Putative Causes Research Articles

New mouse model for inducing and evaluating unilateral vestibular deafferentation syndrome

BackgroundUnilateral vestibular deafferentation syndrome (uVDS) holds a particular place in the vestibular pathology domain. Due to its suddenness, the violence of its symptoms that often result in emergency hospitalization, and its associated original neurophysiological properties, this syndrome is a major source of questioning for the otoneurology community. Also, its putative pathogenic causes remain to be determined. There is currently a strong medical need for the development of targeted and effective countermeasures to improve the therapeutic management of uVDS. New methodsThe present study reports the development of a new mouse model for inducing and evaluating uVDS. Both the method for generating controlled excitotoxic-type peripheral vestibular damages, through transtympanic administration of the glutamate receptors agonist kainate (TTK), and the procedure for evaluating the ensuing clinical signs are detailed. Comparison with existing methodsThrough extensive analysis of the clinical symptoms characteristics, this new animal model provides the opportunity to better follow the temporal evolution of various uVDS specific symptoms, while better appreciating the different phases that composed this syndrome. ResultsThe uVDS evoked in the TTK mouse model displays two main phases distinguishable by their kinetics and amplitudes. Several parameters of the altered vestibular behaviour mimic those observed in the human syndrome. ConclusionThis new murine model brings concrete information about how uVDS develops and how it affects global behaviour. In addition, it opens new opportunity to decipher the etiopathological substrate of this pathology by authorizing the use of genetically modified mouse models.

A Practical Guide for Treatment of Pain in Patients with Systemic Mast Cell Activation Disease

Systemic mast cell activation disease (MCAD, a subclass of mastocytosis), which has a prevalence of around 17% (at least in the German population), is characterized by accumulation of genetically altered dysfunctional mast cells with abnormal release of these cells’ mediators. Since mast cells affect functions in potentially every organ system, often without causing abnormalities in routine laboratory or radiologic testing, this disease has to be considered routinely in the differential diagnosis of patients with chronic multisystem polymorbidity of a generally inflammatory and allergic theme. Pain in its different manifestations is a common symptom in MCAD found in more than three-quarters of the MCAD patients. Because of the specific mast cell-related causes of pain in MCAD it should be treated specifically, if possible, deduced from their putative mast cell mediator-related causes. As yet, there is no official guideline for treatment of MCAD at all. The present review focuses on mast cell mediator-induced acute and chronic pain and the current state of analgesic drug therapy options in MCAD. Due to the high prevalence of MCAD, many physicians are often faced with the issue of pain management in MCAD patients. Hence, our practical guide should contribute to the improvement of patient care.

Understanding processes at the origin of species flocks with a focus on the marine Antarctic fauna.

Species flocks (SFs) fascinate evolutionary biologists who wonder whether such striking diversification can be driven by normal evolutionary processes. Multiple definitions of SFs have hindered the study of their origins. Previous studies identified a monophyletic taxon as a SF if it displays high speciosity in an area in which it is endemic (criterion 1), high ecological diversity among species (criterion 2), and if it dominates the habitat in terms of biomass (criterion 3); we used these criteria in our analyses. Our starting hypothesis is that normal evolutionary processes may provide a sufficient explanation for most SFs. We thus clearly separate each criterion and identify which biological (intrinsic) and environmental (extrinsic) traits are most favourable to their realization. The first part focuses on evolutionary processes. We highlight that some popular putative causes of SFs, such as key innovations or ecological speciation, are neither necessary nor sufficient to fulfill some or all of the three criteria. Initial differentiation mechanisms are diverse and difficult to identify a posteriori because a primary differentiation of one type (genetic, ecological or geographical) often promotes other types of differentiation. Furthermore, the criteria are not independent: positive feedbacks between speciosity and ecological diversity among species are expected whatever the initial cause of differentiation, and ecological diversity should enhance habitat dominance at the clade level. We then identify intrinsic and extrinsic factors that favour each criterion. Low dispersal emerges as a convincing driver of speciosity. Except for a genomic architecture favouring ecological speciation, for which assessment is difficult, high effective population sizes are the single intrinsic factor that directly enhances speciosity, ecological diversity and habitat dominance. No extrinsic factor appeared to enhance all criteria simultaneously but a combination of factors (insularity, fragmentation and environmental stability) may favour the three criteria, although the effect is indirect for habitat dominance. We then apply this analytical framework to Antarctic marine environments by analysing data from 18 speciose clades belonging to echinoderms (five unrelated clades), notothenioid fishes (five clades) and peracarid crustaceans (eight clades). Antarctic shelf environments and history appear favourable to endemicity and speciosity, but not to ecological specialization. Two main patterns are distinguished among taxa. (i) In echinoderms, many brooding, species-rich and endemic clades are reported, but without remarkable ecological diversity or habitat dominance. In these taxa, loss of the larval stage is probably a consequence of past Antarctic environmental factors, and brooding is suggested to be responsible for enhanced allopatric speciation (via dispersal limitation). (ii) In notothenioids and peracarids, many clades fulfill all three SF criteria. This could result from unusual features in fish and crustaceans: chromosome instability and key innovations (antifreeze proteins) in notothenioids, ecological opportunity in peracarids, and a genomic architecture favouring ecological speciation in both groups. Therefore, the data do not support our starting point that normal evolutionary factors or processes drive SFs because in these two groups uncommon intrinsic features or ecological opportunity provide the best explanation. The utility of the three-criterion SF concept is therefore questioned and guidelines are given for future studies.

Abstract 2860: 89Zr-trastuzumab immunoPET imaging to monitor src status after treatment in HER2 breast cancer

Abstract Around 30% of patients with early stage breast cancer have recurrent disease due to resistance. One of the putative causes of acquired resistance can be attributed to the hyperactivation of Src kinases, which are particularly stabilized by aberrant HER2 signaling, which itself is overexpressed in nearly 30% of early stage breast cancers. HER2 downstream effectors include Src, thus, we aim to investigate the use of a HER2-specific monoclonal antibody positron emission tomography (PET) probe, 89Zr-trastuzumab, to monitor Src therapy via surrogate HER2 expression. We hypothesize that 89Zr-trastuzumab can be a surrogate imaging tool for monitoring Src treatment. 89Zr-trastuzumab was previously developed and has demonstrated excellent specificity for breast cancer. Dasatinib-treated HER2+ BT474 (IC50 ~ 1.3 μM), SK-BR-3 (IC50 ~ 5.5 µM), and JIMT-1 (IC50 ~ 8 μM) breast cancer cells lines have shown a negative correlation between treatment and HER2. We next conducted tumorigenic studies using athymic nu/nu mice bearing JIMT-1 or BT474 xenografts, which were treated for 7 or 14 days with dasatinib (75 mg/kg/day). At the end of each treatment, the tumors were subsequently imaged with 18F-FDG, and then followed by 89Zr-trastuzumab, 24 h later. Images were analyzed using AsiPro (show version here and decay-corrected to the time of injection. Tumor uptake values were measured by drawing volumes-of-interest, expressed as % injected dose per gram (%ID/g) of tissue on regions showing the most binding. 18F-FDG did not demonstrate a predictive response in both treated and placebo tumors, whereas, HER2 PET was able to show differential response in treated vs. control cohorts. In JIMT-1 mice imaged with 89Zr-trastuzumab, a two-fold difference (p < 0.01) was observed between untreated and both treated groups (8.0 ± 1.5 %ID/g, n= 7 vs. 7-day treated: 3.8 ± 1.5 %ID/g, n=4, and 14-day-treated: 4.5 ± 1.3 %ID/g, n= 4) mice imaged with 89Zr-trastuzumab. Confirmatory ex vivo western blots of JIMT-1 tumors have shown a decrease in HER2 as treatment time increases, with a decrease in p-Src (Y-416). In vivo studies with BT-474 xenografts are currently underway. Immunohistochemistry on excised tumors, as well as treatment on a metastatic patient-derived xenograft model is currently in progress. We have shown that HER2 PET can potentially be a surrogate marker of Src treatment. The findings from this study potentially afford a powerful tool to non-invasively detect and monitor changes in Src-targeted therapy in man at the early phases of treatment. Citation Format: Brooke N. McKnight, Nerissa T. Viola-Villegas. 89Zr-trastuzumab immunoPET imaging to monitor src status after treatment in HER2 breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2860. doi:10.1158/1538-7445.AM2017-2860

Abstract 4474: Functional characterization of alternatively spliced GSN in head and neck cancer

Abstract PURPOSE: Analyze the functional role of alternative splicing in HPV-related (HPV+) Head and Neck Squamous Cell Carcinoma (HNSCC) biology and oncogenesis. HPV+ HNSCC incidence is rapidly increasing, while its oncogenic mechanisms, demographic, and clinico-pathological characteristics are largely uncharacterized and discrete from those of HPV- HNSCC. Investigation into the molecular biology of HPV pathogenesis in HNSCC may help with creation of more specific clinical tools for diagnosis and treatment, which are limited and lack accuracy. Alternative Splicing Events (ASEs), post-transcriptional modifications resulting in variant transcripts leading to alternative protein isoforms, have been under-investigated as putative causes of gene expression dysregulation in HPV+ HNSCC. Using RNA-Seq data, a genome-wide ASE discovery analysis was performed on a JHU HPV+ HNSCC cohort, which has expanded HPV+ HNSCC transcriptional characterization by identifying novel ASE biomarkers implicated in carcinogenic progression. Outlier Gene Set Analysis (OGSA) was used to identify genes with high rates of ASE and results were validated in TCGA samples from seven different cancer types. To verify the functional implications of differential isoform expression, vectors for knock-in and knock-down of WT-GSN and ASE-GSN isoforms were developed to evaluate changes in cell proliferation and invasion. ASE-GSN/WT-GSN ratio outliers were identified by OGSA in tumor samples from discovery JHU HPV+ HNSCC cohort, as well as in TCGA public domain data from HNSCC, Lung Squamous Cell Carcinoma, Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma, Bladder Urothelial Carcinoma, Breast Invasive Carcinoma, and Colon Adenocarcinoma. The increased ASE-GSN/WT-GSN ratio was also observed in a cohort of HPV+ Head and Neck Cancer Cell lines. SCC61 and UM-SCC-22B HNSCC cell lines were used to evaluate the function GSN ASEs in vitro. Overall expression levels of both GSN isoforms inversely correlated with cell proliferation whereas the high ratio of ASE-GSN to WT-GSN correlated with increased cellular invasion of stroma in vitro. Additionally, a decrease or increase in expression of one isoform is seen to cause a compensatory increase or decrease in the other isoform respectively. ASE-GSN outliers correlated with HPV+ cancers indicate that expression ratio between GSN isoforms is related to tumor behavior, which is reinforced in vitro where WT-GSN is inversely correlated and ASE-GSN is directly correlated to with cell invasion. These relationships suggest that both overall expression and balance between the two GSN isoforms are mediating factors in cell death or proliferation, while shorter ASE-GSN is more likely to indicate progression to cancer. Finally, we propose a mechanism for the role of GSN isoforms in apoptosis and metastasis, in which trends in expression between WT- and ASE-GSN isoforms can alternatively affect rates of apoptosis or metastatic ability. Citation Format: Dylan Z. Kelley, Emily L. Flam, Theresa Guo, Craig Bohrson, Michael Considine, Ludmila V. Danilova, Justin A. Bishop, Chi Zhang, Wayne M. Koch, David Sidransky, William Westra, Sarah Wheelan, Liliana Florea, Elana J. Fertig, Joseph A. Califano, Daria A. Gaykalova. Functional characterization of alternatively spliced GSN in head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4474. doi:10.1158/1538-7445.AM2017-4474

Analysis of cytopenia in geriatric inpatients.

Peripheral blood dyscrasias in older patients are repeatedly seen in geriatric clinical practice; however, there is substantial lack of data about the epidemiology, possible causes and treatment options in this patient group. Proton pump inhibitors (PPI) are extensively used in older patients and associated with leukopenia. The primary objective of this study was the assessment of encoded cytopenia prevalence in ageriatric patient cohort and the secondary objective was the assessment of putative causes and the analysis of PPI administration in patients with cytopenia. Retrospective evaluation of patients admitted to the geriatric department of aGerman urban hospital between 2010 and 2012. Electronic patient data were screened for encoded diagnosis of cytopenia according to the International Classification of Diseases (ICD)10. Inclusion criteria were ICD code D69.0-9 and/or D70.0-7, age ≥60years and exclusion criteria were no ICD code D69.0-9 and/or D70.0-7 and age <60years. Out of 9328 screened inpatients 54patients remained for analysis. Study parameters included hemoglobin (Hb), red blood cell count (RBC), leucocytes, platelets, mean cell volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), presence of leukopenia (<4000/µl), presence of thrombocytopenia (<140,000/µl) and presence of anemia according to the World Health Organization (WHO). Substitution of blood products, medication with PPI and potential causes for dyscrasias were evaluated based on electronic patient records. The mean age was 78.3± 6.5years (27females, 27males), anemia was seen in 78%, leukopenia was encoded in13% and thrombocytopenia in 44.4%. In most of the patients no substitution of blood products was documented. In most of the patients (20.4%) cytopenia was attributed to either heparin-induced thrombocytopenia (HIT) or hemato-oncologic (20.4%) diseases, followed by drug association in 18.5%. In 70.8% of the study patients PPIs were administered but the indication for PPI administration remained unclear in 20.4%. The results encourage accurate assessment of blood dyscrasias and appropriate documentation as well as indication check for PPI treatment in geriatric inpatients.

Cell-free DNA in a three-dimensional spheroid cell culture model: A preliminary study

BackgroundInvestigating the biological functions of cell-free DNA (cfDNA) is limited by the interference of vast numbers of putative sources and causes of DNA release into circulation. Utilization of three-dimensional (3D) spheroid cell cultures, models with characteristics closer to the in vivo state, may be of significant benefit for cfDNA research. MethodsCfDNA was isolated from the growth medium of C3A spheroid cultures in rotating bioreactors during both normal growth and treatment with acetaminophen. Spheroid growth was monitored via planimetry, lactate dehydrogenase activity and glucose consumption and was related to isolated cfDNA characteristics. ResultsChanges in spheroid growth and stability were effectively mirrored by cfDNA characteristics. CfDNA characteristics correlated with that of previous two-dimensional (2D) cell culture and human plasma research. Conclusions3D spheroid cultures can serve as effective, simplified in vivo-simulating “closed-circuit” models since putative sources of cfDNA are limited to only the targeted cells. In addition, cfDNA can also serve as an alternative or auxiliary marker for tracking spheroid growth, development and culture stability. Biological significance3D cell cultures can be used to translate “closed-circuit” in vitro model research into data that is relevant for in vivo studies and clinical applications. In turn, the utilization of cfDNA during 3D culture research can optimize sample collection without affecting the stability of the growth environment. Combining 3D culture and cfDNA research could, therefore, optimize both research fields.

Modulation of natriuretic peptide receptors in human adipose tissue: molecular mechanisms behind the “natriuretic handicap” in morbidly obese patients

The B-type natriuretic peptide (BNP) hormone plays a crucial role in the regulation of cardiovascular and energy homeostasis. Obesity is associated with lowcirculating levels of BNP, a condition known as "natriuretic handicap." Recent evidences suggest an altered expression of BNP receptors-both the signaling natriuretic peptide receptors (NPR)-A and the clearance NPR-C receptor-in adipose tissue (AT) as one of the putative causes of natriuretic handicap. The current study aims at clarifying the molecular mechanisms behind the natriuretic handicap, focusing on NPR modulation in the AT of obese and control subjects. The study enrolled 34 obese and 20 control subjects undergoing bariatric or abdominal surgery, respectively. The main clinical and biochemical parameters, including circulating BNP, were assessed. In visceral (VAT) and subcutaneous AT (SAT) samples, collected during surgery, the adipocytes and stromal vascular fraction (SVF) expression of NPR-A and NPR-C and the SVF secretion of interleukin 6 (IL-6) were determined. Both VAT and SAT from obese patients expressed a lower NPR-A/NPR-C ratio in adipocytes and the SVF secreted a higher level of IL-6, compared with the controls. Moreover, NPR-A/NPR-C ratio expressed by VAT and SAT adipocytes negatively correlated with body mass index, insulin, the Homeostasis Model Assessment of Insulin resistance, and IL-6 secreted by SVF, and the expression of the clearance receptor NPR-C, in both the VAT and SAT adipocytes, showed a negative correlation with circulating BNP. Overall, insulin resistance/hyperinsulinemia and AT inflammation (ie, high level of IL-6) are the major determinants of the lower NPR-A/NPR-C ratio in adipocytes, thus contributing to the natriuretic handicap in obese subjects.

Yield of Brain MRI in Clinically Diagnosed Epilepsy in the Kingdom of Bhutan: A Prospective Study

BackgroundPeople with epilepsy (PWE) in low- and middle-income countries may not access the health resources that are considered optimal for epilepsy diagnosis. The diagnostic yield of magnetic resonance imaging (MRI) has not been well studied in these settings. ObjectivesTo report the diagnostic yield of brain MRI and identify clinical associations of abnormal MRI findings among PWE in a neurocysticercosis-endemic, resource-limited setting and to identify the proportion and putative structural brain causes of drug-resistant epilepsy. MethodsPWE were prospectively enrolled at the Jigme Dorji Wangchuck National Referral Hospital in Bhutan (2014-2015). Each participant completed clinical questionnaires and a 1.5-Tesla brain MRI. Each MRI was reviewed by at least 1 radiologist and neurologist in Bhutan and the United States. A working definition of drug-resistant epilepsy for resource-limited settings was given as (a) seizures for >1 year, (b) at least 1 seizure in the prior year, and (c) presently taking 2 or more antiepileptic drugs (AEDs). Logistic regression models were constructed to test the cross-sectional association of an abnormal brain MRI with clinical variables. FindingsA total of 217 participants (125 [57%] female; 54 [25%] < 18 years old; 199 [92%] taking AEDs; 154 [71%] with a seizure in the prior year) were enrolled. There was a high prevalence of abnormal brain MRIs (176/217, 81%). Mesial temporal sclerosis was the most common finding (n = 115, 53%, including 24 children), exceeding the number of PWE with neurocysticercosis (n = 26, 12%, including 1 child) and congenital/perinatal abnormalities (n = 29, 14%, including 14 children). The number of AEDs (odds ratio = .59, P = .03) and duration of epilepsy (odds ratio = 1.11, P = .02) were significantly associated with an abnormal MRI. Seizure in the prior month was associated with the presence of mesial temporal sclerosis (odds ratio = .47, P = .01). A total of 25 (12%) participants met our definition of drug-resistant epilepsy, with mesial temporal sclerosis (n = 10), congenital malformations (n = 5), and neurocysticercosis (n = 4) being the more common findings. ConclusionsThe prevalence of abnormalities on brain MRI for PWE in resource-limited settings is high as a result of a diffuse range of etiologies, most commonly mesial temporal sclerosis. Drug-resistant epilepsy accounted for 12% of the referral population in a conservative estimation.

The theoretical link between voltage loss, reduction in field enhancement factor, and Fowler-Nordheim-plot saturation

With a large-area field electron emitter, when an individual post-like emitter is sufficiently resistive, and current through it is sufficiently large, then voltage loss occurs along it. This letter provides a simple analytical and conceptual demonstration that this voltage loss is directly and inextricably linked to a reduction in the field enhancement factor (FEF) at the post apex. A formula relating apex-FEF reduction to this voltage loss was obtained in the paper by Minoux et al. [Nano Lett. 5, 2135 (2005)] by fitting to numerical results from a Laplace solver. This letter derives the same formula analytically, by using a “floating sphere” model. The analytical proof brings out the underlying physics more clearly and shows that the effect is a general phenomenon, related to reduction in the magnitude of the surface charge in the most protruding parts of an emitter. Voltage-dependent FEF-reduction is one cause of “saturation” in Fowler-Nordheim (FN) plots. Another is a voltage-divider effect, due to measurement-circuit resistance. An integrated theory of both effects is presented. Both together, or either by itself, can cause saturation. Experimentally, if saturation occurs but voltage loss is small (&amp;lt;20 V, say), then saturation is more probably due to FEF-reduction than voltage division. In this case, existing treatments of electrostatic interaction ("shielding") between closely spaced emitters may need modification. Other putative causes of saturation exist, so the present theory is a partial story. Its extension seems possible and could lead to a more general physical understanding of the causes of FN-plot saturation.

Physiology of the scalp

Human scalp reveals aremarkable difference to hairy skin of other body regions. The structure of the scalp offers effective protection against water loss and contributes to thermoregulation due to significant vascularization. The scalp is innervated by the trigeminus nerve. The majority of the population complain about "sensitive" scalp. However, sensory, regional, ethnic or pathologic parameters contributing to "sensitive" scalp are largely unknown. Quantitative sensory tests and superficial topical application test of irritant substances were performed. Extremely high thresholds to thermal and mechanical stimuli were recorded at the vertex compared to occipital or temporal scalp regions, or the hairy skin of other body regions such as trunk or distal extremities. Pronounced scalp hypersensitivity can be found in Asians as compared to Caucasians and under pathologic conditions, such as alopecia, dandruff, or migraine. Putative causes of "sensitive" scalp can be afacilitated transdermal penetration of irritant substances, altered neuronal innervation or receptor expression, or peripheral and central sensitization of sensory afferents.

The effect of recrystallization of aqueous solutions of metal sulfates on the acid–base balance

A comparative study of pH values in recrystallized aqueous solutions of manganese, copper, and iron sulfates and the calculated values for the pH levels of similar solutions obtained directly from solid salts has been performed for the first time. The trend for the positive deviation of the pHrec values from the pHcalc values has been established. The positive deviation was the strongest in the case of iron sulfate solutions. The sample of the recrystallized aqueous solution (recrystallized water) consisted of water obtained by melting of the initial aqueous solution after freezing of 60% of the solution (by volume). Four hypotheses were proposed to explain the positive deviation of pHrec from pHcalc: (1) low СО2 concentration; (2) more efficient adsorption of anions on the freezing front compared to that of cations; (3) mechanochemical and radiation chemical processes that occur upon the melting of ice; and (4) Fe2+-catalyzed decomposition of hydrogen peroxide formed in the recrystallized solution. Analysis of the putative causes showed that the fourth hypothesis provided the most adequate explanation of the difference between pHrec and pHcalc. The formation of hydrogen peroxide in recrystallized water is proposed to occur due to the recombination of OH radicals formed during the radiation chemical processes.

Diurnal surface activity of the Ciscaucasian hamster (Mesocricetus raddei) in the field

Monitoring daily activity patterns of animals in the field and in captivity often revealed different results. For example, Syrian hamsters Mesocricetus auratus showed diurnal surface activity in the field, but nocturnal patterns in the laboratory. The aim of our study was to monitor the field activity of a close relative of this species, the Ciscaucasian hamster Mesocricetus raddei from Dagestan. Using the radio-frequency identification (RFID) technique we were able to obtain field data from 20 Ciscaucasian hamsters. In contrast to the data obtained in captivity where the Ciscaucasian hamsters were active mainly at night, they showed a diurnal activity pattern in the field, i.e. animals were observed on surface between sunrise and sunset. Discussing the putative causes of the different activity patterns, we are focusing especially on ecological constraints and advantages. Also, we stress the necessity of more field observations.

Behavioral and psychological outcomes for dogs sold as puppies through pet stores and/or born in commercial breeding establishments: Current knowledge and putative causes

A review of 7 published studies and 1 anecdotal report involving dogs born in high-volume commercial breeding establishments and sold to the consumer directly via the Internet or indirectly through retail pet stores revealed an increased incidence of behavioral and emotional problems that cause distress in adulthood compared with dogs from other sources, especially noncommercial breeders. The most consistent finding among studies is an increase in aggression, which is most commonly directed toward the dog's owners and family members but also to unfamiliar people, and other dogs. Increased fear was also identified in response to unfamiliar people, children, other dogs, nonsocial stimuli, and when taken on walks. Undesirable behaviors related to separation and/or attention seeking and a heightened sensitivity to touch have been reported. Because of how dogs sold through pet stores and/or born in commercial breeding establishments are bred, housed, weaned, transported, handled, homed, and raised, potential contributing factors for these reported outcomes are numerous. Some key factors include genetics, early-life stimulus deprivation (inadequate stimulus exposure, inappropriate or lack of social exposure), stress (prenatal maternal stress and postnatal early-life adversity), early weaning and maternal separation, transport and pet-store-related factors, and owner-related factors such as inadequate knowledge and experience with dogs as well as different levels of commitment to the pet dog. All published studies suggest a role for major stressors during puppy development from the prenatal stage through adolescence in the development of many behavioral problems. Accordingly, for any dog breeding operation, a standard of care that adequately redresses the welfare of the mother and pups and the risk of later behavior problems attendant with early stress and distress need to be formulated and followed in a manner supported by the emerging data.

A Literature Review of Senile Pruritus: From Diagnosis to Treatment.

Pruritus occurs frequently in the elderly population, but it has not been fully defined and its precise prevalence is unknown. Putative causes of pruritus are numerous and patients' quality of life can be greatly reduced. In the absence of any specific cause, a diagnosis of senile pruritus is proposed, suggesting that itch is secondary to ageing. However, the relationship of pruritus with skin ageing and/or neuronal ageing and/or immune ageing is poorly understood. Many treatments have been reported, but are poorly effective. This paper reviews the literature on senile pruritus, in order to improve understanding of this condition and to enable improved diagnosis and treatment.

China’s Troubled Waters: Maritime Disputes in Theoretical Perspective, by Steve Chan. Cambridge: Cambridge University Press, 2016. 250 pp. US$99.99 (cloth).

<i>China’s Troubled Waters: Maritime Disputes in Theoretical Perspective</i>, by Steve Chan. Cambridge: Cambridge University Press, 2016. 250 pp. US$99.99 (cloth).

Piecing together a different picture: A host of new studies on autism have begun decoding the longstanding puzzle of its causes.

“A developmental disability that hinders the normal functioning of the brain, affecting, in varying degrees, communication skills and social interaction. Repetitive behaviours, and different ways of learning, paying attention, or reacting to things are often distinctive signs”. This standard definition of autism fails to describe the complexity of a condition that ranges in its manifestations from severe intellectual impairment to superior cognitive skills, like in the Asperger syndrome. To comprise such diversity, autism disorders are now covered under the umbrella term “autism spectrum disorder” (ASD). In most cases, ASD manifests during the first 5 years of life, with boys significantly more likely to be diagnosed than girls. ASD usually goes together with several other problems that frequently include anxiety, sleep disorders, or epilepsy. No cure exists; treatment, such as speech therapy, just attempts to alleviate specific deficits of autistic patients. > Nothing is simple in autism. Even the real number of people affected is uncertain. Nothing is simple in autism. Even the real number of people affected is uncertain. The US CDC estimates that about 1 in 68 (or 1.5%) of children in the USA are living with ASD (http://www.cdc.gov/ncbddd/autism/data.html). The WHO has a more conservative estimate, last revised in January this year, of 1 in 160 children, based on a larger set of epidemiological surveys (http://www.who.int/mediacentre/factsheets/autism-spectrum-disorders/en/). Needless to say, most studies were conducted in developed countries, and the prevalence of ASD in many low‐ and middle‐income countries remains largely unknown. > Along the years, many potential causes have been indicated, including genetic and environmental factors, exposure to toxins during pregnancy, wide gaps between parent ages, and so on Although the general consensus is that prevalence rates are increasing globally, this point is debated too. Some analyses indicate that a large percentage of the increase in ASD owes to improved awareness and …

Mitochondrial DNA hyperdiversity and its potential causes in the marine periwinkle Melarhaphe neritoides (Mollusca: Gastropoda).

We report the presence of mitochondrial DNA (mtDNA) hyperdiversity in the marine periwinkle Melarhaphe neritoides (Linnaeus, 1758), the first such case among marine gastropods. Our dataset consisted of concatenated 16S-COI-Cytb gene fragments. We used Bayesian analyses to investigate three putative causes underlying genetic variation, and estimated the mtDNA mutation rate, possible signatures of selection and the effective population size of the species in the Azores archipelago. The mtDNA hyperdiversity in M. neritoides is characterized by extremely high haplotype diversity (Hd = 0.999 ± 0.001), high nucleotide diversity (π = 0.013 ± 0.001), and neutral nucleotide diversity above the threshold of 5% (πsyn = 0.0677). Haplotype richness is very high even at spatial scales as small as 100m2. Yet, mtDNA hyperdiversity does not affect the ability of DNA barcoding to identify M. neritoides. The mtDNA hyperdiversity in M. neritoides is best explained by the remarkably high mutation rate at the COI locus (μ = 5.82 × 10−5 per site per year or μ = 1.99 × 10−4 mutations per nucleotide site per generation), whereas the effective population size of this planktonic-dispersing species is surprisingly small (Ne = 5, 256; CI = 1,312–3,7495) probably due to the putative influence of selection. Comparison with COI nucleotide diversity values in other organisms suggests that mtDNA hyperdiversity may be more frequently linked to high μ values and that mtDNA hyperdiversity may be more common across other phyla than currently appreciated.

Is Constipation More Common in Overweight Children?

Introduction: Obesity rates have doubled in children and quadrupled in adolescents in the past 30 years. The most important causes of obesity are sedentary life style and low fiber, high calorie diet which are also putative causes of constipation. Some studies have found a correlation between constipation and obesity while others have been inconclusive. In an effort to understand this relationship we undertook a study to assess if overweight children have a higher risk of constipation as compared to normal weight children. Methods: A retrospective review of charts of new patients seen at Pediatric GI Clinic between 1/1/11 and 12/31/12 was performed. All charts were reviewed with an emphasis on growth curves and final diagnoses. Growth status of each patient was assessed by using standard WHO charts for weight, height, and body mass index (BMI) for age and sex. Obesity was defined as a BMI above 95th percentile, overweight between 85-95th percentile, normal between 3rd-85th percentile and failure to thrive (FTT) below 3rd percentile. For statistical analysis, overweight and obese patients were combined into one single group (overweight). The final diagnosis of constipation was made by the attending Pediatric Gastroenterologist. The incidences of constipation in the overweight and FTT were compared to that in normal children using chi-squared analysis. Results: Out of 1876 (M:F, 941:935) new patients seen during study period, 295 patients were overweight (M:F 146:149, 225 obese), 211 FTT and 1370 normal weight (other GI complaints/diagnoses). Observed and expected incidence of constipation in these groups is reported in Table 1. The data was analyzed by chi-squared analysis for significance of incidence of constipation in three individual categories: obese, normal, and failure to thrive (FTT). There was a higher incidence of constipation in overweight patients than expected (a chi-square value of 25. 137 and a p-value < 0.05, CI 95%). Furthermore, the data showed that the incidence of constipation was lower than expected in FTT (a chi-square value of 15.2 and a p-value < 0.05, CI 95%).Table 1: The Observed vs Expected Total Number of Constipated and Non-Constipated PatientsConclusion: There is a higher incidence of constipation in overweight children. Children with FTT have lower incidence of constipation. An analysis of nutrition and exercise habits amongst patients with constipation, obesity, overweight, and failure to thrive could further reveal associations between constipation and BMI.

Deep narrow territories

It is always a pleasure to observe Peter Harrison deploying his skill in argument, repertoire of apt quotations, and power of analogy. One argument joins so neatly to the next that it seems willful to withhold assent from the conclusion. Having enjoyed the sparkle, however, it is well to consider how it is achieved. The principal means, conspicuous also in Harrison’s earlier book, The Bible: Protestantism and the Rise of Science (1998), is a very tight focus: the argument, repertoire, and analogies achieve their effect by operating in a domain of inquiry restricted more severely, and sometimes less justifiably, than might at first appear. In The Bible, Harrison argued that by doing away with allegorical readings of scripture, Protestants liberated ‘‘the study of the natural world from the specifically religious concern of biblical interpretation.’’ The fruit of this liberation fell far from the tree. ‘‘The literalist mentality initiated by the Protestant reformers, and sponsored by their successors [was]....by far the most significant [cause]...for the rise of modern science.’’ With this bold extrapolation, Harrison made the reformers’ method of sola scriptura serve as a motive, indeed, the main motive, for the very diverse sorts of thinking and acting required to investigate and apply natural knowledge. The ascendency of bible reading over all other putative causes of the Scientific Revolution, however brilliantly argued, is not very convincing. The Territories of Science and Religion follows the method of sola scriptura to conclude that clashes between the territories could not have occurred before the Reformation. In England they started even later, in the last third of the nineteenth century. Harrison’s crisp argument for this chronology is quickly summarized. He observes that ‘‘science’’ and ‘‘religion’’ (SR that before the watershed they had to do with the internal cultivation of the individual and after it with a public confession