A method for extraction of exosomes from breast tumour cells and characterisation by transmission electron microscopy.
Exosomes can not only be used as markers of tumour metastasis but also be used for noninvasive diagnosis of clinical diseases, which holds significant medical research value. Observing the morphology and distribution of exosomes using transmission electron microscopy (TEM) is highly significant. In this study, we use breast tumour cell (MDA-MB-231) exosomes as a representative sample and focus on the extraction and purification of exosomes, as well as the investigation of optimal conditions for the observation of exosomes using TEM. Through comparative tests, we obtained the optimal dilution concentration and staining method for TEM of exosomes, the best dilution concentration is 100 times, the best negative staining time is 1.5 min. Western blotting and scanning electron microscopy (SEM) confirmed the presence of exosome. The particle size experiment shows that the size of exosomes is mainly distributed between 60 nm and 150 nm. This method provides a reference for TEM sample preparation of subcellular structures and small molecular biomaterials other than exosomes.


Biochemical characterization of a SET and transposase fusion protein, Metnase: its DNA binding and DNA cleavage activity.
Metnase (SETMAR) is a SET and transposase fusion protein that promotes in vivo end joining activity and mediates genomic integration of foreign DNA. Recent studies showed that Metnase retained most of the transposase activities, including 5'-terminal inverted repeat (TIR)-specific binding and assembly of a paired end complex, and cleavage of the 5'-end of the TIR element. Here we show that R432 within the helix-turn-helix motif is critical for sequence-specific recognition, as the R432A mutation abolishes its TIR-specific DNA binding activity. Metnase possesses a unique DNA nicking and/or endonuclease activity that mediates cleavage of duplex DNA in the absence of the TIR sequence. While the HTH motif is essential for the Metnase-TIR interaction, it is not required for its DNA cleavage activity. The DDE-like motif is crucial for its DNA cleavage action as a point mutation at this motif (D483A) abolished its DNA cleavage activity. Together, our results suggest that Metnase's DNA cleavage activity, unlike those of other eukaryotic transposases, is not coupled to its sequence-specific DNA binding.


Psychosocial impact of the COVID-19 pandemic and shielding in adults and children with early-onset neuromuscular and neurological disorders and their families: a mixed-methods study.
To describe and evaluate the psychosocial impact of the COVID-19 pandemic and measures to reduce the risk of transmission on patients with early-onset neuromuscular and neurological disorders (NMDs) and their families. A mixed-methods study in which data were collected between 17 September 2020 and 31 December 2020 using a semi-structured telephone questionnaire developed specifically to meet research aims, and were analysed using quantitative methods and qualitative inductive thematic analysis. Forty questionnaires were completed by patients with NMDs (eg, muscular dystrophies, spinal muscular atrophy) or their parent. 70% (n=28) of patients were male, aged 2-48 years. 90% (n=36) were wheelchair users; 72.5% (n=29) required long-term non-invasive or tracheostomy ventilation. Strict adherence to risk mitigation strategies, for example, shielding, were reported at the start of the pandemic. Over half continued some or all measures after official limitations were relaxed. 67.5% (n=27) reported changes to personal care assistance arrangements including temporary cessation of outside carers. Three themes were identified: (1) Concern regarding the health impact of COVID-19; (2) Perceptions of strategies to prevent SARS-CoV-2 transmission; (3) Psychological impact of the COVID-19 pandemic. The level and pervasiveness of frequently reported negative psychological effects, for example, anxiety and fear fluctuated, and were related to the perceived risk of COVID-19, concern about attending hospital, and perceived lack of access to intensive care management if severe COVID-19 infection occurred. Support, particularly from family and healthcare services, were considered to have positive psychosocial effects. Measures to reduce transmission of COVID-19 have greatly affected patients with NMDs and their families. For most, negative psychosocial impacts have and will continue to improve, but this may depend on the incidence of further pandemic waves. Consistent, up-to-date and accessible information on clinical outcomes and risk mitigation must be provided to support patients' physical and mental well-being.


Bioluminescence, photophysical, computational and molecular docking studies of fully conformationally restricted enamine infraluciferin.
A new rationally designed fully rotationally restricted luciferin has been synthesised. This synthetic luciferin, based upon the structure of infraluciferin, has two intramolecular H-bonds to reduce degrees of freedom, an amine group to enhance ICT process, and an alkenyl group to increase π-conjugation. In the spectroscopic measurements and computational calculations, enamine luciferin showed more red-shifted absorption and fluorescence emission than LH2 and iLH2. With PpyWT luciferase enamine luciferin gave bioluminescence at 564 nm which is similar to LH2 at 561 nm. Further investigation by docking studies revealed that the emission wavelength of enamine luciferin might be attributed to the unwanted twisted structure caused by Asp531 within the enzyme. With mutant luciferase FlucRed, the major emission peak was shifted to 606 nm, a distinct shoulder above 700 nm, and 21% of its spectrum located in the nIR range.


[Study on safflower yellow for injection based on cell degranulation and acute anaphylactoid model].
This paper was aimed to establish screening methods of anaphylactoid reaction caused by safflower yellow for injection based on RBL-2 H3 cell degranulation model and mice model for acute anaphylactoid reaction,and evaluate the hypersensitivity caused by safflower yellow for injection from different batches. An in vitro cell model was used to keep the cells stimulated for an hour with different batches of safflower yellow for injection as the drug group,serum-free MEM medium as negative control group and 30 mg·L-1 C48/80 as positive control group respectively. The supernatant was then absorbed,and neutral red staining technique was used to detect the effect of safflower yellow injection on the degranulation of RBL-2 H3 cells with the positive cell rate of degranulation as the indicator.An in vivo model was established to validate the experimental results,and mice model for acute anaphylactoid reaction and ELISA method were adopted to detect the plasma histamine content,and screen the hypersensitivity caused by safflower yellow for injection at the animal level by using plasma histamine content as a test index. The results of the neutral red staining experiments showed that the positive control C48/80 could cause cell degranulation,and most of the cells were deeply stained. There was significant difference in positive cell rate between different batches of safflower yellow and positive control group. In the mice model for acute anaphylactoid reaction,it was found that the positive control C48/80 significantly increased the histamine content in the plasma of mice,while the safflower yellow in each batch did not cause a significant increase in plasma histamine( P<0. 000 1). The mechanism of anaphylactoid reaction is relatively complicated. This study was mainly based on the release of histamine and other active substances by degranulation of mast cells. No significant degranulation reaction of RBL-2 H3 cells induced by safflower yellow for injection was detected,nor was the plasma histamine level significantly increased in mice from the in vitro and in vivo aspects.


T102C polymorphism of the serotonin (5-HT) 2A receptor gene in Turkish children with cerebral infarct.
Platelet-dependent thromboembolism is an underlying mechanism in the pathogenesis of stroke. 5-HT2A receptor gene is expressed in human platelet, coronary artery (blood vessels) and brain. A polymorphism T102C at the 5-HT2A receptor gene was found that may possibly affect the 5-HT2A receptor function. As there is no existing data on T102C variant of 5-HT2A receptor gene in pediatric stroke, we aimed to study in this case-control study whether there is a association between this polymorphism and pediatric stroke. 111 patients (10 months-18 years old) with cerebral infarct and 79 healthy control was included to our study. Polymerase chain reaction (PCR) of the T102C alteration at the 5-HT 2A receptor gene was performed. Our data revealed that, 5-HT (2A) receptor T102C polymo phism was not associated with pediatric stroke in our population either alone or in combination with underlying pro thrombotic factors. However, this needs to be clarified with further studies.


Rehabilitation Methods for Patients with Geographic Atrophy due to Age-Related Macular Degeneration and Effects of Rehabilitation on Quality of Life.
The purpose of the study is to evaluate the low vision rehabilitation methods and to investigate the effect of visual rehabilitation on quality of life in patients with low vision due to geographic atrophy from age-related macular degeneration (ARMD). The better-seeing eye of 78 patients with geographic atrophy due to ARMD were included in the study. Sociodemographic characteristics, ophthalmological examination findings, and preferred low vision aids for near and distant were recorded. Fifty-seven patients who preferred to use a low vision aid device in daily life were considered as a rehabilitation group, whereas 21 patients who did not use any device were considered as a control group. The National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) was applied to all patients at the initial examination and at least 6 months after the initial examination. In the rehabilitation group, statistically significant increases were found in the overall composite score, and general vision, near and distance activities, social functioning, mental health, role difficulties, and dependency subscale scores of the NEI-VFQ-25 quality of life scale after low vision rehabilitation (p=0.009 for general vision, p < 0.001; for others). In the control group, there was no statistically significant change in any of the subscale scores or the overall score of the scale (p > 0.05). All patients (n = 78) were recommended to use at least one low vision aid for near vision. Hyperocular glasses were recommended for 77 patients (98.72%), magnifiers for 15 patients (19.23%), electro-optical devices for 2 patients (2.56%), and telemicroscope for one patient (1.28%). Furthermore, 17 patients (21.8%) were prescribed more than one low vision aids. However, for distance vision, only 29 patients (37.18%) received a recommendation for a low vision aid. Low vision patients with ARMD-related geographic atrophy should meet with low vision aids as soon as possible and should be included in low vision rehabilitation programs.


Hope and the brain: Trait hope mediates the protective role of medial orbitofrontal cortex spontaneous activity against anxiety.
As a central research topic in the field of positive psychology, hope refers to an individual's goal-oriented expectations that include both agency thinking (i.e., the motivation to initiate and sustain actions to achieve goals) and pathway thinking (i.e., the capacity to find ways toward goals). Evidence from many previous studies has shown the role of hope in protecting against anxiety. However, little is known about the neurobiological basis of hope and the underlying mechanism that how hope reduces anxiety in the brain. Here, we employed fractional amplitude of low-frequency fluctuations (fALFF) to investigate these issues in 231 high school students using resting-state functional magnetic resonance imaging (RS-fMRI). The whole-brain correlation analyses revealed that higher trait hope was related to lower fALFF in the bilateral medial orbitofrontal cortex (mOFC), which is involved in reward-related processing, motivation production, problem solving and goal-directed behaviors. Furthermore, mediation analyses suggested that trait hope acted as a mediator in the association between mOFC spontaneous activity and anxiety. These results persisted even after adjusting for the effects of positive and negative affect. Overall, this study provides the first evidence for functional brain substrates underlying trait hope and reveals a potential mechanism that trait hope mediates the protective role of spontaneous brain activity against anxiety.


Macular injury resulting from a high-powered tank laser telemetry device.
A high-powered laser is an essential part of a modern military rangefinder; however, this paper presents three cases with macular injury resulting from a high-powered tank laser telemetry device. All injuries occurred when another user deliberately pointed the telemetry target unit at the patient's eyes. The devastating effect of this high-powered laser resulted in a permanent foveal scar in the second patient and a macular hole formation in the third patient. This report emphasises that education plays a primary role in preventing accidental laser injuries. Using general guidelines and safety regulations will prevent accidental macular injuries.


[Assessment of the left atrial myxoma with gated blood pool SPECT].
A left atrial myxoma with 4 cm diameter was studied with ECG gated Tc-99m blood pool SPECT. The long axial images showed the myxoma to protrude through the mitral valve (MV) during diastole and to come back into the atrium in systole. No significant atrial dilatation due to the MV obstruction was detected. The ejection fraction calculated from the SPECT images was 10% lower than that of the conventional method viewed from LAO 45 degrees. Nuclear medicine gave an additional information for the assessment of the hemodynamics.


Epigenetics, an early event in the modulation of gene expression by inositol hexaphosphate in ethylnitrosourea exposed mouse lungs.
Mechanisms of anticancer effects of inositol hexaphosphate are not fully understood. Epigenetic changes are the early changes in tumorigenesis. DNA methyl transferases, methyl CpG binding proteins, methyl CpG DNA binding domain protein, and histone deacetylases are the major molecules involved in epigenetics. We have shown the effects of IP6 at the molecular level in mouse lungs before the tumor is developed. After 3 mo of ENU exposure, there was no tumor formation, but there was hyperplasia and lymphocytic infiltration in the lungs. Inflammation and DNA damage repair enzymes COX-2 and MLH1 appear to be upregulated, whereas tumor suppressor gene p16 was downregulated by ENU. On the other hand, ENU exposure more or less upregulated the epigenetic events such as the expressions of DNMT1, MeCP2, MBD1, and HDAC1. This alteration was reduced by IP6 administration. Results were supported by modulation of global DNA methylation and the modulation of promoter CpG methylation of p16, MLH1, and COX-2 genes. Hence, this study indicates the possible role of epigenetics at the early stage of tumor development and in the regulation of gene expression by IP6 before the onset of ENU-induced lung tumors.


Inhibition, tissue distribution and hormonal specificity of a promoter-binding factor for the uteroglobin gene.
We had earlier reported a uteroglobin promoter-binding factor in nuclei from progesterone-stimulated rabbit endometrium that was inhibited by a factor in nuclei without progesterone stimulation or from non-target tissues (Rider, V., and Bullock, D.W., 1988. Biochem. Biophys. Res. Comm. 156, 1368-1375). In the course of purification of the inhibitory activity, the effect was shown to be due to contaminating genomic DNA. The inhibitor was destroyed by treatment with DNase I and resisted phenol-chloroform extraction. Fractionation of nuclear extracts on columns of DEAE-Sepharose separated the inhibitor and revealed the presence of binding activity in unstimulated or estrogen-treated endometrium, as well as in liver, lung and ovary. The tissue and hormonal specificity of the promoter binding factor is thus less restricted than recently reported.


Contralateral replacement of pacemaker and leads following laser sheath extraction and concomitant stenting for superior vena cava syndrome.
We present the case of a 61-year-old man with a history of pacemaker implantation 15 years ago. The patient was admitted with chronic swelling of the right hemithorax and inflammation of the old incision scar after the replacement of pacemaker generator 6 years ago. The patient also presented symptoms and clinical signs of superior vena cava syndrome. Computed tomography of the thorax showed obstruction of the superior vena cava. The surgical procedure consisted of extraction of infected pacemaker system with excimer laser technique followed by dilatation and stenting of the SVC and finally implantation of a new permanent pacemaker system on the left side.


Modulation of the receptive fields of midbrain neurons elicited by thalamic electrical stimulation through corticofugal feedback.
The ascending and descending projections of the central auditory system form multiple tonotopic loops. This study specifically examines the tonotopic pathway from the auditory thalamus to the auditory cortex and then to the auditory midbrain in mice. We observed the changes of receptive fields in the central nucleus of the inferior colliculus of the midbrain evoked by focal electrical stimulation of the ventral division of the medial geniculate body of the thalamus. The receptive field of an auditory neuron was characterized by five parameters: the best frequency, minimum threshold, bandwidth, size of receptive field, and average spike number. We found that focal thalamic stimulation changed the parametric values characterizing the recorded collicular receptive fields toward those characterizing the stimulated thalamic receptive fields. Cortical inactivation with muscimol prevented the development of the collicular plasticity induced by focal thalamic stimulation. Our data suggest that the intact colliculo-thalamo-cortico-collicular loops are important for the coordination of sound-guided plasticity in the central auditory system.


Copper induction of lignin-modifying enzymes in the white-rot fungus Trametes trogii.
Trametes trogii, a white rot basidiomycete involved in wood decay worldwide, produces several ligninolytic enzymes, laccase being the dominant one, with higher titers than those reported for most other white rot fungi studied up to date. The effect of copper on in vitro production of extracellular ligninolytic activities was studied. CuSO(4)·5H(2)O concentrations from 1.6 μM to 1.5 mM were tested in a synthetic medium with glucose 20 g/L and asparagine 3 g/L. The addition of copper (up to 1 mM) did not affect growth but strongly stimulated ligninolytic enzyme production; faster decolorization of the polymeric dye Poly R-478 was observed as well. Maximal production of manganese peroxidase, laccase, and glyoxal oxidase [1.28 U/mL, 93.8 U/mL (with a specific activity of 720 U/mg protein), and 0.46 U/mL respectively] was attained with 1 mM CuSO(4)·5H(2)O. However, higher copper concentrations inhibited growth and notably decreased manganese peroxidase production, although they did not affect laccase secretion. Laccase activity in the culture filtrate was maximal at 50 C and pH 3.4, and the enzyme was completely stable at pH 4.4 and above, and at 30 C for up to 5 d. Denaturing polyacrylamide gel electrophoresis of extracellular culture fluids showed two laccase activity bands (mol wt 38 and 60 kDa respectively). The pattern of isoenzyme production was not affected by medium composition but differed with culture age.


Chromosome-scale assemblies of S. malaccense, S. aqueum, S. jambos, and S. syzygioides provide insights into the evolution of Syzygium genomes.
Syzygium is a large and diverse tree genus in the Myrtaceae family. Genome assemblies for clove (Syzygium aromaticum, 370 Mb) and sea apple (Syzygium grande, 405 Mb) provided the first insights into the genomic features and evolution of the Syzygium genus. Here, we present additional de novo chromosome-scale genome assemblies for Syzygium malaccense, Syzygium aqueum, Syzygium jambos, and Syzygium syzygioides. Genome profiling analyses show that S. malaccense, like S. aromaticum and S. grande, is diploid (2n = 2x = 22), while the S. aqueum, S. jambos, and S. syzygioides specimens are autotetraploid (2n = 4x = 44). The genome assemblies of S. malaccense (430 Mb), S. aqueum (392 Mb), S. jambos (426 Mb), and S. syzygioides (431 Mb) are highly complete (BUSCO scores of 98%). Comparative genomics analyses showed conserved organization of the 11 chromosomes with S. aromaticum and S. grande, and revealed species-specific evolutionary dynamics of the long terminal repeat retrotransposon elements belonging to the Gypsy and Copia lineages. This set of Syzygium genomes is a valuable resource for future structural and functional comparative genomic studies on Myrtaceae species.


Cervical thymic cyst: report of a case and review of the literature.
Congenital thymic cysts are uncommon and often misdiagnosed as either branchial cleft or cystic hygromas. This paper presents the case of a male child, who presented with a soft, fluctuating mass in the left side of his neck. Surgical excision revealed an ectopic thymic cyst. The histopathologic features of thymic cysts are diagnostic. Ectopic thymic tissue may be an infrequent finding, but it should be included in the differential diagnosis of neck masses, especially in children.


The effects of hormone implants on serum lipoproteins and steroid hormones in bilaterally oophorectomised women.
Serum lipoproteins were measured over a period of 6 months in 14 oophorectomised women treated with oestrogen implants (50 mg oestradiol-17 beta) and 17 oophorectomised women treated with oestrogen/testosterone implants (50 mg oestradiol-17 beta, 100 mg testosterone). Both types of implant caused only minimal changes in lipoprotein metabolism. Low density lipoprotein (LDL) cholesterol decreased with both types of implant and high density lipoprotein (HDL) cholesterol rose with the oestrogen implants. HDL subfractions were also measured. The oestrogen implants caused a transient rise in HDL2 cholesterol levels at 2 months and a slower rise in HDL3 cholesterol. The oestrogen/testosterone implants had no effect on HDL fractions. The results indicate that hormone implants do not cause the profound changes in lipoproteins associated with oral hormone therapy.


Tyrosine sulfation in precursors of collagen V.
Radioactive labeling of p-collagens V, collagens V, and, to a small extent, of procollagen V occurred when [35S]sulfate was incubated with tendons or primary tendon cell cultures, or blood vessels and crops of 17- to 19-day-old chick embryos, or with lung slices from neonatal rats. Most or all of this label is in the form of 1 or more sulfated tyrosine residues/chain of p alpha 1(V), alpha 1(V), p alpha 1'(V), alpha 1'(V), p alpha 2(V), and alpha 2(V), and it remains attached through purification by dialysis, ammonium sulfate precipitation, CsCl-GdnCl2 equilibrium buoyant density and velocity sedimentations, ion-exchange chromatography, and sodium dodecyl sulfate gel electrophoresis. Radioactive tyrosine sulfate was identified in alkaline hydrolysates of these collagen V chains, after labeling the tissues with either [35S]sulfate or [3H]tyrosine, by electrophoretic and chromatographic comigration with a tyrosine sulfate standard. Tunicamycin A1, which inhibits the attachment of N-linked complex carbohydrate, did not interfere with the sulfation process. The tyrosine sulfate is located in a noncollagenous domain, which is probably adjacent to the amino end of the collagen helix, and is retained throughout the physiological proteolytic processing of procollagens V. After digestion with Staphylococcus aureus V8 protease, 35S-labeled p alpha 1(V) and alpha 1(V) chains gave the same map of labeled peptides, and this differed from the map given by p alpha 1'(V) and alpha 1'(V) chains. Little sulfation of p alpha 2(V) and alpha 2(V) chains occurs. The implications of these observations for the structure and properties of procollagens V and their derivatives are considered.


UK laboratory intercomparison on internal dosimetry.
A laboratory intercomparison for internal dose assessment from a variety of intake scenarios is described. This is the first UK intercomparison using the revised ICRP Human Respiratory Tract and biokinetic models. Four United Kingdom laboratories participated and six cases were assessed. Overall, the agreement in internal dose assessments between laboratories was considered satisfactory with 79% of the assessed committed effective doses, e(50), for cases within a band of +/- 40% of the median value. The range (highest/lowest) in e(50) estimated by the laboratories was smallest (1.2) for a case involving inhalation of 137Cs. The range was greatest (6.0) for a case involving a wound with, and possible inhalation of, 238Pu, 239Pu and 241Am; the variation between laboratories in assessment of intakes could not be considered to be satisfactory in this case. Judgements on the most appropriate data to use in estimating intakes, choice of parameter values for use with the ICRP models and allowing for the effects of treatment with DTPA were important sources of variability between laboratories.


Incidence and Cost of Ankle Sprains in United States Emergency Departments.
Ankle sprains represent a common injury in emergency departments, but little is known about common complications, procedures, and charges associated with ankle sprains in emergency departments. There will be a higher incidence of ankle sprains among younger populations (≤25 years old) and in female patients. Complications and procedures will differ between ankle sprain types. Lateral ankle sprains will have lower health care charges relative to medial and high ankle sprains. Descriptive epidemiological study. Level 3. A cross-sectional study of the 2010 Nationwide Emergency Department Sample was conducted. Outcomes such as charges, complications, and procedures were compared using propensity score matching between lateral and medial as well as lateral and high ankle sprains. The sample contained 225,114 ankle sprains. Female patients sustained more lateral ankle sprains (57%). After propensity score adjustment, lateral sprains incurred greater charges than medial ankle sprains (median [interquartile range], $1008 [$702-$1408] vs $914 [$741-$1108]; P < 0.01). Among complications, pain in the limb (1.92% vs 0.52%, P = 0.03), sprain of the foot (2.96% vs 0.70%, P < 0.01), and abrasion of the hip/leg (1.57% vs 0.35%, P = 0.03) were more common in lateral than medial ankle sprain events. Among procedures, medial ankle sprains were more likely to include diagnostic radiology (97.91% vs 83.62%, P < 0.01) and less likely to include medications than lateral ankle sprains (0.87% vs 2.79%, P < 0.01). Hospitalizations were more common following high ankle sprains than lateral ankle sprains (24 [6.06%] vs 1 [0.25%], P < 0.01). Ankle sprain emergency department visits account for significant health care charges in the United States. Age- and sex-related differences persist among the types of ankle sprains. The health care charges associated with ankle sprains indicate the need for additional preventive measures. There are age- and sex-related differences in the prevalence of ankle sprains that suggest these demographics may be risk factors for ankle sprains.


[The number of salivary corpuscles in healthy subjects and in patients with periodontal disease].
Using a standardized, well reproducible method, the authors determined the number of salivary corpuscles in 174 healthy subjects (1,400 to 2,000/microliter) and 109 patients with periodontal disease (5,300 to 7,800/microliter). The statistical analysis revealed a highly significant difference between the two groups. There was a positive correlation between the number of salivary corpuscles and periodontal disease indices.


Solyman Brown, a giant of dentistry and its poet laureate.
Solyman Brown 1790-1876), widely hailed by his contemporary literary establishment as the poet laureate of dentistry," was one of the visionary individuals who put dentistry on a true foundation of professionalism. He was instrumental in the formation of the first dental journal, the first national dental society, and the first dental school. He is one of the true giants in the history of American dentistry. Solyman Brown's great-great-grandson was cleaning out his father's attic and found a trunk full of original material relating to his illustrious forebear. Among the wealth of material was a notebook kept by Solyman's daughter, Augusta. Selections from it give us a rare personal insight into the type of person this significant dental historical figure was, as well as the struggles he endured in his hard life.


Spatial and Temporal Distributions of Organophosphate Ester Concentrations from Atmospheric Particulate Matter Samples Collected across Houston, TX.
Atmospheric particulate matter (PM) samples were collected from four ground-based sites located in the Houston, TX (September 21-28, 2013) and were analyzed for 12 organophosphate esters (OPEs; current-use plasticizers and flame retardants). Samples analyzed included daytime, nighttime, and 24 h PM of <2.5 μm aerodynamic diameter (PM2.5) and total suspended particulate (TSP) samples. PM2.5 and TSP atmospheric ΣOPE concentrations varied over an order of magnitude and were statistically significantly different between urban and suburban and industrial sites. Additionally, significant temporal variability was also identified; for example, daytime atmospheric concentrations of 2-ethylhexyl diphenyl phosphate (EHDPP; 610 ± 220 pg m-3) measured in TSP samples were significantly higher than nighttime concentrations (280 ± 180 pg m-3; p = 0.03). Detailed discussions of the spatial and temporal distribution are given for Tris-(1-chloro-2-propyl) phosphate (TCiPP), EHDPP, tri-n-butyl phosphate (TnBP), and triphenyl phosphate (TPhP). Correlations to bulk measurements of carbonaceous PM including organic carbon, elemental carbon, and water-soluble organic carbon were used to understand potential sources and urban atmospheric transport. These results highlight the fundamental complexity associated with assessing OPE atmospheric concentrations across a large urban landscape and specific knowledge gaps at the intersection of consumer products and safety with environmental and human health.


Financial viability, benefits and challenges of employing a nurse practitioner in general practice.
This case study examines the financial viability, benefits and challenges of employing a primary healthcare (PHC) nurse practitioner (NP) in a bulk-billing healthcare cooperative in the Australian Capital Territory. There are few empirical case reports in the Australian literature that demonstrate financial sustainability of this type of healthcare professional in primary healthcare. This case study demonstrates that the costs of employing a PHC-NP in general practice are offset by direct and indirect Medicare billings generated by the PHC-NP, resulting in a cost-neutral healthcare practitioner. The success of this model relies on bidirectional collaborative working relationships amongst general practitioners and NPs. PHC-NPs should have a generalist scope of practice and specialist expertise in order to maximise their utility within the general practice environment.


Handicapped retroviral vectors efficiently transduce foreign genes into hematopoietic stem cells.
Retroviral vectors, designated handicapped, are described. These are genetically defective viruses that allow transfer of nonselectable genes under the transcriptional control of internal promoters. The basic handicapped vector (pHHAM) is derived from Harvey, Abelson, and Moloney murine retroviruses. It contains a 327-base-pair deletion in the 3' long terminal repeat that spans enhancer and promoter sequences. The deletion is successfully transferred to the 5' long terminal repeat after reverse transcription of viral RNA, yielding a provirus incapable of synthesizing viral transcripts. HHAM viruses containing the mouse c-myc gene under the control of immunoglobulin kappa chain gene regulatory elements, along with a selectable gene (neo) driven by a weak promoter (tk), were stably transmitted to cultured mouse B cells. The donor c-myc gene was transcribed from the kappa promoter in these cells. Helper-free virus-producing cell lines were generated at titers favorable for the efficient introduction of HHAM viruses, even without selection, into hematopoietic stem cells from mouse bone marrow. When returned to unirradiated congenic recipient mice, the cells were capable of long-term reconstitution of the myeloid and lymphoid lineages of W/Wv mutants and the lymphoid system of scid mutants.


Molecular cloning and analysis of the human Tec protein-tyrosine kinase.
Mouse Tec is a non-receptor type protein-tyrosine kinase and is highly expressed in many hematopoietic cell lines. To investigate the roles of the Tec kinase in the human hematopoietic system, we isolated cDNAs encoding the human Tec kinase. The human tec cDNAs can encode a peptide of 631 amino acid residues with a calculated molecular mass of 73,624. The predicted human Tec protein is highly homologous to those of the members of the Tec family including mouse Tec type IV (94% homology), mouse Tsk/Itk (60%), and human Btk (57%). The homology between human Tec and other members of the Tec family can be observed not only in the Src homology 3 (SH3), SH2, and kinase domains, but also in the N-terminal unique domain. Northern blot analysis demonstrated that the major transcripts of tec could be detected at 2.6 kb and 3.6 kb in a wide range of human hematopoietic cell lines including myeloid, B-, and T-cell lineages. Interestingly, high expression of the tec gene could be detected in all of the three patients examined with myelodysplastic syndrome. The human tec gene was mapped by fluorescence in situ hybridization (FISH) to chromosome 4p12.


[Biomechanical studies of various forms of symphysis banding].
The banding operation with polydioxanone cord for pelvis stabilization after accidental symphysis rupture is altogether reliable. Nevertheless in some cases we saw later on non satisfying reduction results in the "pelvis-half-joints"--especially in intense dislocation of the iliosacral amphiarthrosis. For improving the stability we investigated the stiffness character of some different methods with four PDS cords varied by four and six drilling holes. By increasing number of holes and short intentions direction knotted cords the stability is growing by 25%.


Chondrodysplasia Punctata: A Case Report of Fetal Warfarin Syndrome.
Chondrodysplasia punctata is abnormal calcification in the cartilage of developing bones and has been seen in association with deranged vitamin K metabolism. Warfarin, an oral anticoagulant acting on vitamin K dependent clotting factors is known to cause chondrodysplasia punctata. Despite the knowledge of the condition the management of patients with prosthetic heart valves might require use of the drug for anticoagulation. Here, we present a case of a fetal warfarin syndrome in a second born child of a 27 year lady under warfarin for prosthetic heart valve. The pregnancy was complicated by polyhydramnios in third trimester and terminated at term by normal vaginal delivery. The baby was well, except for facial dysmorphism in the form of depressed nasal bridge, narrow nares and suspected left choanal atresia. Radiograph revealed stippled ephiphysis of vertebra, femora and humera supporting diagnosis of fetal warfarin syndrome. The baby did not develop any perinatal complication and was discharged home.


A technique for the measurement of visceral fat by ultrasonography: comparison of measurements by ultrasonography and computed tomography.
This study aims to create a method of calculating intra-abdominal visceral fat volume by using ultrasound (US). The visceral fat volume measured by US was evaluated by comparison with the volume measured by computed tomography (CT). Eighty-seven patients (52 males and 35 females) were enrolled in this study. Both US and CT were performed, and the visceral fat volume was measured. Both the distance and thickness of the parameters in US were measured as follows: 1) the distance between the internal surface of the abdominal muscle and the splenic vein, 2) the distance between the internal surface of the abdominal muscle and the posterior wall of aorta on the umbilicus, and 3) the thickness of the fat layer of the posterior right renal wall. The equation was calculated as follows: [visceral fat volume]=-9.008+1.191x[distance between the internal surface of the abdominal muscle and the splenic vein (mm)]+0.987x[distance between the internal surface of the abdominal muscle and the posterior wall of the aorta on the umbilicus (mm)]+3.644x[thickness of the fat layer of the posterior right renal wall (mm)]. There was a good correlation between the visceral fat volume calculated by the above equation and the volume by CT described (r=0.860, p<0.0001). The measurement of the visceral fat volume using US provided results as effectively as CT, and it was proven to be a useful method.


Zika Virus Meningoencephalitis in an Immunocompromised Patient.
The World Health Organization considers the Zika virus (ZIKV) outbreak in the Americas a global public health emergency. The neurologic complications due to ZIKV infection comprise microcephaly, meningoencephalitis, and Guillain-Barré syndrome. We describe a fatal case of an adult patient receiving an immunosuppressive regimen following heart transplant. The patient was admitted with acute neurologic impairment and experienced progressive hemodynamic instability and mental deterioration that finally culminated in death. At autopsy, a pseudotumoral form of ZIKV meningoencephalitis was confirmed. Zika virus infection was documented by reverse trancriptase-polymerase chain reaction, immunohistochemistry, and immunofluorescence and electron microscopy of the brain parenchyma and cerebral spinal fluid. The sequencing of the viral genome in this patient confirmed a Brazilian ZIKV strain. In this case, central nervous system involvement and ZIKV propagation to other organs in a disseminated pattern is quite similar to that observed in other fatal Flaviviridae viral infections.


Recent advances in medical management of hepatocellular carcinoma.
Transcatheter arterial therapies for hepatocellular carcinoma (HCC) have developed during the last decade. A fine powder formulation of cisplatin and the new platinum agent miriplatin became standard medicines in addition to anthracyclines in transcatheter arterial chemoembolization (TACE) in Japan. Recent prospective and retrospective studies supported the usefulness of platinum agents as a chemotherapeutic at the time of varied TACE therapy. Although balloon-occluded TACE is an effective therapy for localized HCC and drug-eluting microspheres seemed to show a higher response rate in certain HCCs, the definite advantages of those procedures still remain uncertain. Intermediate stage HCC, or Barcelona Clinic Liver Cancer stage B, is regarded as a heterogeneous category with a wide spectrum of tumors and patients, and several subclassifications of the stage have been proposed to show different prognoses; there are also different recommended therapies in each subgroup. Authors have subclassified patients based on combinations of tumor size, tumor number, and liver function, with or without performance status. Because of differences of available medical resources and techniques in treatment procedures between countries, the most ideal and useful subgrouping remains inconclusive at present. Recently, a few systemic chemotherapies proved to be effective for advanced stage HCC in phase III studies: lenvatinib as the first line of therapy, and regorafenib, cabozantinib, and ramucirumab as second-line therapy. Other molecular-targeted and immune-oncological medicines are expected to follow in the near future. Some studies have suggested an advantage of early introduction of molecular-targeted therapy for TACE-resistant HCC in the intermediate stage.


Initial Experience of Single-port Laparoscopic Multivisceral Resection for Locally Advanced Colon Cancer.
The aim of this study was to evaluate the safety and feasibility of single-port laparoscopic multivisceral resection (SLMVR) for locally advanced colon cancer invading or adhering to neighboring organs. A prospectively collected database was collated of all patients undergoing SLMVR from January 2011 to December 2014. Short-term and long-term oncologic outcomes were evaluated. Thirty consecutive patients who underwent SLMVR were identified and the completion rate was 90%. The R0 resection rate of the primary tumor was 100%. The postoperative complication rate was 23.3%. The 3-year relapse-free survival rates of patients with stage II (n=11) and stage III (n=12) disease were 80.8% and 54.6%, respectively. The 5-year overall survival rates of patients with stage II and stage III disease were 75.8% and 75.0% over a median follow-up of 42 months, respectively. SLMVR for locally advanced colon cancer is safe and feasible in selected patients.


The effects of different thermal environments on the physiological and psychological responses of firefighters to a training drill.
Little is known about the impact of thermoregulatory demands on cardiovascular and psychological responses of firefighters during firefighting activities. This study examined selected responses to a training drill in different thermal environments. Male firefighters (n = 16) were randomly assigned to perform a simulated ceiling overhaul task for 16 min in either a neutral (13.7 degrees C) or hot (89.6 degrees C) condition while wearing standard firefighting turnout gear. Physiological and psychological measures were assessed before, after 8 min and 16 min of firefighting activity, and following a 10-min recovery period. The variables assessed included heart rate (HR), tympanic temperature (Ttymp), lactate level (LAC), blood glucose level, ratings of perceived exertion (RPE), perceptions of respiration, thermal sensations (TS) and state anxiety (SA). Significant increases were seen for HR, Ttymp, LAC, RPE and SA, with the increases being much greater following the hot condition. Recovery was significantly slower following work in the hot condition. These findings suggest that the addition of a live fire (a common situation for firefighters) contributes to increased cardiovascular and psychological strain at a standardized workload.


COVID-19 symptoms are reduced by targeted hydration of the nose, larynx and trachea.
Dehydration of the upper airways increases risks of respiratory diseases from COVID-19 to asthma and COPD. We find in human volunteer studies involving 464 human subjects in Germany, the US, and India that respiratory droplet generation increases by up to 4 orders of magnitude in dehydration-associated states of advanced age (n = 357), elevated BMI-age (n = 148), strenuous exercise (n = 20) and SARS-CoV-2 infection (n = 87), and falls with hydration of the nose, larynx and trachea by calcium-rich hypertonic salts. We also find in a protocol of exercise-induced airway dehydration that hydration of the airways by calcium-rich salts increases oxygenation relative to a non-treatment control (P < 0.05). In a random control study of COVID-19 positive subjects (n = 40), thrice-a-day delivery of the calcium-rich hypertonic salts (active) suppressed respiratory droplet generation by 51% ± 11% and increased oxygen saturation over three days of treatment by 48.08% ± 9.61% (P < 0.001), while no changes were observed in the nasal-saline control group. Self-reported symptoms significantly declined in the active group and did not decline in the control group. Hydration of the upper airways appears promising as a non-drug approach for reducing risks of respiratory diseases such as COVID-19.


Interfacial Shish-Kebabs Lengthened by Coupling Effect of In Situ Flexible Nanofibrils and Intense Shear Flow: Achieving Hierarchy To Conquer the Conflicts between Strength and Toughness of Polylactide.
The challenge of hitherto elaborating a feasible pathway to overcome the conflicts between strength and toughness of polylactide (PLA) still remains among academia and industry. In the current work, a unique hierarchal structure of flexible poly(butylene adipate-co-terephthalate) (PBAT) in situ nanofibrils integrating with abundant PLA shish-kebabs as a strong building block was disclosed and expresses its capability to conquer this dilemma. Substantially simultaneous enhancement on tensile strength, impact strength, and elongation at break could be achieved up to 91.2 MPa, 14.9 KJ/m2, and 15.7%, respectively, compared with pure PLA (61.5 MPa, 4.3 KJ/m2, and 6.2%). Through investigating the phase (and crystalline) morphology and molecular chain behavior in the PLA/PBAT system, the formation mechanism of this structure facilitated by a coupling effect of PBAT flexible phase and shear flow was definitely elucidated. The dispersed phase of PBAT would be more inclined to existing as a fibrillar form within the PLA matrix benefiting from low interfacial tension. Interestingly, this phase morphology with large specific surface area changes the crystallization behavior of PLA significantly, once introducing an intense shear flow (∼103 s-1), in situ shear-formed nanofibrils of PBAT would show strong coupling effect with shear flow on PLA crystallization: they can not only induce abundant shish-kebabs of PLA at its interfaces, which possesses lengthened shish and more densely arranged kebabs, but also further retard the relaxation of PLA chains through hysteretic relaxation of its PBAT phase, which can effectively prevent the collapse of established shish. Of immense significance is this particular hierarchical-architecture composed by flexible nanofibers (PBAT) and rigid shish-kebabs (PLA), which provides significant guidance for the simultaneous reinforcement and toughness of polymer materials.


Sec15 interacts with Rab11 via a novel domain and affects Rab11 localization in vivo.
Sec15, a component of the exocyst, recognizes vesicle-associated Rab GTPases, helps target transport vesicles to the budding sites in yeast and is thought to recruit other exocyst proteins. Here we report the characterization of a 35-kDa fragment that comprises most of the C-terminal half of Drosophila melanogaster Sec15. This C-terminal domain was found to bind a subset of Rab GTPases, especially Rab11, in a GTP-dependent manner. We also provide evidence that in fly photoreceptors Sec15 colocalizes with Rab11 and that loss of Sec15 affects rhabdomere morphology. Determination of the 2.5-A crystal structure of the C-terminal domain revealed a novel fold consisting of ten alpha-helices equally distributed between two subdomains (N and C subdomains). We show that the C subdomain, mainly via a single helix, is sufficient for Rab binding.


Development of cardiovascular function in the marine gastropod Littorina obtusata (Linnaeus).
The molluscan cardiovascular system typically incorporates a transient extracardiac structure, the larval heart, early in development, but the functional importance of this structure is unclear. We documented the ontogeny and regulatory ability of the larval heart in relation to two other circulatory structures, the true heart and the velum, in the intertidal gastropod Littorina obtusata. There was a mismatch between the appearance of the larval heart and the velum. Velar lobes appeared early in development (day 4), but the larval heart did not begin beating until day 13. The beating of the larval heart reached a maximum on day 17 and then decreased until the structure itself disappeared (day 24). The true heart began to beat on day 17. Its rate of beating increased as that of the larval heart decreased, possibly suggesting a gradual shift from a larval heart-driven to a true heart-driven circulation. The true heart was not sensitive to acutely declining P(O(2)) shortly after it began to beat, but increased in activity in response to acutely declining P(O(2)) by day 21. Larval heart responses were similar to those of the true heart, with early insensitivity to declining P(O(2)) (day 13) followed by a response by day 15. Increased velum-driven rotational activity under acutely declining P(O(2)) was greatest in early developmental stages. Together, these findings point to cardiovascular function in L. obtusata larvae being the result of a complex interaction between velum, larval and true heart activities, with the functions of the three structures coinciding but their relative importance changing throughout larval development.


Serum Soluble Semaphorin 4D is Associated with Left Atrial Diameter in Patients with Atrial Fibrillation.
The aim of this study was to evaluate the serum soluble semaphorin 4D (sSema4D) in patients with atrial fibrillation and to investigate the relationship of serum sSema4D with left atrial diameter (LAD). We studied a total of 113 patients who were subdivided into paroxysmal and non-paroxysmal (included persistent and permanent) atrial fibrillation groups, respectively. Another 55 subjects without atrial fibrillation were enrolled as the healthy control group. Serum levels of soluble semaphorin 4D (Sema4D) were measured in all subjects using the enzyme-labeled immunosorbent assay method. We also evaluated the coagulation parameters and left atrial diameters. Patients with paroxysmal and non-paroxysmal atrial fibrillation had significantly higher sSema4D level compared with controls (8.50±2.19 ng/mL and 9.30±2.28 ng/mL vs. 6.56±1.27 ng/ml, P<0.05). Serum sSema4D concentrations were elevated in patients with non-paroxysmal atrial fibrillation compared to those with paroxysmal atrial fibrillation (P<0.001). The level of sSema4D was positively correlated with LAD (r=0.606, P<0.001). Multivariate logistic regression analysis revealed that serum sSema4D, LAD, male sex, heart rate, hypertension, and coronary artery disease were associated with atrial fibrillation (P<0.05). Serum sSema4D levels are increased in patients with atrial fibrillation and are independently associated with atrial remodeling.


Drugs prescribed for Phelan-McDermid syndrome differentially impact sensory behaviors in shank3 zebrafish models.
Background: Altered sensory processing is a pervasive symptom in individuals with Autism Spectrum Disorders (ASD); people with Phelan McDermid syndrome (PMS), in particular, show reduced responses to sensory stimuli. PMS is caused by deletions of the terminal end of chromosome 22 or point mutations in Shank3. People with PMS can present with an array of symptoms including ASD, epilepsy, gastrointestinal distress, and reduced responses to sensory stimuli. People with PMS are often medicated to manage behaviors like aggression and/or self-harm and/or epilepsy, and it remains unclear how these medications might impact perception/sensory processing. Here we test this using zebrafish mutant shank3ab PMS models that likewise show reduced sensory responses in a visual motor response (VMR) assay, in which increased locomotion is triggered by light to dark transitions. Methods: We screened three medications, risperidone, lithium chloride (LiCl), and carbamazepine (CBZ), prescribed to people with PMS and one drug, 2-methyl-6-(phenylethynyl) pyridine (MPEP) tested in rodent models of PMS, for their effects on a sensory-induced behavior in two zebrafish PMS models with frameshift mutations in either the N- or C- termini. To test how pharmacological treatments affect the VMR, we exposed larvae to selected drugs for 24 hours and then quantified their locomotion during four ten-minute cycles of lights on-to-off stimuli. Results: We found that risperidone normalized the VMR in shank3 models. LiCl and CBZ had no effect on the VMR in any of the three genotypes. MPEP reduced the VMR in wildtype (WT) to levels seen in shank3 models but caused no changes in either shank3 model. Finally, shank3 mutants showed resistance to the seizure-inducing drug pentylenetetrazol (PTZ), at a dosage that results in hyperactive swimming in WT zebrafish. Conclusions: Our work shows that the effects of drugs on sensory processing are varied in ways that can be highly genotype- and drug-dependent.


The extracellular domain of the epidermal growth factor receptor. Studies on the affinity and stoichiometry of binding, receptor dimerization and a binding-domain mutant.
The binding of epidermal growth factor (EGF) or an EGF-like growth factor to the EGF receptor is the initial event which leads to receptor activation, and consequently the induction of cell growth. In order to study this binding interaction in detail, we produced the extracellular domain of the EGF receptor (EGFR) using the baculovirus expression system. Affinity-labeling and Western-blot analyses revealed that the baculovirus-infected insect cells secrete active EGFR extracellular domain relatively efficiently, however a significant amount of inactive EGFR extracellular domain is retained within the cells. The apparent dissociation constant (Kd) of the secreted EGFR extracellular domain for EGF and transforming growth factor alpha (TGF-alpha), as determined using an immobilized receptor binding assay, was approximately 200 nM. Interestingly, this Kd value is 30-40-fold lower than that of the full-length EGFR derived from detergent-solubilized A431 cell membranes. The stoichiometry of binding of the EGFR extracellular domain to EGF and TGF-alpha was examined by band-shift analysis on non-denaturing PAGE and was estimated to be 1:1. We have also shown, using sedimentation equilibrium analysis, that ligand binding induces significant dimerization of the EGFR extracellular domain. Finally, we carried out site-specific mutagenesis on the EGFR extracellular domain in order to define the ligand-binding region. We identified amino acid residues which are close to the binding site since they are common to the epitopes of several ligand-competitive monoclonal antibodies. However, these residues do not contribute directly to ligand binding since the affinity of the mutated EGFR extracellular domain for EGF and TGF-alpha was unaffected.


Direct Isolation of Seamless Mutant Bacterial Artificial Chromosomes.
Seamless (i.e., without unwanted DNA sequences) mutant bacterial artificial chromosomes (BACs) generated via recombination-mediated genetic engineering (recombineering) are better suited to study gene function compared to complementary DNA (cDNA) because they contain only the specific mutation and provide all the regulatory sequences required for in vivo gene expression. However, precisely mutated BACs are typically rare (∼1:1,000 to 1:100,000), making their isolation quite challenging. Although these BACs have been classically isolated by linking the mutation to additional genes, i.e., selectable markers, this approach is prone to false positives and is labor-intensive because it requires the subsequent removal of the selectable marker. We created Founder Principle-driven Enrichment (FPE), a method based on the population genetics "founder principle," to directly isolate rare mutant BACs, without any selectable marker, from liquid cultures via the polymerase chain reaction (PCR). Here, we provide a detailed description of FPE, including protocols for BAC recombineering and PCR screening. © 2017 by John Wiley & Sons, Inc.


Oxygenation With a Single Portable Pulse-Dose Oxygen-Conserving Device and Combined Stationary and Portable Oxygen Delivery Devices in Subjects With COPD.
Portable oxygen devices simplify and facilitate patient therapy. This study was designed to compare S(pO2) and patient satisfaction with a portable oxygen concentrator or a combined system consisting of a fixed device with continuous-flow oxygen dispensation and a portable device with pulse dispensation for ambulation. This crossover trial assessed 25 subjects with COPD (92% men, mean age of 72.2 ± 7.4 y, mean FEV1 of 34.14 ± 12.51% of predicted) at 4 hospitals in Madrid. All subjects had previously used the combined system, consisting of a fixed oxygenation system and a portable system for ambulation, with 16 (64%) using stationary and portable concentrators and 9 (36%) using a stationary reservoir and portable liquid oxygen bag. Oxygenation settings at rest and while walking were determined at baseline. Subjects were maintained on the previous combined system for 1 week and then switched to the portable oxygen concentrator for 1 week. Mean S(pO2) over 24 h was calculated using the software in the oximeter, and compliance was monitored (Visionox). Low S(pO2) (< 90%) was significantly more frequent during use of the portable concentrator alone than with the combined system (37.1% vs 18.4%, P < .05). The portable system alone was preferred by 43% of subjects, and the combined system was preferred by 36%, whereas 21% were not sure. Subjects preferred using a single portable oxygenation system both at home and during ambulation. Portable systems alone, however, did not supply the same levels of oxygenation as the combination of fixed and portable systems. Before the widespread adoption of portable systems as a single device, additional studies are needed to determine best-practice protocols for adjustment of daytime and nighttime oxygenation settings. (ClinicalTrials.gov registration NCT02079753).


Contemporary techniques of da Vinci SP radical prostatectomy: multicentric collaboration and expert opinion.
The da Vinci SP robot consists of an innovative single port trocar that houses a flexible camera and three biarticulated arms, which minimizes the number of incisions to assess the surgical site, allowing a less invasive procedure. However, due to its recent release in the market, the current literature reporting SP-RARP is still restricted to a few centers. In this scenario, after performing a literature search with all available techniques of SP-RARP, our objective is to report a multicentric opinion of referral centers on different techniques to approach SP-RARP. The SP literature is provided by only a few centers due to the limited number of this new console in the market. Five different approaches are available: transperitoneal, extraperitoneal, Retzius-Sparing, transperineal and transvesical. None of the current studies describe long-term functional or oncological outcomes. However, all approaches had satisfactory operative performance with minimum complication rates. Several techniques of SP-RARP have been reported in the literature. We performed a multicentric collaboration describing and illustrating the most challenging steps of this surgery. We believe that the details provided in this article are useful teaching material for new centers willing to adopt the SP technology.


Corticostriatal Plastic Changes in Experimental L-DOPA-Induced Dyskinesia.
In Parkinson's disease (PD), alteration of dopamine- (DA-) dependent striatal functions and pulsatile stimulation of DA receptors caused by the discontinuous administration of levodopa (L-DOPA) lead to a complex cascade of events affecting the postsynaptic striatal neurons that might account for the appearance of L-DOPA-induced dyskinesia (LID). Experimental models of LID have been widely used and extensively characterized in rodents and electrophysiological studies provided remarkable insights into the inner mechanisms underlying L-DOPA-induced corticostriatal plastic changes. Here we provide an overview of recent findings that represent a further step into the comprehension of mechanisms underlying maladaptive changes of basal ganglia functions in response to L-DOPA and associated to development of LID.


Sorption kinetics and isotherm modelling of imidacloprid on bentonite and organobentonites.
Bentonite was modified by quaternary ammonium cations viz. cetytrimethylammonium (CTA), cetylpyridinium (CP), rioctylmethylammonium (TOM) and pcholine (PTC) at 100% cation exchange capacity of bentonite and was characterized by X-ray diffraction, CHNS elemental analyser and Fourier transform infrared spectroscopy. The sorption of imidacloprid on organobentonites/bentonite was studied by batch method. Normal bentonite could adsorb imidacloprid only upto 19.31-22.18% while all organobentonites except PTC bentonite (PTCB), enhanced its adsorption by three to four times. Highest adsorption was observed in case of TOM bentonite (TOMB) (76.94-83.16%). Adsorption kinetic data were fitted to pseudo-first-order, pseudo-second-order and intraparticle diffusion models. For normal bentonite data were best fitted to pseudo-first-order kinetic, while for organobentonites fitted to pseudo-second-order kinetics. Sorption data were analysed using Freundlich, Langmuir, Temkin and Dubinin-Radushkevich isotherm models. Data were well fitted to Freundlich adsorption isotherm. Product of Freundlich adsorption constant and heterogeneity parameter (Kf.1/n) was in following order: TOMB (301.87) > CTA bentonite (CTAB) (152.12) > CP bentonite (CPB) (92.58) > bentonite (27.25). Desorption study confirmed hysteresis and concentration dependence. The present study showed that the organobentonite could be a good sorbent for removal of imidacloprid from natural water sample also. Percentage adsorption and Distribution coefficient (mL g-1) value of different adsorbent was in following order: TOMB (74.85% and 297.54) > CTAB (55.78% and 126.15) > CPB (45.81% and 84.55) > bentonite (10.65% and 11.92).


The purification and characterization of glutaryl-coenzyme A dehydrogenase from porcine and human liver.
Glutaryl-CoA dehydrogenase, a multifunctional enzyme responsible for dehydrogenation and decarboxylation of glutaryl-CoA to crotonyl-CoA, has been purified 1,680-fold from porcine liver mitochondria. The purified porcine enzyme has a subunit molecular weight of 47,800 and a native molecular weight of 190,500. Porcine glutaryl-CoA dehydrogenase catalyzed the conversion of [1,5-14C]glutaryl-CoA to [14C] crotonyl-CoA and 14CO2 in a 1:1:1 ratio. The porcine enzyme has Km values for electron transfer flavoprotein and glutaryl-CoA of 1.1 and 3.3 microM, respectively, and turnover numbers of 860 mol of electron transfer flavoprotein/min/mol of glutaryl-CoA dehydrogenase and 327 mol of glutaryl-CoA/min/mol of glutaryl-CoA dehydrogenase. Human glutaryl-CoA dehydrogenase has been purified 1,278-fold from human liver mitochondria. The purified human enzyme has a subunit molecular weight of 58,800 and a native molecular weight of 256,000. Human glutaryl-CoA dehydrogenase showed a reaction of only partial identity when compared to porcine glutaryl-CoA dehydrogenase by Ouchterlony double immunodiffusion analysis using antiserum raised against and monospecific for porcine glutaryl-CoA dehydrogenase.


Surgical Site Infection Rate Drops to 0% Using a Vacuum-Assisted Closure in Contaminated/Dirty Infected Laparotomy Wounds.
Wound site infections increase costs, hospital stay, morbidity, and mortality. Techniques used for wounds management after laparotomy are primary, delayed primary, and vacuum-assisted closures. The objective of this study is to compare infection rates between those techniques in contaminated and dirty/infected wounds. Eighty-one laparotomized patients with Class III or IV surgical wounds were enrolled in a three-arm randomized prospective study. Patients were allocated to each group with the software Research Randomizer® (Urbaniak, G. C., & Plous, S., Version 4.0). Presence of infection was determined by a certified board physician according to Centers for Disease Control's Criteria for Defining a Surgical Site Infection. Twenty-seven patients received primary closure, 29 delayed primary closure, and 25 vacuum-assisted closure, with no exclusions for analysis. Surgical site infection was present in 10 (37%) patients treated with primary closure, 5 (17%) with primary delayed closure, and 0 (0%) patients receiving vacuum-assisted closure. Statistical significance was found between infection rates of the vacuum-assisted group and the other two groups. No significant difference was found between the primary and primary delayed closure groups. The infection rate in contaminated/dirty-infected laparotomy wounds decreases from 37 and 17 per cent with a primary and delayed primary closures, respectively, to 0 per cent with vacuum-assisted systems.


Truncation of the mismatch repair protein PMS2 during the neoplastic transformation of human breast epithelial cells in vitro.
Alterations in known mismatch repair (MMR) genes have been found in many cancers, such as in hereditary non-polyposis colorectal cancer syndrome (HNPCC), in addition to specific oncogenes and tumor suppressor gene abnormalities. We have reported mutations in the MMR genes hMSH2, hPMS2 and hMSH6 in human breast epithelial cell lines (HBEC). In hPMS2 we found a non-sense mutation in codon number 471 by a change of glutamine (CAG) by stop codon (TAG). We also observed that the number of alterations in the MMR genes were increased during the process of cell transformation and tumorigenesis. In the present study in order to determine the role of those mutations in the hPMS2 protein activity, we used western blotting, protein truncation test and MMR activity assay employing HBEC cells. In the transformed and tumor cell lines we found a premature hPMS2 protein truncated by western blot and also we confirmed this truncation using the protein truncation test. In the MCF-10F cells only the wild-type hPMS2 protein was detected. The functionality of the hPMS2-truncated protein was confirmed by DNA heteroduplex assay using specific mispaired bases in mutated M13mp2 phages. DNA repair activity was proficient in MCF-10F cells, partially deficient in the transformed cells, and totally deficient in the tumor cell lines. In order to test whether the MMR deficiency was due to an hPMS2 defect, we used the wild-type heterodimer hPMS2-hMLH1 in the in vitro reaction of DNA heteroduplex assay; we have found that addition of the hPMS2 wild-type increased the MMR repair efficiency by more than 40% in tumor cell lines. Our results indicate that a protein-truncating mutation in the hPMS2 gene had occurred during the process of cell transformation and tumorigenesis and that truncation is related to a lack of function of this gene.


Clinical trial: comparative effectiveness of cephalexin plus trimethoprim-sulfamethoxazole versus cephalexin alone for treatment of uncomplicated cellulitis: a randomized controlled trial.
Community-associated methicillin-resistant S. aureus (CA-MRSA) is the most common organism isolated from purulent skin infections. Antibiotics are usually not beneficial for skin abscess, and national guidelines do not recommend CA-MRSA coverage for cellulitis, except purulent cellulitis, which is uncommon. Despite this, antibiotics targeting CA-MRSA are prescribed commonly and increasingly for skin infections, perhaps due, in part, to lack of experimental evidence among cellulitis patients. We test the hypothesis that antibiotics targeting CA-MRSA are beneficial in the treatment of cellulitis. We performed a randomized, multicenter, double-blind, placebo-controlled trial from 2007 to 2011. We enrolled patients with cellulitis, no abscesses, symptoms for <1 week, and no diabetes, immunosuppression, peripheral vascular disease, or hospitalization (clinicaltrials.gov NCT00676130). All participants received cephalexin. Additionally, each was randomized to trimethoprim-sulfamethoxazole or placebo. We provided 14 days of antibiotics and instructed participants to continue therapy for ≥1 week, then stop 3 days after they felt the infection to be cured. Our main outcome measure was the risk difference for treatment success, determined in person at 2 weeks, with telephone and medical record confirmation at 1 month. We enrolled 153 participants, and 146 had outcome data for intent-to-treat analysis. Median age was 29, range 3-74. Of intervention participants, 62/73 (85%) were cured versus 60/73 controls (82%), a risk difference of 2.7% (95% confidence interval, -9.3% to 15%; P = .66). No covariates predicted treatment response, including nasal MRSA colonization and purulence at enrollment. Among patients diagnosed with cellulitis without abscess, the addition of trimethoprim-sulfamethoxazole to cephalexin did not improve outcomes overall or by subgroup. NCT00676130.


An ultrasmall PVP-Fe-Cu-Ni-S nano-agent for synergistic cancer therapy through triggering ferroptosis and autophagy.
Photothermal therapy (PTT) is an emerging field where photothermal agents could convert visible or near-infrared (NIR) radiation into heat to kill tumor cells. However, the low photothermal conversion efficiency of photothermal agents and their limited antitumor activities hinder the development of these agents into monotherapies for cancer. Herein, we have fabricated an ultrasmall polyvinylpyrrolidone (PVP)-Fe-Cu-Ni-S (PVP-NP) nano-agent via a simple hot injection method with excellent photothermal conversion efficiency (∼96%). Photothermal therapy with this nano-agent effectively inhibits tumor growth without apparent toxic side-effects. Mechanistically, our results demonstrated that, after NIR irradiation, PVP-NPs can induce ROS/singlet oxygen generation, decrease the mitochondrial membrane potential, release extracellular Fe2+, and consume glutathione, triggering autophagy and ferroptosis of cancer cells. Moreover, PVP-NPs exhibit excellent contrast enhancement according to magnetic resonance imaging (MRI) analysis. In summary, PVP-NPs have a high photothermal conversion efficiency and can be applied for MRI-guided synergistic photothermal/photodynamic/chemodynamic cancer therapy, resolving the bottleneck of existing phototherapeutic agents.


Precise location of linear epitopes on the capsid surface of feline calicivirus recognized by neutralizing and non-neutralizing monoclonal antibodies.
We report the generation, characterization and epitope mapping of a panel of 26 monoclonal antibodies (MAbs) against the VP1 capsid protein of feline calicivirus (FCV). Two close but distinct linear epitopes were identified at the capsid outermost surface (P2 subdomain) of VP1, within the E5'HVR antigenic hypervariable region: one spanning amino acids 431-435 (PAGDY), highly conserved and recognized by non-neutralizing MAbs; and a second epitope spanning amino acids 445-451 (ITTANQY), highly variable and recognized by neutralizing MAbs. These antibodies might be valuable for diagnostic applications, as well as for further research in different aspects of the biology of FCV.


Patient samples for measuring primary care physician performance: who should be included?
In measuring patients' experiences with individual primary care physicians (PCPs), the reliability and validity of data based on samples of "established" patients of a physician's panel have been shown. However, as large-scale initiatives seek the least costly approach to obtaining these data, little is known about the implications of expanding samples to include any patient who has seen the physician in the relevant time period. A brief validated patient questionnaire was administered to a random sample of patients visiting 67 PCPs in California between January and October 2005. We evaluated the concordance between administrative and patient-reported information on whether the physician was the patient's PCP. Response rates, data quality, and experiences reported by confirmed "established" patients were compared with those of "unestablished" patients. Administrative data designating patients as established to a PCP were highly concordant with patient self-report (96.5%). For unestablished patients, concordance was considerably lower (40.0%). Response rates among established patients were higher than those of patients believed to be unestablished (35.5% vs. 22.2%). Compared with established patients of a PCP's practice, unestablished patients reported significantly less favorable experiences with the doctor (interaction quality, P < 0.001; health promotion, P < 0.001; access, P < 0.001; integration, P < 0.05). The ranking of individual physicians differed for established and unestablished patient samples. Initiatives measuring patients' experiences with individual primary care physicians will achieve different results (response rates, physician scores) if samples include any patient who has seen the physician versus those whom administrative data indicate as established members of the physician's panel.


Effects of zinc and copper on adhesion and hemagglutination of Prevotella intermedia and Prevotella nigrescens.
This study investigated the mechanism of protein attachment to the surface of the putative periodontal pathogens Prevotella intermedia and Prevotella nigrescens in artificial gingival crevicular fluid, and ways to increase protein attachment to the bacterial cells. The effects of cations on protein attachment, bacterial adhesion, and hemagglutination were examined, and cation-binding components on both bacterial species were identified. The presence of cations, especially zinc, copper and cerium, increased attachment of human serum proteins to both bacterial species. In contrast, the presence of hydrophobic inhibitors or sugars had little effect. Protein attachment was reduced by heat treatment of the bacterial cells. Pretreatment of bacteria with human serum proteins inhibited adhesion of both species to buccal epithelial cells and hemagglutination. These effects were enhanced by the presence of zinc and copper during pretreatment. Using a chelating column, specific zinc- and copper-binding proteins were identified on the surfaces of both bacterial species.


Screening a small molecule library to identify inhibitors of NF-κB inducing kinase and pro-labor genes in human placenta.
The non-canonical NF-κB signaling (RelB/p52) pathway drives pro-labor genes in the human placenta, including corticotropin-releasing hormone (CRH) and cyclooxygenase-2 (COX-2), making this a potential therapeutic target to delay onset of labor. Here we sought to identify small molecule compounds from a pre-existing chemical library of orally active drugs that can inhibit this NF-κB signaling, and in turn, human placental CRH and COX-2 production. We used a cell-based assay coupled with a dual-luciferase reporter system to perform an in vitro screening of a small molecule library of 1,120 compounds for inhibition of the non-canonical NF-κB pathway. Cell toxicity studies and drug efflux transport MRP1 assays were used to further characterize the lead compounds. We have found that 14 drugs have selective inhibitory activity against lymphotoxin beta complex-induced activation of RelB/p52 in HEK293T cells, several of which also inhibited expression of CRH and COX-2 in human term trophoblast. We identified sulfapyridine and propranolol with activity against CRH and COX-2 that deserve further study. These drugs could serve as the basis for development of orally active drugs to affect length of gestation, first in an animal model, and then in clinical trials to prevent preterm birth during human pregnancy.


The role of endothelial lipase in lipid metabolism, inflammation, and cancer.
Endothelial lipase (LIPG) plays a critical role in lipoprotein metabolism, cytokine expression, and the lipid composition of cells. Thus far, the extensive investigations of LIPG have focused on its mechanisms and involvement in metabolic syndromes such as atherosclerosis. However, recent developments have found that LIPG plays a role in cancer. This review summarizes the field of LIPG study. We focus on the role of LIPG in lipid metabolism and the inflammatory response, and highlight the recent insights in its involvement in tumor progression. Finally, we discuss potential therapeutic strategies for targeting LIPG in cancer, and the therapeutic potential of LIPG as a drug target.


Effects of Giardia lamblia Colonization and Fenbendazole Treatment on Canine Fecal Microbiota.
The gut microbiota (GM) is the sum of hundreds of distinct microbial species that can equal or outnumber their host'ssomatic cells. The GM influences a multitude of physiologic and immunologic processes in the host, and changes in the GM have been shown to alter the phenotypes of animal models. Previous studies using rodents have also shown that the composition of the GM is affected by many factors, including diet, husbandry, housing, and the genetic background of the animals. However, limited information exists about factors that may modulate GM in other laboratory species, such as dogs. We sought to eliminate sporadic Giardia colonization of dogs using fenbendazole (FBZ), an antiprotozoal widely used in biomedical research dog colonies. Concerns that FBZ could have inadvertent effects on the canine GM led us to assess GM over the course of treatment. FBZ (50 mg/kg) was given orally to all dogs in 3 different facilities (n = 19 to 25) for 10 consecutive days. Fecal samples were obtained 2 d before the initiation of treatment, on the last day of treatment, and 2 wk after the completion of treatment. Targeted 16S rRNA gene sequencing was used to analyze fecal microbiota. All dogs were clinically normal throughout the sample collection period. Statistical analyses of data showed significant differences between dogs housed in the 3 different facilities, further emphasizing the effect of housing and husbandry factors on the GM. However,negligible differences were seen between time points, indicating that FBZ did not significantly alter the canine GM. Comparison of the GM of Giardia lamblia positive and negative dogs revealed no significant difference between the 2 groups. These findings suggest that FBZ can be used therapeutically in dogs with minimal impact on the GM. Furthermore, the presence ofG. lamblia in clinically normal animals may not be sufficient to influence the normal canine microbiota.


Fluid-membrane tethers: minimal surfaces and elastic boundary layers.
Thin cylindrical tethers are common lipid bilayer membrane structures, arising in situations ranging from micromanipulation experiments on artificial vesicles to the dynamic structure of the Golgi apparatus. We study the shape and formation of a tether in terms of the classical soap-film problem, which is applied to the case of a membrane disk under tension subject to a point force. A tether forms from the elastic boundary layer near the point of application of the force, for sufficiently large displacement. Analytic results for various aspects of the membrane shape are given.


Arthrobacter aurescens TC1 atrazine catabolism genes trzN, atzB, and atzC are linked on a 160-kilobase region and are functional in Escherichia coli.
Arthrobacter aurescens strain TC1 metabolizes atrazine to cyanuric acid via TrzN, AtzB, and AtzC. The complete sequence of a 160-kb bacterial artificial chromosome clone indicated that trzN, atzB, and atzC are linked on the A. aurescens genome. TrzN, AtzB, and AtzC were shown to be functional in Escherichia coli. Hybridization studies localized trzN, atzB, and atzC to a 380-kb plasmid in A. aurescens strain TC1.


Differential expression of endothelin-2 along the mouse intestinal tract.
Endothelin (ET)-2, an ET family peptide, is highly expressed in intestine. However, the specific distribution and function of ET-2 remain unknown. We elucidated the expression profile and localization of ET-2 in mouse gastrointestinal tract. Real-time PCR analysis revealed that ET-2 gene expression in the gastrointestinal tract of healthy animals was relatively high in the colon. Immunohistochemical analysis revealed ET-2-like immunoreactivity mainly in epithelial cells of the mucosa throughout the intestinal tract of healthy animals. Intracellularly, ET-2 was concentrated close to the basement membrane of intestinal epithelial cells. A weak ET-2-like immunoreactivity was also localized to some neurofibers and the myenteric plexus of the muscle layer, coexpressing with vasoactive intestinal peptide. ET-2-like immunoreactivity was also detected at Brunner's glands of the duodenum and follicle-associated epithelium of Peyer's patch. In contrast, ET-1-like immunoreactivity was uniformly distributed in epithelial cells. In dextran sulfate sodium (DSS)-induced colitis, colonic ET-2 was upregulated during the late stage of DSS treatment. These results suggest that in intestinal epithelial cells ET-2 could be secreted into the lamina propria and the dome region in Peyer's patch, and that it might modulate immune cells in these sites for mucosal defense.


TEM-71, a novel plasmid-encoded, extended-spectrum beta-lactamase produced by a clinical isolate of Klebsiella pneumoniae.
TEM-71, a novel extended-spectrum beta-lactamase from a Klebsiella pneumoniae clinical isolate, had an isoelectric point of 6.0 and a substrate profile showing preferential hydrolysis of cefotaxime over ceftazidime. It differed from TEM-1 by two substitutions, Gly238Ser and Glu240Lys, and was under the control of the strong P4 promoter.


Morphofunctional studies of the glomerular wall in mice lacking entactin-1.
The architecture of the basement membranes is essential for proper function. This architecture is based on interactions among its components, which assemble in a complex network. Entactin-1 appears to be the mastermind of this assembling. In entactin-1-null transgenic mice, immunocytochemistry established the absence of entactin-1 in the glomerular basement membrane, and morphological thickening of this membrane was demonstrated. This prompted us to investigate the organization of other components of the glomerular basement membrane in the transgenic animals. The distribution of type IV collagen and laminin remained unchanged, whereas that of anionic charges was significantly altered. We also evaluated the impact of the absence of entactin-1 on cell relays by studying the alpha(3)- and the alpha(v)-integrins along the endothelial and epithelial glomerular cell plasma membranes. Only the density of alpha(v) was found to be increased. Finally, the filtration properties of the glomerular wall were evaluated by revealing endogenous albumin distribution across the basement membrane. This was altered in transgenic animals, suggesting changes in permselectivity properties. Entactin-1 appears to be an essential component in basement membranes because its absence appears to modify the molecular organization leading to alterations in functional properties.


[Hepatotoxicity of antitubercular drugs. Apropos of 28 cases].
Anti-tuberculosis have up set the out come of this disease, however undesirable effects principally hepatotoxicity remain of ten the ransom of this therapy success. 28 observations of hepatic toxicity in allergology pneumo service in military hospital of Tunisia during 5 years (1996-2001) among 321 tuberculosis. Hepatic toxicity is observed in 28 cases, 21 cytolytics, 3 cholostatics and 4 mixt. 3 decease (death) have been observed. Anti-tuberculosis medicament (medecine) are sometimes this cause of many undesirable effect, which can be miner or major and even cases of decease. So that we have to take an interest at time and specialty to follow recommandation which are more and more codified.


An Absent Lingual Frenulum in a Non-syndromic Premature Infant.
The lingual frenulum (LF) is a fold of tissue that connects the tongue to the oral cavity's floor. Abnormal frenula are associated with speech alterations. The absence of the LF is associated with Ehler's Danlos syndrome (EDS). In this case report, we present a premature infant incidentally found to have an absent lingual frenulum, with recurrent desaturations during feeding. The desaturations were believed to be due to the absent lingual frenulum, but they resolved after one month without treatment and were then attributed to apnea of prematurity. Whole exome sequence showed no genetic disorders. The infant is now doing well with no interventions. An absent lingual frenulum warrants molecular genetic testing for EDS. However, it does not warrant any treatment; special considerations are only required during intubation.


Comparative vascular effects of midazolam and lorazepam administered during cardiopulmonary bypass.
We studied the comparative vascular effects of midazolam and lorazepam in 52 patients undergoing elective cardiac surgery procedures. After administration of fentanyl, 100 micrograms/kg intravenously, the patients were randomized to receive either midazolam 0.1 mg/kg (M high), midazolam 0.05 mg/kg (M low), lorazepam 0.1 mg/kg (L high), lorazepam 0.05 mg/kg (L low), or placebo during cardiopulmonary bypass (CPB). Compared to the placebo and L-low groups, the high-dose midazolam group had more effect on systemic vascular resistance (SVR) starting at 5 min after study drug administration (P < 0.02). M high and L high required increased quantities of phenylephrine administered to maintain the mean arterial pressure (MAP) at > 50 mm Hg after rewarming and removal of the aortic cross-clamp (P < 0.03). In the first 12 h of the postoperative period, the M-high group required phenylephrine (PHE) infusion for hypotension associated with decreased SVR more often than placebo (8/10 vs 1/11 patients, P < 0.008, chi 2). In conclusion, midazolam more effectively attenuated the increase in SVR that occurred during CPB than patients receiving either placebo or lorazepam. The hemodynamic effects from a single dose of 0.1 mg/kg midazolam administered at the start of CPB may persist into the postoperative period.


Proposal of Lactococcus garvieae subsp. bovis Varsha and Nampoothiri 2016 as a later heterotypic synonym of Lactococcus formosensis Chen et al. 2014 and Lactococcus formosensis subsp. bovis comb. nov.
Currently, Lactococcus garvieae contains two subspecies: L. garvieae subsp. bovis and L. garvieae subsp. garvieae. In a study by Varsha and Nampoothiri, high pheS (99.7 %) and rpoA (99.6 %) sequence similarities indicated that L. garvieae subsp. bovis and Lactococcus formosensis probably have a close taxonomic relationship; low pheS (92.2 %) and rpoA (97.8 %) sequence similarities and relatively low DNA-DNA hybridization value (75.8 %) indicated that L. garvieae subsp. bovis and L. garvieae subsp. garvieae probably represent two different species. In the present study, the taxonomic relationships between L. garvieae subsp. bovis, L. garvieae subsp. garvieae and L. formosensis were re-examined based on sequence analyses of 16S rRNA, pheS, recA, rpoA and rpoB genes, average nucleotide identity (ANI), digital DNA-DNA hybridization (dDDH) values, average amino acid identity (AAI), fatty acid methyl ester analysis and phenotypic characterization. L. garvieae subsp. bovis LMG 30663T exhibited 97.3 % ANI, 78.3 % dDDH and 96.4 % AAI values to L. formosensis NBRC 109475T, higher than the threshold for species demarcation (95-96, 70 and 95-96 %, respectively), indicating that L. garvieae subsp. bovis LMG 30663T and L. formosensis NBRC 109475T belong to the same species. L. garvieae subsp. bovis LMG 30663T had 91.2 % ANI, 43.3 % dDDH and 92.9-93.0% AAI values with the type strain of L. garvieae subsp. garvieae, indicating that they represent two different species. Because L. formosensis has been proposed and validated before L. garvieae subsp. bovis, L. garvieae subsp. bovis is transferred to L. formosensis as L. formosensis subsp. bovis comb. nov. The type strain of L. formosensis subsp. bovis is BSN307T (=DSM 100577T=MCC 2824T=KCTC 21083T=LMG 30663T). The type strain of L. formosensis subsp. formosensis is 516T (=NBRC 109475T=BCRC 80576T).


Fishing for vaccines against Vibrio cholerae using in silico pan-proteomic reverse vaccinology approach.
Cholera, an acute enteric infection, is a serious health challenge in both the underdeveloped and the developing world. It is caused by Vibrio cholerae after ingestion of fecal contaminated food or water. Cholera outbreaks have recently been observed in regions facing natural calamities (i.e., earthquake in Haiti 2010) or war (i.e., ongoing civil war in Yemen 2016) where healthcare and sanitary setups have been disrupted as a consequence. Whole-cell oral cholera vaccines (OCVs) have been in market but their regimen efficacy has been questioned. A reverse vaccinology (RV) approach has been applied as a successful anti-microbial measure for many infectious diseases. With the aim of finding new protective antigens for vaccine development, the V. cholerae O1 (biovar eltr str. N16961) proteome was computationally screened in a sequential prioritization approach that focused on determining the antigenicity of potential vaccine candidates. Essential, accessible, virulent and immunogenic proteins were selected as potential candidates. The predicted epitopes were filtered for effective binding with MHC alleles and epitopes binding with greater MHC alleles were selected. In this study, we report lipoprotein NlpD, outer membrane protein OmpU, accessory colonization factor AcfA, Porin, putative and outer membrane protein OmpW as potential candidates qualifying all the set criteria. These predicted epitopes can offer a potential for development of a reliable peptide or subunit vaccine for V. cholerae.


Scanning electron microscopic observation of immunohistochemically specified stromal cells following the NaOH maceration method.
The present study introduces a method combining immunohistochemistry and a chemical digestion technique applied to scanning electron microscopy (SEM) observation of chemically characterized stromal cells in the myenteric layer of the monkey small intestine. The whole-mount preparation containing the myenteric layer was treated immunohistochemically using small colloidal gold and silver enhancements for detecting a certain type of stromal cell, followed by an alkali maceration method to remove extracellular matrices covering the immunohistochemically stained cells. This method permitted direct SEM observation of the stained stromal cells. Secondary and backscatter electron images clearly visualized the ultrastructure of the immunohistochemically-stained cells and the spatial distribution of the reaction products, respectively. It is suggested that the present method is useful for demonstrating the three-dimensional organization of the chemically specified stromal cells.


Severe right ventricular hypertrophy in hypertrophic cardiomyopathy: Serious symptoms, complex surgical procedures, and poor prognosis in Fuwai Hospital.
Severe right ventricular hypertrophy (SRVH) in hypertrophic cardiomyopathy (HCM) is rare. We studied the clinical characteristics and prognosis of 36 patients with HCM and SRVH in a Chinese cohort. Patients with HCM and SRVH were enrolled between 2013 and 2017. The clinical characteristics, treatment therapies, and clinical outcomes of the 36 patients were retrospectively studied and compared with those of 128 patients without SRVH. Patients in the group with SRVH were younger than those in the group without SRVH (27.58±15.09 years vs 40.34±13.21 years, respectively; p<0.001). Patients with SRVH had more serious clinical symptoms and a higher New York Heart Association functional class than those without SRVH. Most patients in the group with SRVH exhibited diffuse RV hypertrophy, and 13 patients presented with biventricular outflow tract obstruction. Maximal left ventricular (LV) wall thickness (27.29±7.95 mm vs 24.33±5.85 mm, respectively; p=0.027) and LV outflow tract gradient (80.83±24.41 mm Hg vs 42.3±5.7 mm Hg, respectively; p=0.000) were significantly greater in patients with SRVH than in those without SRVH. A total of 30 patients in the group with SRVH underwent surgical correction. During a median follow-up period of 48 months, six patients with SRVH reached primary clinical endpoints (four sudden cardiac deaths, one heart failure-related death, and one heart transplantation), whereas only two deaths occurred in the patients without SRVH. We conclude that patients with HCM and SRVH exhibit serious symptoms and have complex surgical requirements and poor clinical outcomes.


Dental utilization by spouses of active duty U.S. military personnel.
This study explores dental utilization and access barriers to dental care for spouses of active duty U.S. military personnel as reported by their military sponsor. It also compares dental utilization of spouses and civilians. The data, collected using self-administered questionnaires between April 1994 and January 1995, are from 5,732 Army, Air Force, Navy, and Marine personnel with nonmilitary spouses. Comparative civilian data are from the most recent (1985-1986) U.S. oral health survey of working adults. Results show that dental utilization of military spouses parallels that of civilians. Cost was the most commonly cited barrier to dental care for spouses. Logistic regression results show that the likelihood of a spouse having seen a dentist within the past year is influenced by insurance status, sex, branch of service of sponsor, and rank of sponsor. Insurance status is the strongest predictor of dental utilization for spouses.


Wing polymorphism in Anaphothrips graminum (Thysanoptera: Thripidae).
Of the 83 species of Anaphothrips in the world (ThripsWiki 2018), only four are recorded in Iran so far (Alavi et al. 2018): A. microptera, A. obscurus, A. sineconus and A. sudanensis. Among these, A. sineconus was collected from Haloxylon persicum (Amaranthaceae) while the others are related to various grasses (Poaceae). Color and structural variation among and within the species of Anaphothrips have been demonstrated by several authors (Kudo, 1989, Nakao 1996, Mound Masumoto 2009). In this paper a fifth species, A. graminum, is recorded from Iran based on specimens collected from grasses. The previously unknown micropterous morph of this species is described, as this is different in color from the macropterous morph, and in contrast to the original description the male has a pore plate on sternite VIII. The terminology used here follows Mound and Masumoto (2009). Most specimens are deposited in the Department of Plant Protection, Shiraz University, Shiraz, Iran (PPSU). Two females (one micropterous and one macropterous) are deposited in ANIC - the Australian National Insect Collection, CSIRO, Canberra.


Intention to accept Bordetella pertussis booster vaccine during pregnancy in Mexico City.
Adult booster vaccination against pertussis can help prevent severe infections in young infants. We examined influences on intention to accept pertussis booster vaccination among pregnant women in Mexico City. We conducted a cross-sectional survey, recruiting convenience samples of pregnant women receiving prenatal care from three public healthcare centers between March and May 2012. Our primary outcome was intention to accept pertussis vaccination during pregnancy. We examined socio-demographic factors, vaccination history, pertussis knowledge, perceptions of vaccine information sources, and other potential influences on vaccine decision-making. A total of 402 pregnant women agreed to participate, of which 387 (96%) provided their intention to accept or decline pertussis vaccination. Among respondents, 57% intended to accept a pertussis booster vaccine if offered, but only 16% had ever heard of pertussis, and only 2% knew someone who had contracted this disease. Over 80% of respondents would accept pertussis vaccination if recommended by an obstetrician-gynecologist. The most frequently selected reasons to refuse pertussis vaccination were concerns that the vaccine might harm the unborn baby or pregnant woman. In multivariate analysis, rating doctors and nurses as good sources of vaccine information, and having ever heard of pertussis, were independently associated with intention to accept pertussis vaccination. Promoting patient awareness about pertussis disease and vaccine safety, and encouraging general practitioners, nurses and obstetricians to recommend pertussis booster vaccine, may increase vaccine uptake among pregnant women.


Panamanian forest mammals as carriers of Salmonella.
Enteric bacteria pathogenic to man were sought in a total of 974 forest mammals collected from a variety of sites in rural and jungle areas of Panamá. The highest incidence of infection among the mammals was observed during the Panamanian dry season, which normally extends from January through April. A minimum of 10 Salmonella serotypes including, three of the Arizona group and Ewardsiella tarda, was isolated. Opossums of the genera Philander, 11 of 54 (20.1%), and Didelphis, 12 of 102 (11.8%) demonstrated high infection rates. One sloth of the genus Choloepus and specimens of two genera of rodents also were infected to varying degrees: 1(11.1%) of 9 Choloepus, 8 (1.1%) of 704 Proechimys and 1 (16.7%) of 6 Diplomys.


Detection of lepromatous leprosy patients shedding M. leprae in nasal droppings by enzyme immunoassay.
Enzyme immunoassays (EIAs) for detection of lepromatous leprosy (LL) patients harbouring M. leprae in nasal mucosa are described. One EIA measures IgM antibodies against the synthetic disaccharide (ND-BSA) residue of phenolic glycolipid I of M. leprae, whereas the other titrates primarily IgG antibodies against sonicate supernatant antigens of Mycobacterium w. (M.w.). Fifty coded leprosy sera were analysed by EIAs under a double blind code. Amongst the 20 LL patients with positive nasal smear, 18 (90%) were positive in EIA based on ND-BSA, in comparison to 19 (95%) in EIA using M.w. antigens. The assays can be performed on fresh serum samples or on blood samples collected on filter paper discs. These assays can be useful for leprosy control programmes.


Distinguishing elastic shear deformation from friction on the surfaces of molecular crystals.
Elastic deformation on the surfaces of molecular crystals can be imaged using a variant of lateral force microscopy in which the tip is scanned parallel to the cantilever axis. The shear force transverse to this direction has a distinctly different origin than the friction force as determined by the tip velocity and temperature dependence of the cantilever torque. An elastic deformation model for the tip-sample interaction predicts the crystallographic anisotropy of the transverse shear contrast, establishing its connection with the relative magnitude of the in-plane elastic tensor components.