same studies have demonstrated that, in contrast to the brain, the optic nerve of goldfish did not show any GFAP immunoreactivity, suggesting that this intermediate filament protein is not expressed in fish optic nerve astrocytes. The present study shows, however, that the monoclonal antibodies to porcine GFAP react with the optic nerve of carp (Cyprinus carpio), another member of the goldfish family. These antibodies to porcine GFAP cross react with rat brain and carp optic nerve, yielding a band of approximately 52 kDa in both species. Northern blot analysis using mouse GFAP DNA probe revealed that carp optic nerve RNA contains two transcripts of 2.3 and 2.1 kb, which hybridize with the mouse GFAP probe. Injury to the carp optic nerve was followed by a decrease of GFAP immunoreactivity from neural tissue and a strong expression around blood vessels and connective tissues. On the basis of these observations and within the limitation of the techniques it is reasonable to conclude that the carp optic nerve expresses GFAP immunoreactivity and that the pattern of expression of this intermediate filament protein is altered after injury. Such an alteration might be relevant to the process of regeneration.


Enzyme-catalyzed dehalogenation of pentachloroethane: why F87W-cytochrome P450cam is faster than wild type.
Under anaerobic conditions, cytochromes P450 can reductively dehalogenate heavily halogenated hydrocarbons, such as one- and two-carbon organic solvents. This catalytic capacity has drawn attention to the potential use of engineered forms of P450s in the remediation of contaminated deep subsurface ecosystems. Loida (1994, PhD Thesis, University of Illinois at Urbana-Champaign, IL) and S.G. Sligar (personal communication) have observed recently that an active-site variant of cytochrome P450cam (F87W) dechlorinates pentachloroethane approximately three times faster than the wild-type enzyme. Molecular dynamics simulations have revealed that the mutant enzyme binding pocket remains smaller, and that pentachloroethane assumes configurations closer to the heme-Fe in the F87W mutant twice as often as in the wild-type enzyme. This result is consistent with a collisional model of dehalogenation, which agrees with experimental observations [Li and Wackett (1993) Biochemistry, 32, 9355-9361] that solutions containing wild-type P450cam dehalogenate pentachloroethane 100 times faster than those containing free heme. The simulations suggest that it is unlikely that Trp87 significantly stabilizes the developing negative charge on the substrate during carbon-halogen bond reduction. The design of improved microbial enzymes that incorporate both steric and electronic effects continues for use in remediating halogenated contaminants in situ.


Inflammatory and cytotoxic potential of the airborne particle material assessed by nasal lavage and cell exposure methods.
Exposure to bioaerosols in moisture-damaged indoor environments has been shown to be a potential health risk. The aim of the present study was to evaluate the inflammatory and cytotoxic potential of airborne particle material using both the nasal lavage (NAL) method and a cell exposure study. A 24-h sample collection for airborne particles was performed using personal sampling and microenvironmental measurements in homes and an 8-h sample collection in the working places of the studied subjects. At the end of the sampling period, the production of nitric oxide, tumor necrosis factor alpha, interleukin (IL)-1 beta, IL-4, and IL-6 was analyzed in the NAL samples of the subjects. The same mediators, excluding IL-4, were measured in the cell culture medium of mouse RAW264.7 macrophages, which were exposed to the pooled filter extracts representing personal, home, and workplace exposure of each individual during the 24 h before the NAL. Samplings were repeated after 2 wk. The subjects were divided into groups of "low exposure" and "high exposure" according to the concentrations of viable fungi, viable bacteria, or total microbial amount in the pooled extract. Cytokine levels in the NAL samples of subjects with high microbial exposure were slightly increased compared to the corresponding values of the subjects with low exposure. Filter samples collected from the subjects with high microbial exposure induced a significant increase in the production of cytokines in the RAW264.7 macrophages, as compared to those from the subjects with low exposure. The within-subject variation was low in all of the cytokine measurements, but the correlation between the studied methods was poor. In conclusion, both of the methods discriminate at the group level between subjects with high and low microbial exposure. Sampling of airborne particle material and exposure of the mammalian cells to the obtained samples seems to be highly applicable in the environmental monitoring, whereas examination of the exposed subjects directly, for example by using the NAL method, is essential when association between exposure and health effects is evaluated.


[Remineralizing methods in early caries lesions - review of the literature].
Early diagnosis and treatment of early caries lesions are the basic tenets of minimally invasive dentistry. One of the key elements of a biological approach is the usage of remineralizing agents, which are great alternative to surgical intervention. This article shows a review of the literature concering materials used for the treatment of early caries lesions.


Characterization of elemental sulfur in isolated intact spinach chloroplasts.
Incubation of intact spinach (Spinacia oleracea L.) chloroplasts in the presence of (35)SO(4) (2-) resulted in the light-dependent formation of a chloroform-soluble sulfur-containing compound distinct from sulfolipid. We have identified this compound as the most stable form (S(8)) of elemental sulfur (S(0), valence state for S = O) by mass spectrometry. It is possible that elemental sulfur (S(0)) was formed by oxidation of bound sulfide, i.e. after the photoreduction of sulfate to sulfide by intact chloroplasts, and released as S(8) under the experimental conditions used for analysis.


Self-assembly of thiophene derivatives on highly oriented pyrolytic graphite: hydrogen bond effect.
In this paper, to elucidate the hydrogen bond effect on the assembly behavior, we studied the assembly structures of two carboxylic substituted thiophene derivatives on highly oriented pyrolytic graphite (HOPG) by scanning tunneling microscopy. Here thiophene-2-carboxylic acid (TCA) and thiophene-2,5-dicarboxylic acid (TDA) were employed. TDA molecules spontaneously adsorb on the HOPG surface and self-organize into a two-dimensional (2D) assembly with well-defined structure. Two types of domain could be observed. Each TDA molecule appears as a round circle with two small faint dots and forms hydrogen bonds with neighbours. Besides monolayer structure, a bilayer structure of TDA adlayer on HOPG was also observed in this research. Remnant TDA molecules adsorb on the monolayer of TDA and bilayer structure is formed. In contrast to TDA, no ordered structure of TCA on HOPG can be observed. TCA molecules have high propensity to form dimers through H-bond between carboxylic groups. But TCA dimer is not stable enough for either adsorption or imaging. Our result provides a new example for understanding hydrogen effect on stabilizing and controlling two-dimensional assembly structure and is helpful for surface nanofabrication and development of electric nanodevices.


Dynamics of parasite communities and interactions between wild and domestic ruminants.
Dynamics of parasite communities and host-parasite-environment interactions can be influenced by different factors. The present note, by discussing some field experiences in wild and domestic ruminant populations, approaches eco-epidemiology of abomasal nematodes in relation with host health and dynamics. Factors possibly playing a role in the host-parasite relationship are discussed, as well as possible use of macroparasites as ecological and sanitary indicators. The above topics are approached in a management perspective, in particular regarding interactions between domestic and wild ruminant populations.


Citral, an inhibitor of retinoic acid synthesis, modifies chick limb development.
Exogenously applied retinoic acid (RA) is known to affect cartilage pattern in developing and regenerating limbs. There are, however, few reports which analyze the participation of endogenous RA in limb pattern formation. Using an organ culture system, we attempted to reduce the concentration of endogenous RA in the developing chick wing buds by the treatment with citral (3,7-dimethyl-2,6-octadienal), an inhibitor of retinoic acid formation. After this treatment, the cultured wing buds were grafted to the stumps of host embryos. These citral-treated limb buds frequently formed truncated cartilage elements and the defects were rescued by simultaneous treatment with an appropriate concentration of RA. These results suggest that endogenous RA plays a role in chick limb bud development.


PI3K-dependent lysosome exocytosis in nitric oxide-preconditioned hepatocytes.
We investigated the signal mediators and the cellular events involved in the nitric oxide (NO)-induced hepatocyte resistance to oxygen deprivation in isolated hepatocytes treated with the NO donor (Z)-1-(N-methyl-N-[6-(N-methylammoniohexyl)amino])diazen-1-ium-1,2-diolate (NOC-9). NOC-9 greatly induced PI3K activation, as tested by phosphorylation of PKB/Akt. This effect was prevented by either 1H-(1,2,4)-oxadiazolo-(4,3)-quinoxalin-1-one, an inhibitor of the soluble guanylate cyclase (sGC), or KT5823, an inhibitor of cGMP-dependent kinase (cGK), as well as by farnesyl protein transferase inhibitor, which blocks the function of Ras GTPase. Bafilomycin A, an inhibitor of the lysosome-type vacuolar H+-ATPase, cytochalasin D, which disrupts the cytoskeleton-dependent organelle traffic, and wortmannin, which inhibits the PI3K-dependent traffic of lysosomes, all abolished the NOC-9-induced hepatocyte protection. The treatment with NOC-9 was associated with the PI3K-dependent peripheral translocation and fusion with the plasma membrane of lysosomes and the appearance at the cell surface of the vacuolar H+-ATPase. Inhibition of sGC, cGK, and Ras, as well as the inhibition of PI3K by wortmannin, prevented the exocytosis of lysosomes and concomitantly abolished the protective effect of NOC-9 on hypoxia-induced pHi and [Na+]i alterations and cell death. These data indicate that NO increases hepatocyte resistance to hypoxic injury by activating a pathway involving Ras, sGC, and cGK that determines PI3K-dependent exocytosis of lysosomes.


Diabetic Ketoacidosis With Acute Metabolic Pancreatitis: Two Serious Cases.
Hypertriglyceridemia is a rare cause of acute pancreatitis (AP), occupying approximately 7% of cases. The triad of acute pancreatitis, hypertriglyceridemia, and diabetes is a rare event, with a higher death rate. We describe two cases of severe acute metabolic pancreatitis discovered in diabetic ketoacidosis. For both patients, all other causes of AP were excluded (including gallstones, hypercalcemia, drugs, and alcohol). A laboratory workup revealed elevated lipasemia (more than three times the normal) and hypertriglyceridemia. Abdominal computed tomography confirmed the diagnosis of severe acute pancreatitis. Fasting, fluid resuscitation, and insulin therapy were initiated in the intensive care unit with good clinical results and progressive improvement in metabolic disorders.


The bonding behavior of calcite to bone.
Plates of calcite (CaCO3) were implanted in rabbit tibiae, and their biocompatibility and bonding ability to bone were studied. The plates were also implanted subfascially in rabbit muscle for 8 weeks, and changes on their surfaces in the body were examined. Contact microradiography and Giemsa surface stain demonstrated direct bonding between calcite and bone without interpositions. The average failure load of the interface between calcite and bone was 4.11 kg, indicating an adequate strength of bonding. However, a Ca-P-rich layer, which formed on the surfaces of other bioactive ceramics in vivo, was not detected by a scanning electron microscope-electron probe x-ray microanalyzer. Scanning electron micrographs of the surface of calcite implanted subfascially for 8 weeks showed marked degradation and a rough surface. However, the surface apatite layer was not detected by thin-film x-ray diffraction analysis and Fourier transform infrared reflection spectroscopy. Calcite is a biodegradable material that bonds to bone without a surface apatite layer. The mechanical bonding provided by the anchoring effect of the newly formed bone into the surface roughness of calcite is considered to be a major factor in calcite-bone bonding.


Thoracic epidural analgesia in a patient with bilateral phaeochromocytoma undergoing coronary artery bypass grafting.
A patient suffering from phaeochromocytoma and coronary artery stenoses needed coronary artery bypass grafting before adrenalectomy. High thoracic epidural analgesia (T1-T2) with bupivacaine and sufentanil in combination with general anaesthesia was used. Plasma adrenaline and noradrenaline concentrations decreased during the period before bypass grafting compared to the baseline value and no important haemodynamic changes were seen during this period. Thoracic epidural analgesia failed to suppress the release of catecholamine during the bypass period. After the operation, the plasma catecholamine concentrations returned to the baseline value. Excellent analgesia (visual analogue scale = 1-2) was achieved with a postoperative epidural, but the plasma catecholamine concentration increased considerably.


Three-dimensional geometric morphometric analysis of the skull of Protoceratops andrewsi supports a socio-sexual signalling role for the ceratopsian frill.
Socio-sexual selection is predicted to be an important driver of evolution, influencing speciation, extinction and adaptation. The fossil record provides a means of testing these predictions, but detecting its signature from morphological data alone is difficult. There are, nonetheless, some specific patterns of growth and variation which are expected of traits under socio-sexual selection. The distinctive parietal-squamosal frill of ceratopsian dinosaurs has previously been suggested as a socio-sexual display trait, but evidence for this has been limited. Here, we perform a whole-skull shape analysis of an unprecedentedly large sample of specimens of Protoceratops andrewsi using a high-density landmark-based geometric morphometric approach to test four predictions regarding a potential socio-sexual signalling role for the frill. Three predictions-low integration with the rest of the skull, significantly higher rate of change in size and shape during ontogeny, and higher morphological variance than other skull regions-are supported. One prediction, sexual dimorphism in shape, is not supported, suggesting that sexual differences in P. andrewsi are likely to be small. Together, these findings are consistent with mutual mate choice or selection for signalling quality in more general social interactions, and support the hypothesis that the frill functioned as a socio-sexual signal in ceratopsian dinosaurs.


Phenolics Profile and Antioxidant Activity Analysis of Kiwi Berry (Actinidia arguta) Flesh and Peel Extracts From Four Regions in China.
The kiwi berry (Actinidia arguta) has been widely studied because of its rich phenolic, flavonoid, and vitamin C contents. Numerous reports have demonstrated that fruit peels contain higher phenolic content and antioxidant activity than that of flesh. In this study, the phytochemical content and antioxidant activities of peel and flesh extracts of six kiwi berries were analyzed from four regions (namely, Dandong, Benxi, Taian, and Tonghua) in China. The antioxidant activity was determined using the peroxyl radical scavenging capacity (PSC) and cellular antioxidant activity (CAA) assays. The phenolic, flavonoid, and vitamin C contents of kiwi berry peel were 10.77, 13.09, and 10.38 times richer than that of kiwi berry flesh, respectively. In addition, the PSC and CAA values of kiwi berry peel were higher than those of kiwi berry flesh. The analysis of the separation and contents of phenolics were performed by the high-performance liquid chromatography (HPLC)-diode-array detectormass spectrometry/mass (DAD-MS/MS) system, and the results illustrated that protocatechuic acid, caffeic acid, chlorogenic acid, and quinic acid were the major phenolic compounds. In conclusion, this study indicated that kiwi berry peel contains a rich source of antioxidants. These data are of great significance for the full development and utilization of kiwi berries in these four regions of China to produce nutraceutical and functional foods.


Fused Filament Deposition of PLA: The Role of Interlayer Adhesion in the Mechanical Performances.
A set of criteria to enhance mechanical performances of standard specimens (Type V, ANSI D368) made of polylactic acid (PLA) were proposed. Fused PLA deposition was conducted with nozzle temperature ranging from 180 to 230 °C and deposition plate temperature ranging from 70 to 110 °C. Optical microscopy, elastic modulus analysis and density measurement allowed emphasizing the effect of temperature field, also measured during the process, on the morphology and the mechanical characteristics of the specimen. Atomic force microscopy revealed a morphology typical of amorphous samples with globular structures. Poor interlayer adhesion was detected in the part of the specimen located at larger distance from the deposition plate, showing an elastic modulus lower than those measured in the central part (220 MPa vs. 500 MPa). The specimen crystallinity degree was below 3%. The molecular weight between entanglements was adopted as a measure of the interlayer molecular diffusion. A successful diffusion and re-entanglement of the polymer melt at the interface was the key to improving mechanical performance. A mathematical model describing the transient heat transfer during the fused PLA deposition and accounting for solidification and the nonisothermal crystallization kinetics was introduced. Simulated temperature evolutions were consistent with the experimental ones. They were related to the mechanical performances, the morphology, and the molecular weight between entanglements of the parts.


Blocking T cell co-stimulation in primary Sjögren's syndrome: rationale, clinical efficacy and modulation of peripheral and salivary gland biomarkers.
There is accumulating evidence that patients with primary Sjögren's syndrome (pSS) display aberrant CD4+ T cell responses, both in the peripheral compartment and in the inflamed salivary glands. CD4+ T cell abnormalities are also critically associated with B cell hyper activation, one of the hallmarks of disease, which is linked with disease severity and evolution to lymphoma. T cell activation and the cross-talk between T and B cells are tightly regulated by the balance between co-stimulatory pathways, such as the interactions between CD80/CD86:CD28, CD40:CD40L and ICOS:ICOSL, and co-inhibitory signals, including the immunoregulatory CTLA-4 protein. Evidence from patients with pSS as well as data from animal models of the disease suggests that these pathways play a critical role in pSS pathogenesis and their targeting could be exploited for therapeutic purposes. In this review, we first summarise the evidence implicating aberrant T cell co-stimulation and co-inhibition in driving the disease before focusing on the results of recent randomised controlled trials (RCTs) with compounds able to block T cell co-stimulation and enhance T cell co-inhibition. Despite a clear biological effect on downstream B cell activation has been observed in patients treated with CTLA-4-Ig (abatacept) and with monoclonal antibodies targeting CD40 and ICOSL, the clinical efficacy of this approach has so far yielded mixed results; while the anti-CD40 monoclonal antibody iscalimab showed significant improvement in systemic disease activity compared to placebo, two large RCTs with abatacept and a phase IIa RCT with an anti-ICOSL monoclonal antibody (prezalumab) failed to reach their primary endpoints. Although the discrepancies between biological and clinical efficacy of targeting T cell co-stimulation on pSS remain unresolved, several factors including drug bioavailability and receptor occupancy, patient stratification based on T-cell related biomarkers and the choice of study outcome are likely to play an important role and form the basis for further work towards the quest for a disease-modifying biologic therapy in pSS.


Causative agents and antimicrobial susceptibilities of urinary tract infections in the northwest of Iran.
The empirical therapy of urinary tract infections (UTI) relies on the predictability of the agents causing UTI and knowledge of their antimicrobial susceptibility patterns. In a prospective study undertaken over a 14-month period, 5136 samples from patients suspected of having a UTI were analyzed, of which 676 were culture-positive. Isolated bacteria were identified by standard tests, and antibiotic susceptibility was determined by disk diffusion method. According to our results, Escherichia coli was the most common etiological agent of UTI (74.6%), followed by Klebsiella spp (11.7%), Staphylococcus saprophyticus (6.4%), and Pseudomonas aeruginosa (2.2%). Analysis of the frequency of isolated bacteria according to the age of the patients revealed that Klebsiella infections are more prevalent in the older age groups (>10 years) and Pseudomonas infections are more prevalent in children and the elderly (<9 years and >60 years). Results of antimicrobial susceptibility analysis for E. coli, as the most prevalent cause of UTI, to commonly used antibiotics are as follows: amikacin (97.8%), gentamicin (97%), ciprofloxacin (94%), nitrofurantoin (87.1%), nalidixic acid (93.7%), trimethoprim-sulfamethoxazole (48.2%), cephalexin (76%), and ampicillin (6.9%). The results show that the antimicrobial resistance patterns of the causes of UTI are highly variable and continuous surveillance of trends in resistance patterns of uropathogens is important.


Mylohyoid Ridge as a Predictor of Available Bone for Implant Placement: A Cone-Beam Computed Tomography (CBCT) Retrospective Observational Study.
The posterior mandibular region, due to the presence of vital structures, poses a high risk during implant placement because of its susceptibility to neurovascular injury and perforation of the lingual cortex. A breach in implant length and available bone height may lead to serious intraoperative and postoperative complications. Prediction of the exact location of the inferior alveolar nerve and submandibular fossa anatomy is a prerequisite for ideal implant placement, which is always not possible with conventional radiographic and clinical techniques. One hundred ten cone-beam computed tomographies (CBCTs) of patients were acquired from the radiological archives of a radiological center in Chennai. DICOM files from CBCT were exported to Bly Sky Plan software. Cross-sections of the second molar and first molar were extracted following the inclusion criteria. The linear dimension between the mandibular canal and mylohyoid ridge and anatomic variables of the submandibular fossa were measured digitally on the left and right sides using software measuring tools. Descriptive statistics were done. The unilateral and bilateral site and gender differences were evaluated. Bone height superior to the mandibular canal was correlated with the submandibular fossa parameters; depth of undercut in the vertical and horizontal directions; and angle of the undercut. The mandibular canal was on average 5.5 mm and 4 mm inferior to the Mylohyoid ridge in the second molar region and first molar region, respectively, with the right and left sides showing no statistically significant difference. The depth of fossa undercut in vertical and horizontal dimensions was higher in the second molar region compared to the first molar region. The height of the deepest point of the undercut in the vertical dimensions showed a positive correlation with the bone available between the mandibular canal and the mylohyoid ridge. Keeping 2 mm of safety factor in consideration, implants can be safely placed up to the mylohyoid ridge in 100% of cases and 2 mm below the mylohyoid ridge in 78.9% of cases in the mandibular second molar region. In keeping with a safety factor of 2 mm, implants can be safely placed up to the mylohyoid ridge in 82.6% of cases and 2 mm below the mylohyoid ridge in 43.1% of cases in the first molar region. A more pronounced undercut was seen in the second molar region than in the first molar region. Deeper fossa undercuts in vertical dimension are associated with more inferior positioning of the mandibular canal.


Heterozygous P0 deficiency protects mice from vincristine-induced polyneuropathy.
Patients with hereditary neuropathies are more susceptible to vincristine (VIN)-induced neuropathy than patients without this comorbidity. The heterozygous P0(+/-) mouse is an animal model of a distinct form of inherited neuropathies. These mice produce only 50% of the major myelin protein protein zero (P0) and display signs of demyelination in motor nerves at 4 months of age. Here we investigated the development of neuropathic signs in P0(+/-) and wild-type (wt) mice after VIN treatment. Neuropathy was induced by daily intraperitoneal injections of VIN (0.5 mg/kg body weight) over 10 days. Behavioral and electrophysiological tests were performed at regular time points. Wt mice developed significant hypersensitivity to heat and mechanical stimuli between days 7 and 38 after the first VIN injection. Surprisingly, P0(+/-) mice did not show sensory or motor signs of neuropathy over the whole testing period. Immunohistochemical analysis showed an increase in macrophage numbers in sciatic nerve sections of wt mice after VIN, whereas P0(+/-) mice had higher baseline levels of macrophages without changes after VIN treatment. Semithin sections revealed a decrease in the number of small-diameter myelinated fibers in the sciatic nerves of wt mice after VIN application, whereas P0(+/-) mice had higher baseline values of this fiber subtype that did not change under treatment. Dorsal root ganglion neurons of both genotypes showed an up-regulation of voltage-gated sodium channel immunoreactivity after VIN application without differences between the genotypes. Thus, the P0(+/-) phenotype seems to be protected against VIN-induced neuropathy. The mechanism of this neuroprotection remains elusive.


In vivo activity of optimized apidaecin and oncocin peptides against a multiresistant, KPC-producing Klebsiella pneumoniae strain.
The peptides Api88 and Onc72 are highly efficient to treat Escherichia coli bacteremia in mice. Here we extended the animal studies to a systemic murine infection model using a multidrug-resistant carbapenemase-producing Klebsiella pneumoniae clinical isolate. When administered intraperitoneally three times at 2.5, 5 and 10 mg/kg bodyweight to CD-1 mice infected with a KPC-producing K. pneumoniae strain, both Api88 and Onc72 reduced the bacterial counts by at least 5 log₁₀ units, indicating that both peptides are active in vivo. Both peptide treatments increased significantly the survival rates and average survival times compared to untreated animals for all doses except for the highest dose of Onc72. This dose reduced the bacterial counts so fast that it most likely induced a sudden release of large amounts of toxic materials from the killed bacteria reducing the survival time even below that of untreated mice. In conclusion, both peptides were efficient in the lethal murine K. pneumoniae infection model, but the treatment protocol (i.e. dose and time points) has to be further optimized based on future pharmacokinetic studies.


Femtosecond fiber laser welding of dissimilar metals.
In this paper, welding of dissimilar metals was demonstrated for the first time, to the best of our knowledge, by using a high-energy high-repetition-rate femtosecond fiber laser. Metallurgical and mechanical properties were investigated and analyzed under various processing parameters (pulse energy, repetition rate, and welding speed). Results showed that the formation of intermetallic brittle phases and welding defects could be effectively reduced. Strong welding quality with more than 210 MPa tensile strength for stainless steel-aluminum and 175 MPa tensile strength for stainless steel-magnesium has been demonstrated. A minimal heat affected zone and uniform and homogenous phase transformation in the welding region have been demonstrated. This laser-welding technique can be extended for various applications in semiconductor, automobile, aerospace, and biomedical industries.


[Modeling the academic performance of medical students in basic sciences and pre-clinical courses: a longitudinal study].
The study of predictors of academic performance is relevant for medical education. Most studies of academic performance use global ratings as outcome measure, and do not evaluate the influence of the assessment methods. To model by multivariate analysis, the academic performance of medical considering, besides academic and demographic variables, the methods used to assess students' learning and their preferred modes of information processing. Two hundred seventy two students admitted to the medical school of the Pontificia Universidad Católica de Chile from 2000 to 2003. Six groups of variables were studied to model the students' performance in five basic science courses (Anatomy, Biology, Calculus, Chemistry and Physics) and two pre-clinical courses (Integrated Medical Clinic I and IT). The assessment methods examined were multiple choice question tests, Objective Structured Clinical Examination and tutor appraisal. The results of the university admission tests (high school grades, mathematics and biology tests), the assessment methods used, the curricular year and previous application to medical school, were predictors of academic performance. The information processing modes influenced academic performance, but only in interaction with other variables. Perception (abstract or concrete) interacted with the assessment methods, and information use (active or reflexive), with sex. The correlation between the real and predicted grades was 0.7. In addition to the academic results obtained prior to university entrance, the methods of assessment used in the university and the information processing modes influence the academic performance of medical students in basic and preclinical courses.


Accounting for the Nutritional Context to Correctly Interpret Results from Studies of Exercise and Sedentary Behavior.
There is a wealth of research lauding the benefits of exercise to oppose cardiometabolic disease such as diabetes, CVD and hypertension. However, in the great majority of these studies, the nutritional context (energy balance, deficit, or surplus) has been ignored, despite its profound effect on responses to both exercise and inactivity. Even a minor energy deficit or surplus can strongly modulate the magnitude and duration of the metabolic responses to an intervention; therefore, failure to account for this important confounding variable obscures clear interpretation of the results from studies of exercise or inactivity. The aim of this review is to highlight key lessons from studies examining the interaction between exercise and sedentary behavior, energy status, and glucose and insulin regulation. In addition to identifying notable problems, we suggest a few potential solutions.


Two novel mutations in the gonadotropin-releasing hormone receptor gene in Brazilian patients with hypogonadotropic hypogonadism and normal olfaction.
Several point mutations in the GnRH receptor gene have been described in an autosomal recessive form of congenital isolated hypogonadotropic hypogonadism (HH). We investigated 17 Brazilian patients (10 males and 7 females) from 14 different families, with HH and normal olfaction. The diagnosis of HH was based on absent or incomplete sexual development after 17 yr of age associated with low or normal levels of LH in both sexes and low levels of testosterone in males and of estradiol in females. All patients presented with a normal sense of smell in an olfactory specific test. The coding region of the GnRH receptor gene was amplified by PCR and directly sequenced. A novel missense mutation, Arg(139)His, located in the conserved DRS motif at the junction of the third transmembrane and the second intracellular loop of the GnRH receptor was identified in the homozygous state in one female with complete HH. The Arg(139)His mutation completely eliminated detectable GnRH-binding activity and prevented GnRH-induced stimulation of inositol phosphate accumulation in vitro. In another family, a new compound heterozygous mutation (Asn(10)Lys and Gln(106)Arg) was identified in four siblings (two males and two females) with partial HH. The Gln(106)Arg mutation, located in the first extracellular loop, has been previously described, and in vitro analysis indicated that the mutant receptor was able to bind GnRH, but with a reduced affinity. The Asn(10)Lys mutation in the extracellular amino-terminal domain of the receptor also reduced the affinity for GnRH in vitro. In this family we also identified a previously described silent polymorphism at amino acid residue 151 in the second intracellular loop that segregated with the two inactivating mutations of the GnRH receptor. This polymorphism was also found in two unrelated patients with sporadic HH without GnRH receptor loss of function mutations. No mutations were identified in the remaining cases. A good correlation between genotype and phenotype was found in our patients. The woman, who is homozygous for the completely inactivating Arg(139)His mutation, has complete HH with undetectable serum basal LH and FSH levels that failed to respond to GnRH stimulation. In addition, the affected patients who are compound heterozygotes for the Asn(10)Lys/Gln(106)Arg mutations, have partial HH with low serum basal LH levels that were responsive to GnRH stimulation. No clinical or hormonal differences were found between HH patients with and without mutations in the GnRH receptor gene, indicating that these data do not contribute to the identification of HH patients with GnRH receptor mutations. In conclusion, we report the first naturally occurring mutation within the conserved DRS motif of the GnRH receptor in a female with complete HH and a novel compound heterozygous mutation (Asn(10)Lys and Gln(106)Arg) in a family with partial HH, increasing the repertoire of the inactivating mutations of the GnRH receptor.


Role of Zika Virus Envelope Protein Domain III as a Target of Human Neutralizing Antibodies.
Zika virus (ZIKV) is a flavivirus that is structurally highly similar to the related viruses, dengue virus (DENV), West Nile virus, and yellow fever virus. ZIKV causes an acute infection that often results in mild symptoms but that can cause severe disease in rare instances. Following infection, individuals mount an adaptive immune response, composed of antibodies (Abs) that target the envelope (E) glycoprotein of ZIKV, which covers the surface of the virus. Groups have studied monoclonal antibodies and polyclonal immune sera isolated from individuals who recovered from natural ZIKV infections. Some of these antibodies bind to domain III of E (EDIII), but the functional importance of these antibodies is unknown. In this study, we aimed to determine if EDIII is a major target of the potent serum neutralizing antibodies present in people after ZIKV infection. By generating a chimeric virus containing ZIKV EDIII in a DENV4 virus backbone, our data show a minor role of EDIII-targeting antibodies in human polyclonal neutralization. These results reveal that while monoclonal antibody (MAb) studies are informative in identifying individual antibody epitopes, they can overestimate the importance of epitopes contained within EDIII as targets of serum neutralizing antibodies. Additionally, these results argue that the major target of human ZIKV neutralizing antibodies resides elsewhere in E; however, further studies are needed to assess the epitope specificity of the neutralizing response at the population level. Identification of the major epitopes on the envelope of ZIKV recognized by serum neutralizing antibodies is critical for understanding protective immunity following natural infection and for guiding the design and evaluation of vaccines.IMPORTANCE Zika virus is a flavivirus that was recently introduced to Latin America, where it caused a massive epidemic. Individuals infected with ZIKV generate an immune response composed of antibodies which bind to the envelope (E) protein. These anti-E antibodies are critical in protecting individuals from subsequent infection. Multiple groups have found that many ZIKV antibodies bind to domain III of E (EDIII), suggesting that this region is an important target of neutralizing antibodies. Here, we generated a chimeric virus containing ZIKV EDIII in a dengue virus backbone to measure ZIKV EDIII-specific antibody responses. We found that while polyclonal ZIKV immune serum contains antibodies targeting EDIII, they constitute only a small fraction of the total population of antibodies that neutralize ZIKV. Further studies are needed to define the main targets on the viral envelope recognized by human neutralizing antibodies, which is critical for guiding the development of ZIKV vaccines.


Ca2+ and pH modulate alternative splicing of exon 5 in NMDA receptor subunit 1.
RT-PCR and intracellular Ca2+ measurements were used to identify factors that modulate alternative splicing of exon 5 in the NMDA receptor transcript encoding NR1, in cultured cerebellar granule neurons. Although cells grown in media containing 5 mM KCl demonstrate compromised survival, they show the predicted developmental transition from NR1a (-exon 5) to NR1b (+exon 5) mRNA expression. This transition was blocked under culture conditions that promote survival; inclusion or exclusion of exon 5 is a reversible process that is sensitive to alterations in Ca2+ and pH. We conclude that alternative splicing of NR1 pre-mRNA transcripts may be regulated by developmental cues that modulate the degree of glutamate receptor activation.


Developing an audiovisual notebook as a self-learning tool in histology: perceptions of teachers and students.
Videos can be used as didactic tools for self-learning under several circumstances, including those cases in which students are responsible for the development of this resource as an audiovisual notebook. We compared students' and teachers' perceptions regarding the main features that an audiovisual notebook should include. Four questionnaires with items about information, images, text and music, and filmmaking were used to investigate students' (n = 115) and teachers' perceptions (n = 28) regarding the development of a video focused on a histological technique. The results show that both students and teachers significantly prioritize informative components, images and filmmaking more than text and music. The scores were significantly higher for teachers than for students for all four components analyzed. The highest scores were given to items related to practical and medically oriented elements, and the lowest values were given to theoretical and complementary elements. For most items, there were no differences between genders. A strong positive correlation was found between the scores given to each item by teachers and students. These results show that both students' and teachers' perceptions tend to coincide for most items, and suggest that audiovisual notebooks developed by students would emphasize the same items as those perceived by teachers to be the most relevant. Further, these findings suggest that the use of video as an audiovisual learning notebook would not only preserve the curricular objectives but would also offer the advantages of self-learning processes.


Synchrotron Radiation Fourier Transform Infrared (SR-FTIR) spectroscopy in exploring ancient human hair from Roman period Juliopolis: Preservation status and alterations of organic compounds.
We explore the preservation status and alterations of organic compounds in Roman period human hairstrandsfrom a specific individual (M196) excavated at Juliopolis (JP). How do these organic compounds present in this c. 2000-year-old human hair compare to those present in modern hair? Alterations to organic compounds in archaeological human hair are caused by biological degradative processes dependent on multifactorial processes acting on the hair since the deposition of a body in a mortuary context. We investigate the type of organic compounds present using Synchrotron Radiation Fourier Transform Infrared (SR-FTIR). Juliopolis (Iuliopolis) is an ancient multiperiod city, located in the Çayırhan district of Nallıhan, northwest of Ankara. The Juliopolis necropolis from which M196 was recovered was in use throughout the Hellenistic, Roman, and Byzantine periods, and yielded over 700 tombs with numerous human remains. One tomb (M196) contained human remains of exceptional preservation status, including substantial amounts of hair. Human hair from archaeological contexts is not only extremely rare, but importantly, has high analytical value, with potential for analysis of diet, geographical origins, ancient DNA, metal exposure, and other aspects of life in a time-resolved manner. These data make significant contributions to the life history of the individual (osteobiography), as well as contribute towards key archaeological questions. As these analyses are in their majority destructive, prior evaluation of the preservation of sufficient amounts of the organic compounds on which many such analyses rely upon is crucial, to avoid unnecessary loss of precious ancient samples. The results of our SR-FTIR analyses at SESAME synchrotron show that keratin in the JP M196 is more degraded in comparison to the modern reference sample. However, the results also point to clear potential for further analyses with techniques relying on organic compound preservation, such as C and N isotopic analyses for diet, and aDNA.


Functional interaction of osteogenic transcription factors Runx2 and Vdr in transcriptional regulation of Opn during soft tissue calcification.
Loss of Abcc6 gene expression was identified to be responsible for dystrophic calcification of the heart (DCC) or vessels after acute injury in several strains of laboratory mice. This calcification shares features with osteogenesis and may involve osteogenic factors. Tissue expression of osteopontin (Opn) and 11 osteogenic transcription factors were studied in vivo in mouse models for DCC and in vitro using luciferase reporter gene assays. Compared with DCC-resistant C57BL/6 mice, a significant increase in Opn transcription was demonstrated in necrotic lesions of both DCC-susceptible C3H/He and B6.C3H(Dyscalc1) congenic mice at day 3 after injury. Significant increases in gene expression were also demonstrated for the transcription factors runt domain-containing transcription factor 2 (Runx2), vitamin D receptor (Vdr), SRY (sex-determining region Y)-box 9 protein, and Nfkb1 in C3H/He mice versus C57BL/6 controls. However, only Runx2 remained significantly increased in the B6.C3H(Dyscalc1) congenic mice, which carry only the Dyscalc1 locus with functional Abcc6 deletion on a C57BL/6 genetic background. Luciferase assay use increased Opn promoter activity, which was demonstrated after overexpression of Runx2. A poly-T stretch insertion was identified to stabilize the binding of Runx2, thus significantly enhancing Opn promoter activity. This Runx2-mediated activation was further enhanced by cotransfection with Vdr. Our data suggest a key role of Runx2 in the regulation of Opn in a model of cardiovascular calcification and demonstrate a synergistic cooperation of Runx2 and Vdr.


Surgery for focal epilepsy in the older patient.
Eight of the 237 patients who received a surgical procedure for intractable partial epilepsy at the Mayo Clinic during a 3-year period were 50 years of age or older. All patients had disabling partial seizures of several years in duration that were refractory to antiepileptic drug medication. A neuroimaging-identified epileptogenic lesion prompted referral for surgical treatment in three patients. The only clinically significant morbidity associated with surgery in the eight patients was a transient exacerbation of an aphasia in a patient who underwent a left frontal corticectomy. All eight patients had a favorable seizure outcome, and three patients postoperatively are seizure free. Epilepsy surgery may be an appropriate alternate therapy for select patients in the sixth decade of life or older with medically refractory partial seizures.


Small bowel graft revascularization in experiment.
The technical aspects of a procedure are most important for the outcome of the experiment. This study was designed to compare two techniques suitable for graft revascularization. The first technique, where the animal is both donor and the recipient, consists of connecting the grafts' vascular anastomoses to the mesenteric vessel bed. In the second technique, one animal is the graft donor and the other is the recipient, with revascularization to the central vessel bed (subrenal inferior vena cava and aorta). Techniques of restoring digestive tract continuity and creation of diagnostic "chimney ileostomy" were identical in both groups. All experimental animals were monitored clinically regularly basis as per protocol (weight, temperature, stoma appearance, output and nature of stools). Blood and biopsy samples were obtained on days 0, 3, 5, 7, 10, 20, and 30. Overall, 43 transplant procedures were performed. The first group included 18 transplants and 66.7% animals had vascular complications. While in the second group (25 transplant procedures), vascular complication rate was only 12% (3 out of 25) perhaps due to technique modifications. Our experiment showed that both types of vascular anastomosis could be used in small bowel transplantation. Connecting the graft vessels to the aorta and inferior vena cava is technically simpler and safer because of fewer subsequent complications (Tab. 1, Fig. 4, Ref. 6). Full Text (Free, PDF) www.bmj.sk.


Genetic analysis of fructose-1,6-bisphosphatase (FBPase) deficiency in nine consanguineous Pakistani families.
Fructose-1,6-bisphosphatase (FBPase) deficiency is a rare inherited metabolic disorder characterized by recurrent episodes of hypoglycemia, ketosis and lactic acidosis. FBPase is encoded by FBP1 gene and catalyzes the hydrolysis of fructose-1,6-bisphosphate to fructose-6-phosphate in the last step of gluconeogenesis. We report here FBP1 mutations in nine consanguineous Pakistani families affected with FBPase deficiency. Nine families having one or two individuals affected with FBPase deficiency were enrolled over a period of 3 years. All FBP1 exonic regions including splicing sites were PCR-amplified and sequenced bidirectionally. Familial cosegregation of mutations with disease was confirmed by direct sequencing and PCR-RFLP analysis. Three different FBP1 mutations were identified. Each of two previously reported mutations (c.472C>T (p.Arg158Trp) and c.841G>A (p.Glu281Lys)) was carried by four different families. The ninth family carried a novel 4-bp deletion (c.609_612delAAAA), which is predicted to result in frameshift (p.Lys204Argfs*72) and loss of FBPase function. The novel variant was not detected in any of 120 chromosomes from normal ethnically matched individuals. FBPase deficiency is often fatal in the infancy and early childhood. Early diagnosis and prompt treatment is therefore crucial to preventing early mortality. We recommend the use of c.472C>T and c.841G>A mutations as first choice genetic markers for molecular diagnosis of FBPase deficiency in Pakistan.


Prevention of taurolithocholate-induced hepatic bile canalicular distortions by HPLC-characterized extracts of artichoke (Cynara scolymus) leaves.
The effects of water-soluble extracts of artichoke (Cynara scolymus L.) leaves on taurolithocholate-induced cholestatic bile canalicular membrane distortions were studied in primary cultured rat hepatocytes using electron microscopy. Artichoke extracts at concentrations between 0.08 and 0.5 mg/ml were able to prevent the formation of bizarre canalicular membrane transformations in a dose-dependent manner when added simultaneously with the bile acid. However, prevention also occurred when the hepatocytes were preincubated with the extracts, indicating that absorption of the bile acid to components of the extracts was not involved. These results demonstrate that artichoke leaf extracts exert a potent anticholestatic action at least in the case of taurolithocholate. This effect may contribute to the overall hepatoprotective influence of this herbal formulation.


Case series: management of immature permanent teeth with pulpal necrosis: a case series.
The management of immature permanent teeth with necrotic pulps has changed in recent years from apexification techniques to regenerative endodontic procedures, which enable apexogenesis with physiologic maturation of the roots. This regenerative technique lies squarely in the endodontist's scope of practice, but children presenting with necrotic immature incisors may pose behavior management problems that endodontists are untrained and unwilling to handle. Treatment of these immature permanent teeth provides an excellent opportunity for collaboration and shared patient management between pediatric dentists and endodontists that can yield optimal clinical outcomes. The purpose of this paper was to report a series of 32 regenerative endodontic cases in 28 children treated at the Nationwide Children's Hospital, Columbus, Ohio. The report describes procedural and patient management issues and the need for a collaborative relationship between pediatric dentists and endodontists in tackling these challenging cases.


Complexity of agonist- and cyclic AMP-mediated downregulation of the human beta 1-adrenergic receptor: role of internalization, degradation, and mRNA destabilization.
Prolonged agonist exposure often induces downregulation of G protein-coupled receptors (GPCRs). Although downregulation of the prototypical beta(2)-adrenergic receptor (beta(2)AR) has been extensively studied, the underlying mechanisms have yet to be resolved. As even less is known about the beta(1)-subtype, we investigated the downregulation of human beta(1)AR stably expressed in Chinese hamster fibroblasts in response to the agonist isoproterenol or the cell-permeable, chlorophenylthio-cAMP (CPT-cAMP). While either effector mediated decreases in both beta(1)AR binding activity and steady-state beta(1)AR mRNA levels, there were significant differences in their actions. Whereas agonist-mediated downregulation of beta(1)AR followed first-order kinetics, that induced by CPT-cAMP was delayed for several hours and approximately 50% of the former. Furthermore, agonist but not CPT-cAMP induced beta(1)AR internalization, and inhibiting internalization also suppressed agonist-mediated downregulation. The latter, however, was more sensitive than the former to agonist concentration (EC(50) of 0.3 vs 48 nM). Thus, at < or =1 nM agonist, downregulation occurred without internalization and with a pattern similar to that mediated by CPT-cAMP. The amounts of beta(1)AR downregulated or internalized were proportional to initial receptor levels but reached saturation at approximately 2 and 3 pmol/mg of protein, respectively. The fate of beta(1)AR protein during downregulation was determined by immunoblotting with anti-C-terminal antibodies. In agonist-treated cells, beta(1)AR protein disappeared with time and without any immunoreactive degradation products. Agonist-mediated downregulation of the human beta(1)AR appears to be a complex process that consists of both agonist- and cAMP-specific components. The former involves both receptor internalization and degradation whereas the latter involves a reduction in receptor mRNA.


Elimination of diaminopeptidase activity in Pichia pastoris for therapeutic protein production.
Yeast are important production platforms for the generation of recombinant proteins. Nonetheless, their use has been restricted in the production of therapeutic proteins due to differences in their glycosylation profile with that of higher eukaryotes. The yeast strain Pichia pastoris is an industrially important organism. Recent advances in the glycoengineering of this strain offer the potential to produce therapeutic glycoproteins with sialylated human-like N- and O-linked glycans. However, like higher eukaryotes, yeast also express numerous proteases, many of which are either localized to the secretory pathway or pass through it en route to their final destination. As a consequence, nondesirable proteolysis of some recombinant proteins may occur, with the specific cleavage being dependent on the class of protease involved. Dipeptidyl aminopeptidases (DPP) are a class of proteolytic enzymes which remove a two-amino acid peptide from the N-terminus of a protein. In P. pastoris, two such enzymes have been identified, Ste13p and Dap2p. In the current report, we demonstrate that while the knockout of STE13 alone may protect certain proteins from N-terminal clipping, other proteins may require the double knockout of both STE13 and DAP2. As such, this understanding of DPP activity enhances the utility of the P. pastoris expression system, thus facilitating the production of recombinant therapeutic proteins with their intact native sequences.


Joint modelling of anaemia and stunting in children less than five years of age in Lesotho: a cross-sectional case study.
Anaemia and stunting remain jointly a serious health issue worldwide especially in developing countries. In Lesotho, their prevalence is high, particularly among children less than 5 years of age. The primary objective was to determine the association between anaemia and stunting, and identify factors relating to both conditions among children younger than 5 years in Lesotho. This cross-sectional study used secondary data from 3112 children collected during the 2014 Lesotho Demographic Health Survey (LDHS). Haemoglobin (Hb) levels were adjusted for altitude and a level less than 11 g per deciliters (11 g/dl) was determined as the cutoff for being anaemic. A child with the height-for-age z score (HAZ) below minus two standard deviations (SD) was considered to have stunting. We linked factors relating to anaemia and stunting using a multivariate joint model under the scope of the generalized linear mixed model (GLMM). The prevalence of anaemia and stunting in children younger than 5 years were 51% and 43% respectively. The multivariate results revealed a strong association between anaemia and stunting. In addition, maternal education, urban vs. rural residence, wealth index and childbirth weight significantly impacted childhood stunting or malnutrition, while having fever and/or diarrhoea was linked to anaemia. Lastly, age was shown to have a significant effect on both stunting and anaemia. Anaemia and stunting or malnutrition showed linked longitudinal trajectories, suggesting both conditions could lead to synergetic improvements in overall child health. Demographic, socio-economic, and geographical characteristics were also important drivers of stunting and anaemia in children younger than 5 years. Thus, children living in similar resources settings as Lesotho could benefit from coordinated programs designed to address both malnutrition and anaemia.


Effect of aging on the tendon structure and tendon-associated gene expression in mouse foot flexor tendon.
To evaluate the biological changes in tendons during the aging process, the present study examined the effect of aging on the tendon structure, distribution of collagen types I and III, and expression of tendon-associated genes, using flexor tendons in a mouse model. Histological assessment of the tendon structure and distribution of collagen types I and III were performed, and the expression of tendon-associated genes was evaluated in flexor digitorium longus tendons of young (8 weeks) and aged (78 weeks) female C57BL/6 mice. The results indicated that the Soslowsky score, based on the analysis of cellularity, fibroblastic changes, and collagen fiber orientation and disruption, was significantly increased, or worsened, in the tendons of the aged group compared with those in the young group. Furthermore, in the aged group, the distribution of type I collagen was decreased and the distribution of type III collagen was relatively increased compared with the young group. Finally, the mRNA expression levels of collagen (type I and type III) and tenogenic markers (Mohawk homeobox, tenomodulin and scleraxis BHLH transcription factor) were significantly decreased in the aged group compared with the young group. The present observations demonstrated that the structure of the tendons, distribution of types I and III collagen and the expression of tendon-associated genes were modulated by aging in the flexor tendon, and that these changes may contribute to the degeneration of tendons in tendinopathy.


Fecal microbiota in client-owned obese dogs changes after weight loss with a high-fiber-high-protein diet.
The fecal microbiota from obese individuals can induce obesity in animal models. In addition, studies in humans, animal models and dogs have revealed that the fecal microbiota of subjects with obesity is different from that of lean subjects and changes after weight loss. However, the impact of weight loss on the fecal microbiota in dogs with obesity has not been fully characterized. In this study, we used 16S rRNA gene sequencing to investigate the differences in the fecal microbiota of 20 pet dogs with obesity that underwent a weight loss program. The endpoint of the weight loss program was individually tailored to the ideal body weight of each dog. In addition, we evaluated the qPCR based Dysbiosis Index before and after weight loss. After weight loss, the fecal microbiota structure of dogs with obesity changed significantly (weightedANOSIM; p = 0.016, R = 0.073), showing an increase in bacterial richness (p = 0.007), evenness (p = 0.007) and the number of bacterial species (p = 0.007). The fecal microbiota composition of obese dogs after weight loss was characterized by a decrease in Firmicutes (92.3% to 78.2%, q = 0.001), and increase in Bacteroidetes (1.4% to 10.1%, q = 0.002) and Fusobacteria (1.6% to 6.2%, q = 0.040). The qPCR results revealed an overall decrease in the Dysbiosis Index, driven mostly due to a significant decrease in E. coli (p = 0.030), and increase in Fusobacterium spp. (p = 0.017). The changes observed in the fecal microbiota of dogs with obesity after weight loss with a weight loss diet rich in fiber and protein were in agreement with previous studies in humans, that reported an increase of bacterial biodiversity and a decrease of the ratio Firmicutes/Bacteroidetes.


Therapeutic Whole-Lung Lavage for Pulmonary Alveolar Proteinosis: A Procedural Update.
Pulmonary alveolar proteinosis is a disease caused by increased accumulation and impaired clearance of surfactant by alveolar macrophages. This narrative review summarizes the role of therapeutic whole-lung lavage in the management of pulmonary alveolar proteinosis. We describe the preprocedural evaluation, indications, and anesthetic considerations, along with step-by step technical aspects of the procedure, postoperative recovery, potential complications, and long-term outcomes.


Hybrid elastic and discrete-particle approach to biomembrane dynamics with application to the mobility of curved integral membrane proteins.
We introduce a simulation strategy to consistently couple continuum biomembrane dynamics to the motion of discrete biological macromolecules residing within or on the membrane. The methodology is used to study the diffusion of integral membrane proteins that impart a curvature on the bilayer surrounding them. Such proteins exhibit a substantial reduction in diffusion coefficient relative to "flat" proteins; this effect is explained by elementary hydrodynamic considerations.


Inhibition of Ataxia-Telangiectasia Mutated and RAD3-Related (ATR) Overcomes Oxaliplatin Resistance and Promotes Antitumor Immunity in Colorectal Cancer.
Although many patients with colorectal cancer initially respond to the chemotherapeutic agent oxaliplatin, acquired resistance to this treatment remains a major challenge to the long-term management of this disease. To identify molecular targets of oxaliplatin resistance in colorectal cancer, we performed an shRNA-based loss-of-function genetic screen using a kinome library. We found that silencing of ataxia-telangiectasia mutated and RAD3-related (ATR), a serine/threonine protein kinase involved in the response to DNA stress, restored oxaliplatin sensitivity in a cellular model of oxaliplatin resistance. Combined application of the ATR inhibitor VE-822 and oxaliplatin resulted in strong synergistic effects in six different colorectal cancer cell lines and their oxaliplatin-resistant subclones, promoted DNA single- and double-strand break formation, growth arrest, and apoptosis. This treatment also increased replicative stress, cytoplasmic DNA, and signals related to immunogenic cell death such as calreticulin exposure and HMGB1 and ATP release. In a syngeneic colorectal cancer mouse model, combined administration of VE-822 and oxaliplatin significantly increased survival by promoting antitumor T-cell responses. Finally, a DNA repair gene signature discriminated sensitive from drug-resistant patients with colorectal cancer. Overall, our results highlight the potential of ATR inhibition combined with oxaliplatin to sensitize cells to chemotherapy as a therapeutic option for patients with colorectal cancer. SIGNIFICANCE: These findings demonstrate that resistance to oxaliplatin in colorectal cancer cells can be overcome with inhibitors of ATR and that combined treatment with both agents exerts synergistic antitumor effects.Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/79/11/2933/F1.large.jpg.


Formation of thiolate-bridged diiron complexes featuring anionic isocyanide originating from the activation of counterions in the outer sphere.
Transition metal isocyanide complexes have attracted increasing attention owing to their versatile applications in catalytic organic transformations. Compared with metal complexes with neutral isocyanide ligands, those featuring anionic isocyanide groups are relatively rare and poorly understood. So far, there has been no report on structurally characterized metal anionic isocyanopentafluorophosphate complexes that may have potential for the development of some unique polymerization reactions. In this paper, we adopt a dicationic thiolate-bridged diiron complex as the reaction platform for the coordination activation and functionalization of cyanide. When treating with KCN, a facile salt metathesis with hexafluorophosphate anions occurred to generate monocyanide or dicyanide species. However, using trimethylsilyl cyanide as the substrate, an unsymmetrical diiron complex bearing a terminal [CNSiMe3] ligand and an anionic [NCPF5]- group derived from the activation of one non-coordinating anion PF6- was obtained in a high yield. Interestingly, due to the lability of the N-Si bond in the [CNSiMe3] ligand, it can play the role of an active site for the interaction with counter anions in the outer sphere. On one hand, this labile ligand can facilitate the activation of the P-F bond in PF6- and the C-B bond in BPh4- to afford structurally characterized thiolate-bridged diiron anionic isocyanopentafluorophosphate and isocyanotriphenylborate complexes, respectively. On the other hand, it can also interact with Lut·HCl to convert into a cyanide ligand stabilized by a hydrogen bonding interaction. This work represents a new synthetic pathway to furnish metal anionic isocyanide complexes.


Anti-inflammatory activities of crocetin derivatives from processed Gardenia jasminoides.
This study was designed to investigate changes of anti-oxidant and anti-nitric oxide (NO) production activities of Gardenia jasminoides (Gj) by roast processing, and anti-inflammatory activities of crocetin derivatives isolated from Gj. In order to evaluate anti-oxidant and anti-inflammatory activities, DPPH radical scavenging activities and inhibitory activities against lipopolysaccharide (LPS)-induced NO production were determined. Then we isolated crocin (1), gentiobiosyl glucosyl crocetin (3), and mono-gentiobiosyl crocetin (4) from the fruit of Gj, and crocetin (2) from the processed fruit of Gj (PGj) by column chromatography. Their structures were based on spectroscopic methods including IR, MS, and NMR (1D and 2D). Then we assayed contents of crocetin derivatives by HPLC analysis. These crocetin derivatives were evaluated the inhibitory activities on NO production in LPS-stimulated macrophage RAW 264.7 cells and expressions of protein and m-RNA of iNOS and COX-2 by western blot analysis and RT-PCR experiment. The DPPH radical scavenging activities were increased and NO productions in LPS-stimulated RAW 264.7 cells were decreased dose-dependently by processing. Crocin contents were decreased and crocetin contents were increased by processing in HPLC analysis. Compounds 1, 2, 3 and 4 reduced NO production in a dose-dependent manner with IC50 values of 58.9 μM (1), 29.9 μM (2), 31.1 μM (3), and 37.6 μM (4) respectively. Crocetin (2) showed the most potent anti-inflammatory activity (IC₅₀ = 29.9 μM), and compound 3 and 4 were firstly measured for inhibitory activities on NO production. Their correlation between structure and activity was not clear but the activity of aglycone type showed the most potent activity. They also suppressed the protein and m-RNA expressions of iNOS and COX-2 in LPS-activated macrophage. These results suggest that anti-oxidant and anti-NO production activities of Gj were increased by processing, and increased anti-inflammatory activities of Gj by processing were due to the increase of crocetin, the aglycone that has greater activity than crocin.


SS-Wrapper: a package of wrapper applications for similarity searches on Linux clusters.
Large-scale sequence comparison is a powerful tool for biological inference in modern molecular biology. Comparing new sequences to those in annotated databases is a useful source of functional and structural information about these sequences. Using software such as the basic local alignment search tool (BLAST) or HMMPFAM to identify statistically significant matches between newly sequenced segments of genetic material and those in databases is an important task for most molecular biologists. Searching algorithms are intrinsically slow and data-intensive, especially in light of the rapid growth of biological sequence databases due to the emergence of high throughput DNA sequencing techniques. Thus, traditional bioinformatics tools are impractical on PCs and even on dedicated UNIX servers. To take advantage of larger databases and more reliable methods, high performance computation becomes necessary. We describe the implementation of SS-Wrapper (Similarity Search Wrapper), a package of wrapper applications that can parallelize similarity search applications on a Linux cluster. Our wrapper utilizes a query segmentation-search (QS-search) approach to parallelize sequence database search applications. It takes into consideration load balancing between each node on the cluster to maximize resource usage. QS-search is designed to wrap many different search tools, such as BLAST and HMMPFAM using the same interface. This implementation does not alter the original program, so newly obtained programs and program updates should be accommodated easily. Benchmark experiments using QS-search to optimize BLAST and HMMPFAM showed that QS-search accelerated the performance of these programs almost linearly in proportion to the number of CPUs used. We have also implemented a wrapper that utilizes a database segmentation approach (DS-BLAST) that provides a complementary solution for BLAST searches when the database is too large to fit into the memory of a single node. Used together, QS-search and DS-BLAST provide a flexible solution to adapt sequential similarity searching applications in high performance computing environments. Their ease of use and their ability to wrap a variety of database search programs provide an analytical architecture to assist both the seasoned bioinformaticist and the wet-bench biologist.


Levels of lead in blood and bone of women giving birth in a Boston hospital.
Blood lead levels declined among Americans aged 1-74 y between the years 1976 and 1991 (National Health and Examination Survey [NHANES III]). In 1990, umbilical cord blood lead levels were surveyed among 223 women who gave birth in a Boston hospital obstetrical service. In a survey conducted 10 y earlier, women had a mean umbilical cord blood lead level of 6.56 micrograms/dl (standard deviation = 3.19). In another subgroup of 41 women who were 1-4 wk postpartum, bone lead levels were surveyed with a K-x-ray fluorescence instrument and by analysis of venous blood lead levels. In 1990, umbilical cord blood lead levels were very low (i.e., mean and median of 1.19 [standard deviation = 1.32] and 1.0 micrograms/dl, respectively. In the subgroup of postpartum women, lead levels in cortical bone (i.e., tibia) and trabecular bone (i.e., patella) were also low (i.e., tibia mean and median of 4.5 [standard deviation = 4.0] and 4 micrograms/g bone mineral, respectively); patella mean and median of 5.8 [standard deviation = 4.5] and 5 micrograms/g, respectively). Maternal age was the only factor associated (i.e., borderline [p=.076]) with bone lead. Umbilical cord blood lead levels among women served by this Boston hospital from 1980 to 1990 declined dramatically, thus paralleling the National Health and Examination Survey. Bone lead levels were also low, but lead remains a reproductive hazard to women in select high-risk groups.


Obesity: surgical management.
Bariatric surgery is a treatment approach for patients for whom multiple attempts at weight loss through lifestyle interventions and/or pharmacotherapy have not been successful. Surgery for obesity management produces greater weight loss than medical therapy alone. Four procedures frequently covered by health insurance are laparoscopic adjustable gastric band, Roux-en-Y gastric bypass, laparoscopic vertical sleeve gastrectomy, and biliopancreatic diversion with or without duodenal switch. Current indications for bariatric surgery include a body mass index of 40 kg/m(2) or greater or a body mass index of 35 kg/m(2) or greater with at least one major obesity-associated comorbid condition. Expected weight loss can range from 37% to 79% of excess weight at 2 years after surgery depending on the procedure. Patients must commit to lifelong adherence to dietary supplementation and monitoring of vitamin levels, because nutritional deficiencies are common.


Correlation between the morning hypertension on ambulatory blood pressure monitoring and the left ventricular mass in children.
Although high morning blood pressure (BP) is known to be associated with the onset of cardiovascular events in adults, data on its effects in children with hypertension are limited. Our retrospective study aimed to define the clinical characteristics of children with morning hypertension (MH) and to determine its associated factors. We reviewed 31 consecutive patients with hypertension, confirmed by the ambulatory blood pressure monitoring (ABPM). We divided these patients into 2 groups: the MH group (n=21, 67.7%), morning BP above the 95th percentile for age and height (2 hours on average after waking up) and the normal morning BP group (n=10, 32.3%). We compared the clinical manifestations, laboratory results, and echocardiographic findings including left ventricular hypertrophy (LVH) between the groups. The early/atrial (E/A) mitral flow velocity ratio in the MH group was significantly lower than that in the normal morning BP group. In addition, LV mass was higher in the MH group than in the normal morning BP group, although the difference was not statistically significant. The age at the time of hypertension diagnosis was significantly higher in the MH group than in the normal morning BP group (P=0.003). The incidence of hyperuricemia was significantly higher in the MH group than in the normal morning BP group. Older patients and those with hyperuricemia are at higher risk for MH. The rise in BP in the morning is an important factor influencing the development of abnormal relaxation, as assessed by echocardiography. Clinical trials with longer follow-up periods and larger sample sizes are needed to clarify the clinical significance of MH.


Localization and retention of mycobacterial antigen in lymph nodes of leprosy patients.
Although leprosy, a chronic disease caused by M. leprae, primarily affects skin and peripheral nerves, pathological changes and granulomas have been observed in lymph nodes which are: (a) present in tuberculoid lymph nodes in the absence of acid-fact bacilli and (b) persistent in lepromatous patients even after prolonged treatment. We detected substantial amounts of mycobacterial antigen in 16 leprous lymph nodes using anti-BCG by the peroxidase anti-peroxidase method. The load and distribution of antigen varied along the spectrum and with the duration of treatment. Tuberculoid and long-term treated lepromatous lymph nodes had a similar distribution of antigen in clusters of cells giving a 'speckled' appearance. The untreated lepromatous had a 'diffuse' staining of antigen in foamy histiocytes whereas lepromatous lesions with a lower bacillary load had a mixed pattern of 'diffuse' and 'speckled'. Antigen was also detected in a number of plasma cells along the spectrum but predominantly in lepromatous lymph nodes. Our observations indicate that: (a) antigen exists in lymph nodes despite prolonged chemotherapy which may be responsible for the persistent granuloma and (b) antigen is not confined to any particular anatomical compartment of the lymph node.


Advancements in the diagnosis and treatment of renal epithelioid angiomyolipoma: A narrative review.
Renal epithelioid angiomyolipoma (EAML) is a unique subtype of angiomyolipoma that contains a variety of cytoplasmic-rich, eosinophilic cytoplasm epithelioid cells in addition to mature adipocytes, hyaline thick-walled vessels, and smooth muscle-like spindle cells. In recent years, increasing evidence has shown that EAML is a potentially malignant tumor. Due to the lack of typical clinical manifestations and imaging features, it is difficult to diagnose before surgery, and the diagnosis mainly depends on postoperative histopathological examination. With the advancement of pathological diagnostic techniques, more EAML cases has been discovered, but clinicians still lack a comprehensive understanding of EAML. This review comprehensively describes some pathological and clinical features of EAML, with special attention to the pathogenesis and treatment of malignant EAML in order to assist with clinical diagnosis and treatment.


Mimicry for all modalities.
Mimicry is a canonical example of adaptive signal design. In principle, what constitutes mimicry is independent of the taxonomic identity of the mimic, the ecological context in which it operates, and the sensory modality through which it is expressed. However, in practice the study of mimicry is inconsistent across research fields, with theoretical and empirical advances often failing to cross taxonomic and sensory divides. We propose a novel conceptual framework whereby mimicry evolves if a receiver perceives the similarity between a mimic and a model and as a result confers a selective benefit onto the mimic. Here, misidentification and/or deception are no longer formal requirements, and mimicry can evolve irrespective of the underlying proximate mechanisms. The centrality of receiver perception in this framework enables us to formally distinguish mimicry from perceptual exploitation and integrate mimicry and multicomponent signalling theory for the first time. In addition, it resolves inconsistencies in our understanding of the role of learning in mimicry evolution, and shows that imperfect mimicry is expected to be the norm. Mimicry remains a key model for understanding signal evolution and cognition, and we recommend the adoption of a unified approach to stimulate future interdisciplinary developments in this fascinating area of research.


Detection of Persistent Viruses by High-Throughput Sequencing in Tomato and Pepper from Panama: Phylogenetic and Evolutionary Studies.
High-throughput sequencing from symptomatic tomato and pepper plants collected in Panama rendered the complete genome of the southern tomato virus (isolate STV_Panama) and bell pepper endornavirus (isolate BPEV_Panama), and almost-complete genomes of three other BPEV isolates. Tomato chlorosis virus, tomato mosaic virus, and impatiens necrotic spot virus were also detected. Analysis of the complete genome of STV and BPEV worldwide isolates revealed nucleotide diversities of 0.004246 and 0.070523, respectively. Bayesian phylogenetic analysis showed two main groups for each virus (I and II), and several subgroups for BPEV (IA, IB, IC, IIA and IIB). Isolate STV_Panama clustered with NC_12-03-08 from USA and Tom3-T from France (99.97% nucleotide identity) in Group I and BPEV_Panama was close to the Canadian isolate BPEV_Ontario (99.66% nucleotide identity) in Subgroup IB. No correlation was observed between geographic and genetic distances for both viruses. Panamanian BPEV isolates were divergent, belonging to Groups I and II (nucleotide identities > 87.33%). Evolutionary analysis showed purifying selection in all encoding regions of both viruses, being stronger in the overlapping region of both STV genes. Finally, recombination was detected in BPEV but not in STV. This is the first report of STV and BPEV in Panama.


Recent advances in structures and applications of coordination polymers based on cyclohexanepolycarboxylate ligands.
Coordination polymers (CPs) are emerging crystalline materials constructed from metal entities and organic ligands through coordination bonds, containing infinite coordination units in one, two, or three dimensions. Here an overview is given of recent structural research based on seven cyclohexanepolycarboxylates (CHPCs), namely cyclohexanecarboxylate, 1,x-cyclohexanedicarboxylate (x = 2, 3, or 4), 1,3,5-cyclohexanetricarboxylate, 1,2,4,5-cyclohexanetetracarboxylate and 1,2,3,4,5,6-cyclohexanehexacarboxylate, showing the effects of the conformation transformation and auxiliary ligands on the dimensionality and the geometric topology of the assemblies generated. The applications of these CPs as platforms for molecular adsorption, luminescent sensing, magnetism, and catalysis are also briefly discussed.


Treatment of nonneovascular idiopathic macular telangiectasia type 2 with intravitreal ranibizumab: results of a phase II clinical trial.
To evaluate the safety and preliminary efficacy of intravitreal ranibizumab for nonneovascular idiopathic macular telangiectasia Type 2. Single-center, open-label Phase II clinical trial enrolling five participants with bilateral nonneovascular idiopathic macular telangiectasia Type 2. Intravitreal ranibizumab (0.5 mg) was administered every 4 weeks in the study eye for 12 months with the contralateral eye observed. Outcome measures included changes in best-corrected visual acuity, area of late-phase leakage on fluorescein angiography, and retinal thickness on optical coherence tomography. The study treatment was well tolerated and associated with few adverse events. Change in best-corrected visual acuity at 12 months was not significantly different between treated study eyes (0.0 ± 7.5 letters) and control fellow eyes (+2.2 ± 1.9 letters). However, decreases in the area of late-phase fluorescein angiography leakage (-33 ± 20% for study eyes, +1 ± 8% for fellow eyes) and in optical coherence tomography central subfield retinal thickness (-11.7 ± 7.0% for study eyes and -2.9 ± 3.5% for fellow eyes) were greater in study eyes compared with fellow eyes. Despite significant anatomical responses to treatment, functional improvement in visual acuity was not detected. Intravitreal ranibizumab administered monthly over a time course of 12 months is unlikely to provide a general and significant benefit to patients with nonneovascular idiopathic macular telangiectasia Type 2.


Clearance of CMV viremia and survival after double umbilical cord blood transplantation in adults depends on reconstitution of thymopoiesis.
Umbilical cord blood grafts are increasingly used as sources of hematopoietic stem cells in adults. Data regarding the outcome of this approach in adults are consistent with delayed and insufficient immune reconstitution resulting in high infection-related morbidity and mortality. Using cytomegalovirus (CMV)-specific immunity as a paradigm, we evaluated the status, mechanism, and clinical implications of immune reconstitution in adults with hematologic malignancies undergoing unrelated double unit cord blood transplantation. Our data indicate that CD8(+) T cells capable of secreting interferon-gamma (IFN-gamma) in a CMV-specific enzyme-linked immunosorbent spot (ELISpot) assay are detectable at 8 weeks after transplantation, before reconstitution of thymopoiesis, but fail to clear CMV viremia. Clearance of CMV viremia occurs later and depends on the recovery of CD4(+)CD45RA(+) T cells, reconstitution of thymopoiesis, and attainment of T-cell receptor rearrangement excision circle (TREC) levels of 2000 or more copies/mug DNA. In addition, overall survival was significantly higher in patients who displayed thymic regeneration and attainment of TREC levels of 2000 or more copies/mug DNA (P = .005). These results indicate that reconstitution of thymopoiesis is critical for long-term clinical outcome in adult recipients of umbilical cord blood transplant. The trial was prospectively registered at http://www.clinicaltrials.gov (NCT00133367).


Disrupted Resting-State Default Mode Network in Betel Quid-Dependent Individuals.
Recent studies have shown that substance dependence (addiction) is accompanied with altered activity patterns of the default mode network (DMN). However, the neural correlates of the resting-state DMN and betel quid dependence (BQD)-related physiopathological characteristics still remain unclear. Resting-state functional magnetic resonance imaging images were obtained from 26 BQD individuals and 28 matched healthy control subjects. Group independent component analysis was performed to analyze the resting state images into spatially independent components. Gray matter volume was examined as covariate with voxel-based morphometry to rule out its effect on the functional results. The severity of BQD was assessed by the BQD Scale (BQDS). We observed decreased functional connectivity in anterior part of the DMN including ventral medial prefrontal cortex, orbital MPFC (OMPFC)/anterior cingulate cortex (ACC). Furthermore, the functional connectivity within the OMPFC/ACC in BQD individuals was negatively correlated with BQDS (p = 0.01, r = -0.49). We reported decreased functional connectivity within anterior part of the DMN in BQD individuals, which provides new evidence for the role of the DMN in the pathophysiology of BQD.


Acute hypernatremia exerts an inhibitory oxytocinergic tone that is associated with anxiolytic mood in male rats.
Anxiety disorders are the most common psychiatric illnesses and are associated with heightened stress responsiveness. The neuropeptide oxytocin (OT) has garnered significant attention for its potential as a treatment for anxiety disorders; however, the mechanism mediating its effects on stress responses and anxiety is not well understood. Here we used acute hypernatremia, a stimulus that elevates brain levels of OT, to discern the central oxytocinergic pathways mediating stress responsiveness and anxiety-like behavior. Rats were rendered hypernatremic by acute administration of 2.0 M NaCl and had increased plasma sodium concentration, plasma osmolality, and Fos induction in OT-containing neurons relative to 0.15 M NaCl-treated controls. Acute hypernatremia decreased restraint-induced elevations in corticosterone and created an inhibitory oxytocinergic tone on parvocellular neurosecretory neurons within the paraventricular nucleus of the hypothalamus. In contrast, evaluation of Fos immunohistochemistry determined that acute hypernatremia followed by restraint increased neuronal activation in brain regions receiving OT afferents that are also implicated in the expression of anxiety-like behavior. To determine whether these effects were predictive of altered anxiety-like behavior, rats were subjected to acute hypernatremia and then tested in the elevated plus maze. Relative to controls given 0.15 M NaCl, rats given 2.0 M NaCl spent more time in the open arms of the elevated plus maze, suggesting that acute hypernatremia is anxiolytic. Collectively the results suggest that acute elevations in plasma sodium concentration increase central levels of OT, which decreases anxiety by altering neuronal activity in hypothalamic and limbic nuclei.


Performance implications of Porter's generic strategies in Slovak hospitals.
The aim of the study was to examine the use of Porter's generic strategies and their effect on performance in the context of the Slovak hospital industry. Using mail survey the study first identified the natural taxonomy of four strategic types of Slovak hospitals, based on their use of Porter's generic strategies in pure form and in combination. Next the study examined whether different strategic types were associated with different levels of organisational performance, while controlling for such variables as size and location, which have been argued to influence the hospital performance. The findings indicate that hospitals which follow a "stuck-in-the-middle" strategy, in general, have superior performance on all used performance measures, while hospitals that place only low emphasis on cost leadership, differentiation and focus, labelled "wait and see" in this study, perform the poorest. The study concludes that the research provided body of knowledge relevant for the Slovak hospital industry, that may be used by hospital managers in the strategy formulation process as well as by the researches in exploring the influence of different contingencies on hospitals' strategic orientation.


Localization and developmental change of indoleamine 2,3-dioxygenase activity in the human placenta.
Previously, we pointed out the importance of the kynurenine metabolism in fetuses and neonates. We examined localization and developmental change of indoleamine 2,3-dioxygenase activity in human placenta. The indoleamine 2,3-dioxygenase was found localized in syncytiotrophoblast in the placenta. The indoleamine 2,3-dioxygenase activity was not detected in placenta in the early stage of gestation. It was first detected at around 14 weeks of gestation, increased rapidly thereafter and was maintained at high levels till near term. The indoleamine 2,3-dioxygenase activity was significantly lower in placenta with retarded intrauterine development. These results suggest the importance of placental indoleamine 2,3-dioxygenase during fetal development.


Mammographically guided fine-needle aspiration biopsy of nonpalpable breast lesions. Can it replace open biopsy?
The authors report the accuracy of mammographically guided fine-needle aspiration (FNA) for the diagnosis of nonpalpable breast lesions in a series of 71 patients. Each mammographically guided FNA was immediately followed by hookwire localization and open biopsy. Seven malignancies were identified cytologically, but three cancers were not detected in aspirates judged to be sufficient for diagnosis. In addition, 19 aspirates were reported to be insufficient, and 4 of these were obtained from lesions histologically proven to be adenocarcinoma. Although no false-positive diagnoses were encountered, the technique demonstrated a sensitivity of 78% and a diagnostic accuracy of 94% for adequate specimens. In this study, mammographically guided FNA was insufficiently sensitive to replace open biopsy for the diagnosis of nonpalpable breast lesions.


Nutrient-intake-level-dependent regulation of intestinal development in newborn intrauterine growth-restricted piglets via glucagon-like peptide-2.
The objective of the present study was to investigate the intestinal development of newborn intrauterine growth-restricted (IUGR) piglets subjected to normal nutrient intake (NNI) or restricted nutrient intake (RNI). Newborn normal birth weight (NBW) and IUGR piglets were allotted to NNI or RNI levels for 4 weeks from day 8 postnatal. IUGR piglets receiving NNI had similar growth performance compared with that of NBW piglets. Small intestine length and villous height were greater in IUGR piglets fed the NNI than that of piglets fed the RNI. Lactase activity was increased in piglets fed the NNI compared with piglets fed the RNI. Absorptive function, represented by active glucose transport by the Ussing chamber method and messenger RNA (mRNA) expressions of two main intestinal glucose transporters, Na+-dependent glucose transporter 1 (SGLT1) and glucose transporter 2 (GLUT2), were greater in IUGR piglets fed the NNI compared with piglets fed the RNI regimen. The apoptotic process, characterized by caspase-3 activity (a sign of activated apoptotic cells) and mRNA expressions of p53 (pro-apoptotic), bcl-2-like protein 4 (Bax) (pro-apoptotic) and B-cell lymphoma-2 (Bcl-2) (anti-apoptotic), were improved in IUGR piglets fed the NNI regimen. To test the hypothesis that improvements in intestinal development of IUGR piglets fed NNI might be mediated through circulating glucagon-like peptide-2 (GLP-2), GLP-2 was injected subcutaneously to IUGR piglets fed the RNI from day 8 to day 15 postnatal. Although the intestinal development of IUGR piglets fed the RNI regimen was suppressed compared with those fed the NNI regimen, an exogenous injection of GLP-2 was able to bring intestinal development to similar levels as NNI-fed IUGR piglets. Collectively, our results demonstrate that IUGR neonates that have NNI levels could improve intestinal function via the regulation of GLP-2.


Respiratory manifestations of systemic disease. 3. Neurologic, skeletal, dermatologic, gynecologic, and gonadal diseases and disorders of pregnancy.
Many nonpulmonary diseases may present with respiratory manifestations or may involve the lungs later in the disease course. The mechanisms by which such involvement occurs are almost as diverse as the diseases themselves and include the following. Hematogenous spread of disease is one of the most common mechanisms of lung involvement, for example, lung involvement by metastatic malignant disease. Cytotoxic factors from another anatomic location may be deposited in the alveolar basement membranes and cause pulmonary damage, for example, pulmonary hemorrhage associated with Goodpasture's syndrome. The pulmonary vasculature may prominently manifest generalized disease, as frequently occurs in Wegener's granulomatosis. Toxins accumulated as a result of disease in another organ system may damage the alveolar capillaries and result in pulmonary edema, as can occur in patients with severe azotemia due to renal failure. Depletion of lung surfactant as a consequence of disease in another organ system may produce alveolar collapse and respiratory failure; a disease that can have this effect is acute pancreatitis. The lungs may be the first organs to exhibit, through unknown mechanisms, underlying systemic diseases such as the collagenoses. Humoral factors released in another anatomic site may cause pulmonary problems; for example, bronchospasm may develop in patients with carcinoid of the intestine as a result of serotonin released by the tumor. Injury to another organ system can produce lung damage by complex mechanisms; an excellent example is the occasional development of neurogenic pulmonary edema in patients with trauma to the brain.(ABSTRACT TRUNCATED AT 250 WORDS)


Premature mitotic entry induced by ATR inhibition potentiates olaparib inhibition-mediated genomic instability, inflammatory signaling, and cytotoxicity in BRCA2-deficient cancer cells.
Poly(ADP-ribose) polymerase (PARP) inhibitors are selectively cytotoxic in cancer cells with defects in homologous recombination (HR) (e.g., due to BRCA1/2 mutations). However, not all HR-deficient tumors efficiently respond to PARP inhibition and often acquire resistance. It is therefore important to uncover how PARP inhibitors induce cytotoxicity and develop combination strategies to potentiate PARP inhibitor efficacy in HR-deficient tumors. In this study, we found that forced mitotic entry upon ATR inhibition potentiates cytotoxic effects of PARP inhibition using olaparib in BRCA2-depleted and Brca2 knockout cancer cell line models. Single DNA fiber analysis showed that ATR inhibition does not exacerbate replication fork degradation. Instead, we find ATR inhibitors accelerate mitotic entry, resulting in the formation of chromatin bridges and lagging chromosomes. Furthermore, using genome-wide single-cell sequencing, we show that ATR inhibition enhances genomic instability of olaparib-treated BRCA2-depleted cells. Inhibition of CDK1 to delay mitotic entry mitigated mitotic aberrancies and genomic instability upon ATR inhibition, underscoring the role of ATR in coordinating proper cell cycle timing in situations of DNA damage. Additionally, we show that olaparib treatment leads to increased numbers of micronuclei, which is accompanied by a cGAS/STING-associated inflammatory response in BRCA2-deficient cells. ATR inhibition further increased the numbers of cGAS-positive micronuclei and the extent of cytokine production in olaparib-treated BRCA2-deficient cancer cells. Altogether, we show that ATR inhibition induces premature mitotic entry and mediates synergistic cytotoxicity with PARP inhibition in HR-deficient cancer cells, which involves enhanced genomic instability and inflammatory signaling.


[Renal cell carcinoma in a horseshoe kidney with abdominal aortic aneurysm: a case report].
We herein report a case of renal cell carcinoma in a horseshoe kidney with an abdominal aortic aneurysm in a 69-year-old man. Radiological examinations showed a left renal tumor, horseshoe kidney and abdominal aortic aneurysm. We performed a left radical nephrectomy with the division of the isthmus and artificial graft through an abdominal transperitoneal approach. Histological findings revealed clear cell type renal cell carcinoma without invasion of the capsule or renal pelvis. Only 31 cases of renal cell carcinoma in a horseshoe kidney have been reported in Japan, and our case is the 32nd. No case with abdominal aortic aneurysm has been reported previously. We assume that abdominal aortic aneurysm was associated with renal cell carcinoma by chance in the horseshoe kidney in this case. The arterial and venous supplies vary from case to case. We emphasize that arteriography and venography are very important preoperative procedures.


[A case of severe bronchospasm under epidural anesthesia with fentanyl].
A case of severe bronchospasm under epidural anesthesia with fentanyl was described. The etiology of the bronchospasm may not have been related to sympathetic nervous blockade, histamine release, or anaphylaxis. In an asthmatic patient, it should be noted that epidural anesthesia with fentanyl could develop bronchospasm.