Cloning and expression in Escherichia coli of tryptophan genes from Streptomyces griseus IMRU 3570.
Two Sau3A fragments of Streptomyces grisues IMRU 3570 were cloned in pBR322 as a vector. One of these clones contained the genetic information needed to complement trpA and trpB mutations in Escherichia coli. The other complements trpA, trpB and trpC mutations in E. coli. Both fragments originated in the same region of the chromosome but the latter is 1 kilobase (kb) longer in the region nearest the tetracycline promoter.


Presumptive GABAergic centrifugal input to the lamprey retina: a double-labeling study with axonal tracing and GABA immunocytochemistry.
The distribution of GABA-like immunoreactivity (GABA-LI) was performed in the lamprey retinopetal system which was previously identified by either anterograde or retrograde axonal tracing methods. This study was carried out at the ultrastructural level for the retina and under both the light and electron microscope for the mesencephalic retinopetal centers (M5 and RMA). The GABA-LI was distributed in about 40% of anterogradely HRP-labeled axon terminals in the inner retina. These made synaptic contacts upon either HRP-labeled ganglion cell dendrites or mostly on GABA-LI or on immunonegative amacrine cell dendrites and somata. The other immunonegative HRP-labeled axon terminals also established synaptic contacts on amacrine cell dendrites and somata. The mesencephalic retinopetal neurons, retrogradely labeled with HRP or [3H]proline, were GABA-LI in 65% of M5 somata and only in 15% of RMA neurons. M5 and RMA retinopetal neurons and dendrites, either GABA-LI or immunonegative, were contacted: (1) asymmetrically by HRP-labeled or unlabeled axon terminals containing rounded synaptic vesicles, always immunonegative and (2) symmetrically by HRP-unlabeled axon terminals containing pleiomorphic synaptic vesicles, which were either GABA-LI or immunonegative. The role of GABA as a putative neurotransmitter in the centrifugal visual system is discussed.


Purification of recombinant proteins by chemical removal of the affinity tag.
The efficient removal of a N- or C-terminal purification tag from a fusion protein is necessary to obtain a protein in a pure and active form, ready for use in human or animal medicine. Current techniques based on enzymatic cleavage are expensive and result in the presence of additional amino acids at either end of the proteins, as well as contaminating proteases in the preparation. Here we evaluate an alternative method to the one-step affinity/protease purification process for large-scale purification. It is based upon the cyanogen bromide (CNBr) cleavage at a single methionine placed in between a histidine tag and a Plasmodium falciparum antigen. The C-terminal segment of the circumsporozoite polypeptide was expressed as a fusion protein with a histidine tag in Escherichia coli purified by Ni-NAT agarose column chromatography and subsequently cleaved by CNBr to obtain a polypeptide without any extraneous amino acids derived from the cleavage site or from the affinity purification tag. Thus, a recombinant protein is produced without the need for further purification, demonstrating that CNBr cleavage is a precise, efficient, and low-cost alternative to enzymatic digestion, and can be applied to large-scale preparations of recombinant proteins.


Pyrazolidine derivatives with heteroaryl urea as dipeptidyl peptidase IV inhibitors.
In the continuation of efforts to modify the structure of our novel DP-IV inhibitors, a series of pyrazolidine derivatives with heteroaryl urea was synthesized and evaluated for their ability to inhibit dipeptidyl peptidase IV (DP-IV).


[Severe injury in the obstetric patient: considerations for anesthesiologist and critical care specialist].
In spite of the high incidence of moderately severe or severe injury in obstetric patients, the anesthesiology literature contains few references to their diagnosis and treatment. We review the epidemiology, etiology, treatment, and prognosis of severe or moderately severe injuries in obstetric patients.


Role of the cellular prion protein in the neuron adaptation strategy to copper deficiency.
Copper transporter 1 (CTR1), cellular prion protein (PrP(C)), natural resistance-associated macrophage protein 2 (NRAMP2) and ATP7A proteins control the cell absorption and efflux of copper (Cu) ions in nervous tissues upon physiological conditions. Little is known about their regulation under reduced Cu availability, a condition underlying the onset of diffused neurodegenerative disorders. In this study, rat neuron-like cells were exposed to Cu starvation for 48 h. The activation of Caspase-3 enzymes and the impairment of Cu,Zn superoxide dismutase (Cu,Zn SOD) activity depicted the initiation of a pro-apoptotic program, preliminary to the appearance of the morphological signs of apoptosis. The transcriptional response related to Cu transport proteins has been investigated. Notably, PrP(C) transcript and protein levels were consistently elevated upon Cu deficiency. The CTR1 protein amount was stable, despite a two-fold increase in the transcript amount, meaning the activation of post-translational regulatory mechanisms. NRAMP2 and ATP7A expressions were unvaried. The up-regulated PrP(C) has been demonstrated to enhance the cell Cu uptake ability by about 50% with respect to the basal transport, and so sustain the Cu delivery to the Cu,Zn SOD cuproenzymes. Conclusively, the study suggests a pivotal role for PrP(C) in the cell adaptation to Cu limitation through a direct activity of ion uptake. In this view, the PrP(C) accumulation observed in several cancer cell lines could be interpreted as a molecular marker of cell Cu deficiency and a potential target of therapeutic interventions against disorders caused by metal imbalances.


Probing compressibility of the nuclear interior in wild-type and lamin deficient cells using microscopic imaging and computational modeling.
Mechanical properties of the cell nucleus play an important role in maintaining the integrity of the genome and controlling the cellular force balance. Irregularities in these properties have been related to disruption of a variety of force-dependent processes in the cell, such as migration, division, growth or differentiation. Characterizing mechanical properties of the cell nucleus in situ and relating these parameters to cellular phenotypes remain challenging tasks, as conventional micromanipulation techniques do not allow direct probing of intracellular structures. Here, we present a framework based on light microscopic imaging and automated mechanical modeling that enables characterization of the compressibility of the nuclear interior in situ. Based entirely on optical methods, our approach does not require application of destructive or contacting techniques and it enables measurements of a significantly larger number of cells. Compressibility, in this paper represented by Poisson's ratio ν, is determined by fitting a numerical model to experimentally observed time series of microscopic images of fluorescent cell nuclei in which bleached patterns are introduced. In a proof-of-principle study, this framework was applied to estimate ν in wild type cells and cells lacking important structural proteins of the nuclear envelope (LMNA(-/-)). Based on measurements of a large number of cells, our study revealed distinctive changes in compressibility of the nuclear interior between these two cell types. Our method allows an automated, contact-free estimation of mechanical properties of intracellular structures. Combined with knockdown and overexpression screens, it paves the way towards a high-throughput measurement of intracellular mechanical properties in functional phenotyping screens.


Histopathological evaluation and redox assessment in blood and kidney tissues in a rabbit contrast-induced nephrotoxicity model.
Contrast-induced nephropathy (CIN) is a leading cause of hospital-acquired acute kidney injury as a result of iodinated contrast-media use for diagnostic purposes. Pathophysiology remains unclear. In the present study iopromide was administered to New Zealand white rabbits without any prior intervention. Oxidative stress was assessed in blood and tissue level at three anatomical kidney areas (medullary, cortical, juxtamedullary). Histopathological evaluation was also performed. Serum creatinine and urea increased in the CIN groups over 25% at two hours after administration and returned to baseline at 48 h. In kidney tissues, a significant reduction (40%) of catalase in renal cortexes of the CIN groups was observed. Necrosis and tubular vacuolization was also noted that correlated with urea and creatinine levels. Lipid peroxidation decreased at 10 h after administration (>45%) and remained low even at 48 h. Plasma protein carbonyls were significantly increased (67%) in 2 h and dropped later. Serum levels of creatinine and urea at 24 and 48 h significantly correlated with the Total Antioxidant Activity and lipid peroxidation, respectively. Oxidative stress is shown to be involved in CIN development in the rabbit, with more pronounced effects to be confined to the cortex and outer stripe of the outer medulla.


Aquaporin-3 is involved in NLRP3-inflammasome activation contributing to the setting of inflammatory response.
Inflammasomes are large immune multiprotein complexes that tightly regulate the production of the pro-inflammatory cytokines, being dependent on cell regulatory volume mechanisms. Aquaporins (AQPs) are protein channels that facilitate the transport of water and glycerol (aquaglyceroporins) through membranes, essential for cell volume regulation. Although these membrane proteins are highly expressed in monocytes and macrophages, their role in the inflammatory process is still unclear. Here, we investigated the role of aquaglyceroporin AQP3 in NLRP3-inflammasome activation by complementary approaches based either on shRNA silencing or on AQP3 selective inhibition. The latter has been achieved using a reported potent gold-based inhibitor, Auphen. AQP3 inhibition or silencing partially blocked LPS-priming and decreased production of IL-6, proIL-1β, and TNF-α, suggesting the possible involvement of AQP3 in macrophage priming by Toll-like receptor 4 engagement. Moreover, AQP3-dependent cell reswelling increased IL-1β release through caspase-1 activation. NLRP3-inflammasome activation induced by reswelling, nigericin, and ATP was also blocked when AQP3 was inhibited or silenced. Altogether, these data point towards AQPs as potential players in the setting of the inflammatory response.


Identification of altered exosomal microRNAs and mRNAs in Alzheimer's disease.
Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by decreased memory and cognitive functions. Exosomes carry a variety of important information such as proteins, lipids, DNA and RNA of mother cells. It is reported that exosomes play critical roles in nervous system physiology and neurodegenerative diseases. However, the functions of exosomes in AD progression are not fully elucidated. In this study, we detected the expression pattern of mRNAs and miRNAs in exosomes derived from the AD and health mice. A total of 1320 mRNAs and 29 miRNAs were differentially expressed in exosomes between the two groups. Subsequently, the downregulation of Chi3l1 and upregulation of Rhog in AD mice were verified by qRT-PCR. Meanwhile, the downregulation of miR-148a-5p and upregulation of miR-27a-5p in AD group were also tested by qRT-PCR. The functions of differentially expressed mRNAs and potential target genes of miRNAs were determined by GO and KEGG analysis. According to the ceRNA hypothesis, we established an integrated ceRNA network of circRNA-lncRNA-miRNA-mRNA. In conclusion, exosomal lncRNAs, mRNAs, circRNAs and miRNAs were identified to participate in the progression of AD which might be possible biomarkers and therapeutic targets for AD.


Long-term care counselor: an electronic decision-support tool.
With the American population aging at a steady pace, the need to help individuals, families, and aging/health care professionals in making often-difficult long-term care decisions is increasing. Finding accurate, impartial information is also critically important, especially information that is personalized to the individual rather than for the general public. The Long-Term Care Counselor (LTCC) is a free and confidential, web-based, decision-support tool developed by The National Council on the Aging (NCOA) to meet this particular need. It is part of the Centers for Medicare and Medicaid Service's (CMS) long-term care information initiative and is found via the official Medicare website at http://www.medicare.gov/ longtermcare/static/ltccounselor.asp. The LTCC helps individuals, caregivers, and professionals to find information relevant to particular circumstances based on the age, health, level of activity, finances, or personal preferences of the person.


Cord blood B cells are mature in their capacity to switch to IgE-producing cells in response to interleukin-4 in vitro.
Neonatal B cells have been considered immature because of their impaired capacity to produce immunoglobulins in response to polyclonal activators in vitro. Here we demonstrate that cord blood mononuclear cells (MNC) produce normal levels of IgE in vitro when cultured in the presence of interleukin-4 (IL-4), indicating that the B cells are mature in their capacity to switch to IgE-producing cells. However, in contrast to adult peripheral blood T cells, cord blood T cells failed to produce detectable levels of IL-4 upon activation by phytohaemagglutinin (PHA) concanavalin A (Con A) or combinations of PHA and the phorbol ester TPA. Interferon-gamma (IFN-gamma) production by cord blood T cells following activation by Con A or PHA was also strongly reduced. However, high levels of IFN-gamma, significantly higher than those produced by adult T cells, were synthesized in response to combinations of PHA and TPA, indicating that IFN-gamma production by cord blood T cells is not intrinsically defective. In contrast, cord blood T cells produced levels of IL-2 that were significantly higher than those obtained by adult T cells tested in parallel. Collectively, our data indicate that the minimal levels of IgE production measured in cord blood (less than 1 U/ml) are not due to immaturity of the cord blood B cells, but may be associated with the failure of cord blood T cells to produce detectable levels of IL-4, which has been shown to be responsible for induction of IgE synthesis both in vitro and in vivo.


Barriers to medication abortion among Massachusetts public university students.
Proposed legislation in Massachusetts would require public university health centers to provide medication abortion services on campus. This study assesses need for these services by investigating current travel time, costs, wait times and insurance acceptance at off-campus, abortion-providing facilities nearest to public universities in Massachusetts. This investigation projected the total number of students obtaining medication abortions each year at the 13 Massachusetts public universities based on campus enrollment figures and age- and state-adjusted medication abortion rates. Using a cross-sectional study design, the research calculated the distance and public transit time from campuses to the nearest abortion-providing facilities. Researchers contacted facilities to determine costs, wait times and insurance acceptance. We estimate 50 to 115 of Massachusetts public university students obtain medication abortion services each month, or 600 to 1,380 each year. Students have to travel between 2 and 42 miles to reach the nearest abortion-providing facility, with a population-weighted average distance of 19 miles each way. Travel time on public transportation to reach the nearest abortion-providing facility takes between 18 and 400 hundred minutes, with a population-weighted average of 103 minutes each way. Average cost of medication abortion was $680, and average wait time to the first available appointment was 8 days. Eight of 13 abortion-providing facilities did not have weekend appointments. All of the nearest abortion-providing facilities in Massachusetts accepted Mass Health, but one nearest facility was out of state and did not. All accepted multiple private insurance plans. College students face cost, scheduling, and travel barriers to abortion care. Offering medication abortion on campus would reduce these barriers. The barriers to medication abortion experienced by students attending public universities in Massachusetts fall particularly hard on female and low-income students, who are disproportionately students of color. Offering medication abortion on campus would reduce these barriers and enhance gender and racial equity on campus.


Expression of bile duct-type cytokeratin in hepatocellular carcinoma in patients with hepatitis C virus and prior hepatitis B virus infection.
The clinicopathologic findings in patients with hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC) positive for biliary markers, those related to the hepatic progenitor cells, were investigated. Cytokeratin (CK) 19 was reactive for HCCs only in patients with prior hepatitis B virus (HBV) infection. The proportions of patients with prior HBV infection and poorly differentiated HCC were significantly higher among those with CK 19-positive HCC than among those with CK 19-negative HCC. Some HCCs develop from the hepatic progenitor cells in patients with HCV infection and prior HBV infection, which may affect the clinicopathologic findings of HCV-related HCCs.


Qualitative, quantitative and temporal study of cutting agents for cocaine and heroin over 9 years.
Forensic laboratories mainly focus on the qualification and the quantitation of the illicit drug under analysis as both aspects are used for judiciary purposes. Therefore, information related to cutting agents (adulterants and diluents) detected in illicit drugs is limited in the forensic literature. This article discusses the type and frequency of adulterants and diluents detected in more than 6000 cocaine specimens and 3000 heroin specimens, confiscated in western Switzerland from 2006 to 2014. The results show a homogeneous and quite unchanging adulteration for heroin, while for cocaine it could be characterised as heterogeneous and relatively dynamic. Furthermore, the results indicate that dilution affects more cocaine than heroin. Therefore, the results provided by this study tend to reveal differences between the respective structures of production or distribution of cocaine and heroin. This research seeks to promote the systematic analysis of cutting agents by forensic laboratories. Collecting and processing data related to the presence of cutting agents in illicit drug specimens produces relevant information to understand and to compare the structure of illicit drug markets.


Irreversibility of chilling injury in zucchini squash (Cucurbita pepo L) could be a programmed event long before the visible symptoms are evident.
Zucchini fruits were subjected to 2.5 or 10 degrees C for 16d, followed by transfer to 20 degrees C for 24h in order to evaluate the relationship between ripening pattern, measured as CO(2) evolution and ethylene (C(2)H(4)) production, and metabolic heat production (q). Chilling injury (CI) visible symptoms were evident after 8d at 2.5 degrees C, but none were recorded on fruits kept at 10 degrees C. In fruits held at 10 degrees C, q, C(2)H(4) production, and CO(2) evolution diminished in the course of 16d, whereas in those at 2.5 degrees C CO(2) evolution showed an early burst peaking at 8d. Both C(2)H(4) production and q also showed a burst at 2.5 degrees C but they started at 4 and 8d, respectively, and peaked at 12d. The results showed that irreversibility of chilling injury in zucchini could occur long before the appearance of visible symptoms, although the metabolic activity accompanying the irreversibility process was not noticeable by isothermal calorimetry.


Comparative morphology of the nervous system in three phylactolaemate bryozoans.
Though some elements of the bryozoan nervous system were discovered 180 years ago, few studies of their neuromorphology have been undertaken since that time. As a result the general picture of the bryozoan nervous system structure is incomplete in respect of details and fragmentary in respect of taxonomic coverage. The nervous system of three common European freshwater bryozoans - Cristatella mucedo, Plumatella repens (both with a horseshoe-shaped lophophore) and Fredericella sultana (with a circular lophophore) had numerous differences in the details of the structure but the general neuroarchitecture is similar. The nervous system of the zooid consists of the cerebral ganglion, a circumpharyngeal ring and lophophoral nerve tracts (horns), both sending numerous nerves to the tentacles, and the nerve plexuses of the body wall and of the gut. A number of the important details (distal branching of the additional radial nerve, pattern of distribution of nerve cells and neurites in the ganglion, etc.) were described for the first time. The number and position of the tentacle nerves in Cristatella mucedo was ascertained and suggestions about their function were made. The revealed distribution of various neuromediators in the nervous system allowed us to suggest functional affinities of some major nerves. Despite the basic similarity, both the ganglion and the lophophore nervous system in Phylactolaemata have a more complex structure than in marine bryozoans (classes Gymnolaemata and Stenolaemata). First of all, their neuronal network has a denser and more complex branching pattern: most phylactolaemates have two large nerve tracts associated with lophophore arms, they have more nerves in the tentacles, additional and basal branches emitting from the main radial nerves, etc. This, in part, can be explained by the horseshoe shape of the lophophore and a larger size of the polypide in freshwater species. The structure of the nervous system in Fredericella sultana suggests that it underwent a secondary simplification following the reduction of the lophophore arms. Colony locomotion in Cristatella mucedo is based on co-ordinated activity of two perpendicular muscle layers of the sole and the plexus of motor neurons sandwiched between them. The trigger of this activity and the co-ordination mechanism remain enigmatic.


Predicting potential reforestation areas by Quercus ilex (L.) species using machine learning algorithms: case of upper Ziz, southeastern Morocco.
The selection of appropriate areas for reforestation remains a complex task because of influence by several factors, which requires the use of new techniques. Based on the accurate outcomes obtained through machine learning in prior investigations, the current study evaluates the capacities of six machine learning techniques (MLT) for delineating optimal areas for reforestation purposes specifically targeting Quercus ilex, an important local species to protect soil and water in upper Ziz, southeast Morocco. In the initial phase, the remaining stands of Q. ilex were identified, and at each site, measurements were taken for a set of 12 geo-environmental parameters including slope, aspect, elevation, geology, distance to stream, rainfall, slope length, plan curvature, profile curvature, erodibility, soil erosion, and land use/land cover. Subsequently, six machine learning algorithms were applied to model optimal areas for reforestation. In terms of models' performance, the results were compared, and the best were obtained by Bagging (area under the curve (AUC) = 0.98) and Naive Bayes (AUC = 0.97). Extremely favorable areas represent 8% and 17% of the study area according to Bagging and NB respectively, located to the west where geological unit of Bathonian-Bajocian with low erodibility index (K) and where rainfall varies between 250 and 300 mm/year. This work provides a roadmap for decision-makers to increase the chances of successful reforestation at lower cost and in less time.


Arm swing asymmetry in Parkinson's disease measured with ultrasound based motion analysis during treadmill gait.
The reduction of arm swing during gait is a frequent phenomenon in patients with early Parkinson's disease (PD). However, the objective quantification of this clinical sign using treadmill-based gait analysis has not been systematically evaluated so far. We simultaneously measured ultrasound based limb kinematics and spatiotemporal gait parameters during treadmill walking at different speeds in 21 early PD patients in Hoehn and Yahr (HY) stage I, 19 patients with bilateral PD in HY stage II and 25 age-matched controls. Both PD groups showed a highly significant reduction of the arm swing amplitude on the more affected body side (MAS). Decomposing total arm swing resulted in a bilateral decrease of arm retroversion in both PD groups, whereas anteversion was normal on the less affected side of the HY I cohort. Early stage patients exhibited a highly significant, almost threefold increase of the arm swing asymmetry index (I(A)) compared with controls. Reduced retroversion on the MAS and increased arm swing I(A) were the independent variables with the closest association to disease status in a multivariate logistic regression analysis. We conclude that ultrasound based motion analysis during treadmill walking allows reliable investigation of asymmetric arm movements in early PD patients which attenuate with ongoing disease. Impaired active arm retroversion seems to be the earliest sign of upper extremity dysfunction in parkinsonian gait. The measurement of limb kinematics during treadmill gait can broaden our methodological line-up for the analysis of complex motor programs in movement disorders.


Effect of acute and chronic acid suppression on plasma gastrin release in the rat.
The mechanisms by which administration of the H+,K(+)-ATPase inhibitor B 831-78 or intragastric perfusion with NaHCO3 induces plasma gastrin release were studied in the rat. Experiments were performed after a washout of residual intragastric contents in fasted animals provided with chronic gastric fistulae. Acute and chronic administration of B 831-78 elevated plasma gastrin dose-dependently up to 5-6 times above control levels, while the increase was only twofold with intragastric NaHCO3 infusion despite similar neutralization of gastric acidity. The profound hypergastrinaemia induced by the H+,K(+)-ATPase inhibitor, after both acute and chronic treatment, was completely prevented or reversed by intragastric perfusion with physiological amounts of acid (0.15 N HCl, 2.5 ml/h). The hypergastrinaemia was, however, largely resistant to high doses of atropine (4.3 mumol/kg) and of the M1 selective muscarinic antagonist telenzepine (10 mumol/kg). In contrast, the modest increase in plasma gastrin induced by gastric perfusion with NaHCO3 was completely suppressed by the high atropine dose and was attenuated by small doses of atropine or telenzepine (0.01 mumol/kg and 1 mumol/kg). These results demonstrate that, in the rat, blockade of the H+,K(+)-ATPase can potently induce gastrin release in the absence of a meal. Moreover, they suggest that interruption of the negative feedback between acid and gastrin release is the main mechanism through which this class of drugs releases gastrin in the rat. Since a similar degree of gastrin release cannot be achieved by alkalinization of gastric contents, additional hormonal or neural regulatory factors may contribute to the drug-induced hypergastrinaemia.


Eurycoma longifolia increases sexual motivation in sexually naive male rats.
The aim of this study is to provide evidence on the aphrodisiac property of Eurycoma longifolia Jack. An electric grid was used as an obstruction in the electrical copulation cage in order to determine how much an aversive stimulus the sexually naive male rat for both the treated with E. longifolia Jack and control groups were willing to overcome to reach the estrous receptive female in the goal cage. The intensity of the grid current was maintained at 0.12 mA and this was the intensity in which the male rats in the control group failed to crossover to reach the goal cage. Results showed that E. longifolia Jack continued to enhance and also maintain a high level of both the total number of successful crossovers, mountings, intromissions and ejaculations during the 9-12th week observation period. In conclusion, these results further enhanced and strengthened the aphrodisiac property of E. longifolia Jack.


Relationship between HIV-RNA load in blood and semen in antiretroviral-naïve and experienced men and effect of asymptomatic sexually transmissible infections.
We assessed the correlation between HIV-RNA viral load in blood (BPVL) and seminal plasma (SPVL) in a cross-sectional cohort of 119 asymptomatic, antiretroviral-naive and experienced HIV-1 subjects (BPVL < 50 copies/mL for minimum 12 months and stable on one drug regimen). The cohort was divided into 3 groups: 2 according to the non nucleoside reverse transcriptase inhibitor (NNRTI) and protease inhibitor (PI) used, and 1 untreated group. At the initial visit, subjects were screened for gonorrhoea, chlamydia and syphilis. Blood was collected for CD4 count, BPVL, and general biochemistry and haematology. Semen was collected concurrently and SPVL determined by the NucliSens HIV-1 QT PCR (BioMerieux, Boxtel, The Netherlands). At a subsequent visit, a second semen sample was obtained and SPVL was repeated for 10 subjects on ART. All NNRTI subjects (n = 36, mean treatment 33 months +/- 14) and PI subjects (n = 45, mean treatment 31 months +/- 25) had BPVL < 50 copies/mL and SPVL < 250 copies/mL at baseline and with repeat sampling. 9/119 subjects (8%) had an asymptomatic STI; 4 cases in the treated groups and 5 in the untreated group. Treated subjects were less likely to have an STI than untreated subjects. In conclusion, asymptomatic STIs had no effect on BPVL or SPVL in either treated group and SPVL remains undetectable over time. STIs minimally increase, or do not increase, SPVL in untreated patients. Our data supports the role of ART in lowering the risk of sexual transmission of HIV-1.


Imaging properties in two-photon excitation microscopy and effects of refractive-index mismatch in thick specimens.
The detrimental effects of a refractive-index mismatch on the image formation in a two-photon microscope were investigated. Point-spread functions (PSF's) were recorded with an oil-immersion objective numerical aperture (NA) of 1.3 and a water-immersion objective NA of 1.2 in an aqueous sample at different depths. For the oil-immersion objective the enlargement of the PSF volume with increasing depth yields an axial and a lateral loss in resolution of approximately 380% and 160%, respectively, at a 90-microm depth in the sample. For the water-immersion objective no resolution decrease was found. Measurements on a thick aqueous biofilm sample shows the importance of matching the refractive index between immersion fluid and sample. With a good match, no loss in image resolution is observed.


Current state of medical-advice-seeking behaviour for symptoms of colorectal cancer: determinants of failure and delay in medical consultation.
There are few data on the factors associated with healthcare-seeking behaviour for symptoms of colorectal cancer. This study describes the determinants of failure and delay in seeking medical advice for rectal bleeding and change in bowel habit. In total, 1592 persons (56-88 years) were randomly selected from the Hunter Community Study and mailed a questionnaire. In all, 18% (60/332) of respondents experiencing rectal bleeding and 20% (39/195) reporting change in bowel habit had never consulted a doctor. The rate of delay (>1 month) for each symptom was 18% and 37%. The reasons for delay included the assumption that the symptoms were not serious or that they were benign. Triggers for seeking medical advice varied. Healthcare-seeking behaviour for rectal bleeding had not significantly improved compared with a previous community-based study. The seriousness of symptoms, importance of early detection and prompt medical consultation must be articulated in health messages to at-risk persons.


Step-by-Step Guide for Synthesis and Delamination of Ti3 C2 Tx MXene.
To advance the MXene field, it is crucial to optimize each step of the synthesis process and create a detailed, systematic guide for synthesizing high-quality MXene that can be consistently reproduced. In this study, a detailed guide is provided for an optimized synthesis of titanium carbide (Ti3 C2 Tx ) MXene using a mixture of hydrofluoric and hydrochloric acids for the selective etching of the stoichimetric-Ti3 AlC2 MAX phase and delamination of the etched multilayered Ti3 C2 Tx MXene using lithium chloride at 65 °C for 1 h with argon bubbling. The effect of different synthesis variables is investigated, including the stoichiometry of the mixed powders to synthesize Ti3 AlC2 , pre-etch impurity removal conditions, selective etching, storage, and drying of MXene multilayer powder, and the subsequent delamination conditions. The synthesis yield and the MXene film electrical conductivity are used as the two parameters to evaluate the MXene quality. Also the MXenes are characterized with scanning electron microscopy, x-ray diffraction, atomic force microscopy, and ellipsometry. The Ti3 C2 Tx film made via the optimized method shows electrical conductivity as high as ≈21,000 S/cm with a synthesis yield of up to 38 %. A detailed protocol is also provided for the Ti3 C2 Tx MXene synthesis as the supporting information for this study.


Metabolomics Signature for Non-Responders to Total Joint Replacement Surgery in Primary Osteoarthritis Patients: The Newfoundland Osteoarthritis Study.
Although total joint replacement (TJR) surgery is considered as the most effective treatment for advanced osteoarthritis (OA) patients, up to one-third of patients reported unfavorable long-term post-operative pain outcomes. We aimed to identify metabolic biomarkers to predict non-responders to TJR using a metabolomics approach. TJR patients were recruited and followed-up at least 1-year post-surgery; TJR outcomes were assessed by Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain and function subscales. Targeted metabolomic profiling was performed on plasma samples collected pre-surgery and pairwise metabolite ratios, as proxies for enzymatic reactions, were calculated. Association tests were performed between each metabolite ratio and non-responders. The metabolome-wide significance was defined as p < 2 × 10-5 . A total of 461 TJR patients due to primary OA were included in the analysis. Fifteen percent of patients were classified as pain non-responders; 16% were classified as function non-responders. Lower baseline WOMAC pain and function scores were significantly associated with pain and function non-responders, respectively (both p < 0.03). Two metabolite ratios were significantly associated with pain non-responders; acetylcarnitine (C2) to phosphatidylcholine acyl-alkyl C40:1 (PC ae C40:1) was five times higher in pain non-responders whereas phosphatidylcholine diacyl C36:4 (PC aa C36:4) to isoleucine was twenty one times lower in pain non-responders than responders (all p ≤ 1.93 × 10-5 ). One metabolite ratio, glutamine to isoleucine, was significantly lower in function non-responders than responders (eight times lower; p = 1.08 × 10-5 ). Three metabolite ratios (C2 to PC ae C40:1, PC aa C36:4, and glutamine to isoleucine) related to inflammation and muscle breakdown could be considered as novel plasma markers for predicting non-responders to TJR and warrant further investigation. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:793-802, 2020.


Cognitive vulnerability in remitted depressed children and adolescents.
The cognitive model by Beck predicts that formerly depressed individuals remain vulnerable for future depressive episodes due to the existence of stable negative schemas. Activation of those schemas may explain high relapse rates in depression. The present study investigated cognitive vulnerability in remitted depressed children and adolescents. A sample of 44 in-patient youngsters was assigned to three groups after completing the CDI at two different times: a never depressed group, a currently depressed group and a remitted depressed group. All participants received a mood induction before they were given a self-referent encoding task. The results indicated that the currently and the remitted depressed groups rated more negative words as self-descriptive than the never depressed group. On the recall task, the never depressed group showed positive information processing compared to the currently depressed and the remitted depressed groups. The currently depressed group also showed a negative recall bias compared to the never depressed group. Implications and limitations of the findings are discussed.


Response to BCNU of spheroids grown from mixtures of drug-sensitive and drug-resistant cells.
Multicellular spheroids were grown from mixtures of rat brain tumor cells sensitive (9L) and resistant (R3) to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). As shown previously, after treatment with 3 microM BCNU, percentages of each cell subpopulation in mixed-cell spheroids were estimated with the sister chromatid exchange (SCE) assay and found to be approximately the same as percentages used to initiate spheroids. The sensitivity of 9L cells in mixed-cell spheroids treated with BCNU, estimated by changes in the number of SCEs induced by treatment, decreased as the percentage of R3 cells increased. When spheroids were disaggregated into single cells before treatment, however, the number of SCEs induced in the 9L population did not decrease but remained at levels similar to those found for spheroids grown from 9L cells only. These data suggest that the cell-cell interactions that influence BCNU sensitivity in mixed cell spheroids depend on three-dimensional intercellular contact. The response of purely 9L, purely R3, and mixed-cell spheroids to BCNU was also determined using the cell survival and spheroid growth delay assays. The surviving fractions of individual spheroids treated with 40 microM BCNU were slightly greater than expected; growth delays found for mixed-cell spheroids were 2-3 days less than expected. These findings suggest that cells in mixed-cell spheroids are more resistant to BCNU than would be predicted from the sensitivities of purely 9L and R3 spheroids.


The impact of staff case manager-case management supervisor relationship on job satisfaction and retention of RN case managers.
A positive relationship between staff RN case managers and their case management supervisor significantly impacts job satisfaction and retention in case managers. Literature review supports the premise that staff need to trust their supervisor and that there is a connection between this trust and job satisfaction. Staff case managers need to have a voice at work and feel empowered, and a supervisor's leadership style can influence job satisfaction and retention in their staff.


Subunit assembly and metabolic stability of E. coli RNA polymerase.
Immunological cross-reaction was employed for identification of proteolytic fragments of E. coli RNA polymerase generated both in vitro and in vivo. Several species of partially denatured but assembled RNA polymerase were isolated, which were composed of fragments of the two large subunits, beta and beta', and the two small and intact subunits, alpha and sigma. Comparison of the rate and pathway of proteolytic cleavage in vitro of unassembled subunits, subassemblies, and intact enzymes indicated that the susceptibility of RNA polymerase subunits to proteolytic degradation was dependent on the assembly state. Using this method, degradation in vivo was found for some, but not all, of the amber fragments of beta subunit in merodiploid cells carrying both wild-type and mutant rpoB genes. Although the RNA polymerase is a metabolically stable component in exponentially growing cells of E. coli, degradation of the full-sized subunits was found in two cases, i.e., several temperature-sensitive E. coli mutants with a defect in the assembly of RNA polymerase and the stationary-phase cells of a wild-type E. coli. The in vivo degradation of RNA polymerase was indicated to be initiated by alteration of the enzyme structure.


TMPfold: A Web Tool for Predicting Stability of Transmembrane α-Helix Association.
Estimating energies of transmembrane (TM) α-helix association is essential for understanding folding of membrane proteins and formation of their functional assemblies. A new physics-based method was developed and implemented in the TMPfold web server for the calculation of the free energy of TM helix association (ΔGasc) in TM α-bundles of known structure. The method was verified using the experimental ΔGasc values for 36 TM complexes, including dimers of 10 glycophorin A mutants. The calculated free energy changes (ΔΔGasc) caused by mutations in TM helices correlated with experimental changes in the stability of 42 mutants of bacteriorhodopsin and 25 mutants of rhomboid protease. TMPfold was applied for evaluation of ΔGasc in 554 PDB structures of 85 seven-helical TM proteins and identification of stable two-helical folding intermediates. The proposed tentative paths of cotranslational helix assembly of several polytopic proteins were consistent with experimental studies of their folding. TMPfold is accessible at (https://opm.phar.umich.edu/tmpfold_server).


Unusual presentation of disseminated cutaneous leishmaniasis due to Leishmania major: case reports of four Iranian patients.
We report four disseminated cutaneous leishmaniasis(DCL) cases referred to leishmaniasis laboratory at the School of Public Health, Tehran University of Medical Sciences with multiple nodular, ulcerative and crusted lesions extended on the face, trunk, and extremities. None of the patients had any complication and historical involvement in their immunological system conditions that suggest as the criteria for DCL. Direct smears of ulcers were positive for Leishmania parasite. The parasite was isolated from the active lesions and identified as Leishmania major (L. major) using PCR-RFLP assay and sequencing analysis.


A new method for tibial torsion measurement by computerized tomography.
Computerized tomography (CT) is the gold standard technique for tibial torsion assessment. This study compared two methods of tibial torsion assessment and proposed a new method, which could be of value in cases of abnormal fibular changes. The CT-scanograms of 60 participants were assessed by using two different techniques, differed in determination of the distal tibial axis. The interobserver reliability was 0.861 and 0.863 in the first and second methods, respectively. The intraobserver reliability in both measurement methods was 0.868. We proposed a reliable method, independent of the fibular midpoint, in assessment of tibial torsion by CT.


The association of TAAAAn repeat polymorphism in sex hormone-binding protein gene with polycystic ovary syndrome in Chinese population.
TAAAAn polymorphism in sex hormone-binding globulin (SHBG) gene has been found to influence the transcriptional activity of SHBG gene in vitro, and several studies have reported variable associations of this polymorphism with serum SHBG levels in women with polycystic ovary syndrome (PCOS). We have investigated the association of TAAAAn polymorphism with PCOS in 187 women with PCOS and 176 controls; in which five alleles (6-10 repeats) and 13 genotypes were found. None of the TAAAAn alleles or genotypes occurred in significant different frequency in PCOS patients compared with controls. Serum SHBG in the PCOS group was significantly lower than those in controls (33.8 +/- 30.2 nmol/l vs. 65.58 +/- 31.12 nmol/l; P < 0.01). Serum SHBG concentrations were similar for patients with PCOS whether they displayed short or long genotypes for TAAAAn (Log10SHBG: 1.46 +/- 0.38 and 1.58 +/- 0.33, respectively). Similar results were observed for controls (Log10SHBG: 1.79 +/- 0.17 and 1.77 +/- 0.14, respectively). In contrast, serum SHBG values were negatively associated with body mass index (BMI) and homeostasis model assessment of insulin resistance in the PCOS group (B = -0.285 and -0.264, respectively; P < 0.01). Thus, the TAAAAn polymorphism in SHBG gene was not a determinant of PCOS in this population of Chinese women, whereas, serum SHBG was significantly associated with BMI and insulin resistance in these PCOS patients.


Characterization of C-X-C chemokine receptor type 5 in the cornea and role in the inflammatory response after corneal injury.
C-X-C chemokine receptor type 5 (CXCR5) regulates inflammatory responses in ocular and non-ocular tissues. However, its expression and role in the cornea are still unknown. Here, we report the expression of CXCR5 in human cornea in vitro and mouse corneas in vivo, and its functional role in corneal inflammation using C57BL/6J wild-type (CXCR5+/+) and CXCR5-deficient (CXCR5-/-) mice, topical alkali injury, clinical eye imaging, histology, immunofluorescence, PCR, qRT-PCR, and western blotting. Human corneal epithelial cells, stromal fibroblasts, and endothelial cells demonstrated CXCR5 mRNA and protein expression in PCR, and Western blot analyses, respectively. To study the functional role of CXCR5 in vivo, mice were divided into four groups: Group-1 (CXCR5+/+ alkali injured cornea; n = 30), Group-2 (CXCR5-/- alkali injured cornea; n = 30), Group-3 (CXCR5+/+ naïve cornea; n = 30), and Group-4 (CXCR5-/- naïve cornea; n = 30). Only one eye was wounded with alkali. Clinical corneal evaluation and imaging were performed before and after injury. Mice were euthanized 4 h, 3 days, or 7 days after injury, eyes were excised and used for histology, immunofluorescence, and qRT-PCR. In clinical eye examinations, CXCR5-/- mouse corneas showed ocular health akin to the naïve corneas. Alkali injured CXCR5+/+ mouse corneas showed significantly increased mRNA (p < 0.001) and protein (p < 0.01 or p < 0.0001) levels of the CXCR5 compared to the naïve corneas. Likewise, alkali injured CXCR5-/- mouse corneas showed remarkably amplified inflammation in clinical eye exams in live animals. The histological and molecular analyses of these corneas post euthanasia exhibited markedly augmented inflammatory cells in H&E staining and significant CD11b + cells in immunofluorescence (p < 0.01 or < 0.05); and tumor necrosis factor-alpha (TNFα; p < 0.05), cyclooxygenase 2 (COX-2; p < 0.0001), interleukin (IL)-1β (p < 0.0001), and IL-6 (p < 0.0001 or < 0.01) mRNA expression compared to the CXCR5+/+ mouse corneas. Interestingly, CXCR5-/- alkali injured corneas also showed altered mRNA expression of fibrotic alpha smooth muscle actin (α-SMA; p > 0.05) and angiogenic vascular endothelial growth factor (VEGF; p < 0.01) compared to the CXCR5+/+ alkali injured corneas. In summary, the CXCR5 gene is expressed in all three major layers of the cornea and appears to influence corneal inflammatory and repair events post-injury in vivo. More studies are warranted to tease the mechanistic role of CXCR5 in corneal inflammation and wound healing.


ZNF671 methylation test in cervical scrapings for cervical intraepithelial neoplasia grade 3 and cervical cancer detection.
Effective triage of high-risk human papillomavirus (hrHPV)+ women is warranted to avoid unnecessary referral and overtreatment. Molecular triage tests have recently begun to impact cervical intraepithelial neoplasia grade 3 (CIN3) or cervical cancer (CC), termed CIN3+, detection. We find that zinc finger protein 671 methylation (ZNF671m) test has superior performance for CIN3+ detection in all single molecular triage tests, including HPV16/18 genotyping, paired box gene 1 methylation (PAX1m), and ZNF671m, in the training set. Using ZNF671m test instead of Thinprep cytologic test (TCT) as a single triage strategy or as a combined triage strategy with HPV16/18 genotyping has achieved comparable sensitivity but higher specificity for CIN3+ detection among 391 hrHPV+ women in the validation set. Little attention has been paid to the women with hrHPV- status but detected CIN3+. We find that the CIN3+ risk after a negative result could be reduced further by triage using ZNF671m in hrHPV- patients.


Comparison of RNA and DNA synthesis, spontaneous and PHa induced, between blood lymphoid cells of newborn infants, older infants, and adults. A study of scintillation counting and by autoradiography.
Spontaneous and PHA induced RNA and DNA synthesis was measured qualitatively by autoradiography and quantitatively by scintillation counting in blood lymphoic cells of newborn infants, older infants, and adults. Spontaneous RNA synthesis was found in transitional cells, large phagocytic lymphoid cells and lymphocytes. Transitional cells and phagocytic lymphoid cells also synthesized DNA spontaneously. Quantitatively, spontaneous RNA synthesis was active in both newborn infants and in adults, but significantly less so in older infants. PHA stimulation for 18 h increased RNA synthesis significantly in blood lymphoid cells of newborn infants and adults, and to a much lesser but still significant degree in older infants. Spontaneous DNA synthesis was significantly greater in newborn infants than in older infants and adults. PHA stimulation for 18 h had no effect on thymidine incorporation in any of the groups studied.


Molecular cloning and characterization of lactate dehydrogenase gene 1 in the silkworm, Bombyx mori.
Lactate dehydrogenase (LDH) catalyzes the reduction of pyruvate into lactate and constitutes a major checkpoint of anaerobic glycolysis. Recently, LDH draws a great deal of attention for its potential to be used as a novel diagnostic and therapeutic target for various diseases, including cancer and malaria. Insect LDHs have been mainly identified from fruit fly and mosquitoes, but not from silkworm. In this study, a novel LDH homologue, designated as BmLDH1, was firstly identified and characterized from the silkworm, Bombyx mori. The BmLDH1 cDNA contains an open reading frame of 996 bp, and encodes a protein of 331 amino acid residues with calculated molecular mass of 36 kDa. Sequence comparison showed BmLDH1 is a highly conserved protein. RT-PCR revealed BmLDH1 is transcripted in all tissues and in all developmental stages tested, indicating its essential roles for silkworm physiology and development. The BmLDH1 gene was subcloned and expressed in E. coli, and was further characterized by Western blot and Mass Spectrometry. The expressed protein contained the LDH activity, and could be inhibited by reduced glutathione in vitro. Immunofluoresence showed that the BmLDH1 was located in the cytoplasm. The cloned BmLDH1 sequence was deposited in the GenBank (accession number EU334850).


Implementing tobacco use treatment guidelines in community health centers in Vietnam.
Vietnam has a smoking prevalence that is the second highest among Southeast Asian countries (SEACs). According to the World Health Organization (WHO), most reductions in mortality from tobacco use in the near future will be achieved through helping current users quit. Yet, largely due to a lack of research on strategies for implementing WHO-endorsed treatment guidelines in primary care settings, services to treat tobacco dependence are not readily available to smokers in low middle-income countries (LMICs) like Vietnam. The objective of this study is to conduct a cluster randomized controlled trial that compares the effectiveness of two system-level strategies for implementing evidence-based guidelines for the treatment of tobacco use in 26 public community health centers (CHCs) in Vietnam. The current study will use a cluster-randomized design and multiple data sources (patient exit interviews, provider and village health worker (VHW) surveys, and semi-structured provider/VHW interviews) to study the process of adapting and implementing clinical practice guidelines in Vietnam and theory-driven mechanisms hypothesized to explain the comparative effectiveness of the two strategies for implementation. CHCs will be randomly assigned to either of the following: (1) training plus clinical reminder system (TC) or (2) TC + referral to a VHW (TCR) for three in person counseling sessions. The primary outcome is provider adherence to tobacco use treatment guidelines. The secondary outcome is 6-month biochemically verified smoking abstinence. The proposed implementation strategies draw on evidence-based approaches and a growing literature that supports the effectiveness of integrating community health workers as members of the health care team to improve access to preventive services. We hypothesize that the value of these implementation strategies is additive and that incorporating a referral resource that allows providers to delegate the task of offering counseling (TCR) will be superior to TC alone in improving delivery of cessation assistance to smokers. The findings of this research have potential to guide large-scale adoption of promising strategies for implementing and disseminating tobacco use treatment guidelines throughout the public health system in Vietnam and will serve as a model for similar action in other LMICs. NCT01967654.


Role of interganglionic synaptic connections in the control of pedal and parapodial movements in Aplysia.
Locomotion in Aplysia can be elicited by food chemosensory, tactile, proprioceptive and nociceptive stimuli. The effects of these stimuli on cerebral B neurons, pleural neurons and motor activity in the foot were examined and the behavioral roles of identified synaptic connections among these neurons investigated in semi-intact preparations. The motor effects of intracellular stimulation of pleural neurons were determined. Sensory stimulation elicited spiking in the left giant cell (LGC) in quiescent preparations. When the LGC was spontaneously bursting, sensory stimulation caused decreased interburst intervals and increased burst durations. Following sensory stimulation, LGC firing was correlated with motor activity in the foot. Intracellular stimulation of the LGC evoked contractions in the foot and parapodia. Pleural neurons which produced EPSPs in the LGC and/or B neurons were excited by sensory stimuli that elicit locomotion and when driven caused contractions in the foot. Pleural neurons which inhibited the B neurons were excited by nociceptive stimuli that inhibit normal locomotion, and were inhibited by tactile and chemosensory stimulation of the tentacles which excited the B neurons and can elicit normal locomotion. Intracellular stimulation of the neurons which produced IPSPs in the B neurons evoked contractions in the posterior foot suggesting a motor function in nociceptive induced withdrawal. The synaptic connections between pleural and cerebral neurons are consistent with the proposed modulatory role of the pleural ganglion in locomotion, and may account for changes in locomotion with result from cerebro-pleural connective lesions.


Induction of PDGF-B in TCA-treated epidermal keratinocytes.
Trichloroacetic acid (TCA) is one of the most widely used peeling agents, and induces full necrosis of the whole epidermis, followed by reconstitution of the epidermis and the matrix of the papillary dermis. The cytotoxic effects of TCA, such as suppressing proliferation of keratinocytes and fibroblasts and protein synthesis by fibroblasts, have already been reported. However, the entire biological mechanism responsible for TCA peeling has yet to be determined. Hypothetical activation effects of TCA treatment on epidermal cells to induce production of growth factors and cytokines are examined, and are compared with its cytotoxic effects in terms of time course and applied TCA concentrations. After various periods of incubation with TCA, viability of Pam212 murine keratinocytes was investigated with MTT assay and dye exclusion assay, and production of growth factors and cytokines with reverse transcription-polymerase chain reaction (RT-PCR). Changes in platelet-derived growth factor (PDGF)-B mRNA expression and protein production in the human skin specimens after TCA application were then examined by RT-PCR and immunohistochemistry, respectively. Incubation with TCA showed cytotoxicity and induced death of Pam212 cells, depending on the incubation period and the TCA concentration. In addition, expressions of PDGF-B, tumor growth factor (TGF)-alpha, TGF- beta1 and vascular endothelial growth factor, which are the growth factors reportedly secreted from keratinocytes during wound healing, were all detected in Pam212 cells after short-term treatment with TCA. Expressions of inflammatory cytokines such as interleukin (IL)-1 and IL-10 were also induced. In TCA-treated NIH-3T3 fibroblasts, in contrast, observed was upregulation of only keratinocyte growth factor, which is reportedly secreted from fibroblasts, as well as the similar cytotoxic effect. In human skin, PDGF-B mRNA expression became significantly upregulated after TCA application, and then immediately downregulated. Immunoreactive PDGF-B in the cytoplasm of keratinocytes became detectable throughout the epidermis after TCA application, reached maximum after the peak of mRNA expression, and then declined significantly over 24 h when the epidermis became completely necrotic. The TCA-treated epidermis acts as a major source of growth factors, including PDGF-B, before undergoing full necrosis. This effect might contribute to a promotion of re-epithelialization and dermal regeneration without wound contraction and scarring.


Molecular cloning and nucleotide sequence of tuna growth hormone cDNA.
cDNA for mRNA of tuna growth hormone (GH) was cloned by screening a cDNA library constructed from tuna pituitary gland poly(A)+ RNA. The nucleotide sequence of cDNA (911 bases) revealed an open reading frame of 615 nucleotides, including a sequence (51 bases) for a possible secretory protein leader peptide. Noncoding regions were found in the nucleotide sequences up- (5'-terminal: 65 bases) and down- (3'-terminal: 231 bases) stream of the open reading frame. An amino-acid sequence deduced from the nucleotide sequence of the cDNA was identical with that determined in the purified tuna GH. Tuna GH was composed of 187 amino acids, and had a calculated molecular weight of 21,275. Amino-acid sequencing showed that there was one possible N-glycosylation site at Asn (Asn-Cys-Thr). Tuna GH showed amino-acid sequence homologies with chum salmon (67%), yellow tail (90%) and with human (32%) growth hormones.


Clean intermittent catheterization revisited.
Catheterization is the insertion of a hollow flexible tube (called a catheter) to drain the urine from the bladder and is probably one of the oldest urologic procedures, dating back 3000 years. Since 1972, when urologist Dr Jack Lapides described a procedure for performing clean intermittent catheterization, this method of bladder management has become lifesaving for an individual who cannot empty their bladder independently. If the bladder is not emptied regularly, elevated storage pressures can develop which may put the upper tracts at risk of subsequent deterioration. Intermittent catheterization (IC) has become the first-line and preferred method of drainage in patients with neurogenic lower urinary tract dysfunction, as long as dexterity or available caregiver support and body habitus allow access. However, over the past 20 years, an evolution has occurred in the technology of catheters used for IC. But with these advances, so has controversy arisen. This article is a review of the current application of IC, the current technology in catheter material and systems, differences between reusable and one-time, or single-use catheters, the cost benefit of doing IC, and the infectious benefit.


Fallopian tube cancer associated with paraneoplastic dermatomyositis -- asymptomatic multivisceral exacerbated dermatomyositis mimicking recurrent widespread malignant disease: case report.
To report an uncommon case of a recurrent episode of primarily paraneoplastic dermatomyositis which was completely disconnected from the initially triggering malignancy and manifested as a silent pure multivisceral exacerbation. A 70-year-old woman presented with a pure multivisceral episode of dermatomyositis without characteristic musculo-cutaneous symptoms one year after successful treatment of fallopian tube carcinoma with complete resolvement of a concomittant paraneoplastic dermatomyositis. The uncommon manifestation of recurrent dermatomyositis involving the lungs, spleen and liver, both adrenal glands and abdominal lymph nodes, mimicked a highly disseminated recurrence of the fallopian tube cancer. Physicians participating in the interdisciplinary tumor board were misled to opt for reinductive chemotherapy. Only histologic diagnosis obtained from multiple biopsies uncovered the inflammatory nature of the disease and spared the patient unneeded chemotherapy. Asymptomatic multivisceral dermatomyositis may mimic metastatic spread of the initially underlying malignancy and may misdirect therapeutic strategies towards inadequate antineoplastic treatment.


Arachnoid cyst with GnRH-dependent sexual precocity and growth hormone deficiency.
The coexistence of gonadotropin-releasing hormone (GnRH)-dependent sexual precocity and growth hormone deficiency in patients with arachnoid cysts is rarely reported, and its pathogenesis is not well recognized. This report describes an 11-year-old female who had a huge intracranial arachnoid cyst with initial symptoms and signs of sexual precocity. Her brain magnetic resonance imaging revealed distorted hypothalamus with a thin and stretched pituitary stalk. After treatment with cysto-peritoneal shunting and gonadotropin-releasing hormone analogue, her puberty was arrested and subnormal growth rate was observed. Catch-up growth was detected after growth hormone therapy. Hence, coexistence of gonadotropin-releasing hormone-dependent sexual precocity and growth hormone deficiency in this patient was confirmed.


Replacement of the ureter by means of a narrowed terminal ileal segment in one stage.
A proximally hypoplastic ureter causing hydronephrosis was replaced by a tightened ileal segment. The operation was performed in one stage and proved to be successful 10 years later.


Oxidative Coupling Approach to Sarpagine Alkaloids: Total Synthesis of (-)-Trinervine, Vellosimine, (+)-Normacusine B, and (-)-Alstomutinine C.
Sarpagine alkaloids are bioactive indole natural products that contain a highly rigid indole-fused 1-azabicyclo[2.2.2]octane, more than 100 members of which have been identified. Herein, a detailed examination of the intramolecular oxidative coupling between a ketone and a Weinreb amide for assembling the complex 1-azabicyclo[2.2.2]octane core structure of sarpagine family alkaloids is described. Precise late-stage manipulations of the ketone and Weinreb amide enable the divergent syntheses of (-)-trinervine, (+)-vellosimine, (+)-normacusine B, and (-)-alstomutinine C. Other notable transformations of the synthesis featured an aza-Achmatowicz/indole cyclization cascade to generate the azabicyclo[3.3.1]nonane structure, a regioselective elimination reaction to form the ethylidene motif embedded in the (+)-vellosimine and (+)-normacusine B structures, and a diastereoselective indole oxidative rearrangement to form the spirooxindole structure in (-)-alstomutinine C.


Causes of morbidity and mortality, with special reference to surgical complications, after early aneurysm operation: a prospective, one-year study from neurosurgical units in Sweden.
In the present prospective study, 6.93 of Sweden's 8.59 million inhabitants (81%) were covered by the five participating centres. All patients with verified aneurysmal SAH admitted between June 1, 1989 and May 31, 1990, were enrolled. Basically, all participating centres have the same management protocol for SAH victims, including ultra-early referral to a neurosurgical unit, followed by pan-angiography and surgery as early as logistically possible. In this presentation, 145 patients who preoperatively were in Hunt & Hess Grades I-III and who underwent surgery for a supratentorial aneurysm within 72 h after the bleed, are evaluated. Eighty-one % (117 patients) made a good recovery. The morbidity was 12% (17 patients) and the mortality 7% (11 patients). The most common cause of unfavorable outcome was surgical complications, which accounted for 8% of the total series (12 patients). A subanalysis of these cases did reveal a positive correlation to higher age and more severe SAH on CAT scan.


The increase in body mass index observed after lung volume reduction may act as surrogate marker of improved health status.
To assess the effects of lung volume reduction surgery (LVRS) on body mass index (BMI). Prospective data was collected on a series of 63 patients undergoing LVRS (bilateral in 22 patients, unilateral in 41 patients). Median age was 58 (41-70) years. The peri-operative effects of LVRS on BMI, lung function and health status (assessed by SF 36 questionnaire) were recorded at 3, 6, 12 and 24 months. We found an overall increase in BMI after LVRS, which was significant up to 2 years. These changes correlated with the changes in FEV1 (R = 0.3, P < 0.01 6 months after LVRS) and diffusing capacity for carbon monoxide (DLCO) (R = 0.5, P < 0.01 6 months after LVRS). At 6 months, when the best results in health status were found, the patients were divided in a responders group (improved SF 36 score) and a non-responders group (same or worse SF 36 score) for each of the 8 domains of the SF 36. In 6 domains the non-responders showed no increase in BMI. In 6 domains the responders showed a significant increase in BMI. LVRS significantly improves postoperative BMI, which correlates with improvements in DLCO and reflects changes in health status.


Management of massive airway hemorrhage associated with extracorporeal membrane oxygenation: A retrospective case series study.
Extracorporeal membrane oxygenation (ECMO) is an important means of treating patients with respiratory failure. Massive airway hemorrhage is a rare complication of ECMO, with high mortality. The aim of this study was to provide a reference for improving the success rate of treatment of this complication by analyzing and summarizing patient clinical data. We searched PubMed, Medline, and EMBASE databases for case reports of massive airway bleeding associated with ECMO from January 2000 to January 2022 and included one case treated at our facility. All patients were disconnected from the ventilator, and the endotracheal tube was clamped during treatment, resulting in complete airway packing for hemostasis. The clinical data of these patients were analyzed. Through searching and further screening, two works of literature reported four cases that met our inclusion criteria. Including our patient's case, five patients were included in this study (four adults and one neonate). The longest ECMO treatment time before bleeding was 14 days, and the shortest was 20 min. In all patients, conservative treatment was ineffective after a major airway hemorrhage. They were disconnected from the ventilator and the tracheal tube was clamped for 13-72 h. The four adult patients underwent bronchial artery embolization in the interventional radiology suite. All patients' bleeding stopped after treatment; they were successfully weaned off ECMO and discharged. Treatment measures to disconnect the ventilator and clamp the endotracheal tube with full support from ECMO are feasible for massive airway bleeding associated with ECMO. Early bronchial arteriography and embolization can prevent rebleeding.


Static field conformal stereotactic radiosurgery: physical techniques.
Lesions in the head that are irregularly shaped or large present challenges for radiosurgical treatment by conventional techniques. Single, large circular fields may treat normal tissue volumes. Multiple shot or multiple isocenter treatment plans provide better conformation to the target than a single field, but may be difficult to plan and/or treat. As an alternative to these conventions, we are developing static field, conformal stereotactic radiosurgery. In this technique a finite number of fixed, shaped, linear accelerator fields are used to irradiate the target. Computer simulations were performed for a four-path arc and fixed field techniques and evaluated with dose distributions and dose volume histograms. Beam geometries are defined with a 3-D treatment planning system with beam's eye view capabilities. Equipment for treatment delivery has been designed, including a head frame/support stand assembly and a method for manufacturing the required custom collimators. Isodose distributions and dose volume histograms show that beam geometries with seven or more fields provide target dose distributions equivalent to the arc treatment, but with small increases in peripheral dose. Dose homogeneity across the target volume increases as the solid angle of irradiation is increased. For a hemispherical target, the four-path arc and shaped, static fields provide equal target coverage while the shaped fields treat a smaller high-dose volume. Shaped, static fields are an alternative to single isocenter arc radiosurgery and result in smaller volumes at high dose. This smaller volume could translate into sparing for normal adjacent tissues that would otherwise be treated.


RNA1 is sufficient to mediate plasmid ColE1 incompatibility in vivo.
The multicopy plasmid ColE1 specifies a small RNA designated RNA1 that has been implicated in copy number control and incompatibility. We have inserted a 148 base-pair ColE1 DNA fragment containing a promoter-less RNA1 gene into a plasmid vector downstream from the tryptophan promoter of Serratia marcesens . The ColE1 RNA1 produced by this plasmid is not functional in vivo due to the presence of 49 nucleotides appended to the 5'-terminus of the wild-type RNA1 sequence. Deletions of these sequences by Bal3l nuclease in vitro and genetic selection for ColE1 incompatibility function in vivo permitted isolation of a plasmid expressing wild-type ColE1 RNA1 initiated properly from the S. marcesens trp promoter. These experiments demonstrate that RNA1 is sufficient to mediate ColE1 incompatibility in vivo. In addition, several plasmids were isolated that contain altered RNA1 genes. These alterations consist of additions or deletions of sequences at the 5'-terminus of RNA1. Analysis of the ability of these altered RNA1 molecules to express incompatibility in vivo suggests that the 5'-terminal region of RNA1 is crucial for its function.


Association of SLC6A4 methylation with early adversity, characteristics and outcomes in depression.
Childhood adversities have been associated with onset and worse clinical presentations of depression. Epigenetic changes may reflect childhood adversities, while their effects on clinical characteristics of depression are unknown. This study aimed to investigate whether epigenetic changes were associated with childhood adversities, pretreatment characteristics, and treatment outcomes in depressive patients. In 108 patients with major depressive disorders, the methylation status in the promoter of gene encoding serotonin transporter (SLC6A4) was measured. Childhood adversities, socio-demographic and clinical characteristics including assessment scales for depression (Hamilton Depression Rating Scale, HAMD), anxiety (Hamilton Anxiety Rating Scale, HAMA), functioning (Social and Occupational Functioning Assessment Scale, SOFAS), disability (World Health Organization Disability Assessment Schedule-12, WHODAS-12), and quality of life (World Health Organization Quality of Life-Abbreviated form, WHOQOL-BREF) were evaluated at baseline. After a 12-week treatment with antidepressants, the assessment scales were reevaluated. To avoid type I error by multiple comparisons, Bonferroni corrections were applied. Higher SLC6A4 promoter methylation status was significantly associated with childhood adversities, worse clinical presentation (family history of depression, higher perceived stress, and more severe psychopathology assessed by SOFAS, WHODAS-12, and WHOQOL-BREF), but was not associated with treatment outcomes after considering multiple comparisons. SLC6A4 methylation status could be a proxy marker for childhood adversities and a clinical biomarker for certain presentations of depression.


Enhanced performance in the determination of ibuprofen 1-β-O-acyl glucuronide in urine by combining high field asymmetric waveform ion mobility spectrometry with liquid chromatography-time-of-flight mass spectrometry.
The incorporation of a chip-based high field asymmetric waveform ion mobility spectrometry (FAIMS) separation in the ultra (high)-performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) determination of the (R/S) ibuprofen 1-β-O-acyl glucuronide metabolite in urine is reported. UHPLC-FAIMS-HRMS reduced matrix chemical noise, improved the limit of quantitation approximately two-fold and increased the linear dynamic range compared to the determination of the metabolite without FAIMS separation. A quantitative evaluation of the prototype UHPLC-FAIMS-HRMS system showed better reproducibility for the drug metabolite (%RSD 2.7%) at biologically relevant concentrations in urine. In-source collision induced dissociation of the FAIMS-selected deprotonated metabolite was used to fragment the ion prior to mass analysis, enhancing selectivity by removing co-eluting species and aiding the qualitative identification of the metabolite by increasing the signal-to-noise ratio of the fragment ions.


Needle in a haystack: Robotic removal of a retroperitoneal wire.
Retroperitoneal foreign bodies are rare indications for exploratory surgery. We present a case of a 19-year-old male with abdominal pain after a fall who was found to have a linear metallic object adjacent to the right ureter and inferior vena cava. Given the patient's pain and discomfort, he elected for robotic exploration of the retroperitoneum, which was carried out successfully with the Da Vinci Si® robot. This case demonstrates the feasibility of robotic retroperitoneal exploration and foreign body retrieval for a very small object.


Marsupialization facilitates eruption of dentigerous cyst-associated mandibular premolars in preadolescent patients.
The purpose of the present study was to investigate the eruption of dentigerous cyst (DC)-associated mandibular premolars after marsupialization in preadolescents. The present study was a retrospective cohort study of preadolescent patients with DCs who were treated as outpatients at the Department of Oral and Maxillofacial Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. For our study, the data from these patients were collected, and the eruption of the premolar teeth, along with related factors, such as the interval to eruption, cusp depth, angulation, cyst size, and eruption space, were analyzed for the cyst group compared with the noncyst control group. The mean age of the patients was 9.1 years. All teeth associated with DCs erupted successfully after marsupialization. The follow-up panoramic radiograph showed that the cysts had disappeared and had been replaced by regenerated bone. The initial panoramic radiograph showed the angulation of the teeth in the cyst group had a significantly larger inclination angle than did the teeth in the noncyst group (P < .05). However, no significant difference was found for cusp depth, root formation, or space measurement. The gender, age, cusp depth, angulation, and eruption space were not factors influencing the eruption of the DC-associated tooth for preadolescent patients in the present study. In addition, the cyst-associated teeth took less time to erupt than the teeth in the control group. The results of the present study suggest that DC-associated mandibular premolars can erupt spontaneously after marsupialization in preadolescents.


Observations on the dynamics of argyrophilic nucleolar material in the nuclei of mice spermatids.
Applying a new silver staining technique it could be shown that in very early spermatids strong argyrophilia in nucleoli is confined to their granular and fibrillar components; fibrillar centers are devoid of silver. During subsequent developmental stages remnants of these nucleolar components are present in the form of intensively silver stained clusters of coiled fibers. As chromatin condensation proceeds, these fibrous structures decrease in size and density and are finally completely absent. The nucleus of the mature sperm contains only the space in which they formerly existed, now silver negative, as the so-called 'nuclear vacuole'.


Prevention by acebutolol of rhythm disorders following coronary bypass surgery.
Seventy-one patients submitted routinely to coronary artery bypass surgery were randomized into 2 groups. Group A (32 patients) received 24 hr after initiation of surgery an intravenous perfusion of 100 mg of acebutolol given over 24 hr (22 cases) or 600 mg administered orally (10 cases). On subsequent days, they received 1200 mg of acebutolol/day orally. Group B (39 patients) was used as control. The groups were comparable in terms of age, sex, severity of coronary disease, preoperative therapy, duration of extracorporeal circulation, aortic clamping time, and immediate postoperative haemodynamic findings. No patient received digitalis. During hospital stay (10 days), 1 group A patient (3%) and 13 group B patients (33%; P less than 0.001) developed a sustained episode of atrial arrhythmia (fibrillation, flutter or atrial ectopic tachycardia). The majority of these rhythm disorders developed between days 2 and 4. On Holter monitoring on days 7-10, malignant ventricular extrasystoles (grades IV and V of Lown's classification) were more frequent in group B (65.2%) than in group A (19.3%; P less than 0.001). Haemodynamic measurements taken at rest performed in 27 patients on days 7-10 (16 patients of group A; 11 of group B). No difference was observed between the two groups. Acebutolol is a safe and efficacious drug for the prevention of arrhythmias following coronary surgery.


Chitynolytic bacteria in water and bottom sediments of two lakes of different trophy.
Chitynolytic bacteria were found in greater numbers in the eutrophic lake Jeziorak than in the mesotrophic lake Jasne. In both lakes higher chitynolytic activity was found in the planktonic than in the benthic bacteria. The capacity of chitin hydrolysis was found in bacteria of the genera Achromobacter, Bacillus, Nocardia, Pseudomonas, of the Flavobacterium-Cytophaga group and of the Enterobacteriaceae family. The chitinases synthesized by the studied bacteria were active in the pH range 5.0--8.0. The majority of the strains tested showed maximum activity at pH 5.0 or 6.0.


Urethral reconstruction in aphallia using transpubic exposure and colonic monti neo-urethra - An addition to the De Castro reconstruction.
Fashioning a functioning neo urethra in a boy with aphallia is one of the unsolved problems in this condition. We present our technique and outcome of operative exposure and neo urethral construction in four aphallic boys. Retrospective study of the records of four aphallic boys operated in the period 2015-2019 was undertaken. The demographics, presentation, operative procedure, current follow up and voiding outcome was noted. The neo phallus was constructed by the De Castro technique in four aphallic boys aged 1-8 years. Pre operative assessment revealed bladder outflow obstruction in two and urinary incontinence in one boy. The operative exposure afforded after reflection of the De Castro flap was utilized in doing a limited pubic symphysiectomy. This exposed the bladder neck, urethra and the urethral termination into the ano rectum. The healthy native urethra was isolated by disconnecting it from its ano rectal termination and a neo urethra constructed from a segment of proximal sigmoid colon by its reconfiguration into a Monti type tube. The neo urethra was then anastomosed to the native urethra and laid within the bed of the De Castro flap so as to reach the neo phallic termination. The De Castro flap was then tabularized over the neo urethra to form the new phallus. Appendicular or ileal Monti Mitrofanoff was also added to the reconstruction. Three boys are voiding well. One boy had a partial bladder outflow obstruction resulting from operative correction of incontinence and is dry on intermittent catheterization. Follow up was for 1-4 years and upper tracts are stable or improved. We describe the transpubic approach to the construction of a reliable neo urethra in the form of a Monti tube from the sigmoid colon in four boys with aphallia. We believe this to be a useful addition to the De Castro procedure for neo phallic construction.


Hierarchical structure and compressive deformation mechanisms of bighorn sheep (Ovis canadensis) horn.
Bighorn sheep (Ovis canadensis) rams hurl themselves at each other at speeds of ∼9 m/s (20 mph) to fight for dominance and mating rights. This necessitates impact resistance and energy absorption mechanisms, which stem from material-structure components in horns. In this study, the material hierarchical structure as well as correlations between the structure and mechanical properties are investigated. The major microstructural elements of horns are found as tubules and cell lamellae, which are oriented with (∼30⁰) angle with respect to each other. The cell lamellae contain keratin cells, in the shape of pancakes, possessing an average thickness of ∼2 µm and diameter of ∼20-30 µm. The morphology of keratin cells reveals the presence of keratin fibers and intermediate filaments with diameter of ∼200 nm and ∼12 nm, respectively, parallel to the cell surface. Quasi-static and high strain rate impact experiments, in different loading directions and hydration states, revealed a strong strain rate dependency for both dried and hydrated conditions. A strong anisotropy behavior was observed under impact for the dried state. The results show that the radial direction is the most preferable impact orientation because of its superior energy absorption. Detailed failure mechanisms under the aforementioned conditions are examined by bar impact recovery experiments. Shear banding, buckling of cell lamellae, and delamination in longitudinal and transverse direction were identified as the cause for strain softening under high strain rate impact. While collapse of tubules occurs in both quasi-static and impact tests, in radial and transverse directions, the former leads to more energy absorption and impact resistance. Bighorn sheep (Ovis canadensis) horns show remarkable impact resistance and energy absorption when undergoing high speed impact during the intraspecific fights. The present work illustrates the hierarchical structure of bighorn sheep horn at different length scales and investigates the energy dissipation mechanisms under different strain rates, loading orientations and hydration states. These results demonstrate how horn dissipates large amounts of energy, thus provide a new path to fabricate energy absorbent and crashworthiness engineering materials.


GENCOV: a Fortran program that generates randomly censored survival data with covariates.
We present a Fortran program for simulating censored survival data with covariates under the assumption of random censoring. The program generates times distributed according to the uniform distribution, the generalized Gamma distribution, the log-normal distribution and Pettitt's generalized logistic distribution with Box-Cox transformation of the time variable. Covariates can be introduced in the definition of the survival time, resulting in the generalized log-gamma, log-normal and Pettitt's regression models. Thereby the program provides the means for generating censored survival data according to parametric versions of three common regression models for censored survival data: the Accelerated Failure Time, the Proportional Hazards and the Proportional Odds models.


Circulating mucopolysaccharide (mucin) in two adults with metastatic adenocarcinoma. Detection in peripheral blood smear and at autopsy.
We report two cases of circulating mucin that occurred in women with metastatic adenocarcinoma. In both women, mucin was seen on the peripheral blood smear, but no difficulties were encountered in counting blood cells. In one patient, the mucin was grossly visible in the plasma and within phagocytes. Systemic mucin thromboses were detected in the second woman whose death was due to cerebral infarcts from multiple mucin emboli.


Supporting breast self-examination in female childhood cancer survivors: a secondary analysis of a behavioral intervention.
To identify intervention targets that will increase the frequency of breast self-examination (BSE) in female survivors of childhood cancer. Secondary data analysis of longitudinal clinical trial data. Outpatient clinic in a children's research hospital. 149 female survivors (aged 12-18 years) a median of 11 years after diagnosis of leukemia or lymphoma (59%) or solid tumor (41%). Paired t tests, Wilcoxon signed rank tests, repeated measures analysis of variance, and analysis of covariance. BSE frequency, health risk perceptions, motivation, and fears or worries. Baseline BSE frequency was the strongest influence on follow-up BSE. Baseline and follow-up age and school grade influenced follow-up BSE. Other influential variables included motivation for behavior change, motivation to commit to health promotion, concern about appearance, and an interaction between the intervention and mother's highest grade level. When baseline BSE frequency and school grade were statistically controlled, diagnosis and significant interactions between grade level and the follow-up measures of the mother's education, general fears about cancer, fears about cancer returning, and perceptions of susceptibility to late treatment effects were significant influences on BSE after intervention. Survivors least likely to perform BSE are fearful about cancer and are not motivated to change health behaviors. Nurses should explore survivors' fears about cancer and late treatment effects to address misconceptions, use modeling techniques with return demonstrations to ensure competency in BSE, and tailor risk information to each survivor's background (socio-economic status, age, development) and cognitive (disease and treatment knowledge, risks) and affective (fears) characteristics to increase BSE motivation.


Sarcopenia influences fall-related injuries in community-dwelling older adults.
This study aimed to determine the relationship between sarcopenia and fall-related injuries in community-dwelling older adults in Korea. The study population comprised 2848 subjects aged 65 years or older who participated in the Korea National Health and Nutrition Examination Surveys during 2010-2011. Sarcopenia was considered to be present if the subject's appendicular skeletal muscle mass divided by his or her weight was less than 29.9% in men, or less than 25.1% in women. The incidence of fall-related injuries during the past year among all the respondents was 4.3%, and the prevalence of sarcopenia was 32.2%. After controlling for sociodemographic variables and morbidity due to chronic diseases, the incidence of fall-related injuries remained significantly elevated among older adults with sarcopenia (odds ratio = 1.61, 95% confidence internal = 1.01-2.54). Sarcopenia should be considered when investigating means of preventing of fall-related injuries in community-dwelling older adults. To prevent falls among these persons, it is vital to implement intervention programs that increase muscle mass.


Potential Owner-Related Risk Factors That May Contribute to Obesity in Companion Dogs in Aotearoa New Zealand.
Approximately a third of all Aotearoa New Zealand (NZ) households include a dog, with 28% of these dogs being overweight or obese, conditions that are associated with many serious health issues. Therefore, healthy weight interventions that focus on the owner's role are of great importance to companion animal welfare in NZ. Accordingly, the present study explores the feeding practices associated with NZ dogs and identifies potential owner-related risk factors contributing to these animals being overweight or obese. The current study used data collected from a survey conducted online in 2019 between January and March of NZ residents over 18. Along with demographic questions, the respondents were asked questions regarding their dog's body condition and diet questions related to the body. Nearly a quarter (26%, n = 609) of the survey participants (n = 2358) owned at least one dog. The current study reported that increasing age range, household income and the number of children increased the likelihood of having a dog while increasing qualification level and living in a town/city decreased the likelihood. The majority of the respondents fed their dog(s) treats (59%) and 85% fed them specialised food bought from a pet shop, veterinary clinic and/or different online sources. Just over a third of the participant (39%) reported that they fed their dog(s) biscuits from the supermarket, 36% fed their dog(s) raw meat, and 34% of respondents fee their dog(s) table scraps/human food. These results suggest that many dog owners feed their dog(s) various food types, making it a challenging task to determine the exact amount required from each type in order not to exceed caloric intake. Disagreement regarding the correct body condition were reported among twenty per cent of the respondents. This finding indicates a knowledge gap among the NZ dog-owning population that may negatively affect their dogs' welfare and wellbeing. Future research into pro-equity approaches to address these issues is needed so that dogs in NZ can live not only a good life but also their best life.


3D printed carboxymethyl cellulose scaffolds for autologous growth factors delivery in wound healing.
This work aims to use carboxymethyl cellulose (CMC) as main structural and functional component of 3D printed scaffolds for healing of diabetic wounds. Differently from previous inks involving small contents in CMC, herein sterile (steam-heated) concentrated CMC solely dispersions (10-20%w/v) were screened regarding printability and fidelity properties. CMC (15%w/v)-citric acid inks showed excellent self-healing rheological properties and stability during storage. CMC scaffolds loaded with platelet rich plasma (PRP) sustained the release of relevant growth factors. CMC scaffolds both with and without PRP promoted angiogenesis in ovo, stem cell migration in vitro, and wound healing in a diabetic model in vivo. Transparent CMC scaffolds allowed direct monitoring of bilateral full-thickness wounds created in rat dorsum. CMC scaffolds facilitated re-epithelialization, granulation, and angiogenesis in full-thickness skin defects, and the performance was improved when combined with PRP. Overall, CMC is pointed out as outstanding component of active dressings for diabetic wounds.


Getting the most out of maths: How to coordinate mathematical modelling research to support a pandemic, lessons learnt from three initiatives that were part of the COVID-19 response in the UK.
In March 2020 mathematics became a key part of the scientific advice to the UK government on the pandemic response to COVID-19. Mathematical and statistical modelling provided critical information on the spread of the virus and the potential impact of different interventions. The unprecedented scale of the challenge led the epidemiological modelling community in the UK to be pushed to its limits. At the same time, mathematical modellers across the country were keen to use their knowledge and skills to support the COVID-19 modelling effort. However, this sudden great interest in epidemiological modelling needed to be coordinated to provide much-needed support, and to limit the burden on epidemiological modellers already very stretched for time. In this paper we describe three initiatives set up in the UK in spring 2020 to coordinate the mathematical sciences research community in supporting mathematical modelling of COVID-19. Each initiative had different primary aims and worked to maximise synergies between the various projects. We reflect on the lessons learnt, highlighting the key roles of pre-existing research collaborations and focal centres of coordination in contributing to the success of these initiatives. We conclude with recommendations about important ways in which the scientific research community could be better prepared for future pandemics. This manuscript was submitted as part of a theme issue on "Modelling COVID-19 and Preparedness for Future Pandemics".


Leg coordination during turning on an extremely narrow substrate in a bug, Mesocerus marginatus (Heteroptera, Coreidae).
The turning movement of a bug, Mesocerus marginatus, is observed when it walks upside-down below a horizontal beam and, at the end of the beam, performs a sharp turn by 180 degrees . The turn at the end of the beam is accomplished in three to five steps, without strong temporal coordination among legs. During the stance, leg endpoints (tarsi) run through rounded trajectories, rotating to the same side in all legs. During certain phases of the turn, a leg is strongly depressed and the tarsus crosses the midline. Swing movements rotate to the same side as do leg endpoints in stance, in strong contrast to the typical swing movements found in turns or straight walk on a flat surface. Terminal location is found after the search through a trajectory that first moves away from the body and then loops back to find substrate. When a leg during stance has crossed the midline, in the following swing movement the leg may move even stronger on the contralateral side, i.e. is stronger depressed, in contrast to swing movements in normal walking, where the leg is elevated. These results suggest that the animals apply a different control strategy compared to walking and turning on a flat surface.


A levator ani midurethral support via single vaginal incision technique to treat stress urinary incontinence: A case report.
Stress urinary incontinence affects more than one-fourth of adult women. The recommended surgical treatment involves the use of a synthetic mesh sling. Upon unsuccessful treatment using a mesh sling or when patients decline mesh, surgical treatments, including an autologous fascia sling, colposuspension, or bulking injections, are used. After unsuccessful treatment using three mesh slings, an autologous fascia sling, and a midurethral bulking agent, a patient was successfully treated with our modified Kelly plication technique. A 51-year-old woman with recurrent stress urinary incontinence had had three previous mesh midurethral sling exposures with complete mesh removals followed by one autologous fascia sling with severe infection. We initially treated her with a set of urethral bulking injections, which was also unsuccessful. She was successfully treated with our modified Kelly plication technique, which plicates levator ani muscles to support the midurethral instead of plicating the vesicovaginal fascia at the bladder neck. The patient remained continent four years after the performance of this technique and had reported no pelvic pain or dyspareunia. The technique is detailed in this paper. This single vaginal incision native tissue technique may be considered when mesh slings or alternative native tissue procedures are not feasible for patients, as in this case.


Characterization of heparanase-induced phosphatidylinositol 3-kinase-AKT activation and its integrin dependence.
Heparanase functions as a heparan sulfate-degrading enzyme and as a ligand for an unidentified signaling receptor(s). Here, several reactions involved in the activation of the PI3K-AKT pathway by latent heparanase were characterized. Protein suppression using specific siRNAs revealed that heparanase-induced phosphorylation of AKT at Ser-473 was RICTOR-mTOR-dependent, whereas ILK and PAK1/2 were dispensable. p110α was the PI3K catalytic isoform preferred by heparanase for AKT activation and cell proliferation because the p110α inhibitor YM024 blocked these processes. Heparanase-induced AKT phosphorylation was low in mouse embryonic fibroblast cells expressing a RAS interaction-defective p110α compared with wild type cells, indicating that RAS has an important role in the PI3K-AKT activation. The response to heparanase was also inefficient in suspension cultures of several cell lines, suggesting a requirement of integrins in this pathway. Adhesion via either αVβ3 or α5β1 promoted heparanase-induced AKT phosphorylation, and a stronger effect was seen when both integrins were engaged. Simultaneous inhibition of FAK and PYK2 using a chemical inhibitor, or suppression of their expression, inhibited heparanase-induced AKT activation and cell proliferation. Stimulation of cells with heparanase enhanced their resistance against oxidative stress- or growth factor starvation-induced apoptosis. These results demonstrate that there is an intimate cross-talk between the heparanase receptor(s) and integrins during induction of the prosurvival PI3K-AKT pathway by heparanase.