Mitochondria-Targeting 1,5-Diazacyclooctane-Spacered Triterpene Rhodamine Conjugates Exhibit Cytotoxicity at Sub-Nanomolar Concentration against Breast Cancer Cells.
1,5-Diazacyclooctane was prepared by a simple synthetic sequence and coupled to pentacyclic triterpenoic acids oleanolic acid, ursolic acid, betulinic acid, platanic acid, and asiatic acid; these amides were activated with oxalyl chloride and reacted with rhodamine B or rhodamine 101 to yield conjugates. The conjugates were screened in SRB assays with various human breast cancer cell lines (MDA-MB-231, HS578T, MCF-7, and T47D) and found to exert cytotoxic activity even at a low concentration. Therefore, for an asiatic acid rhodamine 101 conjugate (28), an IC50 = 0.60 nM was determined and found to induce apoptosis in MDA-MB-231 and HS578T cells. Extra experiments showed the compound to act as a mitocan and to induce inhibition of proliferation or growth arrest in MDA-MB-231 cells at lower doses followed by an induction of apoptosis at higher doses. Furthermore, differential responses to proliferation inhibition and apoptosis induction may explain differential sensitivity of mammary cell lines to compound 28.


P53 and heat shock protein 70 expressions in colorectal adenocarcinoma.
To examine the localization and over expression of heat shock protein 70 (HSP70) and p53 in patients with colorectal cancer and compared it with control tissue (including normal colon tissue). This was a retrospective study of 60 patients with colorectal adenocarcinoma at the Jordan University of Science and Technology (JUST), Irbid, Jordan from 1997 to 2000. The Pathology Department at JUST is the chief provider of surgical pathology services in the north of Jordan. It receives specimens from both government and private hospitals. Immunohistochemistry was the technique of choice. The HSP70 was over expressed more highly in colorectal cancers than in the control tissue. Immuno-histochemistry showed that over expression of HSP70 had no statistically significant difference with any of the different prognostic factors assessed, mainly the grade and the stage. The p53 was over expressed in 60% of the cases. Control tissue (normal colon) was negative, p53, cell-cycle-related oncogene product, was strongly over expressed in the nuclei of the cancer cells of the cancer tissue. We found no significant difference in terms of size, patient age, lymph node state, and stage. The rate of expression was significantly less in high grade tumors than in intermediate and low grade ones. The strong expression however, may be valuable in estimating a prognosis for patients with colo-rectal carcinoma.


Harlequin (hlq) and short blue root (sbr), two Arabidopsis mutants that ectopically express an abscisic acid- and auxin-inducible transgenic carrot promoter and have pleiotropic effects on morphogenesis.
Plant growth and development is regulated by complex interactions among different hormonal, developmental and environmental signalling pathways. Isolation of mutants in these processes is a powerful approach to dissect unknown mechanisms in regulatory networks. The plant hormones abscisic acid (ABA) and auxin are involved in vegetative, developmental and environmental growth responses, including cell division and elongation, vascular tissue differentiation and stress adaptation. The uidA (beta-glucuronidase; GUS) reporter gene driven by the carrot (Daucus carota) late embryogenesis-abundantDc3 promoter in transgenic Arabidopsis thaliana seedlings is ABA-inducible in the root zone of elongation and vasculature. We show here that the ABA-insensitive2-1 mutation (abi2) reduces ABA-inducible Dc3-GUS expression in these root tissues. Dc3-GUS expression is also induced in root cortex cells by indole-3-acetic acid. We mutagenized, with ethyl methane sulfonate, 5100 M1 abi2/abi2 homozygous plants of a line that carries two independent Dc3-GUS reporter genes and screened M2 clonal lines for ABA-inducible Dc3-GUS expression in roots. We isolated two novel single-gene nuclear mutants, harlequin (hlq) and short blue root (sbr), that ectopically express Dc3-GUS in roots and have pleiotropic effects on morphogenesis. The hlq mutant expresses Dc3-GUS in a checkered pattern in epidermis of roots and hypocotyls, accumulates callose and has deformed and collapsed epidermal cells and abnormal and reduced root hairs and leaf trichomes. It (hlq) is also dwarfed, skotomorphogenic and sterile. The sbr mutant is a seedling-lethal dwarf that over-expresses Dc3-GUS in the root and has radially swollen epidermal cells in the root and hypocotyl, supernumerary cell number in the root cortex and epidermis, abnormal vasculature, and abnormal epidermal cell patterning in cotyledons and leaves. It (sbr) also exhibits a semidominant root phenotype of reduced growth and lateral root initiation. The hlq and sbr mutants are not rescued by exogenous application of plant growth regulators. The hlq and sbr mutants do not require the abi2-1 mutant gene for their phenotypes and map to chromosome III and I, respectively. Further characterization of the hlq and sbr phenotypes and genes may provide insights into the relationship of hormone- and stress-regulated gene expression to morphogenesis and plant growth.


[Follow-up examinations after removal of titanium plates coated with anodic titanium oxide ceramic].
Transformation of the titanium metal surface with titanium oxides produced in various ways belongs among the most up-to-date procedures. The authors as pioneers in this field (e.g. Nobel Biocare TiUnite surface), have been utilizing for more than 15 years dental root implants and fixing elements (for mandibular osteosynthesis) coated with titanium oxide ceramics, produced by anodic oxidation and thermal treatment. The aim of this work was to assess the extent to which a titanium oxide ceramic coating influences the fate of plates applied for osteosynthesis within the human body. During a 5-year period (1995-1999), 108 of 1396 titanium oxide ceramic plates had to be removed for various reasons: loosening of the plate [47], osteomyelitis [25], a palpable swelling and tenderness [21] at the request of the patient for psychological reasons (13) or breaking of the plate [2]. When these 108 plates were removed, it was not possible to detect metallosis in even a single case; nor was there any tissue damage that could be attributed to the surface of the plates, whereas the literature data indicate that such damage is relatively frequent in the environment of traditional titanium fixing elements. The present investigation confirms the favourable properties of the titanium oxide ceramic surface.


Role of nerve growth factor and its TRKA receptor in normal ovarian and epithelial ovarian cancer angiogenesis.
In normal ovarian function a controlled angiogenesis is essential. Several growth factors are involved in this process, such as the vascular endothelial growth factor (VEGF) and nerve growth factor (NGF). The angiogenesis process in the normal ovary is a tightly controlled process that occurs in each ovarian cycle. Also, angiogenesis is critical for ovarian cancer development and it is responsible for tumor spread, metastasis and its peritoneal dissemination. Ovarian cancer is the fifth leading cause of cancer death in women and it is distinguished as the most lethal gynecologic cancer. In recent years angiogenesis has been given considerable attention in order to identify targets for developing effective anti-tumor therapies. Several molecules have been reported to promote angiogenesis, such as platelet-derived growth factor (PDGF) and its receptors, the angiopoietin/Tie ligand/receptor system and fibroblast growth factor (FGF). Primarily, VEGF has been identified to play key roles in driving angiogenesis. The above-mentioned molecules are candidate drug targets. Used in combination with other treatments, anti-angiogenic therapies have managed to reduce disease progression. The present review is focused in NGF and its high affinity receptor tyrosine kinase A (TRKA). The expression of VEGF, proliferation and the angiogenesis process in ovarian cancer is importantly induced by NGF, among other molecules.


Cellular peripheral neural tumors (neurofibromas) in children and adolescents: a clinicopathological and immunohistochemical study.
Nine examples of a cellular peripheral neural tumor (CPNT) were identified in a review of 139 peripheral nerve sheath neoplasms in children, which included 60 neurofibromas and 16 malignant peripheral nerve sheath tumors. The mean age at diagnosis of these nine patients was 7 years, with six presenting in the first decade of life and four were noted at birth. The male:female ratio was 0.5. Topographically, the tumors were located in the extremities, 4; head and neck, 3; and trunk, 2. One or another stigmata of von Recklinghausen's neurofibromatosis (VRN) was present in four patients. After initial resection, seven children remained well, but two developed a recurrence; the histology was identical to the original tumor in one case but overt malignant transformation had occurred in the second. This case was the only tumor-related death in this series. The CPNT was a circumscribed but nonencapsulated mass measuring 1.8-7.5 cm in greatest dimension in the subcutaneous and deep soft tissues and had a compact spindle cell pattern, occasional mitoses, and minor foci of typical neurofibroma. Immunohistochemical staining revealed vimentin expression in all seven cases, Leu-7 in six, myelin basic protein and S-100 protein in five, desmin in one, and actin in none. In contrast to neurofibroma and malignant peripheral nerve sheath tumors, CPNT tended to occur earlier, either congenitally or in the first decade, and slightly more commonly in females. The anatomical distribution and pattern of immunoreactivity were similar to neurofibroma. However, the cellularity and mitotic activity of these neoplasms were sufficiently disquieting as to raise concerns about the prognosis, and in one case, the tumor behaved in an unequivocally malignant fashion. When a peripheral neural tumor with the pathologic features described in this study is encountered, wide excision and careful clinical follow-up are recommended.


Phase-transfer-catalyzed asymmetric S(N)Ar reaction of α-amino acid derivatives with arene chromium complexes.
Although phase-transfer-catalyzed asymmetric S(N)Ar reactions provide unique contribution to the catalytic asymmetric α-arylations of carbonyl compounds to produce biologically active α-aryl carbonyl compounds, the electrophiles were limited to arenes bearing strong electron-withdrawing groups, such as a nitro group. To overcome this limitation, we examined the asymmetric S(N)Ar reactions of α-amino acid derivatives with arene chromium complexes derived from fluoroarenes, including those containing electron-donating substituents. The arylation was efficiently promoted by binaphthyl-modified chiral phase-transfer catalysts to give the corresponding α,α-disubstituted α-amino acids containing various aromatic substituents with high enantioselectivities.


[Temporal variation of background atmospheric CO2 and CH4 at Mount Waliguan, China].
In this paper, the continuous (2009-2010) measurement of atmospheric CO2 and CH4 from the Mount Waliguan Baseline Observatory of Western China are presented. The results show that about 17% of CO2 observations are classified as polluted due to more frequently influence of regional emission on local measurement in summer time. The mean concentration of CO2 measured at the period of 2009 to 2010 was 390.72 x 10(-6) which was 17.4 x 10(-6) higher than that measured from 1995 to 2008, and the median concentration of CH4 was 1851.11 x 10(-9) which was 16 x 10(-9) higher than that from 2002 to 2006, which implied that the regional emission of CO2 and CH4 was continuously increased. The unavailable data were filled by back propagation neural network (BPNN) and optimized by genetic algorithm (GA) , which were analyzed by the Fourier analysis of time series of air temperature, wind speed, concentration of CO2 and CH4. At the daily time scale, strong spectrum peak occurred and concentration recorded at periods of 12 and 24 hours,due to the daily sun activity changes. At the monthly time scale, the spectrum gap occurred in CO2 concentration at periods of 30 day suggesting that the effect of meteorological and phenological factors on the variation of CO2 concentration was insignificant.


A mouse juvenile or adult slice with preserved functional nigro-striatal dopaminergic neurons.
The effect of endogenous dopamine on the activity of target neurons recorded with patch clamp or Ca2+ imaging techniques in slices has been studied to date with intra-striatal stimuli. Yet, this approach is severely handicapped by the nonphysiological and nonspecific stimulation of local neurons and fibers within the striatum. We now report a new juvenile and adult mouse slice preparation in which a component of the nigro-striatal dopaminergic pathway is preserved in its entirety, from cell bodies to axon terminals. This tilted parasagittal slice (380-400 microm) just medial to the subthalamic nucleus contains functional nigro-striatal neurons as assessed by morphological examination of tyrosine hydroxylase positive cell bodies and axons, combined with electrochemical assays of dopamine release in the striatum in response to stimulation of the substantia nigra pars compacta. The nigro-striatal slice constitutes a suitable in vitro preparation to determine the impact of endogenously released dopamine on target neurons of the striatum.


Medical muddle.
Nanette Gartrell, MD, is a psychiatrist and researcher whose investigations have documented the mental health and psychological well-being of lesbian, gay, bisexual, and transgender (LGBT) people over the past four decades. Nanette is the principal investigator of an ongoing longitudinal study of lesbian families in which the children were conceived by donor insemination. Now in its 27th year, this project has been cited internationally in the debates over equality in marriage, foster care, and adoption. Previously on the faculty at Harvard Medical School and the University of California, San Francisco, Nanette is currently a Visiting Distinguished Scholar at the Williams Institute, UCLA School of Law. In 2013, Nanette received the Association of Women Psychiatrists Presidential Commendation Award for "selfless and enduring vision, leadership, wisdom, and mentorship in the fields of women's mental health, ethics, and gender research." At the age of 63, Nanette experienced a 3 ½ month period of intractable, incapacitating dizziness for which there was never a clear diagnosis.


CRISPR/Cas9-mediated knockout of CD47 causes hemolytic anemia with splenomegaly in C57BL/6 mice.
CD47 (integrin-associated protein), a multi-spanning transmembrane protein expressed in all cells including red blood cells (RBCs) and leukocytes, interacts with signal regulatory protein α (SIRPα) on macrophages and thereby inhibits phagocytosis of RBCs. Recently, we generated a novel C57BL/6J CD47 knockout (CD47 -/- hereafter) mouse line by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease, and here report their hematological phenotypes. On monitoring their birth and development, CD47 -/- mice were born viable with a natural male-to-female sex ratio and normally developed from birth through puberty to adulthood without noticeable changes in growth, food/water intake compared to their age and sex-matched wild-type littermates up to 26 weeks. Hematological analysis revealed a mild but significant reduction of RBC counts and hemoglobin in 16 week-old male CD47 -/- mice which were aggravated at the age of 26 weeks with increased reticulocyte counts and mean corpuscular volume (MCV), suggesting hemolytic anemia. Interestingly, anemia in female CD47 -/- mice became evident at 26 weeks, but splenomegaly was identified in both genders of CD47 -/- mice from the age of 16 weeks, consistent with development of hemolytic anemia. Additionally, helper and cytotoxic T cell populations were considerably reduced in the spleen, but not in thymus, of CD47 -/- mice, suggesting a crucial role of CD47 in proliferation of T cells. Collectively, these findings indicate that our CD47 -/- mice have progressive hemolytic anemia and splenic depletion of mature T cell populations and therefore may be useful as an in vivo model to study the function of CD47.


Clipping of complex aneurysms with fenestration tubes: application and assessment of three types of clip techniques.
Straight fenestrated clips can be stacked to create tubes that reconstruct origins of branch arteries at the necks of complex aneurysms. Three variations of fenestration tubes were conceived and applied to a consecutive series of patients with aneurysms to determine their usefulness. Antegrade fenestration tubes are built with stacked, straight fenestrated clips with an open tube transmitting the efferent artery with forward blood flow. Retrograde fenestration tubes are also built with stacked, straight fenestrated clips, but the fenestration tube is closed with a nonfenestrated clip to reverse the direction of blood flow into the efferent artery that exits from the base of the tube. Aneurysm dome fenestration tubes encircle the dome rather than transmitting an efferent artery and use the tips of stacked, straight fenestrated clips to reconstruct the aneurysm neck. During a 2-year period in which 465 aneurysms were treated microsurgically, 25 patients had 26 aneurysms clipped with fenestration tubes (antegrade tubes, 15 aneurysms; retrograde tubes, four aneurysms; dome tubes, seven aneurysms). Angiographically, 92% of the aneurysms were completely eliminated with no branch artery occlusions. Neurologically, good outcomes were observed in 84% of the patients (Glasgow Outcome Scale score 5 or 4) with 96% either improved or unchanged. Clip reconstruction with fenestration tubes is another microsurgical technique that can be used to treat aneurysms with large or giant-sized efferent arteries that adhere to the aneurysm and/or unusual branch anatomy. Fenestration tubes are safe and effective, and they are used more frequently than expected for what might seem to be a subtle technical nuance. This technique relies on intraoperative adjuncts such as temporary clipping, careful application of clips to preserve the patency of branch arteries, and technology to detect inadvertent branch occlusions.


TIM-3/Galectin-9 and CD160 expression in salivary adenoid cystic carcinoma.
Investigating T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), Galectin 9 (Gal-9), CD160 expression and tumor-infiltrating lymphocytes (TILs) and correlation with clinicopathological characteristics of salivary adenoid cystic carcinoma (SACC). Sixty cases of SACC were detected by immunohistochemical staining to evaluate TIM-3, Gal-9, and CD160 expression and analyze the correlation between TIM-3, Gal-9, CD160 expression and clinicopathologic features by rank-sum test. The association of TILs with TIM-3, Gal-9, and CD160 expression in SACC stromal was done by Chi-square test. TIM-3 and CD160 overexpression were correlated with recurrence of SACC (p = 0.029, p = 0.007, respectively). High Gal-9 expression was correlated with pathological classification (p = 0.018). The average percentage of TILs was 18.2% in SACC and most of TILs were more likely to occur in minor salivary glands (p = 0.038). Pairwise positive correlations were observed between the expression of TIM-3, Gal-9, and CD160 in tumor cells as well as in TILs, respectively. Low density of TILs was characteristic of the SACC microenvironment, with upregulation of TIM-3, Gal-9, and CD160 all occurring. However, TIM-3, Gal-9, and CD160 expression in the stromal dependent on the number of TILs represent potential therapeutic targets in SACC.


Fetal hemoglobin, sickling, and sickle cell disease.
Increased numbers of F cells and large amounts of Hb F/F cell appear to produce clinical benefit in rare variants of sickle cell disease and probably in more commonly encountered patients. Fetal hemoglobin interferes with polymerization of Hb S in vitro, but laboratory studies carried out with homogeneous hemoglobin solutions are inadequate models of events in vivo, because RBCs are heterogeneous in their MCHC and Hb F content. Studies of hemoglobin switching in sheep, in tissue culture, and then in baboons led to use of 5-azacytidine for induction of increased Hb F synthesis in SS patients. Drug trials were successful but the theory that led to them was not. An alternate theory, not without flaws, led to the use of hydroxyurea. Chronic administration of the drug can lead to very impressive increases in Hb F synthesis and apparent clinical benefit. It is not clear that such clinical benefit is real rather than a placebo effect. Nor is it entirely clear that all of the effect of hydroxyurea can be related to increased production of F cells and increased F/F cell. Controlled clinical trials and studies of the properties of RBCs from treated patients may answer those questions. It is also likely that they will not only raise still other questions but probably show that our current understanding of the biology and treatment of sickle cell disease is far from complete.


Dendritic excitability and calcium signalling in the mitral cell distal glomerular tuft.
The processing of odour information starts at the level of the olfactory glomerulus, where the mitral cell distal dendritic tuft not only receives olfactory nerve sensory input but also generates dendrodendritic output to form complicated glomerular synaptic circuits. Analysing the membrane properties and calcium signalling mechanisms in these tiny dendritic branches is crucial for understanding how the glomerular tuft transmits and processes olfactory signals. With the use of two-photon Ca2+ imaging in rat olfactory bulb slices, we found that these distal dendritic branches displayed a significantly larger Ca2+ signal than the soma and primary dendrite trunk. A back-propagating action potential was able to trigger a Ca2+ increase throughout the entire glomerular tuft, indicative of the presence of voltage-gated Ca2+ conductances in all branches at different levels of ramification. In response to a train of action potentials evoked at 60 Hz from the soma, the tuft Ca2+ signal increased linearly with the number of action potentials, suggesting that these glomerular branches were able to support repetitive penetration of Na+ action potentials. When a strong olfactory nerve excitatory input was paired with an inhibition from mitral cell basal dendrites, a small spike-like fast prepotential was revealed at both the soma and distal primary dendrite trunk. Corresponding to this fast prepotential was a Ca2+ increase confined locally within the glomerular tuft. In summary, the mitral cell distal dendritic tuft possesses both Na+ and Ca2+ voltage-dependent conductances which can mediate glomerular Ca2+ responsiveness critical for dendrodendritic output and synaptic plasticity.


A specific amino acid formula prevents alcoholic liver disease in rodents.
Chronic alcohol consumption promotes mitochondrial dysfunction, oxidative stress, defective protein metabolism, and fat accumulation in hepatocytes (liver steatosis). Inadequate amino acid metabolism is worsened by protein malnutrition, frequently present in alcohol-consuming patients, with reduced circulating branched-chain amino acids (BCAAs). Here we asked whether dietary supplementation with a specific amino acid mixture, enriched in BCAAs (BCAAem) and able to promote mitochondrial function in muscle of middle-aged rodents, would prevent mitochondrial dysfunction and liver steatosis in Wistar rats fed on a Lieber-DeCarli ethanol (EtOH)-containing liquid diet. Supplementation of BCAAem, unlike a mixture based on the amino acid profile of casein, abrogated the EtOH-induced fat accumulation, mitochondrial impairment, and oxidative stress in liver. These effects of BCAAem were accompanied by normalization of leucine, arginine, and tryptophan levels, which were reduced in liver of EtOH-consuming rats. Moreover, although the EtOH exposure of HepG2 cells reduced mitochondrial DNA, mitochondrial transcription factors, and respiratory chain proteins, the BCAAem but not casein-derived amino acid supplementation halted this mitochondrial toxicity. Nicotinamide adenine dinucleotide levels and sirtuin 1 (Sirt1) expression, as well as endothelial nitric oxide (eNOS) and mammalian/mechanistic target of rapamycin (mTOR) signaling pathways, were downregulated in the EtOH-exposed HepG2 cells. BCAAem reverted these molecular defects and the mitochondrial dysfunction, suggesting that the mitochondrial integrity obtained with the amino acid supplementation could be mediated through a Sirt1-eNOS-mTOR pathway. Thus a dietary activation of the mitochondrial biogenesis and function by a specific amino acid supplement protects against the EtOH toxicity and preserves the liver integrity in mammals. NEW & NOTEWORTHY Dietary supplementation of a specific amino acid formula prevents both fat accumulation and mitochondrial dysfunction in hepatocytes of alcohol-consuming rats. These effects are accompanied also by increased expression of anti-reactive oxygen species genes. The amino acid-protective effects likely reflect activation of sirtuin 1-endothelial nitric oxide synthase-mammalian target of rapamycin pathway able to regulate the cellular energy balance of hepatocytes exposed to chronic, alcoholic damage.


Sexuality during pregnancy.
While pregnancy is a major event in most couples' lives, there are no predictable changes in sexuality. Fears of damaging the fetus are unwarranted, even during the last weeks of pregnancy, provided there are no complications. Meticulous episiotomy technique can lessen the healing time and allow the couple to resume mutually satisfying intercourse sooner.


Tissue-specific regulation of canine intestinal and hepatic phenol and morphine UDP-glucuronosyltransferases by beta-naphthoflavone in comparison with humans.
UDP-glucuronosyltransferases (UGTs) are regulated in a species- and tissue-dependent manner by endogenous and environmental factors. The present study was undertaken to further our knowledge about regulation of UGTs in dogs, a species widely used in preclinical safety evaluation. beta-Naphthoflavone (BNF) was selected as a known aryl hydrocarbon receptor agonist and antioxidant-type inducer. The latter group of inducers is intensively investigated as dietary chemoprotectants against colon cancer. Dog UGTs were investigated in comparison with related human UGTs by examples, (i) expression of dog UGT1A6, the first sequenced dog phenol UGT, and (ii) morphine UGT activities, responsible for intestinal and hepatic first-pass metabolism of morphine. The following results were obtained: (i) dog UGT1A6 was found to be constitutively expressed in liver and marginally increased by BNF treatment. Expression was low in small intestine but ca. 6-fold higher in colon than for example in jejunum. Conjugation of 4-methylumbelliferone, one of the substrates of dog UGT1A6, was also enhanced 7-fold in colonic compared to jejunal microsomes. (ii) Compared to the corresponding human tissues, canine 3-O- and 6-O-morphine UGT activities were found to be >10-fold higher in dog liver and ca. 10-fold lower in small intestinal microsomes. Small intestinal morphine and 4-hydroxybiphenyl UGT activities appeared to be moderately (2- to 3-fold) induced by oral treatment with BNF. (iii) In contrast to dogs, morphine UGT activities were found to be similar in homogenates from human enterocytes and liver. The results suggest marked differences in tissue-specific regulation of canine vs. human hepatic and intestinal phenol or morphine UGTs.


Nurse practitioner acceptance in private Ob/Gyn practice.
One of the ways to help meet consumer demands is by utilizing nurse practitioners in private Ob/Gyn practices. To determine consumer satisfaction with a nurse practitioner in one such practice, patients were asked to fill out a response sheet after their examination by the nurse practitioner. The 86 patients felt they received a good medical examination and would be willing to see the nurse practitioner on future visits. The physician and nurse practitioner found this change in delivery of health care to be a positive experience for both patient and professional.


Protective efficacy of a recombinant bacterial artificial chromosome clone of a very virulent Marek's disease virus containing a reticuloendotheliosis virus long terminal repeat.
Marek's disease virus (MDV), an alphaherpesvirus, causes Marek's disease (MD), a lymphoproliferative disease in poultry characterized by T-cell lymphomas, nerve lesions, and mortality. Vaccination is used worldwide to control MD, but increasingly virulent field strains can overcome this protection, driving a need to create new vaccines. Previous studies revealed that insertion of reticuloendotheliosis virus (REV) long terminal repeat (LTR) into a bacterial artificial chromosome (BAC) clone of a very virulent strain of MDV, Md5, rendered the resultant recombinant virus, rMd5 REV-LTR BAC, fully attenuated in maternal antibody positive (Mab+) chickens at passage 40. In the current study, the protective efficacy of rMd5 REV-LTR BAC was evaluated. First, passage 70 was identified as being fully attenuated in maternal antibody negative chickens and chosen as the optimal passage level for use in protective efficacy studies. Second, three protective efficacy trials were conducted comparing the rMd5 REV-LTR p70 BAC to the CVI988/Rispens vaccine. Groups of Mab+ and Mab- 15I5 × 71 chickens were vaccinated in ovo at 18 days of embryonation or intra-abdominally at day of hatch, and challenged at 5 days post-hatch with the vv+MDV strain 686. Vaccination at day of hatch and in ovo with rMd5 REV-LTR p70 BAC protected chickens against MDV-induced bursa and thymic atrophy, but did not provide the same level of protection against MD tumours as that afforded by the commercial vaccine, CVI988/Rispens.


Nutritional quality of extruded kidney bean (Phaseolus vulgaris L. var. Pinto) and its effects on growth and skeletal muscle nitrogen fractions in rats.
The influence of extrusion cooking on the protein content, amino acid profile, and concentration of antinutritive compounds (phytic acid, condensed tannins, polyphenols, trypsin, chymotrypsin, alpha-amylase inhibitors, and hemagglutinating activity) in kidney bean seeds (Phaseolus vulgaris L. var. Pinto) was investigated. Growing male rats were fed diets based on casein containing raw or extruded kidney beans with or without methionine supplementation for 8 or 15 d. Rates of growth, food intake, and protein efficiency ratio were measured and the weight of the gastrocnemius muscle and the composition of its nitrogenous fraction was determined. Extrusion cooking reduced (P < 0.01) phytic acid, condensed tannins, and trypsin, chymotrypsin, and (alpha-amylase inhibitory activities. Furthermore, hemagglutinating activity was abolished by extrusion treatment. Protein content was not affected by this thermal treatment. Rats fed raw kidney bean lost BW rapidly and the majority died by 9 d. Pretreatment of the beans by extrusion cooking improved food intake and utilization by the rats and they gained BW. Supplementation of extruded kidney bean with methionine further enhanced (P < 0.01) food conversion efficiency and growth. However, BW gains and muscle composition still differed (P < 0.01) from those of rats fed a high-quality protein.


Stabilising the newborn for transfer - basic principles.
Rural general practitioners involved in obstetric service delivery may have occasion to support the sick or premature newborn requiring transfer. This should be achievable for short periods of time in most rural hospitals. This article discusses the planning priorities, and the equipment and skills required for care of the sick or premature newborn. With careful planning, attention to detail, and maintenance of clinical skills and equipment, newborn intensive care can be provided in most hospitals for brief periods while awaiting the arrival of the retrieval team.


Antibody adsorption on the surface of water studied by neutron reflection.
Surface and interfacial adsorption of antibody molecules could cause structural unfolding and desorbed molecules could trigger solution aggregation, resulting in the compromise of physical stability. Although antibody adsorption is important and its relevance to many mechanistic processes has been proposed, few techniques can offer direct structural information about antibody adsorption under different conditions. The main aim of this study was to demonstrate the power of neutron reflection to unravel the amount and structural conformation of the adsorbed antibody layers at the air/water interface with and without surfactant, using a monoclonal antibody 'COE-3' as the model. By selecting isotopic contrasts from different ratios of H2O and D2O, the adsorbed amount, thickness and extent of the immersion of the antibody layer could be determined unambiguously. Upon mixing with the commonly-used non-ionic surfactant Polysorbate 80 (Tween 80), the surfactant in the mixed layer could be distinguished from antibody by using both hydrogenated and deuterated surfactants. Neutron reflection measurements from the co-adsorbed layers in null reflecting water revealed that, although the surfactant started to remove antibody from the surface at 1/100 critical micelle concentration (CMC) of the surfactant, complete removal was not achieved until above 1/10 CMC. The neutron study also revealed that antibody molecules retained their globular structure when either adsorbed by themselves or co-adsorbed with the surfactant under the conditions studied.


In silico prediction of deleterious non-synonymous SNPs in STAT3.
STAT3, a pleiotropic transcription factor, plays a critical role in the pathogenesis of autoimmunity, cancer, and many aspects of the immune system, as well as having a link with inflammatory bowel disease. Changes caused by non-synonymous single nucleotide polymorphisms (nsSNPs) have the potential to damage the protein's structure and function. We identified disease susceptible single nucleotide polymorphisms (SNPs) in STAT3 and predicted structural changes associated with mutants that disrupt normal protein-protein interactions using different computational algorithms. Several in silico tools, such as SIFT, PolyPhen v2, PROVEAN, PhD-SNP, and SNPs&GO, were used to determine nsSNPs of the STAT3. Further, the potentially deleterious SNPs were evaluated using I-Mutant, ConSurf, and other computational tools like DynaMut for structural prediction. 417 nsSNPs of STAT3 were identified, 6 of which are considered deleterious by in silico SNP prediction algorithms. Amino acid changes in V507F, R335W, E415K, K591M, F561Y, and Q32K were identified as the most deleterious nsSNPs based on the conservation profile, structural conformation, relative solvent accessibility, secondary structure prediction, and protein-protein interaction tools. The in silico prediction analysis could be beneficial as a diagnostic tool for both genetic counseling and mutation confirmation. The 6 deleterious nsSNPs of STAT3 may serve as potential targets for different proteomic studies, large population-based studies, diagnoses, and therapeutic interventions.


Central penetrating keratoplasty with peripheral intrastromal tuck.
To report a new technique of central penetrating keratoplasty with peripheral intrastromal tuck in cases of chronic corneal decompensation. Central penetrating keratoplasty with peripheral intrastromal tuck was undertaken in 8 eyes with chronic corneal decompensation. An 8-mm donor tissue with a peripheral flange of 2 mm was fashioned and sutured into a 7.5-mm recipient bed having a 2-mm intralamellar pocket. Sixteen full-thickness interrupted sutures were passed. Suture removal was possible as early as 1 month, and all sutures were removed by 4 months. Postoperative BCVA improved to 20/40 to 20/80 in all eyes, and astigmatism was <4 diopters in all cases at the last follow-up, which ranged from 5 to 16 months. Central penetrating keratoplasty with peripheral intrastromal tuck is a useful technique because it allows early suture removal and visual rehabilitation.


Human mature adipocytes express albumin and this expression is not regulated by inflammation.
Our group investigated albumin gene expression in human adipocytes, its regulation by inflammation and the possible contribution of adipose tissue to albumin circulating levels. Both inflamed and healthy subjects provided adipose tissue samples. RT-PCR, Real-Time PCR, and Western Blot analysis on homogenates of adipocytes and pre-adipocytes were performed. In sixty-three healthy subjects and fifty-four micro-inflamed end stage renal disease (ESRD) patients circulating levels of albumin were measured by nephelometry; all subjects were also evaluated for body composition, calculated from bioelectrical measurements and an thropometric data. A clear gene expression of albumin was showed in pre-adipocytes and, for the first time, in mature adipocytes. Albumin gene expression resulted significantly higher in pre-adipocytes than in adipocytes. No significant difference in albumin gene expression was showed between healthy controls and inflamed patients. A significant negative correlation was observed between albumin levels and fat mass in both healthy subjects and inflamed ESRD patients. In the present study we found first time evidence that human adipocytes express albumin. Our results also showed that systemic inflammation does not modulate albumin gene expression. The negative correlation between albumin and fat mass seems to exclude a significant contributing role of adipocyte in plasma albumin.


Antiepileptic drugs as prophylaxis for post-craniotomy seizures.
The incidence of seizures following supratentorial craniotomy for non-traumatic pathology has been estimated to be 15% to 20%; however, the risk of experiencing a seizure may vary from 3% to 92% over a five-year period. Postoperative seizures can precipitate the development of epilepsy; seizures are most likely to occur within the first month of cranial surgery. The use of antiepileptic drugs (AEDs) administered pre- or postoperatively to prevent seizures following cranial surgery has been investigated in a number of randomised controlled trials. To determine the efficacy and safety of AEDs when used prophylactically in people undergoing craniotomy and to examine which AEDs are most effective. We searched the Cochrane Epilepsy Group's Specialized Register (September 2012), the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library Issue 9, 2012), and MEDLINE (1946 to September 2012). No language restrictions were imposed. Randomised controlled trials of people with no history of epilepsy who were undergoing craniotomy for either therapeutic or diagnostic reasons were included. Trials with adequate randomisation methods and concealment were included; these could either be blinded or unblinded parallel trials. No minimum treatment period was stipulated, trials using active drugs or placebo as a control group were included. Two review authors (JP and JG) independently selected trials for inclusion and carried out data extraction and risk of bias assessments. Any disagreements were resolved through discussion. Outcomes investigated included the number of patients experiencing seizures (early - occurring within first week following craniotomy and late - occurring after first week following craniotomy), the number of deaths and the number of people experiencing disability and adverse effects. Due to the heterogeneous nature of the trials, data from the trials were not combined in a meta-analysis; the findings of the review are presented in narrative format. Six RCTs (N = 1398) were eligible for inclusion within the review with publication dates ranging between 1983 and 1999. Two trials compared a single AED (phenytoin) with a placebo. One three-arm trial compared two AEDs (carbamazepine, phenytoin) with no treatment. A second three-arm trial compared phenytoin, phenobarbital and no treatment. Two other trials were head-to-head trials of AEDs (phenytoin vs. valproate and zonisamide vs. phenobarbital). Of the four trials comparing AEDs with controls only one trial reported a significant difference between AED treatment and controls for early seizure occurrence. All other comparisons were non-significant. Of the head-to-head trials, none reported statistically significant differences between treatments for either early or late seizures. One head-to-head trial showed an increase in the number of deaths following one AED treatment compared to another AED treatment. Incidences of adverse effects of treatment were poorly reported, no significant differences between treatment groups were found due to the limited number of reported occurrences. There is little evidence to suggest that AED treatment administered prophylactically is effective or not effective in preventing post-craniotomy seizures. The current evidence base is limited due to the differing methodologies employed in the trials and inconsistencies in reporting of outcomes. Further evidence from good-quality, contemporary trials is required in order to assess the effectiveness of prophylactic AED treatment compared to control groups or other AEDs in preventing post-craniotomy seizures properly.


The role of magnetic resonance imaging in detecting intestinal fibrosis in Crohn's disease.
One of the main challenges for clinicians dealing with Crohn's disease (CD) is to distinguish between inflammation and fibrosis, two sequential steps in the evolution of the intestinal disease. While inflammation is amenable to medical treatment, stricturing disease can only benefit from surgery. Magnetic resonance imaging (MRI) is a widely used tool in the diagnosis and assessment of CD. Recent data suggest that this technique might help in discriminating between fibrosis and inflammation and as such potentially guide medical decisions. In this review we will first highlight the main diagnostic techniques for CD. We will then briefly review the main mechanisms of fibrosis in this condition and the use of MRI in CD focusing on findings predictive of fibrosis and fibrotic evolution.


Tumor necrosis is an important hallmark of aggressive endometrial cancer and associates with hypoxia, angiogenesis and inflammation responses.
Tumor necrosis is associated with aggressive features of endometrial cancer and poor prognosis. Here, we investigated gene expression patterns and potential treatment targets related to presence of tumor necrosis in primary endometrial cancer lesions. By DNA microarray analysis, expression of genes related to tumor necrosis reflected multiple tumor-microenvironment interactions like tissue hypoxia, angiogenesis and inflammation pathways. A tumor necrosis signature of 38 genes and a related patient cluster (Cluster I, 67% of the cases) were associated with features of aggressive tumors such as type II cancers, estrogen receptor negative tumors and vascular invasion. Further, the tumor necrosis signature was increased in tumor cells grown in hypoxic conditions in vitro. Multiple genes with increased expression are known to be activated by HIF1A and NF-kB. Our findings indicate that the presence of tumor necrosis within primary tumors is associated with hypoxia, angiogenesis and inflammation responses. HIF1A, NF-kB and PI3K/mTOR might be potential treatment targets in aggressive endometrial cancers with presence of tumor necrosis.


Brigatinib vs alectinib in crizotinib-resistant advanced anaplastic lymphoma kinase-positive non-small-cell lung cancer (ALTA-3).
Crizotinib is highly efficacious and more tolerable than chemotherapy for ALK+ non-small-cell lung cancer (NSCLC), but its progression-free survival benefit and intracranial efficacy have limitations. Head-to-head comparisons of next-generation ALK inhibitors in patients with ALK+ NSCLC progressing on crizotinib will contribute toward optimizing survival. This international, Phase III, randomized, open-label study (ALTA-3) will therefore assign patients with locally advanced or metastatic ALK+ NSCLC progressing on crizotinib to receive either brigatinib 180 mg qd (7-day lead-in at 90 mg qd) or alectinib 600 mg twice daily. The primary end point is progression-free survival as assessed by a blinded Independent Review Committee; the key secondary end point is overall survival. Clinical trial registration number: NCT03596866 (ClinicalTrials.gov).


Evaluation of the Acetone and Aqueous Extracts of Mature Stem Bark of Sclerocarya birrea for Antioxidant and Antimicrobial Properties.
We assayed the antimicrobial activity of acetone and aqueous extracts of the stem bark of Sclerocarya birrea on some selected bacteria and fungi species including; Streptococcus pyogenes, Plesiomonas shigelloides, Aeromonas hydrophila, Salmonella typhimurium, Cryptococcus neoformans, Candida glabrata, Trichosporon mucoides, and Candida krusei using both agar well diffusion and minimum inhibitory concentration (MIC) assays. Based on the levels of activity, the acetone extract was examined for total polyphenolic content, radical scavenging and antioxidant activities. Total phenols of the extract were determined spectrophotometrically. The antioxidant activity was determined by the DPPH, ABTS and reducing power. All the bacteria and fungi species were susceptible to the plant extracts. The acetone extract was the most active for the bacterial species with MIC (0.156-0.625 mg/mL) while the aqueous extract was the most active for the fungi species with MIC (0.3125-1.25 mg/mL). The polyphenolic compounds were found as 27.2 mg/g tannic acid equivalent, 25.2 mg/g quercetin equivalent, 9.1 mg/g quercetin equivalent for phenols, flavonoid and flavonols respectively. The acetone extract exhibited a remarkable ability to scavenge radicals, strong reducing ability and a potential source of natural antioxidants. Both the acetone and aqueous extracts of S. birrea may provide a target for drug discovery.


Laser scanning in vivo confocal microscopy reveals reduced innervation and reduction in cell density in all layers of the keratoconic cornea.
The exact pathophysiological processes underlying keratoconus remain an enigma. In this study, laser scanning in vivo confocal microscopy (IVCM) was used to define further the microstructural abnormalities in the keratoconic cornea and to establish the relationship with disease severity. This was a prospective, cross-sectional study comparing 52 subjects with keratoconus and 52 age-matched control subjects. Assessment included demographics, history, slit lamp biomicroscopy, computerized corneal tomography, and laser scanning IVCM. Significantly lower cell densities (in cells per square millimeter, mean +/- SD) were observed in keratoconus corneas than in normal ones: basal epithelial cells, 4340.6 +/- 595.2 vs. 5777.6 +/- 958.2 (P < 0.001), anterior keratocytes, 523.6 +/- 206.4 vs. 859.7 +/- 219.1 (P < 0.001), posterior keratocytes, 240.4 +/- 64.5 vs. 330.6 +/- 52.3 (P < 0.001), and endothelial cells 2412.2 +/- 339.5 vs. 2845.6 +/- 313.0 (P < 0.001). Subbasal nerve fiber density was 52.7% lower in keratoconus corneas than in the control (P < 0.001). Basal epithelial cell density (P = 0.001), subbasal nerve fiber density (P = 0.015), and anterior keratocyte density (P < 0.001) correlated with severity of disease. Lower subbasal nerve density also correlated with younger age at diagnosis (r = 0.397, P = 0.004). Severe disease was associated with diagnosis at a younger age (P = 0.023), a history of eye rubbing (P = 0.025), and Maori or Pacific Island ethnicity (P = 0.001). Significant microstructural abnormalities were identified at every level of the keratoconic cornea and were related to disease severity. IVCM offers a potential insight into the pathophysiology of the microstructural changes in keratoconus.


Investigation of (Fe,Co)NbB-Based Nanocrystalline Soft Magnetic Alloys by Lorentz Microscopy and Off-Axis Electron Holography.
The relationship between microstructure and magnetic properties of a (Fe,Co)NbB-based nanocrystalline soft magnetic alloy was investigated by analytical transmission electron microscopy (TEM). The microstructures of (Fe0.5Co0.5)80Nb4B13Ge2Cu1 nanocrystalline alloys annealed at different temperatures were characterized by TEM and electron diffraction. The magnetic structures were analyzed by Lorentz microscopy and off-axis electron holography, including quantitative measurement of domain wall width, induction, and in situ magnetic domain imaging. The results indicate that the magnetic domain structure and particularly the dynamical magnetization behavior of the alloys strongly depend on the microstructure of the nanocrystalline alloys. Smaller grain size and random orientation of the fine particles decrease the magneto-crystalline anisotropy and suggests better soft magnetic properties which may be explained by the anisotropy model of Herzer.


Mineral content of Chinese medicinal herbs used as diuretic treatments for Taiwanese children.
Eighteen major, minor and trace elements in 12 Chinese medicinal herbs commonly consumed by Taiwanese Children as diuretics were determined by instrumental neutron activation analysis (INAA). Dried and powdered herb samples were irradiated in a neutron flux of ca. 2 x 10(12) n/cm2 s under separate short and long irradiation schemes. Lichen (IAEA-336) was used as the reference standard, and tomato leaves (NIST-SRM 1570a) were employed for cross-checking the accuracy of the results. INAA was shown to be a reliable multi-element analytical method for determining the content of both toxicologically and nutritionally important minerals in Chinese medicinal herbs. Determined elements were present in the dried herbs in concentrations ranging from 10(4) to 10(-3) microg/g. The mineral contents and the maximum daily intake values of the tested herbs were compared with published values and with the recommended daily intakes for Taiwanese children as specified by the World Health Organization.


Biopsy of Onchocerca nodules in the Igbos of Nigeria.
The occurrence of Onchocerca nodules in the Igbos of Nigeria was studied in a biopsy series. There were 90 cases in approximately 5,000 biopsies received in 4.5 years at a central laboratory. The prevalence of infection in males was 1.5 times that of females. The two youngest patients were both aged 3 years; the oldest was approximately 70 years. Of the 79 single-site biopsies, 14 (17.7%) nodules occurred on the head and 65 (82.3%) were done elsewhere. The nodules above the waist line totalled 42 (53.2%) and those below it 37 (46.8%). The favorite location was the forehead cephalically and the hip caudally. Eyelid and breast nodules were also noteworthy.


Photochemistry of allyloxybenzophenones: a pseudo-Paternò-Büchi rearrangement accompanied by hydrogen transfer induced 1,5-cyclization.
The solution photochemistry of the ortho allyloxy-substituted benzophenone has been investigated in detail. Product analysis revealed formation of a diastereomeric mixture of dihydrobenzofuran derivatives by cyclization via a short-lived intermediate 1,5-biradical and an unusual acetal by a pseudo-Paternò-Büchi rearrangement. The latter reaction pathway was supported by means of laser flash photolysis, where a long-lived intermediate with a maximum absorption band at 380 nm was observed. Besides, theoretical calculations (TD-DFT) of this UV-transient resulted in a band with maximum intensity at 390 nm showing a good correlation between experimental results and theoretical calculations. For comparison, the meta-substituent substrate was also investigated showing preferred triplet-triplet energy transfer.


Bacterial Pesticidal Protein Mpp51Aa1 Delivered via Transgenic Citrus Severely Impacts the Fecundity of Asian Citrus Psyllid, Diaphorina citri.
The Asian citrus psyllid (ACP) Diaphorina citri vectors the causative agent of citrus greening disease that has the capacity to decimate citrus production. As an alternative and more sustainable approach to manage D. citri than repeated application of chemical insecticides, we investigated the potential use of the bacteria-derived pesticidal protein, Mpp51Aa1, when delivered by transgenic Citrus sinensis cv. Valencia sweet orange or Citrus paradisi cv. Duncan grapefruit. Following confirmation of transcription and translation of mpp51aa1 by transgenic plants, no impact of Mpp51Aa1 expression was seen on D. citri host plant choice between transgenic and control Duncan grapefruit plants. A slight but significant drop in survival of adult psyllids fed on these transgenic plants was noted relative to those fed on control plants. In line with this result, damage to the gut epithelium consistent with that caused by pore-forming proteins was only observed in a minority of adult D. citri fed on the transgenic Duncan grapefruit. However, greater impacts were observed on nymphs than on adults, with a 40% drop in the survival of nymphs fed on transgenic Duncan grapefruit relative to those fed on control plants. For Valencia sweet orange, a 70% decrease in the number of eggs laid by adult D. citri on transgenic plants was noted relative to those on control plants, with a 90% drop in emergence of progeny. These impacts that contrast with those associated with other bacterial pesticidal proteins and the potential for use of Mpp51Aa1-expressing transgenic plants for suppression of D. citri populations are discussed. IMPORTANCE Pesticidal proteins derived from bacteria such as Bacillus thuringiensis are valuable tools for management of agricultural insect pests and provide a sustainable alternative to the application of chemical insecticides. However, relatively few bacterial pesticidal proteins have been used for suppression of hemipteran or sap-sucking insects such as the Asian citrus psyllid, Diaphorina citri. This insect is particularly important as the vector of the causative agent of citrus greening, or huanglongbing disease, which severely impacts global citrus production. In this study, we investigated the potential of transgenic citrus plants that produce the pesticidal protein Mpp51Aa1. While adult psyllid mortality on transgenic plants was modest, the reduced number of eggs laid by exposed adults and the decreased survival of progeny was such that psyllid populations dropped by more than 90%. These results provide valuable insight for potential deployment of Mpp51Aa1 in combination with other control agents for the management of D. citri.


Differences in body composition and growth persist postnatally in fetuses diagnosed with severe compared to mild fetal growth restriction.
Fetal growth restriction (FGR) is most commonly diagnosed in pregnancy if the estimated fetal weight (EFW) is < 10th%. Those with abnormal Doppler velocimetry, indicating placental insufficiency and pathological FGR, demonstrate reduced fat and lean mass compared to both normally growing fetuses and FGR fetuses with normal Dopplers. The aim of this study was to determine how severity of FGR and abnormal Doppler velocimetry impacts neonatal body composition. Among a cohort of fetuses with an EFW < 10th%, we hypothesized that those with abnormal Dopplers and/or EFW < 3rd% would have persistent reductions in lean body mass and fat mass extending into the neonatal period compared to fetuses not meeting those criteria. A prospective cohort of FGR fetuses with an estimated fetal weight (EFW) < 10th% was categorized as severe (EFW < 3rd% and/or abnormal Dopplers; FGR-S) versus mild (EFW 3-10th%; FGR-M). Air Displacement Plethysmography and anthropometrics were performed at birth and/or within the first 6-8 weeks of life. FGR-S versus FGR-M were born one week earlier (P = 0.0024), were shorter (P = 0.0033), lighter (P = 0.0001) with smaller weight-for-age Z-scores (P = 0.0004), had smaller head circumference (P = 0.0004) and lower fat mass (P = 0.01) at birth. At approximately 6-8 weeks postmenstrual age, weight, head circumference, and fat mass were similar but FGR-S neonates were shorter (P = 0.0049) with lower lean mass (P = 0.0258). Doppler velocimetry abnormalities in fetuses with an EFW < 10th% identified neonates who were smaller at birth and demonstrated catch-up growth by 6-8 weeks of life that favored fat mass accretion over lean mass and linear growth.


A Comparison of CT-Guided Bone Biopsy and Fluoroscopic-Guided Disc Aspiration as Diagnostic Methods in the Management of Spondylodiscitis.
Percutaneous tissue biopsy is a key step in the diagnosis and management of spondylodiscitis, often utilizing CT-guided bone biopsy or fluoroscopic-guided disc aspirations. Our objective was to compare radiation exposure, procedure time, sedation requirement & yield between the two modalities. 103 patients in 2 cohorts underwent fluoroscopic-guided disc aspirations (n = 47) or CTguided bone biopsy (n = 46) for diagnosis of spondylodiscitis. Patient and imaging data were gathered to ensure matched cohorts. Interventional and post-procedural data included radiation exposure, procedure time, complications, and microbiological details. Yield was calculated using MRI findings as the gold standard for infection. There were no significant differences between cohorts in demographics, symptom duration, or pre-procedure antibiotics use. CT-guided bone biopsy required more general anesthesia (26% vs 0%, P < 0.001), had longer radiation exposure time (60 ± 24s vs 2 ± 3s, P < 0.001), radiation dose (114.4 ± 71.6 mGy vs 70.4 ± 147.2 mGy), and procedure time (62 ± 14m vs 31 ± 23m, P < 0.001) than fluoroscopic-guided disc aspirations. There was no significant difference in yield (34% vs 32%, P = 0.661), and it was not affected by antibiotic use. Both modalities have similar utility in isolating causative organisms in suspected cases of spondylodiscitis. Our results suggest that increased radiation exposure, longer procedure time, and increased anesthesia use are relative disadvantages of CT-guided biopsy without an increase in yield. Controlled trials may be beneficial in determining the optimal choice in different scenarios.


Whole-body calorimetry studies in adult men. 2. The interaction of exercise and over-feeding on the thermic effect of a meal.
Studies have claimed that an enhancement of the thermic effect of a meal (TEM) is an important adaptive mechanism to account for energy wastage during over-feeding. Eight healthy normal-weight young men were studied during 1 week on a weight-maintenance diet and again during 1 week when they were over-fed by 50% with fat. During each experimental week, the subject occupied a whole-body indirect calorimeter at 26 degrees for two separate periods of 36 h. The periods differed in the amount of exercise they contained. The thermic responses to the identical meals were measured during rest on one occasion and during exercise on a bicycle ergometer on the other. On the maintenance diet the absolute TEM (kJ/min) was 1.51 (SD 0.42) at rest and 1.31 (SD 0.75) during exercise (no significant difference). The equivalent values (kJ/min) on the over-feeding diet were 2.2 (SD 0.48) and 1.97 (SD 0.64) (no significant difference). The absence of a significant interaction between TEM and exercise was also demonstrated by the fact that the effect of over-feeding on total 24 h energy expenditure was unaffected by the subject's level of physical activity while in the calorimeter. In conclusion, the present study has provided no evidence to support the hypothesis that TEM is enhanced during exercise.


Nonlinear behavior of search strategies for identifying relevant orthodontic articles.
The communication process with electronic literature databases permitting a freetext search is prone in part to nonlinear, chaotic behavior. Even very minor changes in the initial conditions (search query) have a dramatic impact on the concluding event (search outcome). This statement has been verified with reference to alterations to the term "orthodontic" in search strategies of published systematic reviews. The results showed that a variation in one letter led to an average of 152 papers, 81% of them of orthodontic relevance, failing to be localized. Yet, the opposite effect, the elimination of papers (-418), was also observed, confirming the underlying nonlinear pattern. Search queries for orthodontic articles should invariably be equipped with the robust truncation "orthodon*." Truncation variants of key words should be used as a matter of principle to verify the outcome.


Protective Value of Aspirin Loading Dose on Left Ventricular Remodeling After ST-Elevation Myocardial Infarction.
Left ventricular (LV) remodeling after ST-elevation myocardial infarction (STEMI) is a complex process, defined as changes of LV volumes over time. CMR feature tracking analysis (CMR-FT) offers an accurate quantitative assessment of LV wall deformation and myocardial contractile function. This study aimed to evaluate the role of myocardial strain parameters in predicting LV remodeling and to investigate the effect of Aspirin (ASA) dose before primary coronary angioplasty (pPCI) on myocardial injury and early LV remodeling. Seventy-eight patients undergoing CMR, within 9 days from symptom onset and after 6 months, were enrolled in this cohort retrospective study. We divided the study population into three groups based on a revised Bullock's classification and we evaluated the role of baseline CMR features in predicting early LV remodeling. Regarding CMR strain analysis, worse global circumferential and longitudinal strain (GCS and GLS) values were associated with adverse LV remodeling. Patients were also divided based on pre-pPCI ASA dosage. Significant differences were detected in patients receiving ASA 500 mg dose before pPCI, which showed lower infarct size extent and better strain values compared to those treated with ASA 250 mg. The stepwise multivariate logistic regression analysis, adjusted for covariates, indicated that a 500 mg ASA dose remained an inverse independent predictor of early adverse LV remodeling. GCS and GLS have high specificity to detect early LV adverse remodeling. We first reported a protective effect of ASA loading dose of 500 mg before pPCI on LV myocardial damage and in reducing early LV adverse remodeling.


Multiple primary cancers in patients with malignant melanoma.
The incidence of second primary tumours not related to the skin in patients with malignant melanoma was determined. This was compared with the corresponding incidence of a first primary tumour in the general age-matched population. The available data indicate that, in patients with malignant melanoma, there exists a predisposition to developing a second unrelated cancer.


Aryl hydrocarbon receptor gene transitions (c.-742C>T; c.1661G>A) and idiopathic male infertility: a case-control study with in silico and meta-analysis.
Aryl hydrocarbon receptor (AHR) is responsible for crucial events in male reproductive biology. Here, the association of the AHR transitions c.-742C>T and c.1661G>A with idiopathic male infertility was investigated in a case-control study, which is followed by a meta-analysis and a bioinformatic investigation. Blood and semen samples were obtained from a total of 135 idiopathic infertile men and 130 healthy controls. Participants were genotyped for the transitions using a PCR-RFLP method. A meta-analysis of five sets of data evaluated the association of c.1661G>A with male infertility, and using an in silico analysis, the possible molecular effects of the transitions predicted. Genotypes and alleles of AHR-c.-742C>T and c.1661G>A polymorphisms were not associated with the risk of male infertility significantly. However, the frequency of C/A haplotype was significantly associated with the increased risk of male infertility, and T/A haplotype was higher among controls significantly. Also, the frequencies of combined genotypes CT/GG, CT/GA and TT/GG were significantly associated with decreased risk of infertility. And, the meta-analysis showed that the AA versus GA/GG recessive model is associated with decreased risk of male infertility among the Iranian population. In silico analysis predicted that c.-742C>T does not alter the binding sites of the proposed transcription factors, but c.1661G>A poses a tolerable structural disturbance in AHR protein. In conclusion, these results showed that AHR c.-742C>T and c.1661G>A transitions separately could not be nominated as a risk or protective factor for male infertility. However, some combined models could affect infertility risk, especially among Iranian men.


Subtrochanteric Fracture of the Femur Accompanying Pre-existing Ipsilateral Osteoarthritis of the Hip Successfully Treated with Intramedullary Nailing in the Lateral Decubitus Position: A Case Report.
A subtrochanteric fracture of the femur accompanying pre-existing osteoarthritis of the ipsilateral hip is rare. A deformity of the hip joint complicates the insertion of the intramedullary nail and varus malreduction is anticipated when surgery is performed on a fracture table with a perineal post. We report a successful case of intramedullary fixation performed in the lateral decubitus position and discuss the importance of avoiding varus and the superiority of the lateral position in surgery.


Proneural genes define ground-state rules to regulate neurogenic patterning and cortical folding.
Asymmetric neuronal expansion is thought to drive evolutionary transitions between lissencephalic and gyrencephalic cerebral cortices. We report that Neurog2 and Ascl1 proneural genes together sustain neurogenic continuity and lissencephaly in rodent cortices. Using transgenic reporter mice and human cerebral organoids, we found that Neurog2 and Ascl1 expression defines a continuum of four lineage-biased neural progenitor cell (NPC) pools. Double+ NPCs, at the hierarchical apex, are least lineage restricted due to Neurog2-Ascl1 cross-repression and display unique features of multipotency (more open chromatin, complex gene regulatory network, G2 pausing). Strikingly, selectively eliminating double+ NPCs by crossing Neurog2-Ascl1 split-Cre mice with diphtheria toxin-dependent "deleter" strains locally disrupts Notch signaling, perturbs neurogenic symmetry, and triggers cortical folding. In support of our discovery that double+ NPCs are Notch-ligand-expressing "niche" cells that control neurogenic periodicity and cortical folding, NEUROG2, ASCL1, and HES1 transcript distribution is modular (adjacent high/low zones) in gyrencephalic macaque cortices, prefiguring future folds.


Time course of expression of neuropeptide Y, calcitonin gene-related peptide, and NADPH diaphorase activity in neurons of the developing murine bowel and the appearance of 5-hydroxytryptamine in mucosal enterochromaffin cells.
Serotoninergic and cholinergic neurons are known to appear earlier in the ontogeny (day E12) of the murine gut than those containing substance P or vasoactive intestinal peptide (day E14). It has also been demonstrated that proliferating neural precursors coexist with mature neurons in developing enteric ganglia. These observations have led to the hypotheses that peptidergic neurons develop later than those that utilize small molecule neurotransmitters and that the activity of early developing neurons may affect the phenotypic expression of coexisting neuroblasts. As a partial test of these hypotheses we studied the phenotypic expression of neurons recognized by antisera to neuropeptide Y (NPY) and calcitonin gene-related peptide (CGRP), and of those visualized by the histochemical demonstration of reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity. NADPH diaphorase activity, which is coexpressed with NPY immunoreactivity in all submucosal and many myenteric neurons, was first found on day E11 in clusters of cells in the dorsal mesogastrium. These cells also expressed neurofilament reactivity and thus were developing along a neuronal lineage. Enteric neurons that expressed NADPH diaphorase activity were visualized in the stomach one day later, on day E12. At this time, NADPH diaphorase-containing cells could no longer be demonstrated in the dorsal mesogastrium. NPY immunoreactivity first appeared in the wall of the bowel on day E12, when it was seen in cells in the presumptive stomach. By day E13, the entire length of the bowel contained NPY-immunoreactive neurons. Cells that displayed NADPH diaphorase activity were found at this time at both ends of the alimentary tract, but did not appear in the ileum until day E18. In contrast, CGRP immunoreactivity could not be detected anywhere in the gut until day E17, but by day E18 all regions of the bowel contained CGRP-immunoreactive neurons. Endogenous 5-HT was first detected at day E16 in mucosal epithelial cells in all segments of the gut except the stomach, where it appeared at day E18. The NPY/NADPH diaphorase set of neurons thus develop before the acquisition of a detectable level of endogenous 5-HT or enteric neural 5-HT receptors (which arise in the foregut at day E14). These observations demonstrate that enteric neurons that express small molecule neurotransmitters do not necessarily develop earlier than peptidergic neurons as a class; however, various types of enteric neurons do appear in a sequential order.(ABSTRACT TRUNCATED AT 400 WORDS)


Health information and promotion in chiropractic clinics.
To explore the current health education behaviors of chiropractors, ascertain their willingness to provide patient counseling, and compare this with topics of interest to chiropractic patients. This study involved a postal survey of 400 randomly selected members of the Chiropractic Association of Australia (35% response rate) and a semistructured interview of 316 patients attending one of 20 purposively selected chiropractic practices. Data were collated, and the current health information practices of chiropractic respondents and their willingness to undertake counseling on various topics was identified and compared with the information interests of participating patients. Particular emphasis was placed on injury prevention both with respect to patient counseling and chiropractic practice risk. Respondents expressed varying degrees of willingness to provide health information on diverse topics, but no clear health education chiropractic practice pattern emerged. Although expressing willingness to undertake counseling, respondents were more likely to provide health information brochures than develop a tailored health promotion contract. Health education topics ranged from exercise (91%) to osteoporosis prevention (23%). Seventy-eight percent of chiropractors were prepared to offer counseling on injury prevention, yet 45% of respondents themselves reported having some work-related injury. Maintenance care failed to emerge as a global term for describing a common core of topics or chiropractic health education practices. This study demonstrated an interest by chiropractors in providing and by chiropractic patients in obtaining health information that extends beyond spinal health. The range of relevant topics covered and modes used for health information transmission in chiropractic practice requires clarification. The prevalence of work-related injury among chiropractors suggests a need to develop safe chiropractic clinical practice protocols.


Effect of tamoxifen treatment at adolescent age on the sexual behaviour and steroid hormone receptor binding of adult female rats.
Hormonal imprinting takes place perinatally, at the first encounter between the target hormone and its developing receptor. However, there is a secondary critical period of imprinting at puberty. In these periods molecules similar to the hormones (members of the same hormone family, antagonists, certain environmental pollutants, etc.) can cause faulty imprinting with lifelong consequences. In the present experiments 5+2 days of tamoxifen treatment (120 microg/day) at adolescent age dramatically (from approx. 40% to 10%) reduced the sexual activity (Meyerson index and lordosis quotient) of female rats, soon after the finishment of the treatment and between four to six weeks after treatment. Similar results were observed in animals neonatally treated with allylestrenol and tamoxifen treated at puberty. Thymic glucocorticoid receptor and uterine estrogen receptor binding capacity were not influenced.


Weirdo19ES is a novel singleton mycobacteriophage that selects for glycolipid deficient phage-resistant M. smegmatis mutants.
The sequencing and bioinformatics analysis of bacteriophages infecting mycobacteria has yielded a large amount of information on their evolution, including that on their environmental propagation on other genera such as Gordonia, closely related to Mycobacterium. However, little is known on mycobacteriophages cell biology such as the nature of their receptor(s) or their replication cycle. As part of our on-going screening for novel mycobacteriophages, we herein report the isolation and genome bioinformatics analysis of Weirdo19ES, a singleton Siphoviridae temperate mycobacteriophage with a 70.19% GC content. Nucleotide and protein sequence comparison to actinobacteriophage databases revealed that Weirdo19ES shows low homology to Gordonia phage Ruthy and mycobacteriophages falling in clusters Q and G and to singleton DS6A.Weirdo19ES also displays uncommon features such as a very short Lysin A gene (with only one enzymatic domain) and two putative HNH endonucleases. Mycobacterium smegmatis mutants resistant to Weirdo19ES are cross- resistant to I3. In agreement with that phenotype, analysis of cell envelope of those mutants showed that Weirdo19ES shares receptors with the transducing mycobacteriophage I3.This singleton mycobacteriophage adds up to the uncommonness of local mycobacteriophages previously isolated by our group and helps understanding the nature of mycobacteriophage receptors.


Care of the patient post-abdominal aortic aneurysm repair.
Caring for the patient post-abdominal aortic aneurysm repair requires knowledge of pathophysiology and surgical procedure. By understanding these, the PACU nurse will be able to set appropriate patient priorities and optimize patient outcome.


Bowlby's children: the forgotten revolution in Australian children's nursing.
Children's hospitals are vastly different today from fifty years ago. Although there have been dramatic changes in treatment and environment, the biggest contrast for patients is the involvement of parents and family in the nursing and care of the children. This change is largely due to the work of two men from Great Britain, Dr John Bowlby and James Robertson, whose research findings changed the way children were nursed to include consideration of their psychological alongside physical needs. This caused a revolution in the nursing of children that spread throughout Australasia. Bowlby and Robertson's work is largely forgotten now, but it forms the basis for the current policy of nursing children within the context of the family. This paper includes excerpts from an Australian oral history collection of twenty-six narratives from former child patients, parents and nurses and the personal papers of Dr Bowlby.


[Immunization of ewes against experimental Brucella melitensis infection: comparison of eleven vaccines].
A comparative study of 11 antibrucellic vaccines has been carried out in the Limousin region of 360 ewe-lambs divided into groups of 30, a 12th group serving as control. After being vaccinated at 8 months, the ewe-lambs were covered between 9 1/2 and 10 1/2 months, then infected at 11 months with Br. melitensis strain 53 H 38 by instillation of 4 x 10(6) germs on the conjonctiva. Six of these vaccines reduced the number of abortions and the excretion of brucella at parturition: three consisted of an inactivated suspension of a virulent S strain of Br. melitensis in oily adjuvant, another consisted of an inactivated suspension of a modified non-agglutinogenic strain of Br. melitensis also in oily adjuvant, the fifth was prepared from Br. melitensis H 105 and Br. abortus B 112 with saponin and the sixth from a strain of Br. melitensis modified with aluminium hydroxide as adjuvant.


Antiproliferative and metabolic effects of metformin in a preoperative window clinical trial for endometrial cancer.
We conducted a preoperative window study of metformin in endometrial cancer (EC) patients and evaluated its antiproliferative, molecular and metabolic effects. Twenty obese women with endometrioid EC were treated with metformin (850 mg) daily for up to 4 weeks prior to surgical staging. Expression of the proliferation marker Ki-67, estrogen receptor (ER), progesterone receptor (PR), adenosine monophosphate-activated protein kinase (AMPK), and downstream targets of the mammalian target of rapamycin (mTOR) pathway were measured by immunohistochemistry. Global, untargeted metabolomics analysis of serum pre- and postmetformin treatment, and matched tumor, was performed. Metformin reduced proliferation by 11.75% (P = 0.008) based on the comparison of pre- and posttreatment endometrial tumors. A total of 65% of patients responded to metformin as defined by a decrease in Ki-67 staining in their endometrial tumors post-treatment. Metformin decreased expression of phosphorylated (p)-AMPK (P = 0.00001), p-Akt (P = 0.0002), p-S6 (51.2%, P = 0.0002), p-4E-BP-1 (P = 0.001), and ER (P = 0.0002) but not PR expression. Metabolomic profiling of serum indicated that responders versus nonresponders to treatment were more sensitive to metformin's effects on induction of lipolysis, which correlated with increased fatty acid oxidation and glycogen metabolism in matched tumors. In conclusion, metformin reduced tumor proliferation in a pre-operative window study in obese EC patients, with dramatic effects on inhibition of the mTOR pathway. Metformin induced a shift in lipid and glycogen metabolism that was more pronounced in the serum and tumors of responders versus nonresponders to treatment.This study provides support for therapeutic clinical trials of metformin in obese patients with EC.


Cortical Activity during Perception of Musical Rhythm; Comparing Musicians and Non-musicians.
This study investigates the effects of musical training on brain activity to violations of rhythmic expectancies. We recorded behavioral and event-related brain potential (ERP) responses of musicians and non-musicians to discrepancies of rhythm between pairs of unfamiliar melodies based on Western classical rules. Rhythm deviations in the second melody involved prolongation of a note, thus creating a delay in the subsequent note; the duration of the second note was consequently shorter because the offset time was unchanged. In the first melody, on the other hand, the two notes were of equal duration. Musicians detected rhythm deviations significantly better than non-musicians. A negative auditory cortical potential in response to the omitted stimulus was observed at a latency of 150-250 ms from where the note should have been. There were no significant differences of amplitude or latency between musicians and non-musicians. In contrast, the N100 and P200 to the delayed note after the omission were significantly greater in amplitude in musicians compared to non-musicians especially in frontal and frontal-central areas. These findings indicate that long term musical training enhances brain cortical activities involved in processing temporal irregularities of unfamiliar melodies.


A novel role of protein kinase C-delta in cell signaling triggered by glutathione depletion.
Current evidence demonstrates that protein kinase C (PKC) belongs to a group of cell-signaling molecules that are sensitive targets for redox modifications and functional alterations that mediate oxidant-induced cellular responses. Our studies have demonstrated that diminished intracellular GSH was associated to inactivation of classic isoforms and increased activity of novel PKCs, and triggered molecular signals important for cell survival. Loss of GSH and oxidative damage are probably an early signaling event in apoptotic death, which is characterized by the activation of PKC-delta. Apoptotic process consequent to GSH depletion was inhibited by rottlerin, a PKC-delta-specific inhibitor, which exerted a negative effect on oxyradical production. Therefore, it may be concluded that PKC-delta activity is related to reactive oxygen species production and is involved in the pathway leading to apoptosis and growth arrest.


Risk factors associated with multiple hospital readmissions.
The percentage of multiple hospital readmissions averages between 21% and 27% in the United States today. The reasons for this readmission rate and, more important, how readmissions can be prevented, are not clear. In this integrative review we examine 13 research articles in an attempt to identify specific factors leading to the readmission of medical patients. Risk factors continually researched throughout the articles were dependence, patient age, stage of illness, length of hospital stay, prior hospitalization, care after discharge, and mobility status. Congestive heart failure and chronic obstructive pulmonary disease were the medical conditions responsible for most readmissions. No single factor was found to universally predict readmission, although several items were found to be statistically significant.


UV-photodegradation of desipramine: Impact of concentration, pH and temperature on formation of products including their biodegradability and toxicity.
Desipramine (DMI) is a widely used tricyclic antidepressant, and it is the major metabolite of imipramine (IMI) and lofepramine (LMI); IMI and LMI are two of the most commonly used tricyclic antidepressants. If DMI enters the aquatic environment, it can be transformed by the environmental bacteria or UV radiation. Therefore, photolysis of DMI in water was performed using a simulated sunlight Xenon-lamp and a UV-lamp. Subsequently, the biodegradability of DMI and its photo-transformation products (PTPs) formed during its UV photolysis was studied. The influence of variable conditions, such as initial DMI concentration, solution pH, and temperature, on DMI UV photolysis behavior was also studied. The degree of mineralization of DMI and its PTPs was monitored. A Shimadzu HPLC-UV apparatus was used to follow the kinetic profile of DMI during UV-irradiation; after that, ion-trap and high-resolution mass spectrometry coupled with chromatography were used to monitor and identify the possible PTPs. The environmentally relevant properties and selected toxicity properties of DMI and the non-biodegradable PTPs were predicted using different QSAR models. DMI underwent UV photolysis with first-order kinetics. Quantum yields were very low. DOC values indicated that DMI formed new PTPs and was not completely mineralized. Analysis by means of high-resolution mass spectrometry revealed that the photolysis of DMI followed three main photolysis pathways: isomerization, hydroxylation, and ring opening. The photolysis rate was inversely proportional to initial DMI concentration. The pH showed a significant impact on the photolysis rate of DMI, and on the PTPs in terms of both formation kinetics and mechanisms. Although temperature was expected to increase the photolysis rate, it showed a non-significant impact in this study. Results from biodegradation tests and QSAR analysis revealed that DMI and its PTPs are not readily biodegradable and that some PTPs may be human and/or eco-toxic, so they may pose a risk to the environment.


Partnerships to Facilitate Total Force Fitness.
Total force fitness (TFF) is a multi-domain framework designed to assess, promote, and sustain human performance optimization across a service member's career arc. During the September 2021, TFF Summit sponsored by the Consortium for Health and Military Performance (CHAMP), a working group explored partnerships that could facilitate effective implementation and sustainment of TFF principles. Many potential partners were identified, both internal and external to the DoD. This report highlights some positions and organizations that provide the highest yield for successfully inculcating TFF across the services.


Defining the Biologically Plausible Taxonomic Domain of Applicability of an Adverse Outcome Pathway: A Case Study Linking Nicotinic Acetylcholine Receptor Activation to Colony Death.
For the majority of developed adverse outcome pathways (AOPs), the taxonomic domain of applicability (tDOA) is typically narrowly defined with a single or a handful of species. Defining the tDOA of an AOP is critical for use in regulatory decision-making, particularly when considering protection of untested species. Structural and functional conservation are two elements that can be considered when defining the tDOA. Publicly accessible bioinformatics approaches, such as the Sequence Alignment to Predict Across Species Susceptibility (SeqAPASS) tool, take advantage of existing and growing databases of protein sequence and structural information to provide lines of evidence toward structural conservation of key events (KEs) and KE relationships (KERs) of an AOP. It is anticipated that SeqAPASS results could readily be combined with data derived from empirical toxicity studies to provide evidence of both structural and functional conservation, to define the tDOA for KEs, KERs, and AOPs. Such data could be incorporated in the AOP-Wiki as lines of evidence toward biological plausibility for the tDOA. We present a case study describing the process of using bioinformatics to define the tDOA of an AOP using an AOP linking the activation of the nicotinic acetylcholine receptor to colony death/failure in Apis mellifera. Although the AOP was developed to gain a particular biological understanding relative to A. mellifera health, applicability to other Apis bees, as well as non-Apis bees, has yet to be defined. The present study demonstrates how bioinformatics can be utilized to rapidly take advantage of existing protein sequence and structural knowledge to enhance and inform the tDOA of KEs, KERs, and AOPs, focusing on providing evidence of structural conservation across species. Environ Toxicol Chem 2023;42:71-87. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.


A novel resistance mutation in eccC5 of the ESX-5 secretion system confers ofloxacin resistance in Mycobacterium tuberculosis.
Fluoroquinolone resistance in Mycobacterium tuberculosis is often conferred by DNA gyrase mutations. However, a substantial proportion of fluoroquinolone-resistant M. tuberculosis isolates do not have such mutations. Ofloxacin-resistant and lineage-matched ofloxacin-susceptible M. tuberculosis isolates underwent WGS. Novel candidate resistance mutations were confirmed by Sanger sequencing and conferral of resistance was assessed via site-directed mutagenesis and allelic exchange. Ofloxacin MIC was determined by resazurin microtitre assay (REMA) and the effects on MICs of efflux pump inhibitors (CCCP, reserpine and verapamil) were determined. Of 26 ofloxacin-resistant isolates, 8 (31%) did not have resistance-conferring DNA gyrase mutations. The V762G mutation in Rv1783 (eccC5, encoding a protein in the ESX-5 membrane complex secretion system) was present on WGS in 8/26 (31%) resistant isolates and 0/11 susceptible isolates (P = 0.005). The mutation was identified in five isolates without DNA gyrase mutations and three isolates with such mutations; it was identified in both European-American and East Asian M. tuberculosis lineages. The ofloxacin MIC increased from 1 to 32 mg/L after introduction of the V762G mutation into M. tuberculosis H37Rv. In this strain with the V762G mutation, ofloxacin MIC did not change in the presence of efflux pump inhibitors. A novel V762G mutation in Rv1783 conferred ofloxacin resistance in M. tuberculosis by a mechanism other than drug efflux. This occurred in a substantial proportion of resistant isolates, particularly those without DNA gyrase mutations.


Upfront Surgery Versus Definitive Radiotherapy: Competing Risk Analyses in Early Stage Oropharyngeal Cancer.
To compare the survival outcomes of early-stage oropharyngeal cancer (OPC) patients treated with upfront surgery versus definitive radiotherapy (RT). Retrospective observational study. Publicly available database. A total of 1877 patients with T1-2N0-1M0 OPC were retrieved from the Surveillance, Epidemiology, and End Results database. Primary endpoints were cancer-specific and noncancer mortalities, which were estimated using cumulative incidence function and compared by Gray's test. Univariate and multivariate Fine-Gray subdistribution hazard models were used to estimate the effects of treatment modality on mortality. Subgroup analyses were performed in propensity-score-matched cohorts. All the analyses were conducted separately in human papillomavirus (HPV)-negative and HPV-positive cohorts. In the HPV-negative cohort, definitive RT was independently associated with increased risk of cancer-specific mortality (adjusted subdistribution hazard ratio [SHR], 2.29; 95% confidence interval [CI], 1.42-3.68; p = .001) and noncancer mortality (adjusted SHR, 2.74; 95% CI, 1.50-5.02; p = .001). In the HPV-positive cohort, definitive RT and upfront surgery could achieve similar cancer-specific and noncancer survival outcomes. Upfront surgery is associated with lower cancer-specific and noncancer mortality in HPV-negative early-stage OPC patients. However, in the setting of HPV-positive early-stage OPC with better prognosis, the 2 treatment modalities have similar efficacy in terms of cancer-specific and noncancer survival outcomes. In the future, carefully designed prospective clinical trials are needed to confirm our findings.


Black grape and garlic extracts protect against cyclosporine a nephrotoxicity.
The aim of this study was to determine if the natural antioxidant foods, dried black grape and garlic, protect against cyclosporine nephrotoxicity. Forty-two Sprague-Dawley rats were given Cyclosporine A (CsA) orally for 10 days, with the antioxidant food supplementation begun 3 days before CsA treatment and continued during the study period (totaling 13 days). In each group (control, CsA alone, CsA plus black grape, CsA plus aqueous garlic extract, aqueous garlic extract alone and black grape alone), there were 7 animals. At the end of the study period, the animals were sacrificed; their kidneys were removed and prepared for biochemical and histopathological investigations. Oxidant (xanthine oxidase enzyme and malondialdehyde) and antioxidant (superoxide dismutase, glutathione peroxidase and catalase enzymes) parameters were measured in the kidney tissues of the groups. Histopathological examinations of the tissues were also performed. It has been found that CsA creates oxidant load to the kidneys through both xanthine oxidase activation and impaired antioxidant defense system, which accelerates oxidation reactions in the kidney tissue. Supplementation with either dried black grape or aqueous garlic extract led to reduced malondialdehyde level in the kidney tissue possibly, by preventing oxidant reactions. In conclusion, the results suggest that impaired oxidant/antioxidant balance may play part in the CsA-induced nephrotoxicity, and some foods with high antioxidant power may ameliorate this toxicity, in agreement with studies with antioxidant vitamins.


Disruptive technology disorder: A past, present, and future neurologic syndrome.
Based upon an analysis of 6 major historical technological advances over the last 150 years, a new syndrome, disruptive technology disorder (DTD), is introduced. DTD describes the human health ailments that accompany the implementation of disruptive technologies. Elevator sickness, railway spine, and bicycle face are representative examples. Though the underlying causative disruptive technologies may differ, many neurologic symptoms (headache, dizziness, weakness) are common to multiple DTDs. Born of technology-driven societal change, DTDs manifest as a complex interplay between biological and psychological symptoms.


AMiCUS-A Head Motion-Based Interface for Control of an Assistive Robot.
Within this work we present AMiCUS, a Human-Robot Interface that enables tetraplegics to control a multi-degree of freedom robot arm in real-time using solely head motion, empowering them to perform simple manipulation tasks independently. The article describes the hardware, software and signal processing of AMiCUS and presents the results of a volunteer study with 13 able-bodied subjects and 6 tetraplegics with severe head motion limitations. As part of the study, the subjects performed two different pick-and-place tasks. The usability was assessed with a questionnaire. The overall performance and the main control elements were evaluated with objective measures such as completion rate and interaction time. The results show that the mapping of head motion onto robot motion is intuitive and the given feedback is useful, enabling smooth, precise and efficient robot control and resulting in high user-acceptance. Furthermore, it could be demonstrated that the robot did not move unintendedly, giving a positive prognosis for safety requirements in the framework of a certification of a product prototype. On top of that, AMiCUS enabled every subject to control the robot arm, independent of prior experience and degree of head motion limitation, making the system available for a wide range of motion impaired users.


Cerebral blood flow changes after brain injury in human amyloid-beta knock-in mice.
Traumatic brain injury (TBI) is an environmental risk factor for Alzheimer's disease (AD). Increased brain concentrations of amyloid-β (Aβ) peptides and impaired cerebral blood flow (CBF) are shared pathologic features of TBI and AD and promising therapeutic targets. We used arterial spin-labeling magnetic resonance imaging to examine if CBF changes after TBI are influenced by human Aβ and amenable to simvastatin therapy. CBF was measured 3 days and 3 weeks after controlled cortical impact (CCI) injury in transgenic human Aβ-expressing APP(NLh/NLh) mice compared to murine Aβ-expressing C57Bl/6J wild types. Compared to uninjured littermates, CBF was reduced in the cortex of the injured hemisphere in both Aβ transgenics and wild types; deficits were more pronounced in the transgenic group, which exhibited injury-induced increased concentrations of human Aβ. In the hemisphere contralateral to CCI, CBF levels were stable in Aβ transgenic mice but increased in wild-type mice, both relative to uninjured littermates. Post-injury treatment of Aβ transgenic mice with simvastatin lowered brain Aβ concentrations, attenuated deficits in CBF ipsilateral to injury, restored hyperemia contralateral to injury, and reduced brain tissue loss. Future studies examining long-term effects of simvastatin therapy on CBF and chronic neurodegenerative changes after TBI are warranted.


Highly Potent, Anthracycline-based Antibody-Drug Conjugates Generated by Enzymatic, Site-specific Conjugation.
Antibody-drug conjugates (ADC) are highly potent and specific antitumor drugs, combining the specific targeting of mAbs with the potency of small-molecule toxic payloads. ADCs generated by conventional chemical conjugation yield heterogeneous mixtures with variable pharmacokinetics, stability, safety, and efficacy profiles. To address these issues, numerous site-specific conjugation technologies are currently being developed allowing the manufacturing of homogeneous ADCs with predetermined drug-to-antibody ratios. Here, we used sortase-mediated antibody conjugation (SMAC) technology to generate homogeneous ADCs based on a derivative of the highly potent anthracycline toxin PNU-159682 and a noncleavable peptide linker, using the anti-HER2 antibody trastuzumab (part of Kadcyla) and the anti-CD30 antibody cAC10 (part of Adcetris). Characterization of the resulting ADCs in vitro and in vivo showed that they were highly stable and exhibited potencies exceeding those of ADCs based on conventional tubulin-targeting payloads, such as Kadcyla and Adcetris. The data presented here suggest that such novel and highly potent ADC formats may help to increase the number of targets available to ADC approaches, by reducing the threshold levels of target expression required. Mol Cancer Ther; 16(5); 879-92. ©2017 AACR.


Morphology of epidermal melanocytes in different stages of mitosis.
The structural changes which take place in epidermal melanocytes during mitosis have been investigated in serial sections of ear skin of C57Bl mice. Melanocytes were DOPA-positive and contained melanosomes in all stages of the mitotic cycle. They retracted their dendrites in prophase so that metaphase cells assumed an almost spherical shape. Signs of new dendrite growth were seen in late telophase and fully developed dendrites first after the division was completed. It is concluded that new epidermal melanocytes are formed in the mouse by division of already differentiated melanocytes.