Renal uptake of different radiolabelled peptides is mediated by megalin: SPECT and biodistribution studies in megalin-deficient mice.
Radiolabelled peptides used for peptide receptor radionuclide therapy are excreted mainly via the kidneys and are partly reabsorbed and retained in the proximal tubular cells. The resulting high renal radiation dose can cause nephrotoxicity, limiting the maximum activity dose and the effectiveness of peptide receptor radionuclide therapy. The mechanisms of kidney reabsorption of these peptides are incompletely understood, but the scavenger receptor megalin has been shown to play a role in the reabsorption of (111)In-octreotide. In this study, the role of megalin in the renal reabsorption of various relevant radiolabelled peptides was investigated. Groups of kidney-specific megalin-deficient mice and wild-type mice were injected with (111)In-labelled somatostatin, exendin, neurotensin or minigastrin analogues. Single photon emission computed tomographic (SPECT) images of the kidneys were acquired and analysed quantitatively, or the animals were killed 3 h after injection and the activity concentration in the kidneys was measured. Megalin-deficient mice showed significantly lower uptake of all studied radiolabelled peptides in the kidneys, ranging from 22% ((111)In-octreotide) to 65% ((111)In-exendin) of uptake in wild-type kidneys. Quantitative analysis of renal uptake by SPECT and ex vivo measurements showed a very good correlation. Megalin is involved in the renal reabsorption of radiolabelled octreotide, octreotate, exendin, neurotensin and minigastrin. This knowledge may help in the design of strategies to reduce this reabsorption and the resulting nephrotoxicity in peptide receptor radionuclide therapy, enabling more effective therapy. Small-animal SPECT is an accurate tool, allowing in vivo quantification of renal uptake and serial measurements in individual mice.


Focus on Phytochemical and Pharmacological Profile of Prunus lycioides (=Amygdalus lycioides).
Prunus lycioides (Spach) C.K. Schneid. (= Amygdalus lycioides Spach.), popularly recognized as "Badam Talkh kuhi", is an endemic species of Iran. It is widely distributed in Central and West Asia, where it plays a role in preventing and controlling soil erosion. Although domestic species of Prunus genus are well known and widely used in food, pharmaceutical, and cosmetic industries, inadequate information about wild species is available so far. As far as Prunus lycioides, it is commonly used by native people in traditional medicine for treating diabetes, inflammatory diseases, and microbial infections. The wild almonds are traditionally exploited for oil extraction due to their antioxidant properties. This review summarizes advances in the studies regarding Prunus lycioides and its pharmacological properties. The aim of the review is to renew the interest in this promising plant, thus stimulating researchers to go further with the study for discovering new bioactive compounds.


Spectrum of CNS disease caused by rapidly growing mycobacteria.
We present a case of a patient with chronic meningoencephalitis caused by Mycobacterium abscessus. We also summarise the clinical features and outcomes of cases of CNS infection caused by rapidly growing mycobacteria that have been described in the literature. Rapidly growing mycobacteria are notorious for causing skin and soft-tissue infections after trauma or surgery, pulmonary disease in patients with cystic fibrosis, and disseminated disease in immunocompromised patients. CNS infection with this organism is extremely rare. Patients usually present with subacute to chronic meningitis, neutrophilic pleocytosis, and have a history of trauma or neurosurgery. The smears are often negative for acid-fast organisms, but may show Gram-positive rods. Treatment requires a long course of two or more antibiotics that have the ability to penetrate the blood-brain barrier, and possibly of steroids as immunomodulatory agents, such as those used in tuberculous meningitis.


Fallot's tetralogy in a European beaver (Castor fiber).
A 20-mo-old, female, 9-kg European beaver (Castor fiber) presented with apathy, reduced appetite of 3-day duration and a grade 5/6 systolic heart murmur. Thoracic radiographs revealed a diffuse broncho-interstitial pattern suspicious for bronchopneumonia. The echocardiographic findings of a hypertrophied right ventricle, ventricular septal defect with overriding aorta, and infundibular pulmonic stenosis were consistent with Fallot's tetralogy. Even though the bronchopneumonia rather than the congenital cardiac defect was considered of primary importance for the presenting clinical signs, the latter was relevant for the decision not to continue any medical treatment. Both disease processes were confirmed on necropsy. Fallot's tetralogy, European beaver, Castor fiber, heart murmur, ultrasound.


Competing docking interactions can bring about bistability in the MAPK cascade.
Mitogen-activated protein kinases are crucial regulators of various cell fate decisions including proliferation, differentiation, and apoptosis. Depending on the cellular context, the Raf-Mek-Erk mitogen-activated protein kinase cascade responds to extracellular stimuli in an all-or-none manner, most likely due to bistable behavior. Here, we describe a previously unrecognized positive-feedback mechanism that emerges from experimentally observed sequestration effects in the core Raf-Mek-Erk cascade. Unphosphorylated/monophosphorylated Erk sequesters Mek into Raf-inaccessible complexes upon weak stimulation, and thereby inhibits cascade activation. Mek, once phosphorylated by Raf, triggers Erk phosphorylation, which in turn induces dissociation of Raf-inaccessible Mek-Erk heterodimers, and thus further amplifies Mek phosphorylation. We show that this positive circuit can bring about bistability for parameter values measured experimentally in living cells. Previous studies revealed that bistability can also arise from enzyme depletion effects in the Erk double (de)phosphorylation cycle. We demonstrate that the feedback mechanism proposed in this article synergizes with such enzyme depletion effects to bring about a much larger bistable range than either mechanism alone. Our results show that stable docking interactions and competition effects, which are common in protein kinase cascades, can result in sequestration-based feedback, and thus can have profound effects on the qualitative behavior of signaling pathways.


[The medical aspects of road traffic traumatism in the Republic of Sakha (Yakutia)].
The article presents the analysis of road traffic accidents on the territory of the Republic of Sakha (Yakutia) during 2000-2010. The input of traffic lethality to statistics of premature death of population is considered. The assessment of the situation concerning the dynamics of road traffic accidents in the Republic is given. The main causes of accidents and issues of quality of provided medical care to victims are discussed.


Highly Sensitive End-Point PCR and SYBR Green qPCR Detection of Phymatotrichopsis omnivora, Causal Fungus of Cotton Root Rot.
Phymatotrichopsis omnivora, the causal pathogen of cotton root rot, is a devastating ascomycete that affects numerous important dicotyledonous plants grown in the southwestern United States and northern Mexico. P. omnivora is notoriously difficult to isolate from infected plants; therefore methods for accurate and sensitive detection directly from symptomatic and asymptomatic plant samples are needed for disease diagnostics and pathogen identification. Primers were designed for P. omnivora based on consensus sequences of the nuclear ribosomal internal transcribed spacer (ITS) region of geographically representative isolates. Primers were compared against published P. omnivora sequences and validated against DNA from P. omnivora isolates and infected plant samples. The primer combinations amplified products from a range of P. omnivora isolates representative of known ITS haplotypes using standard end-point polymerase chain reaction (PCR) methodology. The assays detected P. omnivora from infected root samples of cotton (Gossypium hirsutum) and alfalfa (Medicago sativa). Healthy plants and other relevant root pathogens did not produce PCR products with the P. omnivora-specific primers. Primer pair PO2F/PO2R was the most sensitive in end-point PCR assays and is recommended for use for pathogen identification from mycelial tissue and infected plant materials when quantitative PCR (qPCR) is not available. Primer pair PO3F/PO2R was highly sensitive (1 fg) when used in SYBR Green qPCR assays and is recommended for screening of plant materials potentially infected by P. omnivora or samples with suboptimal DNA quality. The described PCR-based detection methods will be useful for rapid and sensitive screening of infected plants in diagnostic laboratories, plant health inspections, and plant breeding programs.


Asthma and urticaria during disodium cromoglycate treatment. A case report.
A 10-year-old asthmatic boy began to suffer from urticarial rash and moderately severe bronchospasm after 8 weeks' treatment with disodium cromoglycate. Initially, DSCG had helped to control his asthmatic attacks, and steroid therapy could be discontinued. Inhalation provocation test with DSCG aerosol, 4 months after stopping DSCG treatment, showed an immediate-type 1 response and urticaria. A repeat provocation test, under antihistaminic cover , failed to produce similar response. When DSCG was withdrawn, urticaria vanished and the child remained symptom-free. Disodium cromoglycate is regarded as the possible aetiological agent.


[The interrelations of the lymphatic capillaries with the nerve structures in the wall of the small intestine].
The character of interrelations of nervous structures and the lymphatic capillary walls has been studied in cats. Under the light microscope twisted nervous fiber terminals of the intestinal neuron dendrites have been revealed around the lymphatic capillaries. Electron microscopical investigation has not revealed any specialized contacts of the nervous terminals and the lymphatic capillary walls. The receptors and terminals of axons do not situate nearer than 10 nm from the latter. According to the structure of synaptic vesicles among the axonal terminals next to the lymphatic capillary walls cholinergic, adrenergic and purinergic ones are described. The influence of the nervous system to the function of the small intestine lymphatic capillaries is mediated via the precapillary space. The neuromediators from the axonal terminals get into it owing to absence of neurolemmocytic membranes around them.


Sequencing analysis of the spinal bulbar muscular atrophy CAG expansion reveals absence of repeat interruptions.
Trinucleotide repeat disorders are a heterogeneous group of diseases caused by the expansion, beyond a pathogenic threshold, of unstable DNA tracts in different genes. Sequence interruptions in the repeats have been described in the majority of these disorders and may influence disease phenotype and heritability. Spinal bulbar muscular atrophy (SBMA) is a motor neuron disease caused by a CAG trinucleotide expansion in the androgen receptor (AR) gene. Diagnostic testing and previous research have relied on fragment analysis polymerase chain reaction to determine the AR CAG repeat size, and have therefore not been able to assess the presence of interruptions. We here report a sequencing study of the AR CAG repeat in a cohort of SBMA patients and control subjects in the United Kingdom. We found no repeat interruptions to be present, and we describe differences between sequencing and traditional sizing methods.


Exploring workplace violence on surgical wards in Sweden: a cross-sectional study.
Workplace violence is a global threat to healthcare professionals' occupational health and safety and the situation has worsened during the COVID-19 pandemic. This study aimed to explore workplace violence directed against assistant and registered nurses working on surgical wards in Sweden. This cross-sectional study was conducted in April 2022. Using a convenience sampling procedure, 198 assistant and registered nurses responded to an online questionnaire developed for this specific study. The questionnaire comprised 52 items and included, among other items, subscales from validated and previously used instruments. Data analysis included descriptive statistics, the chi-square test, and independent-samples t-test. The most frequently reported type of workplace violence was humiliation (28.8%), followed by physical violence (24.2%), threats (17.7%), and unwanted sexual attention (12.1%). Patients and patients' visitors were reported as the main perpetrators of all kinds of exposure. Additionally, one third of the respondents had experienced humiliation from colleagues. Both threats and humiliation showed negative associations with work motivation and health (p < 0.05). Respondents classified as working in a high- or moderate-risk environment were more frequently exposed to threats (p = 0.025) and humiliation (p = 0.003). Meanwhile, half of the respondents were unaware of any action plans or training regarding workplace violence. However, of those who indicated that they had been exposed to workplace violence, the majority had received quite a lot or a lot of support, mainly from colleagues (range 70.8-80.8%). Despite a high prevalence of workplace violence, and especially of humiliating acts, there appeared to be low preparedness within the hospital organizations to prevent and/or handle such incidents. To improve these conditions, hospital organizations should place more emphasis on preventive measures as part of their systematic work environment management. To help inform such initiatives, it is suggested that future research should focus on the identification of suitable measures regarding different types of incidents, perpetrators, and settings.


Failure location prediction by finite element analysis for an additive manufactured mandible implant.
In order to reconstruct a patient with a bone defect in the mandible, a porous scaffold attached to a plate, both in a titanium alloy, was designed and manufactured using additive manufacturing. Regrettably, the implant fractured in vivo several months after surgery. The aim of this study was to investigate the failure of the implant and show a way of predicting the mechanical properties of the implant before surgery. All computed tomography data of the patient were preprocessed to remove metallic artefacts with metal deletion technique before mandible geometry reconstruction. The three-dimensional geometry of the patient's mandible was also reconstructed, and the implant was fixed to the bone model with screws in Mimics medical imaging software. A finite element model was established from the assembly of the mandible and the implant to study stresses developed during mastication. The stress distribution in the load-bearing plate was computed, and the location of main stress concentration in the plate was determined. Comparison between the fracture region and the location of the stress concentration shows that finite element analysis could serve as a tool for optimizing the design of mandible implants.


A late surviving Pliocene seal from high latitudes of the North Atlantic realm: the latest monachine seal on the southern margin of the North Sea.
The family of true seals, the Phocidae, is subdivided into two subfamilies: the southern Monachinae, and the northern Phocinae, following the subfamilies' current distribution: extant Monachinae are largely restricted to the (sub-)Antarctic and the eastern Pacific, with historical distributions of the monk seals of the genus Monachus in the Caribbean, the Mediterranean and around Hawaii; and Phocinae to the northern temperate and Arctic zones. However, the fossil record shows that Monachinae were common in the North Atlantic realm during the late Miocene and early Pliocene. Until now, only one late Pliocene record is known from the Mediterranean, Pliophoca etrusca from Tuscany, Italy, but none from farther north in the North Atlantic. We present the description of one partial phocid humerus collected in the early 20th century from the Antwerp area (Belgium), with an assessment of its stratigraphic origin using data from the literature. The studied humerus was recovered during construction works at the former Lefèvre dock in the Antwerp harbour (currently part of the America dock). Combining the information associated to the specimen with data from the literature and from local boreholes, the upper Pliocene Lillo Formation is ascertained as the lithological unit from which the specimen originates. Morphologically, among other features the shape of the deltopectoral crest and the poor development of the supinator crest indicates a monachine attribution for this specimen. The development of the deltopectoral crest is closer to the condition in extant Monachinae than in extinct Monachinae. The presented specimen most likely represents a monachine seal and a literature study clearly shows that it came from the latest early to late Pliocene Lillo Formation. This would be the first known monachine specimen from the latest early to late Pliocene of the North Sea, and more broadly from the northern part of the North Atlantic realm. This humerus differs from the humerus of P. etrusca and suggests a higher diversity of Monachinae in the latest early to late Pliocene than previously assumed.


A histone demethylase is necessary for regeneration in zebrafish.
Urodele amphibians and teleost fish regenerate amputated body parts via a process called epimorphic regeneration. A hallmark of this phenomenon is the reactivation of silenced developmental regulatory genes that previously functioned during embryonic patterning. We demonstrate that histone modifications silence promoters of numerous genes involved in zebrafish caudal fin regeneration. Silenced developmental regulatory genes contain bivalent me(3)K4/me(3)K27 H3 histone modifications created by the concerted action of Polycomb (PcG) and Trithorax histone methyltransferases. During regeneration, this silent, bivalent chromatin is converted to an active state by loss of repressive me(3)K27 H3 modifications, occurring at numerous genes that appear to function during regeneration. Loss-of-function studies demonstrate a requirement for a me(3)K27 H3 demethylase during fin regeneration. These results indicate that histone modifications at discreet genomic positions may serve as a crucial regulatory event in the initiation of fin regeneration.


Population Encoding With Hodgkin-Huxley Neurons.
The recovery of (weak) stimuli encoded with a population of Hodgkin-Huxley neurons is investigated. In the absence of a stimulus, the Hodgkin-Huxley neurons are assumed to be tonically spiking. The methodology employed calls for 1) finding an input-output (I/O) equivalent description of the Hodgkin-Huxley neuron and 2) devising a recovery algorithm for stimuli encoded with the I/O equivalent neuron(s). A Hodgkin-Huxley neuron with multiplicative coupling is I/O equivalent with an Integrate-and-Fire neuron with a variable threshold sequence. For bandlimited stimuli a perfect recovery of the stimulus can be achieved provided that a Nyquist-type rate condition is satisfied. A Hodgkin-Huxley neuron with additive coupling and deterministic conductances is first-order I/O equivalent with a Project-Integrate-and-Fire neuron that integrates a projection of the stimulus on the phase response curve. The stimulus recovery is formulated as a spline interpolation problem in the space of finite length bounded energy signals. A Hodgkin-Huxley neuron with additive coupling and stochastic conductances is shown to be first-order I/O equivalent with a Project-Integrate-and-Fire neuron with random thresholds. For stimuli modeled as elements of Sobolev spaces the reconstruction algorithm minimizes a regularized quadratic optimality criterion. Finally, all previous recovery results of stimuli encoded with Hodgkin-Huxley neurons with multiplicative and additive coupling, and deterministic and stochastic conductances are extended to stimuli encoded with a population of Hodgkin-Huxley neurons.


Azithromycin reduces the viability of human bronchial smooth muscle cells.
The macrolide antibiotic azithromycin has an antiproliferative and autophagic effect on rabbit tracheal smooth muscle cells (SMCs). The purpose of this study is to investigate the effect of azithromycin on human bronchial SMCs. Human bronchial SMCs were treated with azithromycin (10(-5) M) in the presence or absence of 10% fetal bovine serum (FBS). Cell number was estimated using the Cell Titer 96 AQ(ueous) One Solution Assay. Induction of autophagy was studied by observation of cell morphology in cells treated or not with the autophagy inhibitor, 3-methyladenine (3-MA), as well as by Lysotracker Red staining of lysosomes. Activation of apoptosis was assessed with flow cytometry after annexin staining. Incubation with azithromycin for 24, 48 or 72 h reduced viability in FBS-deprived cells, as well as cells cultured in FBS-containing medium. Azithromycin treatment resulted in the formation of cytoplasmic vacuoles that could not be prevented by 3-MA. Furthermore, 3-MA did not reverse the effect of azithromycin on the viability of SMCs. There was an increase in the number of lysosomes in cells treated with azithromycin. Finally, azithromycin increased the percentage of early apoptotic cells. In conclusion, azithromycin reduces the viability of human bronchial SMCs possibly by leading to apoptotic cell death.


Global population structure of the Serratia marcescens complex and identification of hospital-adapted lineages in the complex.
Serratia marcescens is an important nosocomial pathogen causing various opportunistic infections, such as urinary tract infections, bacteremia and sometimes even hospital outbreaks. The recent emergence and spread of multidrug-resistant (MDR) strains further pose serious threats to global public health. This bacterium is also ubiquitously found in natural environments, but the genomic differences between clinical and environmental isolates are not clear, including those between S. marcescens and its close relatives. In this study, we performed a large-scale genome analysis of S. marcescens and closely related species (referred to as the 'S. marcescens complex'), including more than 200 clinical and environmental strains newly sequenced here. Our analysis revealed their phylogenetic relationships and complex global population structure, comprising 14 clades, which were defined based on whole-genome average nucleotide identity. Clades 10, 11, 12 and 13 corresponded to S. nematodiphila, S. marcescens sensu stricto, S. ureilytica and S. surfactantfaciens, respectively. Several clades exhibited distinct genome sizes and GC contents and a negative correlation of these genomic parameters was observed in each clade, which was associated with the acquisition of mobile genetic elements (MGEs), but different types of MGEs, plasmids or prophages (and other integrative elements), were found to contribute to the generation of these genomic variations. Importantly, clades 1 and 2 mostly comprised clinical or hospital environment isolates and accumulated a wide range of antimicrobial resistance genes, including various extended-spectrum β-lactamase and carbapenemase genes, and fluoroquinolone target site mutations, leading to a high proportion of MDR strains. This finding suggests that clades 1 and 2 represent hospital-adapted lineages in the S. marcescens complex although their potential virulence is currently unknown. These data provide an important genomic basis for reconsidering the classification of this group of bacteria and reveal novel insights into their evolution, biology and differential importance in clinical settings.


Rituximab Monotherapy or in Combination with Bendamustine Is Not Inferior to Rituximab-CHOP Regimen in the Treatment of Patients with Splenic Marginal Zone Lymphoma in the Real Life.
Splenic marginal zone lymphoma (SMZL) is a rare lymphoma belonging to the marginal zone lymphoproliferative disorders. Usually, SMZL occurs with indolent presentation and, when required, the standard of care is represented by rituximab-based regimens. No direct comparison of different rituximab-based combinations and polychemotherapy regimens has been conducted to date. In a monocentric cohort of 68 SMLZ patients, we showed that rituximab in monotherapy or in combination with bendamustine, compared with rituximab associated with the polychemotherapy cycle cyclophosphamide, hydroxydaunorubicin, vincristine and prednisolone (CHOP), resulted in a higher 5-year progression-free survival (91.3 ± 9% and 75 ± 15.7% vs. 30.8 ± 12.1%, p < 0.001). Platelets at diagnosis <100 ×109/L (p = 0.034, HR = 4.3) and transformation into diffuse large B-cell lymphoma (p = 0.031, HR = 4.3) were associated with a lower overall survival.


The human core exosome interacts with differentially localized processive RNases: hDIS3 and hDIS3L.
The eukaryotic RNA exosome is a ribonucleolytic complex involved in RNA processing and turnover. It consists of a nine-subunit catalytically inert core that serves a structural function and participates in substrate recognition. Best defined in Saccharomyces cerevisiae, enzymatic activity comes from the associated subunits Dis3p (Rrp44p) and Rrp6p. The former is a nuclear and cytoplasmic RNase II/R-like enzyme, which possesses both processive exo- and endonuclease activities, whereas the latter is a distributive RNase D-like nuclear exonuclease. Although the exosome core is highly conserved, identity and arrangements of its catalytic subunits in different vertebrates remain elusive. Here, we demonstrate the association of two different Dis3p homologs--hDIS3 and hDIS3L--with the human exosome core. Interestingly, these factors display markedly different intracellular localizations: hDIS3 is mainly nuclear, whereas hDIS3L is strictly cytoplasmic. This compartmental distribution reflects the substrate preferences of the complex in vivo. Both hDIS3 and hDIS3L are active exonucleases; however, only hDIS3 has retained endonucleolytic activity. Our data suggest that three different ribonucleases can serve as catalytic subunits for the exosome in human cells.


Immediate implant placement without bone grafting: a retrospective study of 110 cases with 5 years of follow-up.
Immediate implant placement not only reduces the number of surgeries necessary but also decreases the treatment time and treatment costs. The purpose of the study was to present the clinical results of 110 cases of immediate implant placement without using graft materials at the end of 5 years. One hundred and ten implants in 72 patients were included in the study. Inclusion criteria for the patients were presence of at least 2 mm of bone beyond the root apex, the absence of acute signs of infection or inflammation in the treatment area, and the absence of systemic pathologies that would contraindicate bone healing around implants. In cases where bone grafting was necessary, the implant was excluded from the study. Healing progressed uneventfully in 105 cases. Four implants were lost as a result of infection in the first 3 months and 1 implant was lost 1 year after the functional loading of the prosthesis. The soft tissue anatomy was clinically acceptable in all patients. The implants that were placed in the extraction sockets of infectious teeth had also acceptable survival rates and clinical success. With proper patient selection, immediate implant placement without bone grafting has predictable survival rates and clinical success.


Reproductive efficiency of captive Chinese- and Indian-origin rhesus macaque (Macaca mulatta) females.
Reproductive and survival records (n=2,913) from 313 Chinese-origin and 365 Indian-derived rhesus macaques at the Tulane National Primate Research Center (TNPRC) spanning three generations were studied. Least-squares analysis of variance procedures were used to compare reproductive and infant survival traits while proportional hazards regression procedures were used to study female age at death, number of infants born per female, and time from last birth to death. Chinese females were older at first parturition than Indian females because they were older when placed with males, but the two subspecies had similar first postpartum birth interval (1st PPBI) and lifetime postpartum birth interval (LPPBI). Females that gave birth to stillborn infants had shorter first postpartum birth intervals (1st PPBI) than females giving birth to live infants. Postpartum birth intervals decreased in females from age 3 to 12 but then increased again with advancing age. Chinese infants had a greater survival rate than Indian infants at 30 days, 6 months, and 1 year of age. Five hundred and forty-three females (80.01%) had uncensored, or true records for age at death, number of infants born per female, and time from the birth until death whereas 135 females (19.91%) had censored records for these traits. Low- and high-uncensored observations for age at death were 3 and 26 years for Chinese, and 3 and 23 years for Indian females. Uncensored number of infants born per female ranged from 1 to 15 for Chinese females and 1 to 18 for Indian females. Each of these traits was significantly influenced by the origin×generation interaction in the proportional hazards regression analyses, indicating that probabilities associated with age at death, number of infants born per female, and time from last birth to death for Chinese and Indian females did not rank the same across generations.


A dynamic Brownian bridge movement model to estimate utilization distributions for heterogeneous animal movement.
1. The recently developed Brownian bridge movement model (BBMM) has advantages over traditional methods because it quantifies the utilization distribution of an animal based on its movement path rather than individual points and accounts for temporal autocorrelation and high data volumes. However, the BBMM assumes unrealistic homogeneous movement behaviour across all data. 2. Accurate quantification of the utilization distribution is important for identifying the way animals use the landscape. 3. We improve the BBMM by allowing for changes in behaviour, using likelihood statistics to determine change points along the animal's movement path. 4. This novel extension, outperforms the current BBMM as indicated by simulations and examples of a territorial mammal and a migratory bird. The unique ability of our model to work with tracks that are not sampled regularly is especially important for GPS tags that have frequent failed fixes or dynamic sampling schedules. Moreover, our model extension provides a useful one-dimensional measure of behavioural change along animal tracks. 5. This new method provides a more accurate utilization distribution that better describes the space use of realistic, behaviourally heterogeneous tracks.


Special Features of COVID-19 in the FMODB: Fragment Molecular Orbital Calculations and Interaction Energy Analysis of SARS-CoV-2-Related Proteins.
SARS-CoV-2 is the causative agent of coronavirus (known as COVID-19), the virus causing the current pandemic. There are ongoing research studies to develop effective therapeutics and vaccines against COVID-19 using various methods and many results have been published. The structure-based drug design of SARS-CoV-2-related proteins is promising, however, reliable information regarding the structural and intra- and intermolecular interactions is required. We have conducted studies based on the fragment molecular orbital (FMO) method for calculating the electronic structures of protein complexes and analyzing their quantitative molecular interactions. This enables us to extensively analyze the molecular interactions in residues or functional group units acting inside the protein complexes. Such precise interaction data are available in the FMO database (FMODB) (https://drugdesign.riken.jp/FMODB/). Since April 2020, we have performed several FMO calculations on the structures of SARS-CoV-2-related proteins registered in the Protein Data Bank. We have published the results of 681 structures, including three structural proteins and 11 nonstructural proteins, on the COVID-19 special page (as of June 8, 2021). In this paper, we describe the entire COVID-19 special page of the FMODB and discuss the calculation results for various proteins. These data not only aid the interpretation of experimentally determined structures but also the understanding of protein functions, which is useful for rational drug design for COVID-19.


Neurological control of human sexual behaviour: insights from lesion studies.
We review the human literature examining the effects of neurological insult on human sexual behaviour. We provide a synthesis of the findings to date, and identify key brain regions associated with specific aspects of human sexual behaviour. These include subcortical and cortical regions, with the mesial temporal lobe and the amygdala in particular being a crucial structure in the mediation of human sexual drive.


Ceruloplasmin, copper ions, and angiogenesis.
The ability to induce new formation of capillaries in the cornea was tested for ceruloplasmin, the copper carrier of serum, for fragments of the ceruloplasmin molecule with and without copper, for heparin, and for glycyl-L-histidyl-L-lysine, bound or not bound to copper ions. Male or female 2- to 3-kg New Zealand White rabbits were used. These experiments were prompted by the previous observation of copper accumulation in the cornea during angiogenesis and by the inability of copper-deficient rabbits to mount an angiogenic response. The results showed that the three different molecules were all able to induce angiogenesis provided that they were bound to copper. Fragments of the ceruloplasmin molecule also induced angiogenesis but only when copper was bound to the peptides. The data are interpreted to indicate that copper ions are involved in the sequence of events leading to angiogenesis and that the carrier molecules may be of quite a different nature.


Mental Health in Pre-Adolescents with Cerebral Palsy: Exploring the Strengths and Difficulties Questionnaire as a Screening Tool in a Follow-Up Study including Multi-Informants.
There is a high prevalence of mental health problems in children with Cerebral Palsy (CP). Still, knowledge regarding the trajectory of mental health problems throughout childhood and differences according to informants is lacking. There is also a need for more knowledge regarding the validity of mental health screening tools. In the present study, we assessed changes in parent-rated mental health problems in a cohort of 36 children with CP from school-starting age to pre-adolescence and differences in mental health problems according to informants. Further, we assessed the validity of the Strengths and Difficulties Questionnaire (SDQ) for psychiatric disorders. The study cohort was assessed using the SDQ and a child psychiatric diagnostic instrument at school-starting age and at pre-adolescence. Mean parental SDQ scores increased significantly for emotional, hyperactivity and total problems. Self-reported impact of mental health problems was significantly lower than parent-reported impact, and parents and pre-adolescents reported significantly higher mean scores than teachers for emotional problems, conduct problems and total problem scores. Validated against psychiatric disorders, the SDQ was satisfactory for screening children with CP for risk of psychiatric disorders at pre-adolescence. We recommend that mental health screening be integrated into the regular follow-up for children with CP.


Patient Commitment and Its Relationship to A1C.
IN BRIEF This study examined the relationship between patient commitment and A1C. Patients completed the Altarum Consumer Engagement (ACE) measure. Multiple A1C values were extracted from medical records for 273 military beneficiaries. Effects were analyzed with generalized linear models. The ACE Commitment subscale was significantly inversely related to A1C trends. Low-commitment patients were more likely to have a high A1C. High-commitment patients were 16% more likely to have an A1C <7.0%; this likelihood increased to 65% over time. The ACE Commitment domain may be a useful clinical tool. Increasing patients' commitment to managing diabetes may improve their A1C over time.


Postoperative obstructing laryngeal edema in patients with diffuse idiopathic skeletal hyperostosis of cervical spine -A report of two cases-.
Two cases were reported in which severe postoperative laryngeal edema were developed after the operation of diffuse idiopathic skeletal hyperostosis (DISH) of cervical spine. In the first case, sudden airway obstruction was developed in the general ward 6 hour after uneventful decompression surgery for osteophyte. In the second patient, an elective preoperative tracheostomy was performed before surgery but the tube could not be removed for 2 months because of laryngeal edema and decreased vocal cord mobility. It should be emphasized that this airway problem can develop during the postoperative as well as the preoperative period, especially in the case of anterior cervical spine surgery.


Using iterative learning to improve understanding during the informed consent process in a South African psychiatric genomics study.
Obtaining informed consent is a great challenge in global health research. There is a need for tools that can screen for and improve potential research participants' understanding of the research study at the time of recruitment. Limited empirical research has been conducted in low and middle income countries, evaluating informed consent processes in genomics research. We sought to investigate the quality of informed consent obtained in a South African psychiatric genomics study. A Xhosa language version of the University of California, San Diego Brief Assessment of Capacity to Consent Questionnaire (UBACC) was used to screen for capacity to consent and improve understanding through iterative learning in a sample of 528 Xhosa people with schizophrenia and 528 controls. We address two questions: firstly, whether research participants' understanding of the research study improved through iterative learning; and secondly, what were predictors for better understanding of the research study at the initial screening? During screening 290 (55%) cases and 172 (33%) controls scored below the 14.5 cut-off for acceptable understanding of the research study elements, however after iterative learning only 38 (7%) cases and 13 (2.5%) controls continued to score below this cut-off. Significant variables associated with increased understanding of the consent included the psychiatric nurse recruiter conducting the consent screening, higher participant level of education, and being a control. The UBACC proved an effective tool to improve understanding of research study elements during consent, for both cases and controls. The tool holds utility for complex studies such as those involving genomics, where iterative learning can be used to make significant improvements in understanding of research study elements. The UBACC may be particularly important in groups with severe mental illness and lower education levels. Study recruiters play a significant role in managing the quality of the informed consent process.


Detection of the inflammatory process in a Behçet's disease-like mouse model using 18F-fluorodeoxyglucose positron emission tomography.
The major role of herpes simplex virus (HSV) type 1 infection in Behçet's disease (BD) immunopathogenesis has been demonstrated and inoculating the earlobes of ICR mice with HSV produced a BD-like mouse model. (18)Ffluorodeoxyglucose positron emission tomography (FDG PET) is widely used for diagnosing numerous human diseases other than malignancies. The aim of our study was to evaluate the inflammatory activities of BD-like symptoms in a HSV type 1-induced BD-like mouse model by small-animal FDG PET. Five HSV-infected ICR mice with BD-like lesions, two asymptomatic HSV-infected mice, and two untreated mice were scanned with microPET, and autopsy specimens were histopathologically assessed to evaluate for infiltration by mixed inflammatory cells. The histopathological evaluation of the inflammatory process in knee and elbow joints significantly correlated with the quantitative assessment of FDG accumulation in the same joints in BD-like ICR mice, HSV-infected asymptomatic mice, and untreated control mice. Small-animal FDG PET clearly detected asymptomatic joint inflammatory processes in both BD-like mice and HSV-infected asymptomatic mice. In addition, genital ulcers and skin ulcers with associated perilesional lymphadenopathies in BD-like models were detected by microPET. However, biodistributed PET-positive images from the stasis of secreted FDG into the bowel lumen could not be distinguished from the inflammatory bowel lesions of BD when compared to FDG uptake in control mice. Our data indicate that FDG PET can non-invasively and quantitatively detect the inflammatory process in an HSV-induced BD-like mouse model.


Defining midwifery autonomy in Belgium: Consensus of a modified Delphi study.
Although there is substantial literature on autonomy of midwifery, the concept remains vague, and what it exactly constitutes is little clear. Attempts to define this have been carried out, but did not result in a communal understanding. The aim of this study therefore was to define a consistent definition of midwifery autonomy in Belgium. A modified Delphi survey with content experts. Critical components of the available definitions on midwifery autonomy were retrieved from the literature, and translated into Dutch and French. An online Delphi panel of content expert assessed components of autonomy in midwifery on clarity and relevance between June and October 2021. From the validated components, a preliminary consolidated definition was generated, which was validated in a final Delphi round. After round one, content experts (n = 27) evaluated 10 out of 17 components to be clear and relevant. Two components were judged inappropriate and therefore removed. After further adaptation four additional components were identified appropriate after the second round, and one component after a third Delphi round. Experts' suggestions for improving the clarity and relevance were taken into account. Finally, experts assessed the preliminary definition. After minor modifications the definition of midwifery autonomy in Belgium was confirmed valid. We established a communal definition of midwifery autonomy in Belgium, the creation of such a definition results in a joint understanding of the concept of midwifery autonomy. If midwives internationally want to successfully achieve autonomy, a clear understanding of the concept of midwifery autonomy is needed. The consensus definition of midwifery autonomy in Belgium comprises 15 components related to midwives' work content, professionalism and relationship with others. Our definition of midwifery autonomy has the potential to encourage an international dialogue, grounded in a common understanding of autonomy, enabling stakeholders in maternity care to strengthen professional midwifery autonomy.


The mechanism of reduction of cell-mediated cytotoxicity in neonatally thymectomized mice.
The effect of neonatal thymectomy at various times after birth (Tx-1, Tx-7) on effector and suppressor T cells responsible for cell-mediated cytotoxicity (CMC) for allogenic antigens was determined. Following in-vitro primary mixed lymphocyte cultures, in the absence of T-cell growth factor (TCGF), alloreactive CMC was not detected in spleen cells of Tx-1 mice, but was detected in spleen cells of Tx-7 mice at as high levels as in those of sham-operated mice. However, in the presence of TCGF, as much alloreactive CMC was detected in spleen cells of Tx-1 mice as in those of Tx-7 mice. Furthermore, TCGF production was not detected in spleen cells of Tx-1 mice but was detected in those of Tx-7 mice. In in-vivo experiments, inhibition of allogeneic tumour growth and CMC in spleen cells showed the same pattern as in in-vitro experiments. These results support the concept that the reduction of CMC in Tx-1 mice might be due to a defect in helper function (TCGF-producing capacity) rather than to a defect in cytotoxic T lymphocytes and/or cytotoxic T lymphocyte precursors. Alloreactive suppressor T cells could not be induced in spleen cells of Tx-1 mice but were induced in spleen cells of Tx-7 mice. Therefore, it was suggested that alloreactive suppressor T cells require the presence of the thymus for 7 days after birth in their development.


Spinal and peripheral analgesic effects of the CB2 cannabinoid receptor agonist AM1241 in two models of bone cancer-induced pain.
The activation of CB(2) receptors induces analgesia in experimental models of chronic pain. The present experiments were designed to study whether the activation of peripheral or spinal CB(2) receptors relieves thermal hyperalgesia and mechanical allodynia in two models of bone cancer pain. NCTC 2472 osteosarcoma or B16-F10 melanoma cells were intratibially inoculated to C3H/He and C57BL/6 mice. Thermal hyperalgesia was assessed by the unilateral hot plate test and mechanical allodynia by the von Frey test. AM1241 (CB(2) receptor agonist), AM251 (CB(1) receptor antagonist), SR144528 (CB(2) receptor antagonist) and naloxone were used. CB(2) receptor expression was measured by Western blot. AM1241 (0.3-10 mg.kg(-1)) abolished thermal hyperalgesia and mechanical allodynia in both tumour models. The antihyperalgesic effect was antagonized by subcutaneous, intrathecal or peri-tumour administration of SR144528. In contrast, the antiallodynic effect was inhibited by systemic or intrathecal, but not peri-tumour, injection of SR144528. The effects of AM1241 were unchanged by AM251 but were prevented by naloxone. No change in CB(2) receptor expression was found in spinal cord or dorsal root ganglia. Spinal CB(2) receptors are involved in the antiallodynic effect induced by AM1241 in two neoplastic models while peripheral and spinal receptors participate in the antihyperalgesic effects. Both effects were mediated by endogenous opiates. The use of drugs that activate CB(2) receptors could be a useful strategy to counteract bone cancer-induced pain symptoms.


[Problems in LC-MS/MS analysis to ensure food safety].
An accurate analysis is required to address various issues concerning the food safety. Many risk factors, such as agricultural chemical residues, residual veterinary drugs, mycotoxins, food additives, and carcinogens produced during food processing may be present in foodstuffs. High-performance liquid chromatograph-tandem mass spectrometry (LC-MS/MS) is an analytical system advanced in terms of sensitivity, selectivity, and flexibility. The system has been widely applied to food analysis. Food contains a wide variety of nutritional components, which may cause interference with food analyses. Then, sample preparation to remove such interference and an appropriate choice of an analytical technique is required. Even with LC-MS/MS, the analytical reliability may be reduced by matrix effects, due to interference from food components. In this review, we summarize issues on using LC-MS/MS to achieve good analyses for the food safety and discuss how to address it. The topic especially focuses on matrix effects.


Are exposure predictions, used for the prioritization of pharmaceuticals in the environment, fit for purpose?
Prioritization methodologies are often used for identifying those pharmaceuticals that pose the greatest risk to the natural environment and to focus laboratory testing or environmental monitoring toward pharmaceuticals of greatest concern. Risk-based prioritization approaches, employing models to derive exposure concentrations, are commonly used, but the reliability of these models is unclear. The present study evaluated the accuracy of exposure models commonly used for pharmaceutical prioritization. Targeted monitoring was conducted for 95 pharmaceuticals in the Rivers Foss and Ouse in the City of York (UK). Predicted environmental concentration (PEC) ranges were estimated based on localized prescription, hydrological data, reported metabolism, and wastewater treatment plant (WWTP) removal rates, and were compared with measured environmental concentrations (MECs). For the River Foss, PECs, obtained using highest metabolism and lowest WWTP removal, were similar to MECs. In contrast, this trend was not observed for the River Ouse, possibly because of pharmaceutical inputs unaccounted for by our modeling. Pharmaceuticals were ranked by risk based on either MECs or PECs. With 2 exceptions (dextromethorphan and diphenhydramine), risk ranking based on both MECs and PECs produced similar results in the River Foss. Overall, these findings indicate that PECs may well be appropriate for prioritization of pharmaceuticals in the environment when robust and local data on the system of interest are available and reflective of most source inputs. Environ Toxicol Chem 2017;36:2823-2832. © 2017 SETAC.


Fundamental structure of Fresnel diffraction: natural sampling grid and the fractional Fourier transform.
Fresnel integrals corresponding to different distances can be interpreted as scaled fractional Fourier transformations observed on spherical reference surfaces. We show that by judiciously choosing sample points on these curved reference surfaces, it is possible to represent the diffracted signals in a nonredundant manner. The change in sample spacing with distance reflects the structure of Fresnel diffraction. This sampling grid also provides a simple and robust basis for accurate and efficient computation, which naturally handles the challenges of sampling chirplike kernels.


Origin and fate of the lysophosphatidylethanolamine in a chain-forming mutant (envC) of Escherichia coli.
The role of phospholipid metabolism in the functioning of the bacterial envelope was investigated in the chain-forming Escherichia coli envC. Lysophosphatidylethanolamine (LPE) which accumulated in this strain during growth was identified as the product of phosphatidylethanolamine (PE) hydrolysis by a phospholipase A1, i.e. 2-acylLPE. Isotopically labelled LPE transferred into intact mutant and parent cells by liposome/bacteria interaction was rapidly reacylated to PE. However, in envC the final PE/LPE ratio was lower than that in the parent, thus showing that the fate of LPE is modified. Crude cell extracts degraded LPE to a lesser extent in envC than in the parent but were unable to promote reacylation activity under our experimental conditions. In both strains, the lysophospholipase activity was neither calcium-dependent nor inhibited by the SH-group inhibitors pHMB or pCMPS, and hydrolysed 1-acylLPE as well as 2-acylLPE. These results indicate the existence of a deacylation-reacylation cycle in E. coli and show that this cycle is perturbed in envC cells, especially at the lysophospholipase step.


Brain injury and protective effects of hypothermia using triphenyltetrazolium chloride in neonatal rat.
Colorless 2,3,5-triphenyltetrazolium chloride (TTC) is reduced by enzymes in functioning mitochondria to a red-colored compound, and has been used to differentiate areas of viable tissue from areas of infarction in adult animals. TTC was used to study the central nervous system protective effects of hypothermia on the neonatal rat exposed to hypoxia and ischemia. The effect of hypothermia on survival and weight gain was also determined. Seven-day-old Wistar rats with right carotid artery ligation were exposed to 3 hours of 8% oxygen and maintained at either 37 degrees C (n = 22) or 30 degrees C (n = 18). The survivors were sacrificed 2 days later and brain slices exposed to TTC. These slices were photographed and the percentage of damage to the right brain was estimated gravimetrically from the stained and unstained areas of enlarged images. The mean weight gains were 4.2 +/- 1.2 gm in the 30 degrees C group and -1.0 +/- 2.8 gm in the 37 degrees C group (P < .001). The survival in the 37 degrees C group was 77% and in the 30 degrees C group 100% (P < .025). The mean percentage damage to the right side of the brain in the 37 degrees C group was 45.5% (range: 0-87.5%); there was no detectable damage in any of the 30 degrees C group pups (P < .0001). In our study, TTC proved to be a rapid and simple method for assessing central nervous system injury in the neonatal rat. This study also confirms that moderate hypothermia is protective against hypoxic-ischemic brain injury.


[The pathology service in medical insurance].
Stabilization of economy is since qua non of the efficient medical insurance the transition to which should be gradual. One of the most important conditions of successful functioning of the insurance companies is a control of treatment and diagnosis in which a leading role belongs to the pathologist. Hence, the pathology service should be independent (creation of bureaus of pathology and institutes of pathology). Qualified experts in pathology should be trained by 4-year specialization.


Haemopoietic transformation by the TEL/ABL oncogene.
Rare, novel forms of activated ABL kinase, the result of a fusion between TEL (or ETV6, a member of the ETS transcription factor family), and the non-receptor tyrosine kinase ABL, have been identified. We have analysed the TEL/ABL fusion protein (type A) cloned from an acute lymphoblastic leukaemia patient. In contrast to a second TEL/ABL fusion (type B) identified in two cases of myeloid leukaemia, the portion of TEL contained in the type A TEL/ABL fusion was smaller and did not contain a potential Grb2 binding site. The type A TEL/ABL cDNA we used in this study encoded a 155 kD protein with elevated tyrosine kinase activity and was responsible for the phosphorylation of a number of proteins in vivo. Its expression in factor-dependent murine haemopoietic precursor cells efficiently converted these cells to factor independence for both survival and growth. These cells continued to express high levels of myc mRNA after growth factor depletion. We also demonstrated that type A TEL/ABL self-associated in stably expressing haemopoietic cells. Although the TEL portion of the TEL/ABL fusion protein has no sequence similarity to that of BCR in the BCR/ABL protein, all forms of these fusion proteins contain a structure implicated in oligomerization. Our results support the conclusion that the protein interaction domain of BCR and TEL, but not the Grb2 binding site, are the important functional components in the activation of ABL kinase in haemopoietic discase.


Traumatic brain injury substantially reduces the conditioned reinforcing effects of environmental cues in rats.
Traumatic brain injury affects millions of people each year and is an established risk factor for addiction. Recent animal studies have causally demonstrated that injuries can increase drug self-administration across a variety of substances. One potential behavioral mediator for this finding is an increased responsivity to drug-associated cues. This endophenotype can be identified by profiling non-drug-related behaviors. The current study evaluated several paradigms (conditioned approach, conditioned reinforcement, extinction from variable interval responding, conditioned facilitation) to determine how rats with a frontal TBI differed in their response to Pavlovian conditioning in response to food-paired cues. Surprisingly, rats with a TBI demonstrated increased goal-tracking in a conditioned approach paradigm and exerted less effort for a conditioned reinforcer. Moreover, they had slightly facilitated extinction (as demonstrated by significantly larger interresponse times) in the face of reinforcer-associated cues. Despite these effects, TBI rats still demonstrated conditioned facilitation to an auditory stimulus. Together, these effects suggest a phenotype in the opposite direction of what might be anticipated. Cues still served a strong discriminative function and altered behavior; however, they did not function as strong conditioned reinforcers for TBI animals. One potential reason for this is that substantial changes to the dopamine system after TBI may reduce the conditioned reinforcing effects of cues, but sensitize the brain to potent drugs of abuse. More research will be needed to determine whether this is the case.


Magnetic resonance imaging with 3-dimensional analysis of left ventricular remodeling in isolated mitral regurgitation: implications beyond dimensions.
Although surgery is indicated in patients with mitral regurgitation (MR) when left ventricular (LV) end-systolic (LVES) dimension is >40 mm, LV ejection fraction may decrease after mitral valve surgery. We hypothesize that significant LV remodeling before surgery is not reflected by standard echocardiographic parameters measured at the base of the heart. Ninety-four patients (age, 54 ± 11 years; 38% female) with degenerative isolated MR underwent cine magnetic resonance imaging with tissue tagging and 3-dimensional analysis. In 51 control subjects (age, 44 ± 14 years; 53% female), the relation between LVES volume (LVESV) and LVES dimension was quadratic, whereas in 94 MR patients, this relation was cubic, indicating a greater increase in LVESV per LVES dimension among MR patients. Moreover, magnetic resonance imaging LVESV from summated serial short-axis slices was significantly greater than LVESV assessed with the Bullet formula in MR patients, attributed to a more spherical remodeling distal to the tips of the papillary muscles (P<0.001). Thirty-five patients underwent mitral valve repair per current guideline recommendations. LV ejection fraction decreased from 61 ± 7% to 54 ± 8% (P<0.0001) and maximum shortening decreased significantly below normal at 1 year postoperatively (P<0.0001). Despite normalization of LV stroke volume and LV end-diastolic volume/mass ratio, there was a persistent significant increase in distal LVES 3-dimensional radius/wall thickness ratio and LVESV index after surgery. Despite apparently preserved LVES dimension, MR patients demonstrate significant spherical mid to apical LVES remodeling that contributes to higher LVESV than predicted by standard geometry-based calculations. Decreased LV strain after surgery suggests that a volumetric analysis of LV remodeling and function may be preferred to evaluate disease progression in isolated MR.


Rheological Properties of Cystic Fibrosis Bronchial Secretion and in Vitro Drug Permeation Study: The Effect of Sodium Bicarbonate.
Cystic fibrosis (CF) is characterized by a thick, sticky mucus responsible for both airway obstruction and resistance to drug diffusion, reducing the effectiveness of drug delivery to the lung. Studies of drug-mucus interaction may be a crucial step in therapeutic management of CF. In the present research, the effect of a saline solution of sodium bicarbonate (100 mM) on sputum viscosity and the permeation properties of ketoprofen lysinate (Klys) from a previously developed dry powder inhaler were evaluated. Rheological measurements were performed using an ARES rotational rheometer (Rheometrics, Inc.) with a parallel plate geometry. The gel fraction, separated from the liquid phase of various sputum samples from CF patients was loaded onto the plate. The elastic (G') and the viscous (G") moduli, tan δ (ratio of G" to G') and η* (complex viscosity) were evaluated as frequency-dependent parameters. Drug permeation across CF sputum from dry powders was studied by means of Franz-type vertical diffusion cells. The experiments were conducted on untreated sputum and on sputum treated with bicarbonate. Rheological studies showed that the elastic modulus (G') was always greater than the viscous modulus (G") and the viscosity decreased with increasing frequency, as for pseudo-plastic fluids. Bicarbonate caused a downward shift of both the elastic and viscous moduli, with a reduction in complex viscosity. As to drug permeation, the untreated sputum slowed down drug dissolution and permeation compared to buffer permeability (control). Permeation studies across CF sputum treated with bicarbonate showed higher Klys dissolution/permeation than untreated sputum. The interesting results confirm the previously reported bicarbonate. effectiveness in CF; this weak base seems to act by decreasing high viscosity of the CF bronchial secretion and, potentially, resulting in better mucus clearance and in fighting pulmonary infections.


[Volumetric determination of cerebral infarction in the acute phase using skull computed tomography without contrast: comparative study of 3 methods].
The determination of lesion volumes in acute cerebral Infarction (CI) has important prognostic and therapeutic implications. Three different methods -computerised technique, linear planimetry and the A.B.C/2 formula - were compared in 27 patients using non-contrast computerised tomography (CT) made at hospital admission (first 48 hours following symptom onset). Two independent observers performed the measurements. The Intra-class correlation coefficients (ICCs) were determined. The three methods exhibited high interrelation. There was a close correlation between the linear planimetry as well as the ABC/2 formula and the computerised method with ICCs of 0.94-0.95 and 0.835-0.90 respectively. Linear planimetry and the ABC/2 formula were also highly correlated, with ICCs between 0.97 and 0.99. In conclusion, the computerised method is probably the most accurate. Linear planimetry and the ABC/2 formula are also reliable, however. The A.B.C/2 formula should be applied routinely to CI volume determination based on its low cost and fast calculation.


Interactions between bioactive ferulic acid and fumed silica by UV-vis spectroscopy, FT-IR, TPD MS investigation and quantum chemical methods.
The interactions of bioactive ferulic acid with fumed silica were studied by UV/vis spectroscopy, FT-IR, TPD MS techniques and quantum chemical methods. It was found that surface complexes may form through phenol or carboxyl group of ferulic acid depending on its coverage value. The structure of surface complexes and mechanisms of the ferulic acid chemosorption on the silica surface are discussed. The kinetic parameters of the chemical reactions on silica surface are calculated. The mechanisms of thermal transformations of the ferulic chemosorbed surface complexes are proposed.


Muscle bioenergetics in obese Zucker rats.
The purpose of this study was to investigate the energetic metabolism in obese Zucker rats, using phosphorus nuclear magnetic resonance spectroscopy at rest and during a 2-Hz muscle stimulation and subsequent recovery. Animals were anesthetized with ketamine (150 mg/kg ip). Fed obese rats and 2-day-fasted obese rats were compared with their normally fed and 2-day-fasted lean litter mates. No differences were found between the two groups for ATP, total creatine, phosphocreatine (PCr), and intracellular pH. Starvation in lean rats resulted in a significant fall in inorganic phosphate (Pi), increased resting ADP level, and decreased PCr and ADP recovery after stimulation. The obese rats exhibited a decreased PCr/Pi and increased ADP at rest and a decreased PCr resynthesis and ADP metabolization rate after stimulation. Muscle stimulation in fasted obese rats induced higher PCr depletion and more pronounced acidosis. These results suggest an in vivo mitochondrial metabolism dysfunction in fasted lean as well as in fed and fasted obese rats.


Prevalence and Associated Factors of Chronic Kidney Disease Among Patients Infected With Human Immunodeficiency Virus in Cameroon.
Chronic kidney disease (CKD) is frequent amongst human immunodeficiency virus (HIV)-positive patients, and screening is not routinely performed in Sub-Saharan Africa due to resource constraints. We aimed to determine the prevalence of CKD and associated factors in HIV-infected patients in Cameroon. A cross-sectional study in Northern Cameroon included HIV-positive patients who attended the HIV clinic. Patients with an estimated glomerular filtration rate less than 60 mL/min/1.73 m² or urinary abnormalities underwent a second measurement 3 months later. Glomerular filtration rate was estimated using the 4-variable Modification of Diet in Renal Disease (MDRD) and the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations. Logistic regression was used to identify factors associated with CKD. We included 709 participants. The median CD4 count was 219 cells/mL. Proteinuria accounted for 34.4%; leukocyturia, 6.9%; and hematuria, 6.1%. Prevalence of CKD was 44.4% (CKD-EPI) and 47.2% (MDRD). Stages 3 to 5 of CKD were documented in 11.6% using the CKD-EPI and 7.5% using the MDRD. Factors associated with CKD were an age greater than 35 years (odds ratio [OR], 104; 95% confidence interval [CI], 1.02 to 1.06), longer duration of HIV (OR, 2.60; 95% CI, 1.53 to 3.95), history of hepatitis B (OR, 3.04; 95% CI, 1.08 to 8.54), and CD4 cells less than 200 cells/mL (OR, 3.64; 95% CI, 2.55 to 5.21). The prevalence of CKD is high among HIV patients in Cameroon. There is a need of implementing measures to encourage early detection of kidney disease in these patients.


Modified photon sieve as a high-performance bifocal and trifocal diffractive optical element.
We demonstrate that by dividing a Fresnel zone plate into a few regions having different periods in the s=r2 coordinate, then replacing the clear zones by a given distribution of pinholes, a so-called modified photon sieve is constructed. The first feature of the element is to increase its diameter without worrying about its feature size as the limiting factor in the fabrication zone plate. Moreover, it is shown that the number of the zones included in each region is an important parameter that has a great impact on handling the number of foci. So, by choosing a suitable relation between the number of the Fresnel zones of the regions, one gets a high-efficiency unifocal or bifocal or even multifocal element depending on the number of the regions and zones. This technique is detailed by making unifocal, bifocal, as well as trifocal modified photon sieves and surveying their focusing properties. Simulation studies are followed by the corresponding experiments to verify them.


Risk Factor Profile in Parkinson's Disease Subtype with REM Sleep Behavior Disorder.
Numerous large-scale studies have found diverse risk factors for Parkinson's disease (PD), including caffeine non-use, non-smoking, head injury, pesticide exposure, and family history. These studies assessed risk factors for PD overall; however, PD is a heterogeneous condition. One of the strongest identifiers of prognosis and disease subtype is the co-occurrence of rapid eye movement sleep behavior disorder (RBD).In previous studies, idiopathic RBD was associated with a different risk factor profile from PD and dementia with Lewy bodies, suggesting that the PD-RBD subtype may also have a different risk factor profile. To define risk factors for PD in patients with or without associated RBD. In a questionnaire, we assessed risk factors for PD, including demographic, medical, environmental, and lifestyle variables of 189 PD patients with or without associated polysomnography-confirmed RBD. The risk profile of patients with vs. without RBD was assessed with logistic regression, adjusting for age, sex, and disease duration. PD-RBD patients were more likely to have been a welder (OR = 3.11 (1.05-9.223), and to have been regular smokers (OR = 1.96 (1.04-3.68)). There were no differences in use of caffeine or alcohol, other occupations, pesticide exposure, rural living, or well water use. Patients with RBD had a higher prevalence of the combined family history of both dementia and parkinsonism (13.3% vs. 5.5% , OR = 3.28 (1.07-10.0). The RBD-specific subtype of PD may also have a different risk factor profile.


Pear blister canker viroid: sequence variability and causal role in pear blister canker disease.
The sequences of several cDNA clones of pear blister canker viroid (PBCVd) P1914T and P47A isolates have been determined. Seven out of eight P1914T clones analysed have a constant sequence which differs at six positions from that of the P2098T isolate reported previously. The remaining P1914T clone (8) has a single nucleotide substitution. The same six changes have been also observed in most of the ten P47A clones sequenced. However, some P47A clones show additional variability in positions on both strands of the central conserved region (CCR) and in another conserved sequence at the left-terminal region. This is the first report of a change affecting the upper strand of a viroid CCR. Reasons why such a change is tolerated are discussed. Infectivity bioassays have demonstrated that PBCVd is the causal agent of PBC disease.


[Ankle arthroscopy in clinical application: a preliminary report].
The results of arthroscopy or arthroscopic synovectomy on 31 ankle joints in 24 cases were reported. All the patients were followed for 2 to 10 months with an average of 6 months. The results were as follows of the 26 rheumatoid arthritis: excellent in 5 ankles (19.2%), good in 8 (30.8%), fair in 7 (26.9%), and poor in 6 (23.6%), of the three traumatic arthritis: good in one and fair in two; of the remaining 2: good in 1 tuberculous synovitis and fair in 1 chronic pyogenic arthritis. The operative procedure, indications and value of ankle arthroscopy together with its advantages were discussed in detail.


Biodegradable hydrogel derived from cellulose acetate and EDTA as a reduction substrate of leaching NPK compound fertilizer and water retention in soil.
To study the behavior of a biodegradable hydrogel derived from cellulose acetate and ethylenediaminetetraacetic dianhydride (EDTAD), as a reduction substrate of NPK fertilizer in soil. The biodegradable hydrogel (HEDTA) was prepared from cellulose acetate (CA) with a degree substitution (DS) 2.5 by esterification crosslinking with EDTAD catalyzed by triethylamine. We systematically investigated the performance of the HEDTA in the reducing NPK (Ammonium, phosphate and potassium) fertilizer leaching. We also compare the percentage of leaching between the HEDTA and commercial fertilizers. To characterize the esterification and crosslinking between CA and EDTAD, FTIR spectroscopy and thermogravimetric analysis (DTG) were employed. The biodegradation experiments were carried out in simulated soil (23% of sand, 23% of cattle manure, 23% of soil and 31% of water) and the HEDTA was tested in the eucalyptus planting during the dry season in the São Paulo state, Brazil. The HEDTA was able to reduce the leaching of fertilizers and improve the performance of eucalyptus seedlings and reduced the mortality of the seedlings. The HEDTA showed to be an excellent substrate for slow release and water-retention in soil, reducer of the fertilizers leaching, in addition being nontoxic, biodegradable in the soil and environmentally-friendly.


Lung function, airway and peripheral basophils and eosinophils are associated with molecular pharmacogenomic endotypes of steroid response in severe asthma.
Asthma is a complex disease with heterogeneous expression/severity. There is growing interest in defining asthma endotypes consistently associated with different responses to therapy, focusing on type 2 inflammation (Th2) as a key pathological mechanism. Current asthma endotypes are defined primarily by clinical/laboratory criteria. Each endotype is likely characterised by distinct molecular mechanisms that identify optimal therapies. We applied unsupervised (without a priori clinical criteria) principal component analysis on sputum airway cells RNA-sequencing transcriptomic data from 19 asthmatics from the Severe Asthma Research Program at baseline and 6-8 weeks follow-up after a 40 mg dose of intramuscular corticosteroids. We investigated principal components PC1, PC3 for association with 55 clinical variables. PC3 was associated with baseline Th2 clinical features including blood (rank-sum p=0.0082) and airway (rank-sum p=0.0024) eosinophilia, FEV1 change (Kendall tau-b R=-0.333 (-0.592 to -0.012)) and follow-up FEV1 albuterol response (Kendall tau-b R=0.392 (0.079 to 0.634)). PC1 with blood basophlia (rank-sum p=0.0191). The top 5% genes contributing to PC1, PC3 were enriched for distinct immune system/inflammation ontologies suggesting distinct subject-specific clusters of transcriptomic response to corticosteroids. PC3 association with FEV1 change was reproduced in silico in a comparable independent 14-subject (baseline, 8 weeks after daily inhaled corticosteroids (ICS)) airway epithelial cells microRNAome dataset. Transcriptomic PCs from this unsupervised methodology define molecular pharmacogenomic endotypes that may yield novel biology underlying different subject-specific responses to corticosteroid therapy in asthma, and optimal personalised asthma care. Top contributing genes to these PCs may suggest new therapeutic targets.


Structural and mechanistic analysis of sialic acid synthase NeuB from Neisseria meningitidis in complex with Mn2+, phosphoenolpyruvate, and N-acetylmannosaminitol.
In Neisseria meningitidis and related bacterial pathogens, sialic acids play critical roles in mammalian cell immunity evasion and are synthesized by a conserved enzymatic pathway that includes sialic acid synthase (NeuB, SiaC, or SynC). NeuB catalyzes the condensation of phosphoenolpyruvate (PEP) and N-acetylmannosamine, directly forming N-acetylneuraminic acid (or sialic acid). In this paper we report the development of a coupled assay to monitor NeuB reaction kinetics and an 18O-labeling study that demonstrates the synthase operates via a C-O bond cleavage mechanism. We also report the first structure of a sialic acid synthase, that of NeuB, revealing a unique domain-swapped homodimer architecture consisting of a (beta/alpha)8 barrel (TIM barrel)-type fold at the N-terminal end and a domain with high sequence identity and structural similarity to the ice binding type III antifreeze proteins at the C-terminal end of the enzyme. We have determined the structures of NeuB in the malate-bound form and with bound PEP and the substrate analog N-acetylmannosaminitol to 1.9 and 2.2 A resolution, respectively. Typical of other TIM barrel proteins, the active site of NeuB is located in a cavity at the C-terminal end of the barrel; however, the positioning of the swapped antifreeze-like domain from the adjacent monomer provides key residues for hydrogen bonding with substrates in the active site of NeuB, a structural feature that leads to distinct modes of substrate binding from other PEP-utilizing enzymes that lack an analogous antifreeze-like domain. Our observation of a direct interaction between a highly ordered manganese and the N-acetylmannosaminitol in the NeuB active site also suggests an essential role for the ion as an electrophilic catalyst that activates the N-acetylmannosamine carbonyl to the addition of PEP.


Improved chromatographic assessment of Chinese medicinal products by multi-chemical classes analysis.
An improved strategy for the quality evaluation of proprietary Chinese medicines (PCM) in complex matrices is presented. It involves the simultaneous separating and displaying of multi-chemical classes and analytes within a single chromatographic elution. The developed fingerprints have the properties of better coverage of multi-chemical classes among the composing herbs. This comprehensive and more facile approach was illustrated for Lemai Keli, a commonly used PCM listed in the Chinese Pharmacopoeia with highly complex analytical matrices. Ten marker compounds belonging to monoterpene glycosides, chalcone glycosides, phenolic acids and phenolic carboxylic aldehyde groups of the analytes together with three common chromatographic peaks were successively separated and displayed within the chromatographic window as a unique HPLC-PAD fingerprint. Rigorous method validation data demonstrated the suitability of the current method for quality assessment purposes in a highly complex PCM matrix. Additional chemometric treatment further demonstrated the uniqueness of the developed fingerprint, with a distinctive chemical profile compared to a highly similar PCM formulation (Kangen Karyu) from which it was considered to be impossible to differentiate when using the conventional single marker approach.


Two new Brazilian isolates of Bacillus thuringiensis toxic to Anticarsia gemmatalis (Lepidoptera: Noctuidae).
Bacillus thuringiensis is a bacterium used for biopesticides production and pest-resistant plants due to the synthesis of protein crystals by cry genes, which are effective in controlling several insect orders such as Lepidoptera. This work aimed at the evaluation and characterisation of two new B. thuringiensis isolates active against A. gemmatalis (Hübner 1818) larvae, which is the soybean major pest. The results showed that Bt117-4 isolate amplified fragments corresponding to cry2 and cry9 genes, and synthesised protein fragments equivalent to 130, 90 and 45 kDa. The Bt3146-4 isolate amplified DNA fragments corresponding to cry9 gene and synthesised protein fragments of 70, 58 and 38 kDa. Transmission electron microscopy revealed the presence of protein crystals in both isolates. CL50 with Cry purified proteins from Bt117-4 and Bt3146-4, corresponded to 0.195 and 0.191 µg larvae-1, respectively. The two B. thuringiensis isolates selected in this study were effective to control velvetbean caterpillar at laboratory conditions. Field tests should be carried on to develop new biopesticides formulation as well for cry genes resource for Anticarsia gemmatalis resistant transgenic plants.


Tetrameres columbicola (Nematoda: Spiruridae) infection of pigeons: ultrastructure of the gravid female in glands of the proventriculus.
The light and electron microscopies of Tetrameres columbicola gravid females in sections of the parasitized proventriculus of pigeons were studied. By light microscopy, the most conspicuous structures in the sectioned parasite were the intestine, ovary, and especially the uterus that contained numerous eggs. By electron microscopy, there was a thick mat of pigment-coated microvilli on the surfaces of the intestinal epithelial cells. The germinal zone of the ovary contained nonmembrane-bound oocytes, but oocytes were confined by a membrane in the growth zone of the ovary. The core of the spermatheca contained oocytes and the periphery harbored sperm. In this location, the unfertilized oocyte had pseudopods; sperm had invaginations of the plasma membrane. After fertilization, there was proliferation of ribosomes within the oocyte. Embryonating eggs in the uterus had thick shells and were partially enveloped by elongations of the uterine epithelial cells. Surfaces of the epithelial cells were pleated and they had electron opaque areas at the points of the pleats. Larvae in eggs had a well developed annulated cuticle and muscular layer. Somatic muscle cells had tailed appendages that protruded into the pseudocoelom. The single layer of cells beneath the hypodermis had lateral processes at the base of the cells that interdigitated with similarly elongated processes of adjacent muscle cells. Striated fibers were present in the central portion of the cells.


Variability of prostate brachytherapy pre-implant dosimetry: a multi-institutional analysis.
To conduct a multi-institutional comparison of prostate brachytherapy pre-implant dosimetry of Pd-103 and I-125. Eight experienced brachytherapists submitted Pd-103 and I-125 monotherapeutic and boost pre-implant dosimetry plans for central review. All 32 plans were calculated using the same transrectal ultrasound volumetric study. Seeds of any strength were acceptable, but were restricted to Theraseed Model 200 (Theragenics Inc., Buford, GA) and Oncura Oncoseed Model 6711 (Oncura, Plymouth Meeting, PA). The dosimetric analysis included evaluation of target volume, target to prostate ratio, target length, number of needles, seed activity, number of seeds, total activity, total activity divided by treatment planning volume, the use of extracapsular seeds, and average treatment margins (defined as the perpendicular distance between the prostate capsule and the 100% isodose line). Prostate coverage was defined in terms of V(100)/V(150)/V(200)/V(300) and D(100)/D(90)/D(50), whereas urethral dosimetry consisted of UV(100)/UV(150)/UV(200) and UD(90)/UD(50). The mean planning target volume to prostate volume ratio varied dramatically (mean 1.29, range 0.99-1.76) with the target length ranging from 3.5 to 4.5 cm. Although the prostate V(100) was >95% in all cases, the V(150) ranged from 29.9% to 92.1% and the V(200) from 6.72% to 52.5%. The urethral V(100) was 100% in all cases with six of the eight brachytherapists limiting the UV(150) to <3%. However, the median urethral dose varied by up to 50%. Treatment margins also varied significantly (average 3.98 mm, range 0.32-7.68 mm). All brachytherapists used extracapsular seeds with five implanting >25% of the seeds in extracapsular locations (range 6.4-58.2%). In addition, significant variability existed in the number of needles, number of seeds, and seed strength. This study highlights the substantial variability that exists regarding target volume, seed strength, dose homogeneity, treatment margins, and extracapsular seed placement, although prostate brachytherapy prescription doses are uniform. The standardization of pre-implant dosimetry is essential for meaningful multi-institutional comparisons of biochemical outcomes and morbidity.


Immediate versus delayed umbilical cord clamping in premature neonates born < 35 weeks: a prospective, randomized, controlled study.
The purpose of this study was to test whether delayed versus immediate cord clamping would result in higher blood pressure (BP) and hematocrit (Hct), and to assess its clinical effects on the neonatal course in premature neonates (< 35 weeks). This was a prospective, masked, randomized, controlled study. Prior to delivery, 35 neonates were randomly assigned to immediate cord clamping (ICC) at 5 to 10 seconds, and a comparable group of 30 neonates were randomly assigned to delayed cord clamping (DCC) at 30 to 45 seconds. Intention-to-treat analyses revealed that the DCC group tended to have higher initial diastolic BP and higher Hct (especially in vaginally delivered neonates). Infants weighing < 1500 g with DCC tended to have higher mean BP, and needed less mechanical ventilation and surfactant compared with ICC neonates. Infants with DCC did not experience more polycythemia (Hct > 60%), but had a trend toward higher bilirubin levels with no differences in the phototherapy needs. DCC seems to be safe and may be beneficial when compared with ICC in premature neonates. However, the differences between the two methods were modest and the clinical relevance needs to be assessed further by larger studies and additional meta-analysis of randomized trials.


The Effects of Sleep on the Commensal Microbiota: Eyes Wide Open?
Sleep dysfunction is an epidemic, the implications of which have a profound impact on a variety of gastrointestinal disease. Recent data suggests a relationship between sleep dysfunction and intestinal dysbiosis, a known proinflammatory driver. This article evaluates the interplay between sleep dysfunction and gastrointestinal health and disease, with a focus on the impact of circadian rhythm disruption on the commensal microbiota.


Taste difference thresholds for monosodium glutamate and sodium chloride in pigtail macaques (Macaca nemestrina) and spider monkeys (Ateles geoffroyi).
The purpose of this study was to determine taste difference thresholds for monosodium glutamate (MSG) and sodium chloride (NaCl) in pigtail macaques (Macaca nemestrina) and spider monkeys (Ateles geoffroyi). Using a two-bottle preference test of brief duration, three animals of each species were presented with four different reference concentrations of 50, 100, 200, and 400 mM of a tastant and tested for their ability to discriminate these from lower concentrations of the same tastant. The just noticeable differences (JNDs), expressed as Weber ratios (DeltaI/I), were found to range from 0.1 to 0.5 for MSG and 0.2 to 0.45 for NaCl in the pigtail macaques, with a significant tendency for higher Weber ratios with higher reference concentrations. In the spider monkeys, JNDs ranged from 0.15 to 0.4 for MSG and 0.1 to 0.25 for NaCl, with Weber ratios staying fairly constant across the reference concentrations tested. Thus, the JNDs were found to be generally similar in both species and to be at least as low as those found in humans for MSG and NaCl, as well as those found in spider monkeys for sucrose. The results support the assumption that both pigtail macaques and spider monkeys may use differences in perceived intensity of MSG and NaCl as a criterion for food selection.


Abnormal activation of the occipital lobes during emotion picture processing in major depressive disorder patients.
A large number of studies have demonstrated that depression patients have cognitive dysfunction. With recently developed brain functional imaging, studies have focused on changes in brain function to investigate cognitive changes. However, there is still controversy regarding abnormalities in brain functions or correlation between cognitive impairment and brain function changes. Thus, it is important to design an emotion-related task for research into brain function changes. We selected positive, neutral, and negative pictures from the International Affective Picture System. Patients with major depressive disorder were asked to judge emotion pictures. In addition, functional MRI was performed to synchronously record behavior data and imaging data. Results showed that the total correct rate for recognizing pictures was lower in patients compared with normal controls. Moreover, the consistency for recognizing pictures for depressed patients was worse than normal controls, and they frequently recognized positive pictures as negative pictures. The consistency for recognizing pictures was negatively correlated with the Hamilton Depression Rating Scale. Functional MRI suggested that the activation of some areas in the frontal lobe, temporal lobe, parietal lobe, limbic lobe, and cerebellum was enhanced, but that the activation of some areas in the frontal lobe, parietal lobe and occipital lobe was weakened while the patients were watching positive and neutral pictures compared with normal controls. The activation of some areas in the frontal lobe, temporal lobe, parietal lobe, and limbic lobe was enhanced, but the activation of some areas in the occipital lobe were weakened while the patients were watching the negative pictures compared with normal controls. These findings indicate that patients with major depressive disorder have negative cognitive disorder and extensive brain dysfunction. Thus, reduced activation of the occipital lobe may be an initiating factor for cognitive disorder in depressed patients.


Synergy between verapamil and other multidrug -resistance modulators in model membranes.
Various cationic lipophilic compounds can reverse the multidrug resistance of cancer cells. Possible interaction between these compounds, which are known as modulators, has been assessed by measuring leakage of Sulphan blue from anionic liposomes, induced both by verapamil alone and by verapamil in combination with diltiazem, quinine, thioridazine or clomipramine. An equation was derived to quantify the permeation doses and Hill coefficients of the drugs and mixtures between them by simultaneous fitting of the experimental data. The interaction was tested by two methods, the competition plot and the isobole method; both showed synergy between verapamil and each of diltiazem, quinine and thioridazine. The dose factor of potentiation for verapamil determined within membranes was 4.0 +/- 0.4 with diltiazem, 3.2 +/-0.4 with quinine and 2.4 +/- 0.3 with thioridazine. The results suggest that the effectiveness of reversing multidrug resistance may be increased with modulators such as verapamil and diltiazem that have a much greater effect in combination than what would be expected from their effects when considered separately.


Differences in Methadone Metabolism by CYP2B6 Variants.
Methadone is a long-acting opioid with considerable unexplained interindividual variability in clearance. Cytochrome P450 2B6 (CYP2B6) mediates clinical methadone clearance and metabolic inactivation via N-demethylation to 2-ethyl-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP). Retrospective studies suggest that individuals with the CYP2B6*6 allelic variant have higher methadone plasma concentrations. Catalytic activities of CYP2B6 variants are highly substrate- and expression-system dependent. This investigation evaluated methadone N-demethylation by expressed human CYP2B6 allelic variants in an insect cell coexpression system containing P450 reductase. Additionally, the influence of coexpressing cytochrome b5, whose role in metabolism can be inhibitory or stimulatory depending on the P450 isoform and substrate, on methadone metabolism, was evaluated. EDDP formation from therapeutic (0.25-1 μM) R- and S-methadone concentrations was CYP2B6.4 ≥ CYP2B6.1 ≥ CYP2B6.5 >> CYP2B6.9 ≈ CYP2B6.6, and undetectable from CYP2B6.18. Coexpression of b5 had small and variant-specific effects at therapeutic methadone concentrations but at higher concentrations stimulated EDDP formation by CYP2B6.1, CYP2B6.4, CYP2B6.5, and CYP2B6.9 but not CYP2B6.6. In vitro intrinsic clearances were generally CYP2B6.4 ≥ CYP2B6.1 > CYP2B6.5 > CYP2B6.9 ≥ CYP2B6.6. Stereoselective methadone metabolism (S>R) was maintained with all CYP2B6 variants. These results show that methadone N-demethylation by CYP2B6.4 is greater compared with CYP2B6.1, whereas CYP2B6.9 and CYP2B6.6 (which both contain the 516G>T, Q172H polymorphism), are catalytically deficient. The presence or absence of b5 in expression systems may explain previously reported disparate catalytic activities of CYP2B6 variants for specific substrates. Differences in methadone metabolism by CYP2B6 allelic variants provide a mechanistic understanding of pharmacogenetic variability in clinical methadone metabolism and clearance.


CD14-mediated alterations in transcription and splicing of endogenous retroviruses after injury.
Increase in systemic levels of lipopolysaccharide (LPS) contributes to the pathogenesis of distant organ injury after burn. Stress signals elicited from burn influence transcriptional activities of mouse endogenous retroviruses (MuERVs) in various distant organs. The involvement of LPS pathways in the burn-mediated regulation of MuERVs in the spleen was investigated in this study. Spleen harbors substantial numbers of tissue macrophages, a key responder to LPS stimulation. Spleen tissues collected from CD14 (LPS receptor) knockout (KO) and wild type (WT) mice after burn were subjected to RT-PCR analysis of MuERV expression. There was a substantial induction of 2 bands and a marked downregulation of a band in CD14 KO mice compared to WT mice after burn. Sequence analysis of these CD14- and burn-dependent bands identified 3 new alternatively spliced and 2 defective env transcripts of MuERVs as well as novel splicing signals. Chromosomal loci of putative MuERVs sharing the unique U3 sequences of these transcripts were mapped by surveying the entire genome of C57BL/6J mice. In addition, coding potentials, transcriptional regulatory elements, and adjacent cellular genes of these putative MuERVs were analyzed. The results from these studies suggest that injury-triggered LPS/CD14 signaling events play roles in the transcriptional regulation of certain MuERVs carrying unique U3 promoter sequences.


Estimation of erythropoietin secretion rate in normal and uremic subjects.
We have developed a method for directly estimating the secretion rate of endogenous erythropoietin (EPO) in human subjects. Carrier-free recombinant human EPO was labeled with 125I by the chloramine T method. Human serum albumin was then added prior to removal of free iodide. Seven normal subjects and nine patients with chronic uremia and anemia were studied. Each subject received a bolus of 125I-EPO according to body size and then a constant infusion for 5 h. Secretion rate was calculated as the rate of infusion divided by serum EPO specific activity given by the protein-bound counts divided by the EPO concentration. Serum EPO concentration was significantly higher in the patients than the controls (mean 30.9 pg/ml vs. 11.1 pg/ml, P < 0.05), although all values were within the normal range. Mean EPO secretion rate was 255 in the patients (range 68-1,101) and 190 pg.kg-1 x h-1 in the normal group (range 52-382). This difference was not significant. As the subjects were in a steady state, EPO disappearance/degradation was the same in patients compared with controls despite a relatively hypoplastic marrow.


Adenomatous polyposis coli alteration in digestive endocrine tumours: correlation with nuclear translocation of beta-catenin and chromosomal instability.
The role of Wnt pathway in digestive endocrine tumours is debated. The aim of this work is to investigate key players in Wnt pathway by a multimodal approach. Sixty cases (49 well-differentiated and 11 poorly differentiated) were investigated for methylation of adenomatous polyposis coli (APC) and E-cadherin promoters, the loss of heterozygosity (LOH) at APC locus and beta-catenin and E-cadherin expression by immunohistochemistry. Tumours showing altered beta-catenin localization were tested for beta-catenin and APC mutations. APC promoter methylation was restricted to gastroduodenal tumours (21 out of 59, 36%), prevalent in poorly differentiated carcinomas (P=0.042) and correlating with aggressive features (high histology grade, P<0.02; tumour death, P=0.026; high fractional allelic loss, P=0.002, in turn correlating with short survival, P=0.017). LOH at APC locus was found in 14 out of 53 cases (26%, 10 gastroduodenal and 4 colorectal), prevalent in poorly differentiated carcinomas (P=0.002) and correlating with histology grade (P=0.012). beta-catenin abnormal expression was found in 41 out of 54 cases (76%), with nuclear staining correlating with APC alteration (P=0.047) and short survival (P=0.006). APC, but not beta-catenin, gene mutations were found (7 out of 35 tumours), 4 of which in the midgut. E-cadherin promoter methylation was rarely detected (2 out of 52 cases), with cytoplasmic expression in 18 out of 43 cases (42%), not correlating with any clinico-pathological feature. In conclusion, Wnt pathway alterations, as represented by abnormal beta-catenin localization, are common events in digestive endocrine tumours, but only nuclear expression correlates with tumour aggressiveness. Though with different alteration mechanisms according to anatomical site, APC plays a major role in Wnt pathway activation and in determining the high chromosomal instability observed in aggressive endocrine carcinomas.


Buzz Factor or Innovation Potential: What Explains Cryptocurrencies' Returns?
Cryptocurrencies have become increasingly popular since the introduction of bitcoin in 2009. In this paper, we identify factors associated with variations in cryptocurrencies' market values. In the past, researchers argued that the "buzz" surrounding cryptocurrencies in online media explained their price variations. But this observation obfuscates the notion that cryptocurrencies, unlike fiat currencies, are technologies entailing a true innovation potential. By using, for the first time, a unique measure of innovation potential, we find that the latter is in fact the most important factor associated with increases in cryptocurrency returns. By contrast, we find that the buzz surrounding cryptocurrencies is negatively associated with returns after controlling for a variety of factors, such as supply growth and liquidity. Also interesting is our finding that a cryptocurrency's association with fraudulent activity is not negatively associated with weekly returns-a result that further qualifies the media's influence on cryptocurrencies. Finally, we find that an increase in supply is positively associated with weekly returns. Taken together, our findings show that cryptocurrencies do not behave like traditional currencies or commodities-unlike what most prior research has assumed-and depict an industry that is much more mature, and much less speculative, than has been implied by previous accounts.


Current methods for the identification of carbapenemases.
Detection of carbapenemases in clinical microbiology labs is a challenging issue. Comparison of the results of susceptibility testing with the breakpoint values of carbapenems is the first step in the screening of carbapenemase producers. To date, screening of carbapenemase-producing (CP) bacteria has been mostly performed by a selective medium. Although these media are practical for the detection of most CP isolates, the inoculated plates have to be incubated overnight. Subsequently, we need the confirmation of the carbapenemase producers present in the culture medium by additional testing [e.g. inhibition studies with liquid or solid media, modified Hodge test (MHT), or gradient strips], which can take up to another 48 hours. Despite the lack of discrimination between the three different classes of carbapenemases (KPC, MBL and OXA) and difficulties in the interpretation of the results, the MHT is usually deemed as the phenotypic reference method for the confirmation of carbapenemase production. Molecular techniques, such as real-time polymerase chain reaction (PCR) assays, in contrast to phenotypic methods that are very time consuming, are faster and allow for the quick identification of carbapenemase genes. These techniques can detect and characterize carbapenemases, including NDM- and KPC-mediated resistance, which is critical for epidemiological investigations. The aim of this review is to gather a summary of the available methods for carbapenemase detection and describe the strengths and weaknesses of each method.