Tumor-induced platelet aggregation and growth promoting factors as determinants for successful tumor metastasis.
Two clones isolated from a metastatic variant of mouse colon adenocarcinoma 26; the high metastatic NL-17 and the low metastatic NL-44, induced similar platelet aggregation in vitro. Heterotypic aggregates of tumor cells and platelets were injected i.v. into mice. The lung colonization potential of the NL-17 was dependent on the extent of tumor cell platelet aggregation. This clearly indicates that the interaction of tumor cells with platelets can lead to enhanced tumor metastasis possibly through more efficient intravascular arrest of the heterotypic aggregates. The interaction of tumor cells with platelets could thus be an important determinant for successful metastasis. NL-44, however, did not form pulmonary metastasis even after the tumor cells formed heterotypic aggregates with platelets, suggesting that tumor metastasis, is dependent on the intrinsic nature of tumor cells. Lung extract enhanced the growth of NL-17 more effectively than that of NL-44. These results suggest that, in addition to the interaction of metastatic cells and platelets, host factors including growth-promoting factors might also have an important role in tumor metastasis.


Swelling Cholesteric Liquid Crystal Shells to Direct the Assembly of Particles at the Interface.
Cholesteric liquid crystals can exhibit spatial patterns in molecular alignment at interfaces that can be exploited for particle assembly. These patterns emerge from the competition between bulk and surface energies, tunable with the system geometry. In this work, we use the osmotic swelling of cholesteric double emulsions to assemble colloidal particles through a pathway-dependent process. Particles can be repositioned from a surface-mediated to an elasticity-mediated state through dynamically thinning the cholesteric shell at a rate comparable to that of colloidal adsorption. By tuning the balance between surface and bulk energies with the system geometry, colloidal assemblies on the cholesteric interface can be molded by the underlying elastic field to form linear aggregates. The transition of adsorbed particles from surface regions with homeotropic anchoring to defect regions is accompanied by a reduction in particle mobility. The arrested assemblies subsequently map out and stabilize topological defects. These results demonstrate the kinetic arrest of interfacial particles within definable patterns by regulating the energetic frustration within cholesterics. This work highlights the importance of kinetic pathways for particle assembly in liquid crystals, of relevance to optical and energy applications.


Increase in intracranial pressure by application of a rigid cervical collar: a pilot study in healthy volunteers.
Rigid cervical collars are known to increase intracranial pressure (ICP) in severe traumatic brain injury (TBI). Cerebral blood flow might decrease according to the Kellie Monroe doctrine. For this reason, the use of the collar in patients with severe TBI has been abandoned from several trauma protocols in the Netherlands. There is no evidence on the effect of a rigid collar on ICP in patients with mild or moderate TBI or indeed patients with no TBI. As a first step we tested the effect in healthy volunteers with normal ICPs and intact autoregulation of the brain. In this prospective blinded cross-over study, we evaluated the effect of application of a rigid cervical collar in 45 healthy volunteers by measuring their optical nerve sheath diameter (ONSD) by transocular sonography. Sonographic measurement of the ONSD behind the eye is an indirect noninvasive method to estimate ICP and pressure changes. We included 22 male and 23 female volunteers. In total 360 ONSD measurements were performed in these 45 volunteers. Application of a collar resulted in a significant increase in ONSD in both the left (β=0.06, 95% confidence interval: 0.05-0.07, P<0.001) and the right eye (β=0.01, 95% confidence interval: 0.00-0.02, P=0.027) CONCLUSION: Application of a rigid cervical collar significantly increases the ONSD in healthy volunteers with intact cerebral autoregulation. This suggests that ICP may increase after application of a collar. In healthy volunteers, this seems to be of minor importance. On the basis of our findings the effect of a collar on ONSD and ICP in patients with mild and moderate TBI needs to be determined.


[The assessment of anxiety in pregnant women in respect of biological, medical and socio-environmental factors].
Anxiety in the perinatal period may lead to serious complications resulting in the pathological course of pregnancy, fear and disturbances of delivery, and postnatal emotional problems. The latter, if unsolved in this period, may contribute to the worsening of relations between partners, between a mother and a child, and within a family. They also deteriorate the psychological well -being of a woman. This thesis was aimed at assessing the occurrence of anxiety in pregnant women throughout each pregnancy trimester, and attempting to explain its biological, medical and social -environmental determinants. The material for this prospective longitudinal study consisted of 181 pregnant residents of Szczecin. Each woman was examined three times during pregnancy, once in each trimester. The research instruments were: a questionnaire of the author's own design and a standardized instrument – Spielberger's State -Trait Anxiety Inventory (STAI). 1. Anxiety level in the course of pregnancy is lowest in the second trimester. 2. In those women who during pregnancy complained of discomfort or worsened mood, and in those who experienced problems with sleeping, higher intensity of anxiety is noted throughout the whole period of pregnancy. 3. Such factors as unintended pregnancy, lack of family support, and disapproval of pregnancy by a partner or the community are associated with suffering from aggravated anxiety in each trimester of pregnancy. 4. Women who have a negative judgement of their housing situation and 􀏐inancial standing are at risk of high levels of anxiety throughout the whole pregnancy. 5. Perinatal education is connected with signi􀏐icantly lesser occurrence of anxiety in each trimester of pregnancy. 6. Failure to have a preventive medical examination prior to pregnancy (woman, man) is related to experiencing signi􀏐icantly higher level of anxiety in the 􀏐irst and third trimesters of pregnancy.


Pectus excavatum in adult women: repair and the impact of prior or concurrent breast augmentation.
Women present with pectus excavatum five times less frequently than men. Adult women may have additional, associated cosmetic factors, including hypoplastic or asymmetric breasts, or prior augmentation. The authors evaluated the impact of prior or concurrent cosmetic breast surgery in an adult female cohort undergoing repair of pectus excavatum deformity. A retrospective review was performed of women (≥18 years old) who underwent pectus excavatum repair at a single institution from January of 2010 to September of 2013. Pectus excavatum repair was performed on 47 women with a median age of 35 years (range, 18 to 63 years). Mean pectus severity index was 6.2 (range, 3.1 to 16). All patients had physiologic symptoms as the primary purpose for seeking repair. Twenty patients (43 percent) presented with existing implants or the desire for implants at the time of repair. Fifteen patients (32 percent) had a history of implant placement including prior breast augmentation (n = 14) and/or pectus implant (n = 4). Concurrent augmentation (n = 5), breast implant exchange (n = 8), and/or removal of chest wall implants (n = 4) was performed during repair. Morbidity included one implant-related hematoma. Complications and hospital stay were not significantly different for patients undergoing primary repair alone versus those with prior or concurrent augmentation. Breast cosmesis was a concern in nearly half of adult women presenting for pectus excavatum repair. The authors' experience suggests neither prior nor concurrent breast augmentation increases the risk of complications in repair. The authors recommend that cosmetic breast surgery be performed concurrently with pectus excavatum repair. Therapeutic, III.


[Iatrogenic pseudoaneurysm of the brachial artery in a 4-month-old child - a case report].
We present a rare case of an iatrogenic pseudoaneurysm of the brachial artery in a 4-month-old child. As there are reports of such lesions in children that undergo invasive treatment such as sophisticated cardiovascular or radiological interventions and have preexisting risk factors, our patients had no history of vessel caniulation in hospital conditions and no systemic problems. A simple ambulatory blood draw caused a tear in the brachial artery wall, that later formed a pseudoaneurysm, a month before parents suddenly noticed mass on their child's arm. A microsurgical repair was performed. A significant increase in endovascular treatment options has been observed over last decades. Successful treatment was a result of fast radiological diagnostic and effective co-operation between paediatric and general vascular surgeons. Our case should raise awareness of paediatric practitioners about iatrogenic trauma of vessels in the differential diagnosis of tumours and vascular lesions.


[Social aspects of the adaptation and human ecology].
Influence of natural factors on humans or human migration into extreme conditions alters relationships between population and environment, therefore functional resources are mobilized. This alteration sometimes induces genotypic and phenotypic inadequacy to new, more strict requirements of natural and occupational factors. That is because evolution develops and genetically fixes in most people various mechanisms providing and supporting biologic balance. Requirements set in favorable environmental conditions for homeostatic systems including immune one are adequate to genotype of most individuals. When those mechanisms are not fixed and/or environmental conditions appear unfavorable the changes occur that could transform into disease.


Toxic Elements in Food: Occurrence, Binding, and Reduction Approaches.
Toxic elements such as mercury, arsenic, cadmium, and lead, sometimes called heavy metals, can diminish mental and central nervous system function; elicit damage to blood composition as well as the kidneys, lungs, and liver; and reduce energy levels. Food is considered one of the main routes of their entry into the human body. Numerous studies have been performed to examine the effects of common food processing procedures on the levels of toxic elements in food. While some studies have reported negative effects of processing, several have shown that processing practices may have a positive effect on the reduction of toxic elements in foodstuffs. A number of studies have also introduced protocols and suggested chemical agents that reduce the amount of toxic elements in the final food products. In this review, the reported methods employed for the reduction of toxic elements are discussed with particular emphasis on the chemical binding of both the organic and inorganic forms of each element in various foods. The molecular groups and the ligands by which the food products bind with the metals and the types of these reactions are also presented.


Association of temporal factors and suicides in the United States, 2000-2004.
The purpose of the study was to examine the association of temporal factors, in particular days of the week and seasons of the year and death from suicide in the United States. Data were pooled from the Multiple Cause of Death Files. Hierarchical logistic regression models were fitted to all deaths occurring in 2000 through 2004 by suicide. The incidence of suicide was significantly higher on Wednesdays, compared to Sunday. Specifically, individuals were 99% more likely to kill themselves on Wednesday than on Sunday. Suicides were more prevalent in the summer months, and they were less likely to occur in winter. The state suicide rate significantly elevated individual suicide risk. The results held even after controlling for the potentially confounding effects of socio-economic and demographic variables at both the individual and state levels. It was concluded that the observed association between seasonality and suicide cannot be discounted as a mere coincidence. Future research ought to focus on integrating individual level data and contextual variables when testing for seasonality effects.


Pharmacy Student Satisfaction and Perceptions Following a Virtual Sterile Compounding Experience.
Sterile compounding education is an essential curricular component across pharmacy schools. A virtual sterile intravenous compounding experience was implemented in place of traditional in-person delivery within the patient care lab course at one college of pharmacy. The objective of this manuscript is to describe student perceptions regarding student preparedness, satisfaction, and preferences after the virtual intravenous sterile compounding experience. Students reviewed a pre-recorded lecture and readings which covered sterile compounding fundamentals. Prior to the class session, students were provided with a kit that included simulated intravenous products. Each class session included approximately 33 students and three instructors via online video conferencing. The class session began with a large-group discussion to clarify questions following review of the videos and readings. Then, the class was divided into three breakout rooms with up to 11 students and one instructor. The instructor led the small group through stepwise sterile compounding procedures and provided feedback to the students. Students then completed an online multiple-choice quiz. A survey assessing student perceptions including preparedness, satisfaction, and preferences regarding the virtual experience was disseminated to students. A Mann-Whitney U analysis was performed to compare the ordinal data. A P-value of 0.05 was used to determine significance. Seventy-two (75%) second-year students and 32 (33.3%) third-year students completed the survey yielding an overall response rate of 54%. The majority of students (66%) reported satisfaction with the virtual compounding experience compared with in-person compounding experience. Fifty-seven percent of students felt the virtual experience prepared them for clinical rotation sterile compounding experiences. Overall, students indicated satisfaction with a virtual intravenous compounding experience.


Efficacy and Safety of the PD-1 Inhibitor Combined with Albumin-Bound Paclitaxel and Nedaplatin in Preoperative Neoadjuvant Therapy of Unresectable Stage III Lung Squamous Cell Carcinoma.
To investigate the efficacy and safety of preoperative neoadjuvant therapy (PD-1 inhibitor plus nab-PTX and nedaplatin) for resectable stage III lung squamous cell carcinoma (SCC) patients. Patients with locally advanced lung SCC (stage IIIA, IIIB) who received PD-1 inhibitor combined with nab-PTX and NED between February 2019 and June 2021 in Weihai Municipal Hospital were included and underwent surgical treatment 4 weeks after 2-4 cycles neoadjuvant therapy. The rate of resection R0, the effective rate, the complete pathological remission rate (pCR) and the rate of major pathological remission (MPR) were observed. A total of 14 initially unresectable male patients with lung SCC were included and received neoadjuvant treatment after evaluation. Nine out of 14 patients (64.3%) experienced treatment-related adverse events (TRAE), among which 8 (57.1%) experienced grade (G) I-II TRAEs including nausea, vomiting, fatigue, constipation, elevated ALT and AST, hyperthyroidism, hypothyroidism, rash, granulocytopenia, and thrombocytopenia, and 1 (7.1%) experienced grade III-V TRAEs (G), including granulocytopenia and atelectasis. Thirteen patients (92.86%) achieved RECIST-assessed partial remission (PR), while 1 patient (7.14%) achieved stable disease (SD) on imaging assessment after neoadjuvant treatment and continued to be progression-free for 26 months. Of the 11 patients who underwent resection, all were alive and recurrence/progression-free. MPR and pCR were observed in 2 (18.18%) and 9 (81.82%), respectively. IHC results exhibited that all NSCLC patients exhibited positive PD-L1 expression (9/14, TPS ≥50% or greater; 5/14, 1% < TPS < 50%). Two were negative for ALK, EGFR, and ros-1, and the rest were not examined for driver oncogene mutation. The neoadjuvant therapy of the PD-1 inhibitor combined with nab-PTX and NED demonstrated remarkable therapeutic efficacy and good safety on stage III lung SCC without increasing the risk of TRAE, mortality and surgery-related complications, or impede surgery feasibility.


[Carbapenem-resistant bacteria transferred to the Netherlands: patients colonised during hospitalisation in Romania].
Three patients were recently transferred to two Intensive Care Units (ICUs) in the Netherlands from two different ICUs in Bucharest, Romania. The patients appeared to be colonised with several, partly identical, carbapenemase-producing bacteria (CPB) after a short hospitalisation in Romania. In this article, we show that it is important to be aware of the possibility that patients are colonised with CPB after a short hospitalisation abroad. This has consequences for infection prevention measurements, but probably also for the discontinuation of selective bowel decontamination in the ICU.


Pearls & Oy-sters: Gait Instability, Jaw Dystonia, and Horizontal Diplopia in a Woman With Anti-Ri Antibodies and Breast Cancer.
A 40-year-old woman was admitted for six months of progressive gait disturbance, lower limb-predominant weakness, stiffness, falls, jaw dystonia, horizontal diplopia, and weight loss. Neurological examination revealed horizontal gaze paresis, limited jaw opening with palpable masseter hypertrophy, and spastic paraparesis with sustained clonus and upgoing plantar responses. MRI revealed T2-hyperintense signal abnormalities in dorsal pons, medulla and upper cervical cord central grey matter extending to C3, without gadolinium enhancement. Cerebrospinal fluid (CSF) showed mildly elevated protein and immunoglobulin (IgG) index with CSF-specific oligoclonal bands. Neural autoantibody testing was positive for anti-Ri in CSF and serum by mouse brain indirect immunofluorescence and immunoblot. Testing for aquaporin 4 (AQP4)-IgG and myelin oligodendrocyte glycoprotein (MOG)-IgG by cell-based assay were negative. The patient received methylprednisolone 1 gram for 5 days and intravenous immunoglobulin 2 grams/kilogram over 2 days with prednisone taper, and botulinum toxin injections for jaw dystonia. PET-CT revealed an enlarged left axillary lymph node with high FDG uptake. Left axillary lymph node biopsy confirmed high-grade, locally invasive breast adenocarcinoma. Neurologic stabilization was documented at two-week follow-up after hospital discharge before modified radical mastectomy. Our case demonstrates a clinical triad highly suggestive of anti-Ri-associated paraneoplastic neurologic syndrome (Ri-PNS): gait instability, jaw dystonia, and horizontal gaze paresis. The more slowly progressive course and poor response to immunotherapy help distinguish it from AQP4-IgG seropositive neuromyelitis optica spectrum disorder (NMOSD) and MOG-IgG associated disease that share similar radiographic features. Early diagnosis, prompt immunotherapy and cancer treatment are paramount for disease stabilization.


Distribution and significance of interstitial fibrosis and stroma-infiltrating B cells in tongue squamous cell carcinoma.
Inflammation and desmoplasia are frequently identified in the tumor microenvironment, and have been demonstrated to be effective modulators of malignant biological events. However, the mechanisms by which the inflammatory microenvironment and interstitial fibrosis interact with one another remain to be elucidated. The present study aimed to investigate the degree of inflammation and interstitial fibrosis in tongue squamous cell carcinoma (TSCC), and how this acts to affect the outcome of TSCC. Tissue samples from 93 cases of TSCC and paired tumor-adjacent non-neoplastic tongue epithelium, as well as 14 cases of epithelial dysplasia, were used. Interstitial collagen fibers were assessed using Masson's trichrome stain. Immunohistochemical identification of cancer-associated fibroblasts (CAFs) and stroma-infiltrating B cells was performed via detection of α-smooth muscle actin (SMA), vimentin, desmin and cluster of differentiation 19 (CD19). The clinicopathological significance and overall survival of the TSCC patients were statistically analyzed. Regularly distributed CAFs and CD19+ B cells were identified in the TSCC stroma, whereas no CAFs or CD19+ B cells were observed in epithelial dysplasia samples or paired tumor-adjacent non-neoplastic tongue epithelium samples. The distribution of interstitial collagen fibers and CAFs was closely associated with the tumor stage of the primary cancer, and high levels of CD19+ B cells together with low CAF infiltration were identified to be associated with favorable prognosis in TSCC. In conclusion, the inflammatory and interstitial fibrotic microenvironments coexist in TSCC, and each has specific effects on disease outcome, individually or perhaps collectively. However, it remains to be determined exactly how the microenvironments affect one another in TSCC.


Glucose-induced electrical activity in the pancreatic beta-cell: effect of veratridine.
Veratridine was used in the presence of glucose to assess the role of the Na pump in the regulation of glucose-induced burst activity. In the presence of 8.4 mM glucose, veratridine elicited a silent hyperpolarization, followed by burst activity. The magnitude of depolarization to plateau potential and the duration of the silent phase were increased. The addition of tetrodotoxin (TTX) restored the pattern of electrical activity to that observed in the absence of veratridine. Similar results were observed when veratridine was used in the presence of 16.7 mM glucose and tetraethylammonium (blocks voltage-dependent potassium permeability). TTX or ouabain blocked the effects of veratridine, and produced depolarization and continuous spike activity. Quinine (blocks Ca-dependent potassium permeability) elicited continuous spike activity in the presence of 16.7 mM glucose. The addition of veratridine induced only a transient return to burst activity, followed by a return to continuous spike activity. These results suggest that an electrogenic Na pump is an important factor in maintaining the transmembrane potential at an optimum level for operation of a voltage- and Ca-sensitive potassium permeability: changes in potassium permeability operating on a background of electrogenic current may be responsible for the voltage transitions associated with burst activity.


Brain iron and metabolic abnormalities in C19orf12 mutation carriers: A 7.0 tesla MRI study in mitochondrial membrane protein-associated neurodegeneration.
Mitochondrial membrane protein-associated neurodegeneration is an autosomal-recessive disorder caused by C19orf12 mutations and characterized by iron deposits in the basal ganglia. The aim of this study was to quantify iron concentrations in deep gray matter structures using quantitative susceptibility mapping MRI and to characterize metabolic abnormalities in the pyramidal pathway using 1 H MR spectroscopy in clinically manifesting membrane protein-associated neurodegeneration patients and asymptomatic C19orf12 gene mutation heterozygous carriers. We present data of 4 clinically affected membrane protein-associated neurodegeneration patients (mean age: 21.0 ± 2.9 years) and 9 heterozygous gene mutation carriers (mean age: 50.4 ± 9.8 years), compared to age-matched healthy controls. MRI assessments were performed on a 7.0 Tesla whole-body system, consisting of whole-brain gradient-echo scans and short echo time, single-volume MR spectroscopy in the white matter of the precentral/postcentral gyrus. Quantitative susceptibility mapping, a surrogate marker for iron concentration, was performed using a state-of-the-art multiscale dipole inversion approach with focus on the globus pallidus, thalamus, putamen, caudate nucleus, and SN. In membrane protein-associated neurodegeneration patients, magnetic susceptibilities were 2 to 3 times higher in the globus pallidus (P = 0.02) and SN (P = 0.02) compared to controls. In addition, significantly higher magnetic susceptibility was observed in the caudate nucleus (P = 0.02). Non-manifesting heterozygous mutation carriers exhibited significantly increased magnetic susceptibility (relative to controls) in the putamen (P = 0.003) and caudate nucleus (P = 0.001), which may be an endophenotypic marker of genetic heterozygosity. MR spectroscopy revealed significantly increased levels of glutamate, taurine, and the combined concentration of glutamate and glutamine in membrane protein-associated neurodegeneration, which may be a correlate of corticospinal pathway dysfunction frequently observed in membrane protein-associated neurodegeneration patients. © 2019 International Parkinson and Movement Disorder Society.


Sustainable Aquafeed Formulations Containing Insect Larval Meal and Grape Marc for the New Zealand Farmed Abalone.
The aquaculture industry has been criticised for the excessive use of fish meal (FM) in feeds due to the utilisation of wild fish in the formulation and the exacerbation of overfishing marine resources. Land-based abalone aquaculture mainly uses commercial feeds (CFs) to promote faster growth, which include FM as a primary protein component. Alternative ingredients, such as insect meal (IM) and grape marc (GM) are potential candidates for FM replacement due to their suitable nutritional profile and sustainable production. This paper reports on a novel nutritional approach for the New Zealand farmed abalone, which replaces FM with IM by 10% and includes a waste by-product (GM) by 30% as a potential prebiotic source. The study was performed in two stages: (a) physico-chemical determination of diets delivered in an alginate matrix (experimental diets) and their stability in seawater compared to CF and (b) evaluation of growth and feed intake for the New Zealand black-foot abalone. There were significant differences between experimental diets and CF in terms of sinking rate, particle weight, and microscopic observations. Water stability of the experimental diets was increased by 50% in 24 and 48 hr compared to CF, producing less solid waste, and potentially reducing cleaning efforts in the farm. The inclusion of IM and GM did not compromise overall animal growth or their feed conversion ratio, however, further evaluation need to be explored in the future research. The findings revealed that the developed encapsulated feeds are a more stable food delivery method for Haliotis iris compared to the CF. Furthermore, both IM and GM can be included in feed formulations as a more sustainable strategy without compromising weight and shell gains in the abalone farming.


Proteasome nuclear activity affects chromosome stability by controlling the turnover of Mms22, a protein important for DNA repair.
To expand the known spectrum of genes that maintain genome stability, we screened a recently released collection of temperature sensitive (Ts) yeast mutants for a chromosome instability (CIN) phenotype. Proteasome subunit genes represented a major functional group, and subsequent analysis demonstrated an evolutionarily conserved role in CIN. Analysis of individual proteasome core and lid subunit mutations showed that the CIN phenotype at semi-permissive temperature is associated with failure of subunit localization to the nucleus. The resultant proteasome dysfunction affects chromosome stability by impairing the kinetics of double strand break (DSB) repair. We show that the DNA repair protein Mms22 is required for DSB repair, and recruited to chromatin in a ubiquitin-dependent manner as a result of DNA damage. Moreover, subsequent proteasome-mediated degradation of Mms22 is necessary and sufficient for cell cycle progression through the G(2)/M arrest induced by DNA damage. Our results demonstrate for the first time that a double strand break repair protein is a proteasome target, and thus link nuclear proteasomal activity and DSB repair.


MicroRNA-93 targets WASF3 and functions as a metastasis suppressor in breast cancer.
Cancer cells with cancer stem cell (CSC) properties initiate both primary tumor formation and metastases at distant sites. Acquisition of CSC properties is highly associated with epigenetic alterations, including those mediated by microRNAs (miRNAs). We have previously established the breast cancer patient-derived tumor xenograft (PDX) mouse model in which CSC marker CD44+ cancer cells formed spontaneous microscopic metastases in the liver. In this PDX mouse, we found that the expression levels of 3 miRNAs (miR-25, miR-93, and miR-106b) in the miR-106b-25 cluster were much lower in the CD44+ human cancer cells metastasized to the liver than those at the primary site. Constitutive overexpression of miR-93 suppressed invasive ability and 3D-organoid formation capacity of breast cancer cells in vitro and significantly suppressed their metastatic ability to the liver in vivo. Wiskott-Aldrich syndrome protein family member 3 (WASF3), a regulator of both cytoskeleton remodeling and CSC properties, was identified as a functional target of miR-93: overexpression of miR-93 reduced the protein level of WASF3 in breast cancer cells and WASF3 rescued the miR-93-mediated suppression of breast cancer cell invasion. These findings suggest that miR-93 functions as a metastasis suppressor by suppressing both invasion ability and CSC properties in breast cancers.


Construction and characterization of a common bean bacterial artificial chromosome library.
We have constructed a common bean (Phaseolus vulgaris L.) bacterial artificial chromosome (BAC) library consisting of 33,792 clones and an estimated 3- to 5-fold coverage of the common bean genome. Leaf nuclei were used as the source for high-molecular-weight DNA, and an endonuclease/methylase competition assay was employed to partially cleave the DNA. The library was screened with a number of nuclear and mitochondrial probes. Each nuclear probe identified at least two BACs with an average insert size of ca. 100 kb. Only 26 clones were identified after hybridizing with mitochondrial probes, indicating contamination with organellar sequences is low. Numerous clones could be identified after screening the library with two repetitive probes flanking the nuclear fertility restorer Fr. Intriguingly, 12 clones appeared to hybridize to both markers, and restriction analysis of these clones revealed that they can be assembled into maximally four contigs, suggesting that these repetitive probes may be useful for the physical mapping of the Fr locus.


Neuropsychiatric assessment as a secondary outcome measure in a multiple sclerosis intravenous immunoglobulin L(IMg) trial.
Multiple sclerosis (MS) patients often suffer from abnormalities of mood including euphoria, depression, anxiety, pathological laughing and crying (PLC), and psychoses. We assessed neuropsychological functions (NF) as secondary outcome measures in relapsing-remitting multiple sclerosis (RRMS) patients treated by intravenous immunoglobulin (IVIg). Forty RR-MS patients (mean age 34.5±2.4; M : F=8: 32) were randomized to receive either IVIg or placebo in a double-blind trial for 2 years. NF evaluation at baseline, 1 and 2 years included the Goldberg Anxiety and Depression scales, the Brief Psychiatric Rating Scale, the Hamilton Anxiety Scale, the Beck Depression Inventory, and the Mini-Mental State Examination. Baseline anxiety, depression and general psychopathology scores were similar for the IVIg and placebo groups. At 1 and 2 years, anxiety, depression and general psychopathology had decreased similarly in both groups compared with baseline. No significant cognitive changes were observed in either group. In the IVIg group PLC emerged in one patient and one patient developed clinically overt depression necessitating antidepressant treatment. In the placebo group, two patients developed a hypomanic episode, and PLC emerged in two patients. IVIg treatment is safe, according to psychiatric outcome measures, and these measures should be routinely used in RR-MS drug trials.


Postnatal development of oxidative enzymes in various rat nephron segments: effect of weaning on different diets.
Activities of fumarase and 3-hydroxyacylCoA dehydrogenase (3-OHDH) were determined in homogenates of rat kidneys between day 21 of gestation and postnatal day 10 and in single isolated nephron segments at postnatal days 16, 21, and 30, and in adult segments. For 3-OHDH activity, main developmental changes were found in proximal convoluted (PCT) and straight tubules (PST) and were characterized by an overshoot of adult level from postnatal days 21 to 30 (59.7 +/- 3.0 and 37.5 +/- 3.4 at day 21 vs. adult values 27.1 +/- 1.5 and 22.7 +/- 1.5 mol.kg dry wt-1.h-1). When rats were precociously weaned on day 16 and fed a diet containing lipid to equal 13% of total caloric intake, a significant decrease in 3-OHDH activity was observed in some parts of the nephron. These changes could be prevented by maintaining early weaned animals on high-fat diet providing 70% of total calories as lipid. Results suggest that changes in fat content of diet during kidney maturation can in part regulate 3-OHDH activity in some nephron segments. Fumarase activity increased 2.6-fold in the medullary thick ascending limb between days 16 and 30; pattern of development was similar to the one reported for Na(+)-K(+)-ATPase activity in this segment. High levels of both enzymes were reached noticeably earlier during development in PCT and PST than in medullary thick ascending limb, which emphasizes metabolic heterogeneity of developing rat kidney nephron.


Sub-microscopic malaria cases and mixed malaria infection in a remote area of high malaria endemicity in Rattanakiri province, Cambodia: implication for malaria elimination.
Malaria microscopy and rapid diagnostic tests are insensitive for very low-density parasitaemia. This insensitivity may lead to missed asymptomatic sub-microscopic parasitaemia, a potential reservoir for infection. Similarly, mixed infections and interactions between Plasmodium species may be missed. The objectives were first to develop a rapid and sensitive PCR-based diagnostic method to detect low parasitaemia and mixed infections, and then to investigate the epidemiological importance of sub-microscopic and mixed infections in Rattanakiri Province, Cambodia. A new malaria diagnostic method, using restriction fragment length polymorphism analysis of the cytochrome b genes of the four human Plasmodium species and denaturing high performance liquid chromatography, has been developed. The results of this RFLP-dHPLC method have been compared to 1) traditional nested PCR amplification of the 18S rRNA gene, 2) sequencing of the amplified fragments of the cytochrome b gene and 3) microscopy. Blood spots on filter paper and Giemsa-stained blood thick smears collected in 2001 from 1,356 inhabitants of eight villages of Rattanakiri Province have been analysed by the RFLP-dHPLC method and microscopy to assess the prevalence of sub-microscopic and mixed infections. The sensitivity and specificity of the new RFLP-dHPLC was similar to that of the other molecular methods. The RFLP-dHPLC method was more sensitive and specific than microscopy, particularly for detecting low-level parasitaemia and mixed infections. In Rattanakiri Province, the prevalences of Plasmodium falciparum and Plasmodium vivax were approximately two-fold and three-fold higher, respectively, by RFLP-dHPLC (59% and 15%, respectively) than by microscopy (28% and 5%, respectively). In addition, Plasmodium ovale and Plasmodium malariae were never detected by microscopy, while they were detected by RFLP-dHPLC, in 11.2% and 1.3% of the blood samples, respectively. Moreover, the proportion of mixed infections detected by RFLP-dHPLC was higher (23%) than with microscopy (8%). The rapid and sensitive molecular diagnosis method developed here could be considered for mass screening and ACT treatment of inhabitants of low-endemicity areas of Southeast Asia.


1 alpha,25-(OH)2-vitamin D3 stimulates the adenylyl cyclase pathway in muscle cells by a GTP-dependent mechanism which presumably involves phosphorylation of G alpha i.
To further understand the mechanism underlying 1,25(OH)2D3 activation of the cAMP pathway, the effect of the hormone on adenylyl cyclase (AC), GTPase and protein kinase A (PKA) activities as well as on the phosphorylation of G alpha i was studied in membranes from chick skeletal muscle cells. The sterol stimulated AC activity in a dose (0.1-10 nM) and time (1-5 min.) dependent fashion, provided GTP (10 microM) was present in the assay. High affinity GTPase activity was unaffected by the hormone. In the absence of GTP or in the presence of Mn2+ (20 mM), 1,25(OH)2D3 effects on AC were abolished. PKA activity was increased (+120%) in cells pretreated (1 nM, 5 min.) with the sterol. Moreover, immunoprecipitation of G alpha i from [32P]-labeled myoblast membranes showed that 5 min. exposure to 1 nM 1,25(OH)2D3 increased (1.5-2 fold) the phosphorylation of its alpha subunit. The present data suggest that in muscle cells, 1,25(OH)2D3 activates AC by a non direct, GTP-dependent action which could imply amelioration of Gi function by sterol-induced alpha i phosphorylation.


Pilot Trial AMC-063: Safety and Efficacy of Bortezomib in AIDS-associated Kaposi Sarcoma.
AIDS-related Kaposi sarcoma is often incompletely controlled, requiring serial therapies. Kaposi sarcoma herpesvirus (KSHV) induces transformation of endothelial cells, where it resides in a predominately latent state. We hypothesized proteasome inhibition would have direct antitumor activity, induce lytic activation of KSHV, and inhibit HIV infectivity, improving control of both Kaposi sarcoma and HIV. The primary objective was determining the MTD of bortezomib in AIDS-Kaposi sarcoma. Secondary objectives included estimating the impact of bortezomib on Kaposi sarcoma response, KSHV plasma DNA copy number (PDCN), and HIV viral loads (VL). A 3+3 dose escalation design was employed evaluating four dose levels of bortezomib (0.75, 1, 1.2, or 1.6 mg/m2) administered intravenously on days 1, 8, and 15 of 28-day cycles in patients with relapsed/refractory (r/r) AIDS-Kaposi sarcoma taking antiretroviral therapy. Seventeen patients enrolled. No dose-limiting toxicities occurred and the MTD was not reached. The most common adverse events included diarrhea, fatigue and nausea. Among 15 evaluable patients, partial response (PR) occurred in nine (60%), with a PR rate of 83% in the 1.6 mg/m2 cohort; the remainder had stable disease (SD). Median time to response was 2.1 months. Median change in KSHV PDCN was significantly different between those with PR versus SD. During cycle 1, seven of 11 evaluable patients had decreases in HIV VL. Bortezomib is well-tolerated and active in AIDS-Kaposi sarcoma. The 60% PR rate is notable given the dose-finding nature of the study in a r/r population. Changes in KSHV PDCN and HIV VL trended as hypothesized.


Generation of Transgenic Energy Cane Plants with Integration of Minimal Transgene Expression Cassette.
Lignocellulosic biomass has the potential to serve as feedstock and direct replacement for petrochemicals in the fuel, chemical, pharmaceutical and material industries. Energy cane has been identified by the U.S. Department of Energy (DOE) as prime lignocellulosic feedstock as it produces record biomass yields and is able to grow on low-value land with reduced inputs. Molecular improvement of energy cane is an essential step toward the development of a high-value crop and may contribute to improved biomass conversion to value added products. Such improvements require a development of an efficient regeneration and transformation system for the vegetatively propagated energy cane varieties. In this report, an efficient biolistic gene delivery protocol for energy canes (genotype L 79-1002 and Ho 00-961) has been established with immature leaf rolls as explants. Embryonic calli, developed approximately 6 weeks after culture initiation and was used as target for biolistic transfer of a minimum expression cassette of P-ubi::nptII::35S polyA derived from plasmid pJFNPTII. Putative transgenic clones of callus were obtained after selection on callus induction medium supplemented with 30 mg l-1 geneticin. Regeneration was carried out on NB medium, which is modified from MS supplemented with 1.86 mg l-1 naphthaleneacetic acid (NAA) and 0.1mg l-1, 6-benzylaminopurine (BAP) and 20mg l-1 paromomycin. Shoots growing on selection media were transferred to hormone free medium with 20 mg l-1 paromomycin. Putative transgenic lines were first analyzed by PCR. Transgene integration was confirmed by Southern blot analysis. ELISA (Enzyme-Linked Immunosorbent Assay) and Immunochromathography assays confirmed transgene expression.


LncRNA DANCR-miR-758-3p-PAX6 Molecular Network Regulates Apoptosis and Autophagy of Breast Cancer Cells.
This study set out to probe into the effects of long non-coding RNA (LncRNA) differentiation antagonizing non-protein coding RNA (DANCR) on apoptosis and autophagy of breast cancer (BC) cells. The expression levels of DANCR, miR-758-3p and paired box 6 (PAX6) in BC tissues and cell lines were detected. The transcription and protein levels of PAX6, apoptosis-related factors (caspase-3, caspase-9, Bax/Bcl-2), and autophagy-related factors (LC3B, Atg5, Beclin-1) in BC cells were detected. The cell proliferation, apoptosis, autophagy and the regulatory relationship between genes and target genes were analyzed. DANCR and PAX6 were up-regulated in BC tissues and cell lines, while miR-758-3p was opposite. Down-regulating DANCR inhibited the malignant proliferation of BC cells and also promoted apoptosis and autophagy, which showed that caspase-3, caspase-9, Bax/Bcl-2, LC3B, Atg5 transcription and protein levels increased, while Beclin-1 transcription and protein levels decreased. DANCR regulated miR-758-3p in a targeted manner, and its over-expression could weaken the anti-cancer effect of miR-758-3p on BC cells. In addition, miR-758-3p also directly targeted PAX6, and knocking down its expression could weaken the inhibitory effect of down-regulating PAK6 on BC cell apoptosis and autophagy. We also found that DANCR acted as a competitive endogenous RNA sponge miR-758-3p, thus regulating the PAX6 expression. DANCR-miR-758-3p-PAX6 molecular network plays a key regulatory role in BC cell apoptosis and autophagy, which may provide reference for treating patients.


Temporal autocorrelation functions for movement rates from global positioning system radiotelemetry data.
Autocorrelation has been viewed as a problem in telemetry studies because sequential observations are not independent in time or space, therefore violating assumptions for statistical inference. Yet nearly all ecological and behavioural data are autocorrelated in both space and time. We argue that there is much to learn about the structure of ecological and behavioural data from patterns of autocorrelation. Such patterns include periodicity in movement and patchiness in spatial data, which can be characterized by an autocorrelogram, semivariogram or spectrum. We illustrate the utility of temporal autocorrelation functions (ACFs) for analysing step-length data from GPS telemetry of wolves (Canis lupus), cougars (Puma concolor), grizzly bears (Ursus arctos) and elk (Cervus elaphus) in western Alberta. ACFs often differ by season, reflecting differences in foraging behaviour. In wilderness landscapes, step-length ACFs for predators decay slowly to apparently random patterns, but sometimes display strong daily rhythms in areas of human disturbance. In contrast, step lengths of elk are consistently periodic, reflecting crepuscular activity.


Causes of Vitamin K Deficiency in Patients on Haemodialysis.
Background: A low vitamin K status is common in patients on haemodialysis, and this is considered one of the reasons for the accelerated atherosclerosis in these patients. The vitamin is essential in activation of the protein Matrix Gla Protein (MGP), and the inactive form, dp-ucMGP, is used to measure vitamin K status. The purpose of this study was to investigate possible underlying causes of low vitamin K status, which could potentially be low intake, washout during dialysis or inhibited absorption capacity. Moreover, the aim was to investigate whether the biomarker dp-ucMGP is affected in these patients. Method: Vitamin K intake was assessed by a Food Frequency Questionnaire (FFQ) and absorption capacity by means of D-xylose testing. dp-ucMGP was measured in plasma before and after dialysis, and phylloquinine (vitamin K1) and dp-ucMGP were measured in the dialysate. Changes in dp-ucMGP were measured after 14 days of protein supplementation. Results: All patients had plasma dp-ucMGP above 750 pmol/L, and a low intake of vitamin K. The absorption capacity was normal. The difference in dp-ucMGP before and after dialysis was -1022 pmol/L (p < 0.001). Vitamin K1 was not present in the dialysate but dp-ucMGP was at a high concentration. The change in dp-ucMGP before and after protein supplementation was -165 pmol/L (p = 0.06). Conclusion: All patients had vitamin K deficiency. The reason for the low vitamin K status is not due to removal of vitamin K during dialysis or decreased absorption but is plausibly due to a low intake of vitamin K in food. dp-ucMGP is washed out during dialysis, but not affected by protein intake to a clinically relevant degree.


Acquisition of LURE-binding activity at the pollen tube tip of Torenia fournieri.
Pollen tube guidance is controlled by multiple complex interactions with the female tissues. Here, we show that pollen tubes of Torenia fournieri are regulated by a stylar tissue in a length-dependent manner to receive and respond to attractant LURE peptides secreted from synergid cells. We developed an immunostaining method to visualize LURE peptides bound at the plasma membrane of the tip region of the pollen tube. Using this method, we found that LURE peptides bound specifically to pollen tubes growing through a cut style. The peptides also bound to pollen tubes growing through a shorter style, which were not competent to respond to these peptides. These observations suggested a possibility that acquisition of the LURE peptide reception ability and acquisition of full competency are separable processes. RNA-Seq suggested that the transcription profile of pollen tubes was affected by both the length of the style and the cultivation period, consistently with physiological changes in binding activity and LURE response ability. The database generated from de novo RNA-Seq of Torenia pollen tubes was shown to be useful to identify pollen tube proteins by mass spectrometry. Our studies provide insight and an effective platform for protein identification to understand pollen tube guidance.


Proper actin ring formation and septum constriction requires coordinated regulation of SIN and MOR pathways through the germinal centre kinase MST-1.
Nuclear DBF2p-related (NDR) kinases constitute a functionally conserved protein family of eukaryotic regulators that control cell division and polarity. In fungi, they function as effector kinases of the morphogenesis (MOR) and septation initiation (SIN) networks and are activated by pathway-specific germinal centre (GC) kinases. We characterized a third GC kinase, MST-1, that connects both kinase cascades. Genetic and biochemical interactions with SIN components and life cell imaging identify MST-1 as SIN-associated kinase that functions in parallel with the GC kinase SID-1 to activate the SIN-effector kinase DBF-2. SID-1 and MST-1 are both regulated by the upstream SIN kinase CDC-7, yet in an opposite manner. Aberrant cortical actomyosin rings are formed in Δmst-1, which resulted in mis-positioned septa and irregular spirals, indicating that MST-1-dependent regulation of the SIN is required for proper formation and constriction of the septal actomyosin ring. However, MST-1 also interacts with several components of the MOR network and modulates MOR activity at multiple levels. MST-1 functions as promiscuous enzyme and also activates the MOR effector kinase COT-1 through hydrophobic motif phosphorylation. In addition, MST-1 physically interacts with the MOR kinase POD-6, and dimerization of both proteins inactivates the GC kinase hetero-complex. These data specify an antagonistic relationship between the SIN and MOR during septum formation in the filamentous ascomycete model Neurospora crassa that is, at least in part, coordinated through the GC kinase MST-1. The similarity of the SIN and MOR pathways to the animal Hippo and Ndr pathways, respectively, suggests that intensive cross-communication between distinct NDR kinase modules may also be relevant for the homologous NDR kinases of higher eukaryotes.


A synthetic decursin analog with increased in vivo stability suppresses androgen receptor signaling in vitro and in vivo.
Targeting androgen receptor (AR) signaling with agents distinct from current antagonist drugs remains a rational approach to the prevention and treatment of prostate cancer (PCa). Our previous studies have shown that decursin and isomer decursinol angelate (DA), isolated from the Korean medicinal herb Angelica gigas Nakai, interrupt AR signaling and possess anti-PCa activities in vitro. In the LNCaP PCa cell model, these pyranoccoumarin compounds exhibit properties distinct from currently used antagonists (e.g., Casodex). However, both are rapidly de-esterified to decursinol, a partial AR agonist. We report here that a synthetic decursin analog, decursinol phenylthiocarbamate (DPTC), has greater in vivo stability than the parent compounds. DPTC-decursinol conversion was undetectable in mice. Furthermore, in LNCaP cells, DPTC decreased prostate specific antigen (PSA) expression, down-regulated AR abundance and mRNA and inhibited AR nuclear translocation. The effect of DPTC on AR and PSA mRNA and protein abundance was also observed in VCaP cells expressing wild type AR. DPTC inhibited growth of both PCa cell lines through G(1) cell cycle arrest and apoptosis, as did decursin and DA. Furthermore, i.p. administration of DPTC for 3 weeks suppressed the expression of AR target genes probasin and Nkx3.1 in mouse prostate glands. Overall, our data suggest that DPTC represents a prototype lead compound for development of in vivo stable and active novel decursin analogs for the prevention or therapy of PCa.


IFN-gamma-mediated survival enables human neutrophils to produce MCP-1/CCL2 in response to activation by TLR ligands.
TLRs are key elements of the pathogen recognition mechanism used by the host immune system. Neutrophils express almost all TLRs, and activation of TLRs, such as TLR2 and TLR4, has been shown to induce the production of proinflammatory cytokines and chemokines, potentially linking innate and adaptive immunity. In the present study, we investigated whether activation of TLRs induces neutrophil production of MCP-1/CCL2, a key mediator involved in the development of adaptive immunity. Activation of neutrophils with LPS, lipoteichoic acid, or N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-[R]-Cys-[S]-Ser-[S]-Lys did not induce significant MCP-1 production and release; however, the Th1 cytokine IFN-gamma dramatically up-regulated MCP-1 production in cells activated with each TLR ligand. The majority of MCP-1 was released between 24 and 48 h of culture, indicating that this is a late event. The effect of IFN-gamma appeared to be due to its antiapoptotic effect, but not priming effect, revealing a biological consequence of IFN-gamma-induced neutrophil survival. Although IFN-gamma failed to protect neutrophils from cell death at a higher dose of LPS, the p38 MAPK inhibitor SB203580 dramatically increased MCP-1 release and neutrophil survival at this LPS concentration. Thus, p38 MAPK plays a previously uncharacterized role in neutrophil function. Taken together, our results indicate that human neutrophils produce MCP-1 in a Th1 microenvironment and this neutrophil-derived MCP-1 potentially amplifies the development of Th1 adaptive responses.


Intracranial brain abscess preceded by orbital cellulitis and sinusitis.
A 17-year-old boy with pyrexia, headache, and frequent drop attacks reported an acute onset of periorbital pain and swelling 1 month previously. Coronal computed tomography (CT) identified an ethmoid sinusitis, which was treated with functional endoscopic sinus surgery and intravenous gentamicin, prostaphylline, and metronidazone. Because of persistent symptoms, the patient returned 1 month later. The CT identified accumulation of debris in both frontal sinuses and a multilobulated lesion over the right frontal lobe. Bicoronal craniotomy was performed, and a mass located in the right frontal lobe was excised; the mass comprised chronic inflammatory tissues without evidence of malignancy. A postoperative brain CT confirmed the absence of a residual mass, and no recurrence or neurologic deficits were noted during the 3-month follow-up period. Intracranial complications cannot be prevented entirely even with the judicious use of antibiotics. Early application of the appropriate imaging modality and institution of aggressive therapy in any patient, not just pediatric patients, to prevent potential long-term disabilities and death are essential.


Reliability of the Pediatric Specific Musculoskeletal Ultrasound Scoring Systems for the Elbow, Wrist, and Finger Joints.
Musculoskeletal ultrasound (MSUS) is increasingly being used in the evaluation of pediatric musculoskeletal diseases. In order to provide objective assessments of arthritis, reliable MSUS scoring systems are needed. Recently, joint-specific scoring systems for arthritis of the pediatric elbow, wrist, and finger joints were proposed by the Childhood Arthritis and Rheumatology Research Alliance (CARRA) MSUS workgroup. This study aimed to assess the reliability of these scoring systems when used by sonographers with different levels of expertise. Members of the CARRA MSUS workgroup attended training sessions for scoring the elbow, wrist, and finger. Subsequently, scoring exercises of B mode and power Doppler (PD) mode still images for each joint were performed. Interreader reliability was determined using 2-way single-score intraclass correlation coefficients (ICCs) for synovitis and Cohen [Formula: see text] for tenosynovitis. Seventeen pediatric rheumatologists with different levels of MSUS expertise (1-15 yrs) completed a 2-hour training session and calibration exercise for each joint. Excellent reliability (ICC > 0.75) was found after the first scoring exercise for all the finger and elbow views evaluated on B mode and PD mode, and for all of the wrist views on B mode. After a second training session and a scoring exercise, the wrist PD mode views reached excellent reliability as well. The preliminary CARRA MSUS scoring systems for assessing arthritis of the pediatric elbow, wrist, and finger joints demonstrate excellent reliability among pediatric MSUS sonographers with different levels of expertise. With further validation, this reliable joint-specific scoring system could serve as a clinical tool and scientific outcome measure.


Positive and negative regulation of the Drosophila immune response.
Insects mount a robust innate immune response against a wide array of microbial pathogens. The hallmark of the Drosophila humoral immune response is the rapid production of antimicrobial peptides in the fat body and their release into the circulation. Two recognition and signaling cascades regulate expression of these antimicrobial peptide genes. The Toll pathway is activated by fungal and many Gram-positive bacterial infections, whereas the immune deficiency (IMD) pathway responds to Gram-negative bacteria. Recent work has shown that the intensity and duration of the Drosophila immune response is tightly regulated. As in mammals, hyperactivated immune responses are detrimental, and the proper down-modulation of immunity is critical for protective immunity and health. In order to keep the immune response properly modulated, the Toll and IMD pathways are controlled at multiple levels by a series of negative regulators. In this review, we focus on recent advances identifying and characterizing the negative regulators of these pathways.


Comment on "A southern tyrant reptile".
Benson et al. (Brevia, 26 March 2010, p. 1613) reported on an Australian tyrannosauroid, represented by a pubis from the late Early Cretaceous of Victoria. However, our examination of this specimen reveals that the critical character used for this referral is not present. We contend that the bone likely belongs to a currently recognized group of Australian theropod or another group not currently known.


Microbiology of animal bite wound infections.
The microbiology of animal bite wound infections in humans is often polymicrobial, with a broad mixture of aerobic and anaerobic microorganisms. Bacteria recovered from infected bite wounds are most often reflective of the oral flora of the biting animal, which can also be influenced by the microbiome of their ingested prey and other foods. Bacteria may also originate from the victim's own skin or the physical environment at the time of injury. Our review has focused on bite wound infections in humans from dogs, cats, and a variety of other animals such as monkeys, bears, pigs, ferrets, horses, sheep, Tasmanian devils, snakes, Komodo dragons, monitor lizards, iguanas, alligators/crocodiles, rats, guinea pigs, hamsters, prairie dogs, swans, and sharks. The medical literature in this area has been made up mostly of small case series or case reports. Very few studies have been systematic and are often limited to dog or cat bite injuries. Limitations of studies include a lack of established or inconsistent criteria for an infected wound and a failure to utilize optimal techniques in pathogen isolation, especially for anaerobic organisms. There is also a lack of an understanding of the pathogenic significance of all cultured organisms. Gathering information and conducting research in a more systematic and methodical fashion through an organized research network, including zoos, veterinary practices, and rural clinics and hospitals, are needed to better define the microbiology of animal bite wound infections in humans.


Indian Ocean tsunami: relationships among posttraumatic stress, posttraumatic growth, resource loss, and coping at 3 and 15 months.
This study examines variables associated with posttraumatic stress symptoms (PTS) and posttraumatic growth among 2 independent samples of survivors following the Indian Ocean tsunami in Khao Lak, Thailand. Participants were exposed to unprecedented horror and loss of life and property. At 3 months participants (N = 248) were living in temporary shelters, and at 15 months a second sample (N = 255) was living in homes built after the tsunami. Prior traumatic experiences, life threat, loss of personal characteristic resources and condition resources, somatic problems, and social support accounted for close to half of the variance in PTS in each sample. At 3 months, emotion-focused coping and concerns about government favoritism also contributed to PTS. At 15 months, lack of prior disaster experience and loss of energy resources also contributed to PTS. Distress was higher among participants surveyed at 3 months than among those surveyed at 15 months. Posttraumatic growth was positively associated with social support and problem-focused coping in both samples. The findings support conservation of resources stress theory ( Hobfoll, 2012 ) and underscore how systemic issues affect mental health. The implications of the findings are discussed, as is the educational International Tsunami Museum designed by the first author to address systemic stressors.


Sugar Metabolism of Scardovia wiggsiae, a Novel Caries-Associated Bacterium.
Scardovia wiggsiae has been detected from caries in children and adolescents and has been suggested to be a caries-associated microorganism. To investigate the cariogenic potential of S. wiggsiae, we examined carbohydrate metabolism and acid productivity, the fluoride sensitivity of carbohydrate metabolism and the mechanism by which fluoride inhibits carbohydrate metabolism, and the acid sensitivity of carbohydrate metabolism in this bacterium. S. wiggsiae metabolized glucose and reduced the environmental pH to 3.5. It mainly produced acetic acid from glucose, together with small amounts of lactic and formic acid. The 50% inhibitory concentration of fluoride for acid production was 8.0 mM at pH 7.0 and 1.5 mM at pH 5.5, which were much higher than those of representative caries-associated bacteria, such as Streptococcus mutans. Metabolomic profiles showed the accumulation of 3-phosphoglycerate and a marked reduction in the pyruvate concentration in the presence of fluoride, suggesting that fluoride inhibits the latter half of glycolysis, including enolase activity. Enolase activity was inhibited by fluoride in S. wiggsiae, but it was more fluoride-tolerant than the enolase activity of S. mutans. Unlike in S. mutans, lactic acid did not inhibit acid production by S. wiggsiae at acidic pH. These results indicate that S. wiggsiae exhibits high acid production and tolerance to fluoride and lactic acid. S. wiggsiae possesses a unique metabolic pathway, the F6PPK shunt, which might allow it to avoid the lactate-formate pathway, including fluoride-sensitive enolase activity, and enable metabolic flow to the fluoride-tolerant acetate pathway. The fluoride tolerance of S. wiggsiae's enolase activity also increases the fluoride tolerance of its carbohydrate metabolism. The lactic acid tolerance of S. wiggsiae's acid production might result in S. wiggsiae having high acidogenic and aciduric potential and make it ecologically competitive in acidic environments, such as caries lesions, where lactic acid predominates.


Short-Time Treatment with Alkali and/or Hot Water To Remove Common Pathogenic and Spoilage Bacteria From Chicken Wing Skin.
Dipping in 10% trisodium phosphate (TSP) at 10°C for 15 s and/or hot water (95°C) for 5 s significantly (P < 0,05) reduced the numbers of live Salmonella typhimurium , Listeria monocytogenes , and Staphylococcus aureus inoculated on the surface of chicken wings. Mean reductions after treatment with TSP (after storage at 10°C or 4°C, respectively) were 93.45% and 62.42% for S. typhimurium , 80.33% and 54.45% for S. aureus , and 39.04% and 81.41 % for L. monocytogenes . Similarly treatment with hot water resulted in reductions of 83.5% and 47.44%, 90.19% and 91.49%, and 68.57% and 77.83%, respectively, for the three bacterial species. The combined effects of TSP and hot water were 94.76% and 99.67%, 84.41 % and 96.68%, and 79.49% and 94.88%. After treatment with TSP, there was always a better recovery of L. monocytogenes when the wings were stored at 10°C compared to 4°C. No similar storage temperature effect on recovery of L. monocytogenes was observed in the absence of TSP. Based on the smell and appearance of uninoculated, fresh chicken wings after treatment with 10% solutions of TSP or Na2CO3 (10°C) and hot water, the control group was always preferred after 1 day of storage, but not after 6 days of storage. Combination treatment with TSP and hot water showed that after 7 days of storage the number of spoilage organisms was 3 log units higher on the control samples than on the treated wings. The combined TSP and hot water treatments were more effective in reducing counts of S. typhimurium , S. aureus , and L. monocytogenes than the combined Na2CO3 and hot water treatment (95°C for 5 s). Changes in subcutaneous temperature as a result of treatment with TSP and hot water treatment were minimal.


Increased Beat-to-Beat Variability of T-Wave Heterogeneity Measured From Standard 12-Lead Electrocardiogram Is Associated With Sudden Cardiac Death: A Case-Control Study.
The prognostic significance of beat-to-beat variability of spatial heterogeneity of repolarization measured from standard 12-lead ECG is not well-understood. We measured the short-term variability of repolarization parameters, such as T-wave heterogeneity in leads V4-V6 (TWH) and QT interval (QT), from five consecutive beats of previously recorded standard 12-lead ECG in 200 victims of unexpected sudden cardiac death (SCD) confirmed to be due to complicated atherosclerotic coronary artery disease (CAD) in medico-legal autopsy and 200 age- and sex-matched controls with angiographically confirmed CAD. The short-term variability of repolarization heterogeneity was defined as the standard deviation (SD) of the measured repolarization parameters. All ECGs were in sinus rhythm, and no premature ventricular contractions were included in the measured segment. TWH-SD and QT-SD were significantly higher in SCD victims than in subjects with CAD (6.9 ± 5.6 μV vs. 3.8 ± 2.6 μV, p = 1.8E-11; 8.3 ± 13.1 ms vs. 3.8 ± 7.1 ms, p = 0.00003, respectively). After adjusting in the multivariate clinical model with factors, such as diabetes, RR interval, and beta blocker medication, TWH-SD and QT-SD retained their significant power in discriminating between the victims of SCD and the patients with CAD (p = 0.00003, p = 0.006, respectively). TWH-SD outperformed QT-SD in identifying the SCD victims among the study subjects (area under the curve in the receiver operating characteristics curve 0.730 vs. 0.679, respectively). Increased short-term variability of repolarization heterogeneity measured from standard 12-lead ECG is associated with SCD.


pH regulation in barnacle muscle fibers: dependence on extracellular sodium and bicarbonate.
Intracellular pH (pHi) regulation was studied in barnacle muscle fibers with pH-sensitive microelectrodes. The cells were acid loaded, and the subsequent recovery of pHi was monitored. The rate of recovery was reduced by one-third when external Na+ ([Na+]o) was replaced by Li+, but recovery was completely abolished when Na+ was replaced by choline or N-methyl-D-glucamine. In other experiments, varying amounts of Na+ were replaced by choline, and the acid extrusion rate, derived from the recovery rate of pHi, was calculated at a single value of pHi, 6.80. The dependence of the acid extrusion rate on [Na+]o could be described by Michaelis-Menten kinetics; at pHo (extracellular) = 8.0 and [HCO3-]o (extracellular) = 10 mM, the apparent Km and Vmax were 59 mM and 1.3 mmol x l(-1) x min-1. When [HCO3-]o was reduced to 2.5 mM at the same pHo, Km did not change significantly, but Vmax was substantially reduced. On the other hand, when pHo was reduced to 7.4 at constant [HCO3-]o, Vmax changed only slightly, but Km increased substantially. In similar experiments, we examined the dependence of the acid extrusion rate on [HCO3-]o. At pHo = 8.0 and [Na+]o = 440 mM, the apparent Km and Vmax were 4.1 mM and 2.1 mmol x 1-1 x min-1. When pHo was reduced to 7.4, Vmax was not altered, but Km substantially increased. The kinetic data are discussed in terms of the role of pHo, [Na+]o, and [HCO3-]o in the pHi-regulating system.


Gastrin induces parathyroid hormone-like hormone expression in gastric parietal cells.
Parietal cells play a fundamental role in stomach maintenance, not only by creating a pathogen-free environment through the production of gastric acid, but also by secreting growth factors important for homeostasis of the gastric epithelium. The gastrointestinal hormone gastrin is known to be a central regulator of both parietal cell function and gastric epithelial cell proliferation and differentiation. Our previous gene expression profiling studies of mouse stomach identified parathyroid hormone-like hormone (PTHLH) as a potential gastrin-regulated gastric growth factor. Although PTHLH is commonly overexpressed in gastric tumors, its normal expression, function, and regulation in the stomach are poorly understood. In this study we used pharmacologic and genetic mouse models as well as human gastric cancer cell lines to determine the cellular localization and regulation of this growth factor by the hormone gastrin. Analysis of PthlhLacZ/+ knock-in reporter mice localized Pthlh expression to parietal cells in the gastric corpus. Regulation by gastrin was demonstrated by increased Pthlh mRNA abundance after acute gastrin treatment in wild-type mice and reduced expression in gastrin-deficient mice. PTHLH transcripts were also observed in normal human stomach as well as in human gastric cancer cell lines. Gastrin treatment of AGS-E gastric cancer cells induced a rapid and robust increase in numerous PTHLH mRNA isoforms. This induction was largely due to increased transcriptional initiation, although analysis of mRNA half-life showed that gastrin treatment also extended the half-life of PTHLH mRNA, suggesting that gastrin regulates expression by both transcriptional and posttranscriptional mechanisms.NEW & NOTEWORTHY We show that the growth factor parathyroid hormone-like hormone (PTHLH) is expressed in acid-secreting parietal cells of the mouse stomach. We define the specific PTHLH mRNA isoforms expressed in human stomach and in human gastric cancer cell lines and show that gastrin induces PTHLH expression via transcription activation and mRNA stabilization. Our findings suggest that PTHLH is a gastrin-regulated growth factor that might contribute to gastric epithelial cell homeostasis.


Stimulation of lipolysis in rat heart myocytes by isoproterenol.
Calcium-tolerant myocytes were isolated from rat hearts. Isoproterenol produced a dose-dependent increase in glycerol output (lipolysis) that could be blocked by propranolol. The presence of glucose in the incubation medium enhanced the release of glycerol from myocytes but had no effect on the decline in triacylglycerol content. No incorporation of radioactivity from [U-14C]glucose into glycerol could be detected. In incubations with isoproterenol, there was a stoichiometric relationship between the glycerol output and the decrease in triacylglycerol levels. The addition of the phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine resulted in an increase in the basal glycerol output and an enhancement of the isoproterenol-stimulated lipolytic rate. Forskolin and 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate also produced a concentration-dependent stimulation of lipolysis in myocytes. Therefore, lipolysis in isolated myocytes must be regulated by adenosine 3',5'-cyclic monophosphate-dependent mechanisms. These results demonstrate that lipolysis can be observed in myocardial cells and that the lipolytic response to isoproterenol cannot be secondary to a physiological (inotropic) response since these myocyte preparations are quiescent.


Developmental plasticity of the structural network of the occipital cortex in congenital blindness.
The occipital cortex is the visual processing center in the mammalian brain. An unanswered scientific question pertains to the impact of congenital visual deprivation on the development of various profiles within the occipital network. To address this issue, we recruited 30 congenitally blind participants (8 children and 22 adults) as well as 31 sighted participants (10 children and 21 adults). Our investigation focused on identifying the gray matter regions and white matter connections within the occipital cortex, alongside behavioral measures, that demonstrated different developmental patterns between blind and sighted individuals. We discovered significant developmental changes in the gray matter regions and white matter connections of the occipital cortex among blind individuals from childhood to adulthood, in comparison with sighted individuals. Moreover, some of these structures exhibited cognitive functional reorganization. Specifically, in blind adults, the posterior occipital regions (left calcarine fissure and right middle occipital gyrus) showed reorganization of tactile perception, and the forceps major tracts were reorganized for braille reading. These plastic changes in blind individuals may be attributed to experience-dependent neuronal apoptosis, pruning, and myelination. These findings provide valuable insights into the longitudinal neuroanatomical and cognitive functional plasticity of the occipital network following long-term visual deprivation.


Subregional efforts to improve childhood cancer care in the Andean countries.
Cancer is one of the leading causes of death in children and adolescents younger than 19 years. An estimated 10 000 deaths are caused by this disease annually in this age group in Latin America and the Caribbean. In high-income countries, the survival of children and adolescents with neoplasms can reach 85%; however, in middle- and low-income countries, despite progress, survival rates are significantly lower (between 10% and 60%). Important inequities exist is survival from childhood cancer that need to be addressed through decisive actions from the health systems. This report describes the work of the ministries of health and the Secretariat of the Andean Health Organization (Organismo Andino de Salud - Convenio Hipólito Unánue (ORAS-CONHU)), to develop the Andean Cancer Prevention and Control Policy, with consideration given to childhood cancers. The policy was based on analysis of the cancer situation in the six Andean countries - Bolivia (Plurinational State of), Colombia, Chile, Ecuador, Peru and Venezuela (Bolivarian Republic of) - between 2015 and 2020, and it was approved in 2022. An in-depth study is currently being carried out on the situation of childhood cancer in the Andean countries.


Gaze direction differentially affects avoidance tendencies to happy and angry faces in socially anxious individuals.
Increasing evidence indicates that eye gaze direction affects the processing of emotional faces in anxious individuals. However, the effects of eye gaze direction on the behavioral responses elicited by emotional faces, such as avoidance behavior, remain largely unexplored. We administered an Approach-Avoidance Task (AAT) in high (HSA) and low socially anxious (LSA) individuals. All participants responded to photographs of angry, happy and neutral faces (presented with direct and averted gaze), by either pushing a joystick away from them (avoidance) or pulling it towards them (approach). Compared to LSA, HSA were faster in avoiding than approaching angry faces. Most crucially, this avoidance tendency was only present when the perceived anger was directed towards the subject (direct gaze) and not when the gaze of the face-stimulus was averted. In contrast, HSA individuals tended to avoid happy faces irrespectively of gaze direction. Neutral faces elicited no approach-avoidance tendencies. Thus avoidance of angry faces in social anxiety as measured by AA-tasks reflects avoidance of subject-directed anger and not of negative stimuli in general. In addition, although both anger and joy are considered to reflect approach-related emotions, gaze direction did not affect HSA's avoidance of happy faces, suggesting differential mechanisms affecting responses to happy and angry faces in social anxiety.


Clinical gait analysis.
Technologies and techniques that have evolved to aid in the assessment of human locomotion and some of the difficulties associated with the clinical application of quantitative gait analysis are examined. The parameters used for the clinical description of gait are outlined, and examples of application of clinical gait analysis are listed. Problems encountered in measuring and assessing gait patterns are discussed. Some commercially available motion measurement systems are described. Future research directions are suggested.


Targeted gene transfer to the brain via the delivery of brain-penetrating DNA nanoparticles with focused ultrasound.
Gene therapy holds promise for the treatment of many pathologies of the central nervous system (CNS), including brain tumors and neurodegenerative diseases. However, the delivery of systemically administered gene carriers to the CNS is hindered by both the blood-brain barrier (BBB) and the nanoporous and electrostatically charged brain extracelluar matrix (ECM), which acts as a steric and adhesive barrier. We have previously shown that these physiological barriers may be overcome by, respectively, opening the BBB with MR image-guided focused ultrasound (FUS) and microbubbles and using highly compact "brain penetrating" nanoparticles (BPN) coated with a dense polyethylene glycol corona that prevents adhesion to ECM components. Here, we tested whether this combined approach could be utilized to deliver systemically administered DNA-bearing BPN (DNA-BPN) across the BBB and mediate localized, robust, and sustained transgene expression in the rat brain. Systemically administered DNA-BPN delivered through the BBB with FUS led to dose-dependent transgene expression only in the FUS-treated region that was evident as early as 24h post administration and lasted for at least 28days. In the FUS-treated region ~42% of all cells, including neurons and astrocytes, were transfected, while less than 6% were transfected in the contralateral non-FUS treated hemisphere. Importantly, this was achieved without any sign of toxicity or astrocyte activation. We conclude that the image-guided delivery of DNA-BPN with FUS and microbubbles constitutes a safe and non-invasive strategy for targeted gene therapy to the brain.


A Pediatric Case of Bacteremia and Possible Cholecystitis Due to Moraxella osloensis.
We encountered a pediatric case of bacteremia and possible cholecystitis due to Moraxella osloensis that was treated successfully. We confirmed the diagnosis with the presence of a high serum titer of the antibody to the organism. Furthermore, 16S rRNA sequencing was performed to identify the bacteria.


Development of diving capacity in emperor penguins.
To compare the diving capacities of juvenile and adult emperor penguins Aptenodytes forsteri, and to determine the physiological variables underlying the diving ability of juveniles, we monitored diving activity in juvenile penguins fitted with satellite-linked time/depth recorders and examined developmental changes in body mass (Mb), hemoglobin concentration, myoglobin (Mb) content and muscle citrate synthase and lactate dehydrogenase activities. Diving depth, diving duration and time-at-depth histograms were obtained from two fledged juveniles during the first 2.5 months after their depature from the Cape Washingon colony in the Ross Sea, Antarctica. During this period, values of all three diving variables increased progressively. After 8-10 weeks at sea, 24-41 % of transmitted maximum diving depths were between 80 and 200 m. Although most dives lasted less than 2 min during the 2 month period, 8-25 % of transmitted dives in the last 2 weeks lasted 2-4 min. These values are lower than those previously recorded in adults during foraging trips. Of the physiological variables examined during chick and juvenile development, only Mb and Mb content did not approach adult values. In both near-fledge chicks and juveniles, Mb was 50-60 % of adult values and Mb content was 24-31 % of adult values. This suggests that the increase in diving capacity of juveniles at sea will be most dependent on changes in these factors.


Activation of JUN in fibroblasts promotes pro-fibrotic programme and modulates protective immunity.
The transcription factor JUN is highly expressed in pulmonary fibrosis. Its induction in mice drives lung fibrosis, which is abrogated by administration of anti-CD47. Here, we use high-dimensional mass cytometry to profile protein expression and secretome of cells from patients with pulmonary fibrosis. We show that JUN is activated in fibrotic fibroblasts that expressed increased CD47 and PD-L1. Using ATAC-seq and ChIP-seq, we found that activation of JUN rendered promoters and enhancers of CD47 and PD-L1 accessible. We further detect increased IL-6 that amplified JUN-mediated CD47 enhancer activity and protein expression. Using an in vivo mouse model of fibrosis, we found two distinct mechanisms by which blocking IL-6, CD47 and PD-L1 reversed fibrosis, by increasing phagocytosis of profibrotic fibroblasts and by eliminating suppressive effects on adaptive immunity. Our results identify specific immune mechanisms that promote fibrosis and suggest a therapeutic approach that could be used alongside conventional anti-fibrotics for pulmonary fibrosis.


Bird remains from an archaeological site on Henderson Island, South Pacific: Man-caused extinctions on an "uninhabited" island.
Long thought never to have been inhabited and to be in a pristine ecological state, Henderson Island (southeast Pacific) is now known to have been colonized and then abandoned by Polynesians. Bones from an archaeological site on the island associated with (14)C dates of approximately 800 and approximately 500 years B.P. include specimens of 12 species of birds, of which 3, a storm-petrel and two pigeons (Nesofregetta fuliginosa, Ducula cf. aurorae or D. pacifica, and Ducula cf. galeata), no longer occur on Henderson, and two others (Puffinus nativitatis and Sula sula) still visit but are not known to breed. The vanished species were presumably exterminated by Polynesians and the biota of Henderson Island can thus no longer be regarded as being in an unaltered state. The prehistoric abandonment of various small, unarable islands by Polynesians may have been due to the depletion of seabirds and pigeons, the only readily available food source. The species of pigeons identified from Henderson are known historically only from distant archipelagos and have never before been found sympatrically. Distributional patterns resulting from man-caused extinctions may give rise to erroneous interpretations of the relationships and evolutionary history of insular organisms. Certain endangered species, such as Ducula galeata, might effectively be preserved by reintroduction to abandoned islands that they occupied before human intervention.


Cytochrome P450 1B1 is required for 7,12-dimethylbenz(a)-anthracene (DMBA) induced spleen cell immunotoxicity.
7,12-Dimethylbenz(a)anthracene (DMBA) is a potent carcinogen that induces immunosuppression of both humoral and cell-mediated immunity in mice and other species. Previous studies have shown that CYP1B1 is required for bone marrow toxicity produced by DMBA in mice. Therefore, the purpose of these studies was to determine whether CYP1B1 was required for spleen cell immunotoxicity. Female C57BL/6N wild-type (WT) and CYP1B1 knockout (-/-) mice were treated with 0, 17, 50, or 150 mg/kg (cumulative dose) DMBA in corn oil by oral gavage once a day for five days. Several immunotoxicological assays were used to assess the effects of DMBA on systemic immunity. These included the in vitro T-dependent antibody response to sheep red blood cells (SRBC) measured using a direct plaque forming cell (PFC) assay, T- and B-cell mitogenesis induced by Con A and LPS, and nonspecific cell-mediated immunity was evaluated using an NK cytotoxicity assay. In addition, lymphocyte subpopulations were measured by flow cytometry using specific cell surface markers. Following five days of DMBA treatment, the body weights and spleen cell surface markers of the WT and CYP1B1 (-/-) mice showed no significant changes. A decrease in NK activity was found at the 50 mg/kg DMBA dose in WT mice, but not in the CYP1B1 (-/-) mice. Interestingly, at the 150 mg/kg dose of DMBA, CYP1B1 null mice had decreased NK activity, whereas WT mice did not. The SRBC PFC response demonstrated that the IgM antibody response was suppressed by DMBA in WT mice in a dose-dependent manner (significant at 50 and 150 mg/kg). However, there were no changes in the SRBC PFC responses in any DMBA test group in the CYP1B1 (-/-) mice. Similarly, while DMBA suppressed B- and T-cell mitogenesis at the 50 and 150 mg/kg dose levels in C57BL/6N WT mice, no effect was seen in CYP1B1 (-/-) mice. Thus, CYP1B1 appears to be critical for the immunosuppression of DMBA in mice, suggesting a role for bioreactive metabolites in the spleen cell immunotoxicity produced by DMBA.


Anti-inflammatory and anti-oxidant mechanisms of an MMP-8 inhibitor in lipoteichoic acid-stimulated rat primary astrocytes: involvement of NF-κB, Nrf2, and PPAR-γ signaling pathways.
Recent evidence suggests that reactive astrocytes play an important role in neuroinflammation and neurodegenerative diseases. Thus, controlling astrocyte reactivity has been suggested as a promising strategy for treating neurodegenerative diseases. In the present study, we investigated whether a matrix metalloproteinase (MMP)-8 inhibitor, M8I, could control neuroinflammation in lipoteichoic acid (LTA)-stimulated rat primary astrocytes. The effects of M8I on the expression of inducible nitric oxide synthase, cytokines, and MMPs were examined in LTA-stimulated rat primary astrocytes by ELISA, RT-PCR, and Western blot analysis. The effects of M8I on reactive oxygen species (ROS) generation and phase II antioxidant enzyme expression were examined by the DCF-DA assay, RT-PCR, and Western blot analysis. The detailed molecular mechanisms underlying the anti-inflammatory and antioxidant effects of M8I were analyzed by the electrophoretic mobility shift assay, the reporter gene assay, Western blot, and RT-PCR analysis. Treatment with LTA, a major cell wall component of Gram-positive bacteria, led to astrocyte activation and induced the expression of inflammatory molecules such as iNOS, COX-2, and pro-inflammatory cytokines. In addition, LTA induced the expression of MMPs such as MMP-1, MMP-3, MMP-8, MMP-9, and MMP-13 in rat primary astrocytes. Based on previous reports showing that MMP-8 plays a role as a proinflammatory mediator in microglia, we investigated whether MMP-8 is also involved in inflammatory reactions of reactive astrocytes. We found that treatment of astrocytes with M8I significantly inhibited LTA-induced expression of iNOS, TNF-α, IL-1β, IL-6, and TLR-2. In addition, M8I inhibited LTA-induced NF-κB, MAP kinase, and Akt activities, while it increased the anti-inflammatory PPAR-γ activities. Moreover, M8I showed antioxidant effects by suppressing ROS production in LTA- or H2O2-stimulated astrocytes. Interestingly, M8I increased the expression of phase II antioxidant enzymes such as hemeoxygenase-1, NQO1, catalase, and MnSOD by modulating the Nrf2/ARE signaling pathway. The data collectively suggest the therapeutic potential of an MMP-8 inhibitor in neuroinflammatory disorders that are associated with astrocyte reactivity.


Temperament in rhesus, long-tailed, and pigtailed macaques varies by species and sex.
Temperament differs among individuals both within and between species. Evidence suggests that differences in temperament of group members may parallel differences in social behavior among groups or between species. Here, we compared temperament between three closely related species of monkey-rhesus (Macaca mulatta), long-tailed (M. fascicularis), and pigtailed (M. nemestrina) macaques-using cage-front behavioral observations of individually housed monkeys at a National Primate Research Center. Frequencies of 12 behaviors in 899 subjects were analyzed using a principal components analysis to identify temperament components. The analysis identified four components, which we interpreted as Sociability toward humans, Cautiousness, Aggressiveness, and Fearfulness. Species and sexes differed in their average scores on these components, even after controlling for differences in age and early-life experiences. Our results suggest that rhesus macaques are especially aggressive and unsociable toward humans, long-tailed macaques are more cautious and fearful, and pigtailed macaques are more sociable toward humans and less aggressive than the other species. Pigtailed males were notably more sociable than any other group. The differences observed are consistent with reported variation in these species' social behaviors, as rhesus macaques generally engage in more social aggression and pigtailed macaques engage in more male-male affiliative behaviors. Differences in predation risks are among the socioecological factors that might make these species-typical behaviors adaptive. Our results suggest that adaptive species-level social differences may be encoded in individual-level temperaments, which are manifested even outside of a social context. Am. J. Primatol. 75:303-313, 2013. © 2012 Wiley Periodicals, Inc.


Acute change of titin at mid-sarcomere remains despite 8 wk of plyometric training.
The purpose of this study was to investigate skeletal muscle changes induced by an acute bout of plyometric exercise (PlyEx) both before and after PlyEx training, to understand if titin is affected differently after PlyEx training. Healthy untrained individuals (N = 11) completed the 1stPlyEx (10 × 10 squat-jumps, 1-min rest). Thereafter, six subjects completed 8 wk of PlyEx, while five controls abstained from any jumping activity. Seven days after the last training session, all subjects completed the 2ndPlyEx. Blood samples were collected before and 6 h and 1, 2, 3, and 4 days after each acute bout of PlyEx, and muscle biopsies 4 days before and 3 days after each acute bout of PlyEx. The 1stPlyEx induced an increase in circulating myoglobin concentration. Muscle sample analysis revealed Z-disk streaming, a stretch or a fragmentation of titin (immunogold), and increased calpain-3 autolysis. After training, 2ndPlyEx did not induce Z-disk streaming or calpain-3 activation. The previously observed post-1stPlyEx positional change of the titin COOH terminus was still present pre-2ndPlyEx, in all trained and all control subjects. Only two controls presented with Z-disk streaming after 2ndPlyEx, while calpain-3 activation was absent in all controls. Eccentric explosive exercise induced a stretch or fragmentation of titin, which presented as a positional change of the COOH terminus. Calpain-3 activation does not occur when titin is already stretched before explosive jumping. Enzymatic digestion results in titin fragmentation, but since an increase in calpain-3 autolysis was visible only after the 1stPlyEx acute bout, fragmentation cannot explain the prolonged positional change.


Release of cAMP from a renal epithelial cell line.
LLC-PK1 is an established porcine renal cell line with epithelial characteristics. Upon hormonal stimulation by vasopressin, LLC-PK1 cells release adenosine 3',5'-cyclic monophosphate (cAMP) into the medium. Release of cAMP is inhibited by the organic anion transport inhibitor probenecid and by cold phosphodiesterase inhibitors and iodoacetate but not by prostaglandins A1 or E1. The kinetics of release are first order, and cAMP analogues do not induce the release of cAMP. When grown on cellulose filters, monolayers of LLC-PK1 have morphological characteristics of transporting epithelia (apical microvilli and intercellular tight junctions) and maintain a transepithelial potential difference. Stimulation of such monolayers by vasopressin elicits probenecid-sensitive release of cAMP into the medium bathing the apical surface. Smaller quantities of cAMP are released from the basolateral surface, but release in this direction is not inhibited by probenecid. In contrast, release of cAMP from the nonepithelial cell line BHK is symmetrical and is symmetrically inhibited by probenecid. Probenecid-sensitive release of cAMP from LLC-PK1 is thus a function of the apical (brush-border) membrane.


A sterol-PI(4)P exchanger modulates the Tel1/ATM axis of the DNA damage response.
Upon DNA damage, cells activate the DNA damage response (DDR) to coordinate proliferation and DNA repair. Dietary, metabolic, and environmental inputs are emerging as modulators of how DNA surveillance and repair take place. Lipids hold potential to convey these cues, although little is known about how. We observed that lipid droplet (LD) number specifically increased in response to DNA breaks. Using Saccharomyces cerevisiae and cultured human cells, we show that the selective storage of sterols into these LD concomitantly stabilizes phosphatidylinositol-4-phosphate (PI(4)P) at the Golgi, where it binds the DDR kinase ATM. In turn, this titration attenuates the initial nuclear ATM-driven response to DNA breaks, thus allowing processive repair. Furthermore, manipulating this loop impacts the kinetics of DNA damage signaling and repair in a predictable manner. Thus, our findings have major implications for tackling genetic instability pathologies through dietary and pharmacological interventions.


Herniation of the mylohyoid muscle.
During ultrasound examinations in patients with dysphagia, deficiencies in the mylohyoid muscle with herniation of the sublingual gland were found, which may be mistaken as soft tissue tumors. Between the years 2001 and 2003, 205 half-heads used in dissection courses were examined to determine the location and contents of these gaps. In 25 of these cadaveric specimens, the hiatus (of variable size) appeared as small fissures between the divided fibers of the mylohyoid. In 18 cases (72%), the sublingual gland slipped through these deficiencies and occurred in the front part of the submandibular triangle. From the inferior surface of the muscle, the submental artery also coursed through the separated muscle fibers. The herniations of 7 specimens (28%) were found without any contents, neither with gland perforation nor with a submental artery. The following study points out the spatial relation of the salivary glands to the floor of the mouth and the clinical significance, and some factors referred to the development of herniations are discussed.


A primary burn wound does not slow the contraction rate of an adjacent excisional wound.
The response to major burn injury includes systemic release of mediators that may have an effect on wound healing. The authors evaluated the effect of a burn injury on the contraction of an excisional wound adjacent to the burn, and the effect of plasma derived from burn-injured animals on the contraction of the fibroblast-populated collagen matrix (FPCM). Nine rats (90-100 days old) under anesthesia received a standardized 40% total body surface area burn to the dorsum, and eight rats (controls) were sham burned. Immediately thereafter all animals had a square (2.25 cm2) of unburned dermis excised from the dorsum, superior to the burn wound. The excisional wound area was measured at 2 to 3-day intervals postoperatively. Plasma was collected from some animals on postburn day 15; the contraction-stimulating ability of burn vs. control plasma was measured in the FPCM. All animals remained free of sepsis. The excisional wound area in all animals decreased to 50% and then 25% of the initial area after approximately 4 and 8 days respectively. The rate of wound contraction (i.e., wound area reduction) did not differ between burn and control animals. Contraction stimulated by 5% plasma in the FPCM (expressed as a percentage of the original matrix area) was 70.2+/-6.4 (standard deviation) mm2 vs. 62.4 +/-3.9 mm2 for burn vs. control rats respectively (p>0.05). Burn injury in this model did not alter the contraction of an excisional wound at an unburned site. There was no significant difference in the contraction-stimulating ability (FPCM model) of plasma from the burned rats compared with plasma from unburned control rats. Burn injury appears to have an inconsequential effect on the contraction of an adjacent wound.


Family, school, and community factors and relationships to racial-ethnic attitudes and academic achievement.
This study examined family, school, and community factors and the relationships to racial-ethnic attitudes and academic achievement among 98 African American fourth-grade children. It has been posited that young people who feel better about their racial-ethnic background have better behavioral and academic outcomes, yet there is a need for more empirical tests of this premise. Psychometric information is reported on measures of parent, teacher, and child racial-ethnic attitudes. Path analysis was used to investigate ecological variables potentially related to children's racial-ethnic attitudes and achievement. Parental education and level of racial-ethnic pride were correlated and both were related to children's achievement though in the final path model, only the path from parental education level was statistically significant. Children whose teachers exhibited higher levels of racial-ethnic trust and perceived fewer barriers due to race and ethnicity evidenced more trust and optimism as well. Children living in communities with higher proportions of college-educated residents also exhibited more positive racial-ethnic attitudes. For children, higher racial-ethnic pride was related to higher achievement measured by grades and standardized test scores, while racial distrust and perception of barriers due to race were related to reduced performance. This study suggests that family, school, and community are all important factors related to children's racial-ethnic attitudes and also to their academic achievement.


[Monaural phasic sensitivity of auditory system neurons in the cat].
During monaural stimulation, studies have been made of impulse reactions in neurons from the cochlear nucleus and inferior colluculus to changes in the phase value (psi) of one of the harmonics in two-tone signals. It was shown that about 70% of the neurons from the cochleaur nucleus with a tonic discharge pattern may sharply change their activity (from maximum one up to a complete inhibition) due to changes in psi value. Unlike the cochlear nucleus units, neurons from the inferior colliculus change their tonic or burst-like activity rather seldomly and to a slight extent. At the same time, their phasic on- and especially off-responses exhibit high sensitivity to changes in psi value.