Sequence heterology and gene conversion at his-3 of Neurospora crassa.
Although sequence heterology clearly reduces crossing over in yeast, conflicting studies suggest that mismatches may increase or decrease gene conversion. To investigate this issue in an additional species, we measured the effect of local sequence heterology on conversion in his-3 of Neurospora crassa. Mismatches close to the cog recombination initiator or within his-3 reduce conversion to 70% and 30% of the homologous level, respectively, while heterologous insertions between his-3 and cog increase conversion by 20%. We suggest that, in both Neurospora and yeast, mismatches reduce the efficiency of the establishment and resolution stages of recombination, but substantial heterology may increase the progress of already established events by preventing repair synthesis from switching between templates. These data provide additional support that recombination at his-3 (and perhaps at yeast hotspots) proceeds by a synthesis-dependent strand-annealing mechanism, during which synthesis can switch templates, with the process being more tolerant of sequence mismatch in Neurospora.


[Melanoma microenvironment-impact of modern therapies].
A considerable proportion of patients with advanced melanoma succumb to metastatic disease despite the initial success of modern therapies. An overview of the melanoma tumor microenvironment with special focus on approved therapies and new innovative strategies is given. Current clinical trials and scientific insights concerning the impact of the tumor microenvironment on progression and therapy of advanced melanoma are reviewed and discussed. The tumor microenvironment with its manifold components and interactions plays a major role in the treatment of malignant melanoma. Innovative new strategies that target an immunosuppressive microenvironment may improve the therapeutic efficacy of current treatment of advanced melanoma.


Age and education as factors associated with medication literacy: a community pharmacy perspective.
Aging implies a higher prevalence of chronic pathologies and a corresponding increase in medication. The correct adherence and use of the medication are prerequisites for reducing risks of disease progression, comorbidity, and mortality. Medication literacy (ML) is the specific ability to safely access and understand the information available concerning medication, and to act accordingly. Currently, there are few specific instruments that ascertain the extent of ML in the general population. The aim of this work was to analyse ML in a large cohort of pharmacy customers. A total of 400 community pharmacy clients were analyzed to assess the level of ML (documental and numeracy) through the validated MedLitRxSE tool. The results showed that out of a total of 400 community pharmacy clients only 136 (34%) had an adequate degree of ML, while the rest of the clients (n = 264; 66%) were adjudged not to have this ability. Statistically significant differences were found between the different age groups in terms of ML (P < 0.001; OR = 0.312; 95% CI: 0.195-0.499), the 51-65 and >65-year age groups having a lower frequency of adequate ML (23.5 and 7.1%, respectively) than the rest of the age groups. A statistically significant increase in adequate ML was observed as the academic level of the clients increased (P < 0.001; OR = 15.403; 95% CI: 8.109-29.257). Multivariate logistic regression confirmed the influence of both variables on ML. An inadequate ML level was found in community pharmacy clients over the age of 51, and also in those with primary or non-formal studies. Our data add to our knowledge about ML, and should pharmacists and other health professionals to adopt new strategies to prevent, or at least reduce, errors in taking medicines, thus avoiding the undesirable effects of any misuse.


The effects of heparin and protamine on resorption of bone particles.
Implantation of bone particles into rats initiates rapid, reproducible resorption that can be quantitated by histomorphometric analysis. Mast cells are found with the mononuclear and multinucleated cells that digest the bone. One of the constituents of mast cell secretory granules, heparin, is known to regulate collagenase activity. This study shows that 1) exogenous heparin stimulated resorption of implanted bone to 141% of control values and that 2) protamine, a heparin antagonist, reduced resorption to rates 50% of controls.


Global approaches to understanding ubiquitination.
Ubiquitination--the linkage of one or more molecules of the protein ubiquitin to another protein--regulates a wide range of biological processes in all eukaryotes. We review the proteome-wide strategies that are being used to study aspects of ubiquitin biology, including substrates, components of the proteasome and ubiquitin ligases, and deubiquitination.


MRA as the Preferred Test for Pulmonary Embolism During the Iodinated Contrast Media Shortage of 2022: A Single-Center Experience.
BACKGROUND. Closure of a GE Healthcare facility in Shanghai, China, in 2022 disrupted the iodinated contrast media supply. Technologic advances have addressed limitations associated with the use of pulmonary MRA for diagnosis of pulmonary embolism (PE). OBJECTIVE. The purpose of this study was to describe a single institution's experience in the use of pulmonary MRA as an alternative to CTA for the diagnosis of PE in the general population during the iodinated contrast media shortage in 2022. METHODS. This retrospective single-center study included all CTA and MRA examinations performed to exclude PE from April 1 through July 31 (18 weekly periods) in 2019 (before the COVID-19 pandemic and contrast media shortage), 2021 (during the pandemic but before the shortage), and 2022 (during both the pandemic and the shortage). From early May through mid-July of 2022, MRA served as the preferred test for PE diagnosis, to preserve iodinated contrast media. CTA and MRA reports were reviewed. The total savings in iodinated contrast media volume resulting from preferred use of MRA was estimated. RESULTS. The study included 4491 examinations of 4006 patients (mean age, 57 ± 18 [SD] years; 1715 men, 2291 women): 1245 examinations (1111 CTA, 134 MRA) in 2019, 1547 examinations (1403 CTA, 144 MRA) in 2021, and 1699 examinations (1282 CTA, 417 MRA) in 2022. In 2022, the number of MRA examinations was four (nine when normalized to a 7-day period) in week 1, and this number increased to a maximum of 63 in week 10 and then decreased to 10 in week 18. During weeks 8-11, more MRA examinations (range, 45-63 examinations) than CTA examinations (range, 27-46 examinations) were performed. In 2022, seven patients with negative MRA underwent subsequent CTA within 2 weeks; CTA was negative in all cases. In 2022, 13.9% of CTA examinations (vs 10.3% of MRA examinations) were reported as having limited image quality. The estimated 4-month savings resulting from preferred use of MRA in 2022, under the assumption of uniform simple linear growth in CTA utilization annually and a CTA dose of 1 mL/kg, was 27 L of iohexol (350 mg I/mL). CONCLUSION. Preferred use of pulmonary MRA for PE diagnosis in the general population helped to conserve iodinated contrast media during the 2022 shortage. CLINICAL IMPACT. This single-center experience shows pulmonary MRA to be a practical substitute for pulmonary CTA in emergency settings.


The effect of decentralisation or chronic hypogastric nerve stimulation in vivo on the innervation and responses of the guinea-pig vas deferens.
The physiological, pharmacological and morphological characteristics of guinea-pig vas deferens supplied by hypogastric nerves rendered inactive by decentralisation were compared with those of vas deferens in which the nerve supply had been chronically stimulated for 3-9 days using implanted electrodes. No change was seen in decentralised preparations prior to 7 days, but from 8-15 days, increased sensitivity to application of noradrenaline in vitro was observed, which was shown to be related to reduced transmitter uptake by nerve terminals as well as to an increase in postjunctional sensitivity; there was also increased fatigability 7-14 days following decentralisation. Continuous stimulation of hypogastric nerves at 2 Hz for 4-8 h daily for 4-8 days resulted in enhanced transmitter uptake and reduced responses to noradrenaline; this was associated with a slight increase in noradrenaline content and a faster adrenergic neuromuscular response with a shorter latency. No appreciable changes in nerve or muscle structure studied by electron microscopy were observed following decentralisation, but there was an increase of between 12.5 and 29.6% in the number of close (less than 100 nm) neuromuscular junctions following chronic stimulation for 8 days.


Linear order of the four BCR-related loci in 22q11.
It has recently been shown the a probe for the 3' end of the BCR gene recognizes a family of four BCR-like genes that map to 22q11. Using a panel of somatic cell hybrids with rearrangement of chromosome 22, we have determined their order within 22q11: BCR-2, BCR4, BCR1, BCR-3, with BCR-2 the most centromere proximal. All of the BCR-like genes map proximal to the 22q11-q12 breakpoint of a t(11;22) in a Ewing sarcoma.


An ultrasensitive biosensor for colorimetric detection of Salmonella in large-volume sample using magnetic grid separation and platinum loaded zeolitic imidazolate Framework-8 nanocatalysts.
Salmonella is the leading risk factor in food safety. Rapid, sensitive and accurate detection of Salmonella is a key to prevent and control the outbreaks of foodborne diseases caused by Salmonella. In this study, we reported a colorimetric biosensor for ultrasensitive detection of Salmonella Typhimurium using a magnetic grid separation column to efficiently separate target bacteria from large volume of sample and platinum loaded zeolitic imidazolate framework-8 (Pt@ZIF-8) nanocatalysts to effectively amplify biological signal. The target Salmonella cells in large volume of sample were first separated and concentrated using the magnetic grid separation column with immune magnetic particle chains, then conjugated with the immune Pt@ZIF-8 nanocatalysts to mimic peroxidase for catalysis of hydrogen peroxide-3,3',5,5'-tetramethylbenzidine, and finally determined by measuring the catalysate at characteristic wavelength of 450 nm. This proposed biosensor was able to separate ∼70% of target Salmonella cells from 50 mL of bacterial sample and quantitatively detect Salmonella from 101 to 104 CFU/mL in 2.5 h with the lower detection limit of 11 CFU/mL. The mean recovery for Salmonella in spiked chicken carcass was about 109.8%. This new magnetic grid separation method was first time reported for efficient separation of target bacteria from very large volume of sample to greatly improve the sensitivity of this biosensor and could be used with various biosensing assays for practical applications in routine detection of foodborne pathogens without any bacterial pre-enrichment.


Collagen synthesis in normal BHK cells and temperature-sensitive chemically transformed BHK cells.
Collagen synthesis in normal BHK 21/cl 13 and chemically transformed temperature sensitive BHK 21/cl 13 cells (Me2N4) was assessed by examination of hydroxyproline formation and collagenase-susceptible protein. The Me2N4 cells lost their ability to synthesize collagen at both permissive and nonpermissive temperatures for transformation. These conclusions were confirmed by polyacrylamide-gel eletrophoresis and CM-cellulose chromatography. Prolyl hydroxylase activity was present in both normal and transformed cells even when no collagen could be demonstrated. The production of noncollagen protein, although decreased in the transformed cell, did not change as drastically as the collagen synthesis.


Amino acid sequence requirements in the epitope recognized by the alpha-tubulin-specific rat monoclonal antibody YL 1/2.
We have characterized the epitope of the rat monoclonal antibody YL 1/2 in detail using synthetic peptides and several alpha-tubulin derivatives. The epitope seems to be provided by the linear sequence spanning the carboxy-terminal residues of tyrosinated alpha-tubulin. By competitive ELISA, dipeptides covering the carboxyl end could be antigenically recognized. Three sites were deduced at the dipeptide level: a negatively charged side chain in the penultimate position followed by an aromatic residue which must carry the free carboxylate group. Experiments with longer peptides point to a further negative charge provided by a carboxylate group on the third residue from the end. Thus the tripeptide Glu-Glu-Tyr was only 5-fold less active than the octapeptide spanning the carboxy-terminal alpha-tubulin sequence. The octapeptide itself showed only a 40-fold lower activity than tyrosinated alpha-tubulin. In line with the emerging epitope requirements of YL 1/2, the Escherichia coli rec A protein, the catalytic subunit of the cyclic AMP-dependent muscle protein kinase as well as performic acid-oxidized actin were recognized by YL 1/2 in immunoblots. These results thus define the sequence requirements within a probably linear epitope and give rise to some general questions concerning experiments where monoclonal antibodies are microinjected into cells in order to assess the contribution of a known antigen to cellular physiology.


TrfA dimers play a role in copy-number control of RK2 replication.
Copy-number regulation of the broad-host-range plasmid RK2 is dependent on the plasmid-encoded initiator protein, TrfA, and the RK2 origin of replication. The handcuffing model for copy-number control proposes that TrfA-bound oris reversibly couple to prevent the further initiation of plasmid replication when the copy number in vivo is at or above the replicon-specific copy number. TrfA mutants have been isolated which allow for oriV replication at elevated copy numbers. To better understand the mechanism of 'handcuffing', the copy-up TrfA(G254D/S267L) mutant was characterized further. In the present study we show by size exclusion chromatography and native gel electrophoresis that unlike wt TrfA which is largely dimeric, purified His6-TrfA(G254D/S267L) is primarily monomeric. In vivo, TrfA33(G254D/S267L) supports replication of an RK2 ori plasmid in trans at a greatly elevated copy number, while in cis the plasmid exhibits runaway replication. However, expression of either of two previously isolated DNA-binding defective TrfA mutants, TrfA33(P151S) or TrfA33(S257F), in a cell transformed with a mini-RK2 replicon encoding TrfA33(G254D/S267L) results in suppression of the runaway phenotype. His6-TrfA(P151S) and His6-TrfA(S257F) purify as dimers, and when expressed in vivo are incapable of supporting RK2 plasmid replication. In contrast, combination of the trfA(P151S) or trfA(S257F) mutation with the trfA(G254D/S267L) mutations results in the expression of mutant TrfA proteins which are mainly monomers and which can no longer restore copy control to replication directed by TrfA33(G254D/S267L) in vivo. On the basis of these findings a handcuffing model is proposed, whereby oriV-bound TrfA monomers are coupled by dimeric TrfA molecules.


Using available clinical information in practice-based research: mining for silver while dreaming of gold.
Social workers in health and mental health settings routinely collect and record enormous quantities of clinical information about clients, psycho-social interventions and client responses to these interventions. Despite its abundance and non-intrusiveness, social work researchers generally have ignored available clinical information, claiming that it is unreliable and subject to too many threats to validity to warrant serious consideration as a data source. Instead, many researchers have advocated "gold standard" experimental studies, employing standardized instruments and prospective data-collection. As a result, the research potential of retrospective studies based on available clinical information has been relatively unexplored and untested. This paper asserts that available clinical information can be converted into valuable retrospective, data-bases for practice-based research studies. Exemplars of such studies in health and mental health settings are provided as are guidelines for their conduct.


Assessing the pharmacodynamic profile of intravenous antibiotics against prevalent Gram-negative organisms collected in Colombia.
This study was designed to simulate standard and optimized dosing regimens for intravenous antibiotics against contemporary populations of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa using MIC distribution data to determine which of the tested carbapenem regimens provided the greatest opportunity for obtaining maximal pharmacodynamic (PD) activity. The isolates studied were obtained from the COMPACT-COLOMBIA surveillance program conducted between February and November 2009. Antimicrobial susceptibility testing was conducted by broth microdilution method according to the CLSI guidelines. Doripenem, imipenem-cilastatin, and meropenem, were the modeled antibiotics. A 5,000 patient Monte Carlo simulation was performed for each regimen and PD targets were defined as free drug concentrations above the MIC for at least 40% of the dosing interval. All carbapenem regimens obtained optimal exposures against E. coli, unlike the other Enterobacteriaceae tested. Against P. aeruginosa, only a prolonged infusion of doripenem exceeded the 90% cumulative fraction of response (CFR) threshold. Worrisomely, no regimens for any of the drugs tested obtained optimal CFR against A. baumannii. For P. aeruginosa intensive care unit (ICU) isolates, CFR was approximately 20% lower for isolates collected in the respiratory tract compared with bloodstream or intra-abdominal for imipenem and meropenem. Noteworthy, all doripenem and meropenem regimens achieved greater than 90% CFR against bloodstream and respiratory isolates of K. pneumoniae. Our data suggests that higher dosing and prolonged infusion of doripenem or meropenem may be suitable for empirically treating ICU P. aeruginosa, while none of the carbapenems achieved optimal cumulative fraction of response against A. baumannii. Standard dosing regimens of all the carbapenems tested achieved optimal CFR against E. coli isolates, but higher carbapenem dosages might be required for empiric treatment of K. pneumoniae, particularly from an intra-abdominal source. Non-standard dosage regimens studied in this modeling should be proven effective in prospective clinical trials.


Facilitating influence of disopyramide on atrial flutter termination by overdrive pacing.
Long-lasting (mean 30 days) type I atrial flutter was treated with overdrive pacing in 30 patients (mean age 69 years) with organic heart disease. To evaluate the effect of pretreatment with disopyramide, the study population was divided in 3 groups of 10 patients each: group A, no disopyramide therapy; group B, intravenous disopyramide (maximum dose 250 mg in 1 hour); and group C, oral disopyramide (400 mg daily for 4 days). There were no differences in baseline cycle length of atrial flutter among the 3 groups before drugs were given. The stimulation protocol included overdrive atrial pacing up to the shortest paced cycle of 150 ms performed at a maximum of 3 atrial sites. Reversion to sinus rhythm occurred in 2 patients in group A, 7 in group B (p less than 0.01) and 5 in group C. Pacing was performed from a mean number of 2.1 sites/patient in group A, 1.2 in group B and 2.0 in group C. Atrial fibrillation occurred in 7, 3 and 4 patients, respectively. Acceleration to a faster form of atrial flutter occurred in 3, 3 and 4 patients, respectively, and reversion to sinus rhythm occurred in all patients who had intravenous disopyramide and in 1 who took the drug orally. The administration of disopyramide before overdrive pacing improved the rate of conversion to sinus rhythm and allowed an easier stimulation protocol with a lower incidence of pacing-induced atrial fibrillation. Disopyramide is beneficial when overdrive atrial pacing is performed for the treatment of long-standing atrial flutter in patients with organic heart disease.


Gaze step distributions reflect fixations and saccades: a comment on.
In three experimental tasks Stephen and Mirman (2010) measured gaze steps, the distance in pixels between gaze positions on successive samples from an eyetracker. They argued that the distribution of gaze steps is best fit by the lognormal distribution, and based on this analysis they concluded that interactive cognitive processes underlie eye movement control in these tasks. The present comment argues that the gaze step distribution is predictable based on the fact that the eyes alternate between a fixation state in which gaze is steady and a saccade state in which gaze position changes rapidly. By fitting a simple mixture model to Stephen and Mirman's gaze step data we reveal a fixation distribution and a saccade distribution. This mixture model captures the shape of the gaze step distribution in detail, unlike the lognormal model, and provides a better quantitative fit to the data. We conclude that the gaze step distribution does not directly suggest processing interaction, and we emphasize some important limits on the utility of fitting theoretical distributions to data.


The magnificent compromise: cortical field evolution in mammals.
The neocortex of mammals is composed of cortical fields that have a unique organization associated with the animal's ecological niche and lifestyle. Each cortical field has a specific pattern of connections with other cortical fields and brain structures, and together they comprise a neocortical network that generates a variety of behaviors. These networks and the behaviors they generate are variable across mammals, and are particularly complex in some species such as humans. Here I discuss the mechanisms that contribute to neocortical organization in mammals, and how this organization has been altered to generate the variability that exists in different lineages.


Formate: A promising electron donor to enhance trichloroethene-to-ethene dechlorination in Dehalococcoides-augmented groundwater ecosystems with minimal bacterial growth.
Various substrates have been used to stimulate habitat microbes in chloroethene-contaminated groundwater, however, the specific efficiency and minimum growth of microbes have rarely been studied. This study investigated the effects of seven substrates on trichloroethene (TCE) dechlorination by augmentation of groundwater with Dehalococcoides mccartyi NIT01 and its contribution to the microbial community. Three out of eight test groups completed dechlorination of 1 mM TCE-to-ethene in varying durations; groundwater supplemented with formate (FOR) required 78 days, whereas the microcosms with lactate (LAC) and citrate (CIT) required approximately twice as long (143 days). The calculated efficiency of how much produced H2 was used in dechlorination indicated a higher efficiency in FOR (36%) compared with LAC (1.9%) or CIT (2.9%). FOR showed lower microbial growth (3.4 × 105 copies/mL) than LAC (1.5 × 106) or CIT (4.4 × 106), and maintained a higher Shannon diversity index (5.65) than LAC (4.97) and CIT (4.30). The rapid and higher H2 transfer efficiency with lower bacterial growth by using formate was attributed to the slightly positive Gibbs free energy identified in H2 production requiring a H2-utilizer, lower carbon in the molecule, and adaptation to metabolic potential of the original groundwater microbiome. Formate is, therefore, a promising electron donor for rapid Dehalococcoides-augmented remediation with minimum bacterial growth. Sequential transferring of the FOR culture successfully maintained TCE-to-ethene dechlorination activity and enriched the members of genera Dehalococcoides (33%), Methanosphaerula (23%), Rectinema (13%), and Desulfitobacterium (5.6%). This suggests that formate is transferred to H2 and acetate, and provided to Dehalococcoides.


Studies on the dynamics of transmission of onchocerciasis in a Sudan-savanna area of North Cameroon IV. The different exposure to Simulium bites and transmission of boys and girls and men and women, and the resulting manifestations of onchocerciasis.
The sojourn times of the human population were monitored over one year at 12 sites in the vicinity of three villages in the Cameroon Sudan-savanna, where the Onchocerca volvulus transmission potentials had been measured one year previously. Boys stayed longer outside the villages, and were exposed 2.1 to 2.7 times more than girls to transmission of onchocerciasis, whereas the exposure of men was similar or only moderately higher than the exposure of women. In boys, the onset of infections and ocular lesions was earlier and the average microfilarial density at the buttock (2.2, 9.4 and 79.3 mff snip-1) was much higher than in girls (0.1, 5.8 and 42.2 mff snip-1 at the three villages respectively). These differences were maintained in the adult population, where the average microfilarial density was 52.3, 80.4 and 183.1 mff snip-1 in men and 15.6, 49.6 and 114.7 mff snip-1 in women. Ocular lesions due to onchocerciasis were found in 5, 13 and 55% of the male population in the three villages, as compared with 2, 3 and 8% of the female population. There was a close relationship between the degree of exposure to the transmission of disease and the resulting microfilarial load in the skin which was not different for the two sexes, and a similar trend was seen for the occurrence of ocular lesions due to onchocerciasis. The influence of an early and heavy infection on the evolution of disease manifestations is discussed.


Health service experiences among adults with hereditary spastic paraparesis or neurofibromatosis type 1.
Persons with rare disorders may experience poorer health services due to limited knowledge about rare disorders among health professionals. Knowledge about how persons with rare disorders perceive health services can help inform service providers to enhance their practices. We conducted a self-report survey among adults with the rare disorders hereditary spastic paraparesis (HSP; n = 108; mean age 57.7 years; 54.2% females) and neurofibromatosis type 1 (NF1, n = 142; mean age = 50.3 years; 62.0% females). Their responses concerning perceived health experiences were compared to healthy controls from the population study HUNT-3 (n = 7,312). Both rare disorder groups reported lower satisfaction, trust, and participation in meetings with their general practitioner and specialist health services. More reported health problems were overall associated with poorer health service experiences. There is a need to identify predictors of health service experiences at the patient and health service provider levels with the aim to tighten the gap between the health experiences of patients with and without rare disorders.


Inhibition of protein arginine deiminase II suppresses retinoblastoma in orthotopic transplantation in mice.
Chemotherapies are used for treating retinoblastoma; however, numerous patients suffer from recurrence or symptoms due to chemotherapy, which emphasizes the need for alternative therapeutic strategies. The present study demonstrated that protein arginine deiminase Ⅱ (PADI2) was highly expressed in human and mouse retinoblastoma tissues due to the overexpression of E2 factor (E2F). By inhibiting PADI2 activity, the expression of phosphorylated AKT was reduced, and cleaved poly (ADP‑ribose) polymerase level was increased, leading to induced apoptosis. Similar results were obtained in orthotopic mouse models with reduced tumor volumes. In addition, BB‑Cl‑amidine showed low toxicity in vivo. These results suggested that PADI2 inhibition has potential clinical translation. Furthermore, the present study highlights the potential of epigenetic approaches to target RB1‑deficient mutations at the molecular level. The current findings provide new insights into the importance of retinoblastoma intervention by managing PADI2 activity according to the treatment of specific inhibitors and depletion approaches in vitro and in orthotopic mouse models.


Patterns of cilia formation in the lower respiratory tract of the dog: a scanning electron microscopic study.
A scanning electron microscopic study of the distribution of ciliated cells in the tracheobronchial tree of 25 dogs whose ages ranged from four hours to six months was carried out. In newborn puppies, only the dorsal wall of the trachea was completely ciliated while the lateral and ventral walls showed patchy ciliation. This pattern of cilia formation persisted until five days of age when the whole tracheal wall was found to be completely ciliated. The bronchus of newborn puppies was uniformly poorly ciliated but almost complete ciliation was achieved by two days. Likewise, large and small bronchioles of newborn animals had few cilia and, although the number of ciliated cells had increased by two days, complete ciliation was never observed regardless of the age of the dog. No ciliated cells were found in the respiratory bronchioles of any of the 25 dogs.


Molecular mechanisms of Salmonella pathogenesis.
The success of a pathogen depends on its capacity to enter a host, circumvent host defense barriers and establish infection. The Gram-negative bacterium Salmonella enterica has evolved different strategies to subvert normal host cellular functions, which allow it to enter into and proliferate within host cells.


Sphincter of Oddi Manometry: Reproducibility of Measurements and Effect of Sphincterotomy in the EPISOD Study.
The reproducibility of sphincter of Oddi manometry (SOM) measurements and results of SOM after sphincterotomy has not been studied sufficiently. The aim of our study is to evaluate the reproducibility of SOM and completeness of sphincter ablation. The recently published Evaluating Predictors and Interventions in sphincter of Oddi dysfunction (EPISOD) study included 214 subjects with post-cholecystectomy pain, and fit the criteria of sphincter of Oddi dysfunction type III. They were randomized into 3 arms, irrespective of manometric findings: sham (no sphincterotomy), biliary sphincterotomy, and dual (biliary and pancreatic). Thirty-eight subjects had both biliary and pancreatic manometries performed twice, at baseline and at repeat endoscopic retrograde cholangiopancreatography after 1-11 months. Sham arm was examined to assess the reproducibility of manometry, and the treatment arms to assess whether the sphincterotomies were complete (elevated pressures were normalized). Biliary and pancreatic measurements were reproduced in 7/14 (50%) untreated subjects. All 12 patients with initially elevated biliary pressures in biliary and dual sphincterotomy groups normalized after biliary sphincterotomy. However, 2 of 8 subjects with elevated pancreatic pressures in the dual sphincterotomy group remained abnormal after pancreatic sphincterotomy. Paradoxically, normal biliary pressures became abnormal in 1 of 15 subjects after biliary sphincterotomy, and normal pancreatic pressures became abnormal in 5 of 15 patients after biliary sphincterotomy, and in 1 of 9 after pancreatic sphincterotomy. Our data suggest that SOM measurements are poorly reproducible, and question whether we could adequately perform pancreatic sphincterotomy.


Quantitative proton magnetic resonance spectroscopic imaging: regional variations in the corpus callosum and cortical gray matter.
To evaluate regional variations of metabolite concentrations in normal adult brain cortical gray matter regions, and the genu and splenium of the corpus callosum, using proton magnetic resonance spectroscopic imaging (MRSI). Quantitative, multislice proton MRSI (TR/TE = 2000/280 msec) was performed in 12 normal human volunteers (age = 39 +/- 6 years, 7 male). Metabolite concentrations in selected cortical gray matter regions and the corpus callosum were estimated using the phantom replacement methodology. Frontal and parietal gray matter (PGM) showed strong differences in choline-containing compound (Cho) concentrations; in particular, Cho was higher in mesial frontal gray matter than in both dorsolateral prefrontal cortex (P < 0.0005) and PGM (P < 0.004). In contrast, both N-acetylaspartate (NAA) and creatine (Cr) were relatively uniformly distributed in the cortical gray matter regions evaluated. Significant metabolic differences were found between the genu and splenium of the corpus callosum. Cho concentrations were significantly higher in genu than splenium (P < 0.005), while Cr was lower (P < 0.004). NAA showed a trend to be higher in the splenium than the genu (P = 0.05). Metabolite concentrations, particularly Cho, showed strong regional variations both within cortical gray matter regions and between the genu and splenium of the corpus callosum. Mesial frontal regions showed the highest Cho signals. Differences in spectra presumably reflect underlying changes in structure and cellular composition. Normal spectral variations should always be considered when evaluating pathology within those brain regions.


Readability of Online Patient Education Materials Related to IR.
To assess the readability of online patient education materials (OPEM) related to common diseases treated by and procedures performed by interventional radiology (IR). The following websites were chosen based on their average Google search return for each IR OPEM content area examined in this study: Society of Interventional Radiology (SIR), Cardiovascular and Interventional Radiological Society of Europe (CIRSE), National Library of Medicine, RadiologyInfo, Mayo Clinic, WebMD, and Wikipedia. IR OPEM content area was assessed for the following: peripheral arterial disease, central venous catheter, varicocele, uterine artery embolization, vertebroplasty, transjugular intrahepatic portosystemic shunt, and deep vein thrombosis. The following algorithms were used to estimate and compare readability levels: Flesch-Kincaid Grade Formula, Flesch Reading Ease Score, Gunning Frequency of Gobbledygook, Simple Measure of Gobbledygook, and Coleman-Liau Index. Data were analyzed using general mixed modeling. On average, online sources that required beyond high school grade-level readability were Wikipedia (15.0), SIR (14.2), and RadiologyInfo (12.4); sources that required high school grade-level readability were CIRSE (11.3), Mayo Clinic (11.0), WebMD (10.6), and National Library of Medicine (9.0). On average, OPEM on uterine artery embolization, vertebroplasty, varicocele, and peripheral arterial disease required the highest level of readability (12.5, 12.3, 12.3, and 12.2, respectively). The IR OPEM assessed in this study were written above the recommended sixth-grade reading level and the health literacy level of the average American adult. Many patients in the general public may not have the ability to read and understand health information in IR OPEM.


Regional differences in water beetle communities networks settling in dystrophic lakes in northern Poland.
The relationships between the species that form the networks in small dystrophic lakes remain poorly recognised. To investigate and better understand the functioning of beetle communities in different ecosystems, we created three network models that we subjected to graph network analysis. This approach displays correlation-based networks of connections (edges) between objects (nodes) by evaluating the features of the whole network and the attributes of nodes and edges in the context of their roles, expressed by centrality metrics. We used this method to determine the importance of specific species in the networks and the interspecific relationships. Our analyses are based on faunal material collected from 25 dystrophic lakes in three regions of northern Poland. We found a total of 104 species representing different ecological elements and functional trophic groups. We have shown that the network of relationships between the biomass of species differs considerably in the three study regions. The Kashubian Lakeland had the highest cohesion and density, while the network in the Suwalki Lakeland was the thinnest and most heterogeneous, which might be related to the fractal structure and the degree of development of the studied lakes. Small-bodied predators that congregated in different clusters with species with similar ecological preferences dominated all networks. We found the highest correlations in the Masurian Lakeland, where we obtained the highest centralisation of the network. Small tyrphophiles typically occupied the central places in the network, while the periphery of the network consisted of clusters with different habitat preferences, including large predators. The species that were most important for network cohesion and density were mainly tyrphophilous species, such as Anacaena lutescens, Hygrotus decoratus, Enochrus melanocephalus and Hydroporus neglectus. The values of attributes determining the role of species in community networks were influenced by both biotic and environmental factors.


Death by apoptosis in salivary glands of females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae).
The salivary glands of females of the tick Rhipicephalus sanguineus at three feeding stages: unfed, engorged, and at day three post-engorgement, were subjected to cytochemical methods of enzymatic analysis and cell viability. Comparing glands at these stages, was observed distinct staining patterns in cells of different types of acini, specially in degenerating types III, II, I, which were affected in this sequence by cell death. This study also revealed changes in: nuclei, staining intensity for acid phosphatase and ATPase activities, and permeability of the plasma membrane. Acid phosphatase activity was inversely proportional to that of ATPase, while ATPase activity was always proportional to membrane integrity. The glands of unfed females exhibited high metabolic activity and cells with intact nucleus and plasma membrane, suggesting that the presence of acid phosphatase detected in these individuals may participate in the normal physiology of some acini, as they were not undergoing degeneration. In acini I and II of engorged females, we observed cells with intact membranes, as well as changes characterized by nuclear changes, decrease in ATPase activity, and stronger acid phosphatase activity. At day three post-engorgement, degeneration progressed to more advanced stages, loss of membrane integrity was observed in most cells (of some type I acini, most type II acini, and all type III acini), as well as prominent nuclear changes, decrease in ATPase activity, and intense acid phosphatase activity, resulting in apoptotic bodies. During the death of cells nuclear changes preceded cytoplasmic ones in the following sequence: nuclear changes, loss of ATPase activity, loss of integrity of the plasma membrane, increase in acid phosphatase activity, and formation of apoptotic bodies. The presence of acid phosphatase with a secondary role (late) during cell death, degrading final cell remnants, characterized this process in the glands of R. sanguineus females as atypical or non-classic apoptosis.


Anatomical Changes of the Peripheral Zone Depending on Benign Prostatic Hyperplasia Size and Their Potential Clinical Implications:A Review for Clinicians.
The inverse relationship between benign prostate hypertrophy and incidence/severity of prostate cancer is well documented in the clinical literature. However, this phenomenon is not well understood. The purpose of this review is to offer an update in the evolving hypothesis of how benign prostate hypertrophy may be protective in prostate cancer. A literature search was conducted on PubMed limited to articles published within the past 10 years with the search criteria of "interaction" AND "benign prostate hypertrophy" AND "prostate cancer" as well as the key words of this paper. Nine articles from the literature search met inclusion criteria. The articles analyze the prostate on parameters of peripheral zone volume, glandular tissue density and prostate capsule thickness. All 9 articles described peripheral zone atrophy and transition zone hypertrophy in benign prostate hypertrophy patients. As the transition zone grows in benign prostate hypertrophy, volume and glandular density of the peripheral zone as well as the prostate capsule undergo significant changes. The disease processes outlined in this review support the hypothesis that the growing transition zone compresses the peripheral zone against the prostatic capsule causing secondary atrophy, apoptosis, and necrosis of the peripheral zone's glandular tissue. If this hypertrophy-induced disease process of glandular tissue atrophy within the peripheral zone is confirmed in future studies, it will have relevant clinical implications on the diagnosis and treatment of benign prostate hypertrophy and prostate cancer.


Purification and properties of fructose-1,6-bisphosphatase of Bacillus subtilis.
Fructose-1,6-bisphosphatase (D-fructose-1,6-bisphosphate 1-phosphohydrase, EC 126.96.36.199) of Bacillus subtilis is a constitutive enzyme that was purified 1000-fold (30% yield) to 80% purity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis where it exhibits a band corresponding to 72,000 daltons. It sediments at 15 S in sucrose density gradients indicating a molecular weight of 380,000, but apparently is very asymmetric. Its activity is irreversibly inactivated in the absence of Mn2+. The enzyme specifically catalyzes dephosphorylation of D-fructose 1,6-bisphosphate with a pH optimum of 8.0. It has 40 to 60% of full activity in the absence of P-enolpyruvate; 20 microM P-enolpyruvate activates it maximally. High concentrations of monovalent cations also activate, NH4+ being most effective. Inhibitors fall into two groups. 1) Nucleoside monophosphates, phosphorylated coenzymes, and polynucleotides inhibit competitively with P-enolpyruvate (AMP (Ki = 2 microM) and dAMP are most effective). 2) The inhibition by nucleoside di- and triphosphates, PPi, and highly phosphorylated nucleotides (guanosine 5'-triphosphate 3'-diphosphate (pppGpp) and adenosine 5'-triphosphate 3'-diphosphate are most effective) is not competed by P-enolpyruvate but is partially overcome by fructose 1,6-bisphosphate (2 microM). Therefore, highly phosphorylated nucleotides (pppGpp and others), produced in over 0.2 mM concentrations upon step down from fast to slow growth rates (Gallant, J., and Lazzarini, R.A. (1976) in Protein Synthesis (McConkey, E.H., ed) Vol. 2, pp. 309-349, Marcel Dekker, Inc., New York), can reduce the conversion rate of fructose 1,6-bisphosphate to fructose 6-phosphate during gluconeogenesis. Comparing glycolytic growth on D-glucose and gluconeogenic growth on L-malate, the intracellular concentrations of fructose 1,6-bisphosphate differ but are both above the Km (13 microM) of the enzyme, those of AMP are similar, whereas those of P-enolpyruvate (0.18 mM versus 1.3 mM) indicate that the enzyme has only 40% of its full activity during glycolysis; nucleotides other than AMP may inhibit additionally. Thus, the futile cycle of fructose 1,6-bisphosphate synthesis and degradation during glycolysis is partially avoided, but the cells are poised for rapid adaptation upon change to gluconeogenic growth conditions.


Accumulation of copper in the liver and hepatic injury in chronic hepatitis C.
Relationships between chronic liver disease and trace metals have not been clearly understood. To examine connections between severity of hepatic fibrosis in chronic hepatitis C and copper, iron and zinc we measured the contents of these metals in liver tissue and serum in the patients. Forty-one patients (26-62 years), 13 with fibrosis representing grade F1, 16 with F2, seven with F3, and five with F4, entered this study. Metals were quantified in needle liver-biopsy specimens by particle-induced X-ray emission. In serum, metals were measured by flameless atomic absorption spectrometry. Hepatic copper content increased with progression of hepatic fibrosis (P < 0.05). The copper content correlated positively with bilirubin (r = 0.466, P = 0.0023), and with type IV collagen (r = 0.402, P = 0.0086) and correlated negatively with albumin (r = -0.404, P = 0.080). However, hepatic iron and zinc contents did not show a significant differences between grades of fibrosis. Copper accumulation in fibrotic livers caused by chronic hepatitis C may contribute to hepatic injury. The real mechanism is not known at present, but excess copper may damage the liver by oxidative stress.


High Mobility Group-Box 1 (HMGB1) levels are increased in amniotic fluid of women with intra-amniotic inflammation-determined preterm birth, and the source may be the damaged fetal membranes.
High Mobility Group Box-1 (HMGB1) is considered a prototype alarmin molecule. Upon its extracellular release, HMGB1 engages pattern recognition receptors and the Receptor for Advanced Glycation End-products (RAGE) followed by an outpouring of inflammatory cytokines, including interleukin (IL)-6. We assayed the amniotic fluid (AF) levels of HMGB1 and IL-6 in 255 women that either had a normal pregnancy outcome or delivered preterm. Immunohistochemistry on fetal membranes was used for cellular localization and validation of immunoassay findings. HMGB1 also was analyzed in amniochorion tissue explants subjected to endotoxin. AF HMGB1 levels are not gestational age regulated but are increased in women with intra-amniotic inflammation and preterm birth. The likely source is the damaged amniochorion, as demonstrated by immunohistochemistry and explant experiments. Our research supports a role for HMGB1 in the inflammatory response leading to preterm birth. As a delayed phase cytokine, in utero exposure to elevated AF HMGB1 levels may have an impact on the newborn beyond the time of birth.


Evaluation of Dynamic CT Scans and Analyzing Its Efficacy Accuracy in Staging of Laryngeal CA: A Prospective Analysis of 30 Patients and Review of Literature.
Laryngeal and hypopharyngeal cancer is the most common cancer in the head and neck. It is the most commonly reported human malignancy, with more than 100,000 cases reported annually worldwide. Thus, exact staging of the tumor is of utmost importance in the decision making of the management of these patients. CT can directly evaluate penetration of tumor into the laryngeal soft tissues and cartilages; whereas laryngoscopy and contrast laryngography can only infer deepseated abnormalities from changes in surface contour of the laryngeal cavity. The main aims of our study were: (1) To assess whether dynamic CT scans increase the diagnostic efficacy for laryngeal and hypopharyngeal cancers and its extent in surrounding structures. (2) To assess the importance of phonation CT scans in evaluation and diagnosis of vocal cord paralysis/fixity and involvement of subglottic region and pre epiglottic space.


Enhanced dye adsorption with conductive polyaniline doped chitosan nanofibrous membranes.
Polyaniline is widely used in the field of electrochemistry due to its excellent electrical conductivity. However, its effectiveness and mechanism of enhancing adsorption property are unclear. Herein, chitosan/polyaniline nanofibrous composite membranes with average diameter ranging from 200 to 300 nm were fabricated by electrospinning technology. The as-prepared nanofibrous membranes exhibited significantly improved adsorption capacity of 814.9 mg/g and 618.0 mg/g towards acid blue 113 and reactive orange dyes, which were 121.8 % and 99.4 % higher than that of pure chitosan membrane. The doped polyaniline promoted the dye transfer rate and capacity due to the enhanced conductivity of the composite membrane. Kinetic data showed that chemisorption was the rate-limiting step, and thermodynamic data indicated the adsorption of the two anionic dyes was spontaneous monolayer adsorption. This study provides a feasible strategy to introduce conductive polymer into adsorbent to construct high performance adsorbents for wastewater treatment.


A stilbene derivative as dual-channel fluorescent probe for mitochondrial G-quadruplex DNA in living cells.
G-quadruplex DNA has attracted the widespread attention as a novel target of anticancer strategy. Herein, two novel stilbene derivatives 2a and 2b were designed and synthesized under mild reaction conditions, and their interactions with G-quadruplex DNA, cytotoxicity, and distribution in living cells were investigated in detail. Both compounds display a low cytotoxicity and the higher affinity to G-quadruplex DNA than to the other secondary structures, including duplex, single-stranded and i-motif DNA, moreover, the affinity of 2b with m-allyl pyridine salt group to G-quadruplex DNA is about 10-fold stronger than that of 2a with p-allyl pyridine salt group. The interactions of the compounds with the promoter G-quadruplexes are enthalpy-driven by an ITC assay. 2a and 2b not only stabilize the G-quadruplex structure but also induce the G-rich sequences (bcl-2, HRCC and KSS) to fold into the mixed-type G-quadruplex in Na+/K+ free Tris-HCl buffer at pH 7.0, and 2b presents the higher stabilization to G-quadruplex than 2a by a FRET-melting assay. 2b presents a dual-emission at 508 and 600 nm and gives a turn-on and stronger and more sensitive fluorescence response over 2a to the promoter (bcl-2, c-kit 2 and c-myc) and mitochondrial (HRCC and KSS) G-quadruplex DNA at both emission wavelengths, moreover, the peak at 508 nm is blue-shifted to 466 nm after binding to DNA. The blue and red dual-channel CLSM images indicate that 2b is mainly distributed in the mitochondrion of living HepG2 cells. The results show that 2b is a potential dual-channel fluorescent probe for mitochondrial G-quadruplex DNA in living cells.


Drosophila melanogaster as a powerful tool for studying insect toxicology.
Insecticides are valuable and widely used tools for the control of pest insects. Despite the use of synthetic insecticides for >50 years, we continue to have a limited understanding of the genes that influence the key steps of the poisoning process. Major barriers for improving our understanding of insecticide toxicity have included a narrow range of tools and/or a large number of candidate genes that could be involved in the poisoning process. Herein, we discuss the numerous tools and resources available in Drosophila melanogaster that could be brought to bear to improve our understanding of the processes determining insecticide toxicity. These include unbiased approaches such as forward genetic screens, population genetic methods and candidate gene approaches. Examples are provided to showcase how D. melanogaster has been successfully used for insecticide toxicology studies in the past, and ideas for future studies using this valuable insect are discussed.


Comparison of cyclic nucleotide specificity of guanosine 3',5'-monophosphate-dependent protein kinase and adenosine 3',5'-monophosphate-dependent protein kinase.
Guanosine 3',5'-monophosphate-dependent protein kinase (cyclic GMP-dependent protein kinase) and adenosine 3',5'-monophosphate-dependent protein kinase (cyclic AMP-dependent protein kinase) exhibited a high degree of cyclic nucleotide specificity when hormone-sensitive triacylglycerol lipase, phosphorylase kinase, and cardiac troponin were used as substrates. The concentration of cyclic GMP required to activate half-maximally cyclic dependent protein kinase was 1000- to 100-fold less than that of cyclic AMP with these substrates. The opposite was true with cyclic AMP-dependent protein kinase where 1000- to 100-fold less cyclic AMP than cyclic GMP was required for half-maximal enzyme activation. This contrasts with the lower degree of cyclic nucleotide specificity of cyclic GMP-dependent protein kinase of 25-fold when histone H2b was used as a substrate for phosphorylation. Cyclic IMP resembled cyclic AMP in effectiveness in stimulating cyclic GMP-dependent protein kinase but was intermediate between cyclic AMP and cyclic GMP in stimulating cyclic AMP-dependent protein kinase. The effect of cyclic IMP on cyclic GMP-dependent protein kinase was confirmed in studies of autophosphorylation of cyclic GMP-dependent protein kinase where both cyclic AMP and cyclic IMP enhanced autophosphorylation. The high degree of cyclic nucleotide specificity observed suggests that cyclic AMP and cyclic GMP activate only their specific kinase and that crossover to the opposite kinase is unlikely to occur at reported cellular concentrations of cyclic nucleotides.


Management of hormonal induced recurrent ulcerative fibrous epulis with osseous metaplasia: A case report.
Understanding the role of hormones in periodontitis is important. Periodontal microscopic surgery approach in the treatment of fibrous epulis is not indicated. Wider flap access with root planning is indicated to control the lesion in one phase. We present a case of a 40-year-old female who presented with a gingival hyperplastic lesion around the maxillary left permanent central and lateral incisors. Patient's medical history reveals a recent pregnancy, hypothyroidism, ulcerative colitis, and schizoaffective disorder. All medical conditions were controlled by medications. The lesion was excised using a minimally invasive periodontal surgical technique, and the biopsy results confirmed a diagnosis of ulcerative fibrous epulis with osseous metaplasia. No curettage or local debridement was done under the assumption that the patient's oral hygiene was satisfactory and due to aesthetic concerns of gingival recession and creation of black triangles. The lesion recurred after 3 months and was removed using a traditional more invasive surgical technique. The patient was followed up for 2 years, and there was no further recurrence due to the complete excision of the lesion in the second surgery and the disappearance of the hormonal-related factors post-pregnancy that could have contributed to the gingival hyperplasia. The unique component of the case we are presenting is the comparison between two different surgical techniques and the conduction versus absence of local periodontal debridement after surgical excision as well as the possible correlation between oral supplements and the calcific nature of the lesion(s) reported. The case we present demonstrates that a more invasive traditional surgical approach together with local periodontal therapy provide an optimum treatment outcome in conjunction with elimination of any associated etiological factors. We also propose that hormones are more important as an etiological factor in developing fibrous epulis lesions than medical conditions and medications.


Asymptomatic Cholelithiasis and Bariatric Surgery: a Comprehensive Long-Term Analysis of the Risks of Biliary Disease in Patients Undergoing Primary Roux-en-Y Gastric Bypass.
Currently, there is little consensus on management of the in situ gallbladder of patients undergoing gastric bypass. Our aim was to evaluate outcomes of selective concomitant cholecystectomy (CCY) and long-term biliary outcomes after Roux-en-Y gastric bypass (RYGB). We performed a retrospective analysis of patients undergoing laparoscopic RYGB (LRYGB) between 2008 and 2018. Chi-square, Fisher's exact, or Wilcoxon rank-sum tests were used to compare outcomes. Concomitant CCY was performed on a selective basis. Three thousand and four patients underwent a RYGB (LRYGB n = 2458, open RYGB n = 546). Fifty-two percent (n = 1670) of patients had undergone CCY at any stage. Thirty-one percent of patients (n = 933) had CCY prior to RYGB, 13% (n = 403) had a concomitant CCY and 13% (n = 214) of the remainder required interval CCY. In the LRYGB subgroup, 29.9% (n = 735) had a prior CCY; 12.9% (n = 202) of those with an in situ gallbladder required interval CCY. Those who underwent concomitant CCY/LRYGB (n = 328) were compared with LRYGB alone (n = 1231). The concomitant CCY group was significantly older and had higher percentage of females, higher preoperative BMI, higher Charlson Comorbidity Index, and a higher medication count. There was no significant difference in BMI nadir, length of stay, complications, or mortality. Interval CCY had a higher incidence of CCY-related complications. Our study suggests a higher percentage of bariatric patients with in situ gallbladders will undergo interval CCY than documented in recently published guidelines. Concomitant CCY can be performed without an increase in length of stay or complications. Interval CCY may be associated with a higher complication rate.


Radical radiation alone versus radical radiation plus microwave hyperthermia for N3 (TNM-UICC) neck nodes: a prospective randomized clinical trial.
Between September 1985 and December 1986, 44 N3 (TNM-UICC) metastatic squamous cell cervical lymph-nodes were randomized to receive conventionally fractionated radical irradiation (RT) to a total dose of 64-70 Gy, or conventionally fractionated radical irradiation plus twice a week local microwave hyperthermia (Ht). The two major end points of this study were (a) local control rates evaluated at 3 months after the end of combined therapy and (b) incidence of acute local toxicity. Thirty-six nodes (82%) were evaluable as of December 1986, at which time there was a premature closure of this study due to ethical reasons. An interim analysis had revealed a statistically significant difference in complete response rates in favor of the combined arm (p = 0.0152). The complete response rates were 82.3% (14/17) for the combined treatment arm versus 36.8% (7/19) for the control irradiation arm, leading to an iso-dose thermal enhancement ratio (TER) value of 2.23. Both arms are comparable in average total RT dose delivered (RT: 67.05 Gy; RT + Ht: 67.85 Gy) and in average maximum node diameter (RT arm: 4.81 cm; RT + Ht: 4.88 cm). Acute local toxicities were similar in irradiated and heated plus irradiated neck regions; only one skin burn was observed. As possible treatment related death, one patient in the RT + Ht arm died 2 months after completion of therapy with a carotid rupture associated with extensive tumor necrosis. These results confirm previous non-randomized reports suggesting that hyperthermia in combination with full dose conventionally fractionated irradiation significantly enhances the chance of early local control of fixed N3 neck nodes without exhibiting an increase of acute local toxicity.


Ophthalmic manifestations of mosaic Down syndrome.
The cognitive and physical stigmata of mosaic Down syndrome (DS) are often considered to be less severe than complete trisomy-21 DS. In contrast to complete trisomy-21 DS, the ophthalmic manifestations in mosaic DS have rarely been reported. The aim of the present study is to report clinically significant ophthalmic abnormalities in a cohort of individuals with mosaic DS. A prospective cross-sectional observational case series was designed to evaluate ophthalmic manifestations of mosaic DS. Individuals with mosaic DS were recruited and examined at the biennial meeting of the International Mosaic Down Syndrome Association. A medical, surgical, and ocular history was obtained. Each subject received a complete eye examination on site, including assessment of visual acuity, alignment, motility, sensory function, accommodation, anterior segment, fundus, and cycloplegic refraction. Seventeen individuals with mosaic DS (mean age, 9 years; range, 6 months to 32 years) underwent eye examinations. Clinically significant refractive errors were present in 41% of the subjects, accommodative insufficiency in 59%, strabismus in 35%, nystagmus in 6%, and cataract in 6%. Ten individuals completed optotype visual acuity testing. Mean LogMAR acuity of the better eye of each subject was 0.2 (20/32 equivalent). Clinically significant ophthalmic disorders are common among children and young adults with mosaic DS. Our findings support regular periodic eye examinations for these individuals.


Venlafaxine extended-release capsules in panic disorder: flexible-dose, double-blind, placebo-controlled study.
Venlafaxine extended-release (ER) has proven efficacy in the treatment of anxiety symptoms in major depression, generalised anxiety disorder and social anxiety disorder. To evaluate the efficacy, safety and tolerability of venlafaxine ER in treating panic disorder. Adult out-patients (n=361) with panic disorder were randomly assigned to receive venlafaxine ER (75-225 mg/day) or placebo for up to 10 weeks in a double-blind study. Venlafaxine ER was not associated with a greater proportion of patients free from full-symptom panic attacks at the final on-therapy evaluation, but was associated with lower mean panic attack frequency and a higher proportion free from limited-symptom panic attacks, higher response and remission rates, and improvements in anticipatory anxiety, fear and avoidance. Adverse events were comparable with those of the drug in depression and anxiety disorders. Venlafaxine ER seems to be effective and well tolerated in the short-term treatment of panic disorder.


Environmental Risks for Systemic Sclerosis.
There is an increasing body of literature suggesting a relationship between environmental factors and the development of systemic sclerosis (SSc). These include occupational exposures, chemical materials, medications, alterations in the microbiome, and dysbiosis. Environmental exposures may impact epigenetic regulation thereby triggering an aberrant immune response resulting in the clinical and serologic phenotype that we diagnose as SSc. Screening and studying putative triggers will not only improve our understanding of the pathogenesis of SSc but also inform the institution for protective measures.


Insight into the role of SO2 on selenium conversion during adsorption by CaO: Theoretical calculation and experimental study.
SO2 can noticeably impact the control of high toxic selenium emissions from flue gas by CaO. Surprisingly, our experiments showed that under certain conditions, SO2 can promote selenium capture by CaO, rather than hinder it. To elucidate the underlying mechanism, a combination of theoretical calculations and experiments was conducted. Thermodynamic equilibrium analysis revealed that gaseous SO2 and solid Ca-S reaction products can promote SeO2 converting to SeO/Se0. The Ca-S products facilitated greater SeO2 conversion compared to SO2. Experimental results demonstrated that selenium adsorption capacity of incompletely sulfurized CaO (CaO with pre-adsorbed SO2) was higher than that of completely sulfurized CaO (Ca-S products), highlighting the importance of adsorption sites of CaO. Density functional theory calculations showed that the pre-adsorbed SO2 hardly affected selenium adsorption energy on the SO2/CaO surface, while completely sulfurized CaO had low selenium adsorption energy, explaining the experimental phenomenon and proving necessary of CaO. Additionally, SeO/Se0 had higher adsorption energy on CaO than SeO2. Overall, the promotion of SO2 on selenium adsorption was primarily affected by two factors: 1) sulfur facilitating SeO2 conversion to SeO/Se0 which can be adsorbed more easily by CaO; 2) sufficient adsorption sites on CaO surface existing for SeO/Se0 adsorption, despite co-adsorption with sulfur.


[Five-component visual analogue scale for assessement of pain status retrospective analysis of outcomes after surgical treatment of degenerative spine disease].
The study was performed for development of more precise criteria for assessment of pain using five-component visual analogue scale. Pain status was studied in 493 patients who underwent lumbar spinal nerve root decompression. Pain scale included the following parameters: pain at the moment, usual pain, maximal pain, minimal pain and overall pain score which was calculated as an average quantity of the previous four parameters. Overall pain score is a more precise criterion for evaluation of postoperative pain status in patients with degenerative lumbar spine disease.


Corticostriatal beta oscillation changes associated with cognitive function in Parkinson's disease.
Cognitive impairment is the most frequent non-motor symptom in Parkinson's disease and is associated with deficits in a number of cognitive functions including working memory. However, the pathophysiology of Parkinson's disease cognitive impairment is poorly understood. Beta oscillations have previously been shown to play an important role in cognitive functions including working memory encoding. Decreased dopamine in motor cortico-striato-thalamo-cortical (CSTC) circuits increases the spectral power of beta oscillations and results in Parkinson's disease motor symptoms. Analogous changes in parallel cognitive CSTC circuits involving the caudate and dorsolateral prefrontal cortex (DLPFC) may contribute to Parkinson's disease cognitive impairment. The objective of our study is to evaluate whether changes in beta oscillations in the caudate and DLPFC contribute to cognitive impairment in Parkinson's disease patients. To investigate this, we used local field potential recordings during deep brain stimulation surgery in 15 patients with Parkinson's disease. Local field potentials were recorded from DLPFC and caudate at rest and during a working memory task. We examined changes in beta oscillatory power during the working memory task as well as the relationship of beta oscillatory activity to preoperative cognitive status, as determined from neuropsychological testing results. We additionally conducted exploratory analyses on the relationship between cognitive impairment and task-based changes in spectral power in additional frequency bands. Spectral power of beta oscillations decreased in both DLPFC and caudate during working memory encoding and increased in these structures during feedback. Subjects with cognitive impairment had smaller decreases in caudate and DLPFC beta oscillatory power during encoding. In our exploratory analysis, we found that similar differences occurred in alpha frequencies in caudate and theta and alpha in DLPFC. Our findings suggest that oscillatory power changes in cognitive CSTC circuits may contribute to cognitive symptoms in patients with Parkinson's disease. These findings may inform the future development of novel neuromodulatory treatments for cognitive impairment in Parkinson's disease.


A vault ribonucleoprotein particle exhibiting 39-fold dihedral symmetry.
Vault is a 12.9 MDa ribonucleoprotein particle with a barrel-like shape, two protruding caps and an invaginated waist structure that is highly conserved in a wide variety of eukaryotes. Multimerization of the major vault protein (MVP) is sufficient to assemble the entire exterior shell of the barrel-shaped vault particle. Multiple copies of two additional proteins, vault poly(ADP-ribose) polymerase (VPARP) and telomerase-associated protein 1 (TEP1), as well as a small vault RNA (vRNA), are also associated with vault. Here, the crystallization of vault particles is reported. The crystals belong to space group C2, with unit-cell parameters a = 708.0, b = 385.0, c = 602.9 angstroms, beta = 124.8 degrees . Rotational symmetry searches based on the R factor and correlation coefficient from noncrystallographic symmetry (NCS) averaging indicated that the particle has 39-fold dihedral symmetry.


Chronic stress exposure may affect the brain's response to high calorie food cues and predispose to obesogenic eating habits.
Exaggerated reactivity to food cues involving calorically-dense foods may significantly contribute to food consumption beyond caloric need. Chronic stress, which can induce palatable "comfort" food consumption, may trigger or reinforce neural pathways leading to stronger reactions to highly rewarding foods. We implemented functional magnetic resonance imaging (fMRI) to assess whether chronic stress influences activation in reward, motivation and executive brain regions in response to pictures of high calorie and low calorie foods in thirty women. On separate lab visits, we also assessed food intake from a snack food buffet and circulating cortisol. In women reporting higher chronic stress (HCS), pictures of high calorie foods elicited exaggerated activity in regions of the brain involving reward, motivation, and habitual decision-making. In response to pictures of high calorie food, higher chronic stress was also associated with significant deactivation in frontal regions (BA10; BA46) linked to strategic planning and emotional control. In functional connectivity analysis, HCS strengthened connectivity between amygdala and the putamen, while LCS enhanced connectivity between amygdala and the anterior cingulate and anterior prefrontal cortex (BA10). A hypocortisolemic signature and more consumption of high calorie foods from the snack buffet were observed in the HCS group. These results suggest that persistent stress exposure may alter the brain's response to food in ways that predispose individuals to poor eating habits which, if sustained, may increase risk for obesity.


Human and animal health risk assessment of metal contamination in soil and plants from Ait Ammar abandoned iron mine, Morocco.
The goal of this paper is to investigate metal pollution in food chain and assess the resulting health risks to native citizens in Ait Ammar village. The results showed that cadmium (Cd), lead (Pb), and copper (Cu) concentrations in animal organs were above the metal concentration safety limit. Nevertheless, soils and plants from mining area were contaminated with iron (Fe), chromium (Cr), zinc (Zn), and Cr, Cu, Zn respectively. Cd concentrations in almost animal organs were higher than the acceptable daily upper limit, suggesting human consumption of this livestock meat and offal may pose a health risk. The estimated intake of Pb and Cd for Ait Ammar population could be a cause of concern because it exceeded the Provisional Tolerable Weekly Intake (PTWI) proposed by Joint Expert Committee on Food Additives (JECFA) in this area. Thus, conducting regular periodic studies to assess the dietary intake of mentioned elements are recommended.


Persistence of MHC DR nonexpression on swine cells by introduction of a mutated MHC class II transactivator gene: a comparison with the effect induced by antisense RNA.
CIITA (class II transactivator) is a coactivator essential for transcription of MHC class II genes. In this study, a construct with a mutated CIITA gene with N-terminal domains depleted was constructed. This mutated CIITA (mCIITA) was able to repress DR and DQ expression in 45.0-60.0% of the mCIITA-transfected clones of swine endothelial cell line PIEC and in B cell line L23, as well as in human cell lines HeLa and Raji. Similarly, 30.0-46.7% of swine cell clones containing the human CIITA antisense RNA also failed to express DR molecules. However, the persistence of the DR repression on the cell lines is quite different. Transfection with mCIITA was persistent for at least 120 days, while with the CIITA antisense RNA, persistence existed for only 35-45 days. To explore the underlying mechanism, Raji cells were transfected with pUHD10-3-mCIITA, a mCIITA-containing, doxycycline-dependent plasmid. The intensity of DR repression is correlated quite well with the efficiencies of the mCIITA expression within the cells in a doxycycline dose-dependent manner. This implicates a competition between the mCIITA and its endogenous full-length counterpart. In addition, we were able to show that purified human CD4 T cells did not respond to the mCIITA-transfected PIECs in xenogeneic mixed lymphocyte endothelial reaction (MLER). The stimulating indices (SI) were only 1.0-1.5, compared with 15.2-18.2 for those transfected with empty vector or an initiation codon-depleted mCIITA that is dysfunctional for protein translation. The results we obtained, especially those for persistent suppression of class II genes, show promise for the possible development of mCIITA-transgenic swine for organ/tissue xenotransplantation.


Changes of CD68, CD163, and PD-L1 tumor expression during high-dose-rate and pulsed-dose-rate brachytherapy for cervical cancer.
We hypothesized that radiation doses delivered with high-dose-rate (HDR) and pulsed-dose-rate (PDR) brachytherapy in patients with cervical cancer could trigger immune stimulation by modulating immune cells in the tumor microenvironment. The objective was to determine CD68, CD163, and PD-L1 expression in biopsies from patients with cervical cancer and to compare the effects of HDR vs. PDR brachytherapy on the expression of these proteins. Nineteen consecutive women (mean age, 55.9 years) with histologically proven cervical cancer scheduled for brachytherapy after standard external beam irradiation therapy combined with platinum-based chemotherapy were included in a prospective study. Core tissue biopsies were obtained before radiochemotherapy (biopsy #1), after completion of radiochemotherapy and before brachytherapy (biopsy #2), and 2 weeks after completion of brachytherapy (biopsy #3). HDR or PDR brachytherapy was delivered according to availability of equipment. CD68, CD163, and PD-L1 immunohistochemical expression was estimated using a quantitative method. Conditional logistic regression models were used to assess the relationship between gene expression and time of biopsy for each brachytherapy technique. In relation to CD68 and CD163, statistically significant relationships between gene expression and biopsy tissue samples were not found in any of the brachytherapy techniques, although there was trend toward downexpression of CD68 and CD163 in biopsies #2 and #3 in the HDR brachytherapy cohort only. There was a significant increase in PD-L1 expression in biopsy #3 also in the HDR cohort as compared with the PDR cohort. Decreased CD68 and CD163 expression did not reach statistical significance, but this trend may have clinical translational potential. Overexpression of PD-L1 in tissue biopsies taken at 14 days in the HDR brachytherapy cohort may tentatively suggest that this time period would be an adequate interval for blockade of the PD-1/PD-L1 axis.


Simultaneous determination of 24 free amino acids in MGC803 cells by hydrophilic interaction liquid chromatography with tandem mass spectrometry.
Amino acids play key roles in cellular protein biosynthesis and energy metabolism pathways. In this study, a simple, rapid and sensitive method was developed for the simultaneous determination of 24 free amino acids in cell samples using hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry (HILIC-MS/MS). Cell samples were deproteinized with methanol/H2O (80:20, v/v) without intricate derivatization process. The analytes were separated on a Waters BEH Amide column (2.1 mm × 100 mm, 1.7 μm), and accomplished within 5 min at a flow rate of 0.2 mL/min. The good linearity was obtained for all analytes (r2 > 0.99) with the limits of quantification from 0.1 to 25 ng/mL. The intra- and inter-day precision ranged from 0.35 to 10.36% and from 2.22 to 9.93%, respectively. The recoveries of most analytes were between 80% and 120% with RSD less than 10.0%. The developed method was then applied to the direct analysis of 24 underivatized amino acids in human gastric cancer cell line MGC803 treated with the antitumor candidate drug J3, and significant differences in the concentration levels of amino acids were also assessed.


Objective sleep measures after endoscopic sinus surgery in patients with chronic rhinosinusitis.
Poor sleep quality is common in chronic rhinosinusitis (CRS). Prior studies have demonstrated improvements in patient-reported measures of sleep quality following endoscopic sinus surgery (ESS). The purpose of this study was to evaluate the effect of ESS on objective sleep parameters and identify any associations between specific objective measures and patient characteristics, comorbidities, and patient-reported outcome measures (PROMs). Adults with CRS undergoing ESS were prospectively enrolled from 4 centers across North America. Any subject previously diagnosed with a known primary sleep disorder was excluded. Objective sleep indices were recorded using a portable sleep diagnostic device preoperatively and postoperatively. Patient-reported outcome instruments were completed including the Pittsburgh Sleep Quality Index (PSQI) and the 22-item Sino-Nasal Outcome Test (SNOT-22). Thirty-six patients (mean age 47 years, 56% male) completed baseline and postoperative sleep studies with mean ± standard deviation (SD) follow-up 9.6 ± 7.7 months. Mean PSQI and SNOT-22 before and after ESS was 10.2 ± 3.9 vs 7.8 ± 4.4 (p = 0.001); and 54.6 ± 14.6 vs 28.5 ± 15.3 (p < 0.001), respectively. Total sleep time, sleep latency, and awakenings after sleep onset did not change following ESS (all p > 0.5) despite improvements in PSQI and SNOT-22. Changes in PSQI did not correlate with comorbidities or objective sleep indices (all p > 0.1). In this multicenter prospective cohort, objective sleep indices were not improved following ESS for CRS despite significant improvements in patient-reported sleep quality and CRS-specific QOL.


An automated data acquisition system for isolated tissue studies.
The automation of an isolated atria assay is described. Data acquisition, operation of the strip chart recorder, data reduction and manipulation and generation of notebook pages showing final EC50's, dose-ratios and local pA2's has been completely automated. The data are acquired via a SYM-1 (6502 CPU) 8-bit single board computer running an assembly language program stored on an EPROM chip. The data from a physiological recorder system are stored by the SYM-1 and, at the conclusion of the experiment, transmitted to a DEC MINC-11 microminicomputer running a sequence of programs in BASIC for the mathematical manipulation of the data and the automatic generation of lab notebook pages. The automated system totally eliminates hand transcription of data, manual plotting of curves and mathematical errors.


In vivo demonstration of focal fibrinolytic activity in abdominal aortic aneurysms.
Disseminated intravascular coagulation in association with arterial aneurysm has been reported in several cases. Reports have suggested a continuous deposition of fibrin and/or platelets at the site of the aneurysm followed by fibrinolysis as responsible for this disorder. To further investigate this theory we studied the in vivo binding of an indium-111-labelled monoclonal antibody against human tissue plasminogen activator (t-PA), a proteolytic enzyme involved in the fibrinolysis. Six patients admitted to the hospital for elective resection of an abdominal aortic aneurysm with a mural thrombus were examined. One day before the operation the antibody was injected intravenously. Scintigrams acquired just prior to operation and tissue samples obtained at the operation revealed an increased t-PA accumulation in the wall of the aneurysm. The study demonstrates in vivo, focally increased fibrinolytic activity in the aneurysm, explaining the coagulopathy observed in these patients.


RNA-Seq revealed the effect of adding different proportions of wheat diet on fat metabolism of Tibetan lamb.
The objective of this study was to evaluate the effect of the level of wheat substitution for corn on the fat metabolism of Tibetan lamb. A total of 90 Tibetan lambs [body weight (BW) of 19.78 ± 2.45 kg] were arranged with three substitution levels of wheat: WC (100 % corn), WL (10 % wheat substitution for corn), and WH (15 % wheat substitution for corn) on a dry matter basis. After the experiment, cry section technology was used to scrutinize the subcutaneous adipose tissue morphology, and genes related to fat metabolism were excavated using high-throughput sequencing technology. According to the study results, fat diameter and fat biovolume of the WL and WH groups were less than the WC group. A total of 506 differentially expressed genes (DEGs) were identified by RNA sequencing (RNA-Seq) technology. Compared with the WC group, 66 DEGs were upregulated and 59 DEGs were downregulated in the WL group, and 179 DEGs were upregulated and 269 DEGs were downregulated in the WH group. The top 20 DEGs were analyzed by Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway, indicating significant differences in the fat metabolism pathway. Five DEGs were randomly screened for quantitative reverse transcription-polymerase chain reaction (qRT-PCR) verification, and the results were consistent with the RNA-Seq results, which proved the accuracy of sequencing. In summary, with the increase in the proportion of supplemental wheat, the fat cells became smaller, and the genes related to fat decomposition were significantly upregulated.


The influence of helium pressure on the reduction induced in field potentials by various amino acids and on the GABA-mediated inhibition in the CA1 region of hippocampal slices in the rat.
In a previous study, it was shown that helium pressure depressed excitatory synaptic transmission mediated by the Schaffer-commissural afferents and increased the intrinsic excitability of pyramidal cells, in the CA1 region of hippocampal slices in the rat. In the present study, the neurochemical bases of these changes was investigated. Various excitatory amino acids were studied under normal and up to 80 atm of helium. At normal pressure, the amino acids tested induced a decrease in the field excitatory postsynaptic potential (EPSP) and antidromic field potential of CA1 pyramidal cells. These changes probably resulted from the well known depolarizing effect of the compounds. Quisqualate is supposed to activate the synaptic receptors of the pathway tested. Since the effect of this amino acid and other agonists were not significantly affected by helium pressure, it is suggested that the depressed hippocampal synaptic potentials under pressure did not result from reduced sensitivity of synaptic receptors. On the other hand, helium pressure enhanced the action of N-methyl-D-aspartate (NMDA) and depressed the GABA-mediated inhibition of CA1 pyramidal cells. Given that the excitability of these neurones is modulated by NMDA-related events and GABA inhibition, these results indicate that both neurochemical systems were probably involved in the helium pressure-induced hyperexcitability of the cells studied.


Breadth of the CD4+ T cell response to Anaplasma marginale VirB9-1, VirB9-2 and VirB10 and MHC class II DR and DQ restriction elements.
MHC class II molecules influence antigen-specific CD4+ T lymphocyte responses primed by immunization and infection. CD4+ T cell responses are important for controlling infection by many bacterial pathogens including Anaplasma marginale and are observed in cattle immunized with the protective A. marginale outer membrane (OM) vaccine. Immunogenic proteins that comprise the protective OM vaccine include type IV secretion system (T4SS) proteins VirB9-1, VirB9-2 and VirB10, candidates for inclusion in a multiepitope vaccine. Our goal was to determine the breadth of the VirB9-1, VirB9-2 and VirB10 T cell response and MHC class II restriction elements in six cattle with different MHC class II haplotypes defined by DRB3, DQA and DQB allele combinations for each animal. Overlapping peptides spanning each T4SS protein were tested in T cell proliferation assays with autologous antigen-presenting cells (APC) and artificial APC expressing combinations of bovine DR and DQ molecules. Twenty immunostimulatory peptides were identified; three representing two or more epitopes in VirB9-1, ten representing eight or more epitopes in VirB9-2 and seven representing seven or more epitopes in VirB10. Of the eight DRA/DRB3 molecules, four presented 15 peptides, which was biased as DRA/DRB3*1201 presented ten and DRA/DRB3*1101 presented four peptides. Four DQA/DQB molecules composed of two intrahaplotype and two interhaplotype pairs presented seven peptides, of which five were uniquely presented by DQ molecules. In addition, three functional mixed isotype (DQA/DRB3) restriction elements were identified. The immunogenicity and broad MHC class II presentation of multiple VirB9-1, VirB9-2 and VirB10 peptide epitopes justify their testing as a multiepitope vaccine against A. marginale.


Activation of IGFBP4 via unconventional mechanism of miRNA attenuates metastasis of intrahepatic cholangiocarcinoma.
Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver malignancy. Although its incidence is lower than that of hepatocellular carcinoma (HCC), ICC has a worse prognosis, and it is more prone to recur and metastasize, resulting in a far greater level of malignancy. Bioinformatics analysis and qRT-PCR were applied to assess the level of miR-122-5p and IGFBP4. Western blot, transwell assays, wound-healing assays, real-time cellular invasion monitoring, in vivo study were applied to explore the function of miR-122-5p and IGFBP4. Dual luciferase reporter assays and chromatin isolation by RNA purification (ChiRP) were applied to explore the regulation of IGFBP4 by miR-122-5p. Using The Cancer Genome Atlas (TCGA) data set, Sir Run Run Shaw hospital data set and bioinformatics analyses, we identified miR-122-5p as a potential tumor suppressor in ICC and validated its suppressive effect in metastasis and invasion of ICC. Transcriptome sequencing, rescue and complement experiments were used to identify insulin-like growth factor binding protein 4 (IGFBP4) as a target of miR-122-5p. The mechanism by which miR-122-5p regulates IGFBP4 was clarified by chromatin separation RNA purification technology, and dual-luciferase reporter assays. We discovered a rare novel mechanism by which miR-122-5p promotes IGFBP4 mRNA transcription by binding to its promoter region. Furthermore, in mouse orthotopic metastasis model, miR-122-5p inhibited the invasion of ICC. In summary, our study revealed a novel mechanism of miR-122-5p and function of the miR-122-5p/IGFBP4 axis in the metastasis of ICC. We also highlighted the clinical value of miR-122-5p and IGFBP4 in inhibiting ICC invasion and metastasis.


Laparoscopic ovariectomy and ovariohysterectomy in llamas and alpacas.
The minimally invasive nature of laparoscopic techniques and shorter convalescent periods have made these techniques increasingly popular for use in New World camelids (llamas and alpacas). This article outlines the instruments and steps needed to perform laparoscopic surgery on the female reproductive tract in llamas and alpacas.


Robotic system for magnetic resonance imaging-guided focused ultrasound treatment of thyroid nodules.
Herein, a robotic system offering Magnetic Resonance-guided Focused Ultrasound (MRgFUS) therapy of thyroid nodules was developed. The robotic system offers linear motion in 2 PC-controlled axes that navigate a 3 MHz single-element focused transducer. The system, through a C-arm structure attaches to the table of Magnetic Resonance Imaging (MRI) scanners and couples to the neck of patients lying in the supine position. The MRI compatibility of the developed system was assessed inside a 3 T scanner. Benchtop and MRI feasibility studies evaluating the heating performance of the system were executed on excised pork tissue and on homogeneous and thyroid model agar-based phantoms. The MRI compatibility of the system was successfully established. Grid sonications executed using robotic motion inflicted discrete and overlapping lesions on the excised tissue, while magnetic resonance (MR) thermometry successfully monitored thermal heating in agar-based phantoms. The developed system was found to be efficient with ex-vivo evaluation. The system can perform clinical MRgFUS therapy of thyroid nodules and other shallow targets after further in-vivo evaluation.


Synaptic vesicle size and number are regulated by a clathrin adaptor protein required for endocytosis.
Clathrin-mediated endocytosis is thought to involve the activity of the clathrin adaptor protein AP180. However, the role of this protein in endocytosis in vivo remains unknown. Here, we show that a mutation that eliminates an AP180 homolog (LAP) in Drosophila severely impairs the efficiency of synaptic vesicle endocytosis and alters the normal localization of clathrin in nerve terminals. Most importantly, the size of both synaptic vesicles and quanta is significantly increased in lap mutants. These results provide novel insights into the molecular mechanism of endocytosis and reveal a role for AP180 in regulating vesicle size through a clathrin-dependent reassembly process.


Understanding the Unique Barriers and Facilitators that Affect Men's Initiation and Retention in HIV Care: A Qualitative Study to Inform Interventions for Men Across the Treatment Cascade in Malawi.
Men in sub-Saharan Africa are underrepresented in antiretroviral therapy (ART) programs. Our secondary analysis of 40 in-depth interviews with Malawian men living with HIV examined barriers and facilitators for ART initiation versus retention. Interviewees included men who never initiated or initiated ART late (initiation respondents, n = 19); and men who initiated ART but were late for an appointment (retention respondents, n = 21). Transcribed interviews were coded using deductive and inductive coding techniques and analyzed using constant comparison methods. Long wait times, frequent facility visits, and insufficient in-clinic privacy were barriers for initiation and retention. Poor knowledge of ART was primarily a barrier for initiation; unexpected travel was a barrier for retention. Key facilitators for initiation and retention included previous positive experiences with health facilities. Having examples of successful men using ART primarily facilitated initiation; support from spouses and male peers facilitated retention. Results may inform interventions to increase men's engagement in ART services.


[The fibrin and plasma-protein content of human thrombi].
The protein composition of cell-free thrombus contained in abdominal aneurysms of the aorta was investigated using PAGE and immunoreactivity after solubilization. 1. The dry weight was 18--29%, with minor differences between various locations. 2. 35--40% of the dry weight was extractable with NaCl/citrate and was identified as albumin (80%) and IgG (15%). Trace amounts of fibrinogen immunoreactive material were present. 3. The NaCl/citrate insoluble part was 80% hydrolyzable with plasmin. The predominant fragment was D-D dimer (from cross-linked fibrin). In addition, fragments Dand E were observed. Reduction of the NaCl/citrate insoluble part with 2-mercaptoethanol resulted in almost complete solubilization. PAGE analysis demonstrated alpha-polmers, gamma-gamma dimers and little free alpha-chains, indicating almost complete cross-linking of fibrin. Thus, the major protein were cross-linked fibrin, albumin and IgG.


Mesenteric lipoma: report of a case with emphasis on US findings.
Mesenteric lipoma (ML) is a relatively rare disease that has been very infrequently reported. We present the clinical data and medical imaging results of an asymptomatic case with ML incidentally detected by sonography (US). On US the lesion was imaged as a well-demarcated hypoechoic mass with multiple linear echoes. The mass also changed location under probe compression. The lesion was imaged as a clearly demarcated non-enhanced homogeneous fat-density mass on CT. Color Doppler US and angiography confirmed the avascular nature of the mass. Knowledge of the US findings in this case is useful for the diagnosis of ML.


Afirma Benign Thyroid Nodules Show Similar Growth to Cytologically Benign Nodules During Follow-Up.
The Afirma gene expression classifier (GEC) is a molecular diagnostic test that has a high negative predictive value for ruling out malignancy in thyroid nodules with indeterminate cytology. Many patients with a cytologically indeterminate and GEC benign (Cyto-I/GEC-B) nodule undergo monitoring instead of diagnostic surgery, but few data describe their follow-up. The objective of the study was to determine the sonographic changes and clinical outcomes for patients with Cyto-I/GEC-B nodules compared with patients with cytologically benign (Cyto-B) nodules. This was a retrospective analysis of consecutive Cyto-I/GEC-B nodules evaluated at Brigham and Women's Hospital compared with Cyto-B nodules. Nodule growth of 20% or greater in two dimensions or of 50% or greater in volume, change in sonographic features, and rates of repeat fine-needle aspiration, thyroidectomy, and malignancy. Ninety-five Cyto-I/GEC-B nodules in 90 patients were identified. Five patients underwent primary surgical resection. Of the remaining 90 nodules, 58 (64.4%) had sonographic follow-up available at a median of 13 months (range 4-40 mo). Cyto-I/GEC-B nodules showed similar growth compared with 1224 Cyto-B nodules using either of the following criteria: 20% or greater in two dimensions (8.6% vs 8.3%, P = .80) or 50% or greater in volume (17.2% vs 13.8%, P = .44). Thyroidectomies were more frequent in the Cyto-I/GEC-B group (13.8% vs 0.9%, P < .0001), but cancer was found in only one patient, with no evidence of persistent disease after initial treatment. Cyto-I/GEC-B nodules demonstrate similar growth to Cyto-B nodules during follow-up. Although Cyto-I/GEC-B nodules were more frequently resected, only one malignancy was found. These data suggest that reassessment of Cyto-I/GEC-B nodules may be performed similarly to those with benign cytology.


A novel prognostic model for patients with sarcomatoid renal cell carcinoma.
To demonstrate sarcomatoid differentiation is an independent prognostic feature for patients with grade 4 renal cell carcinoma (RCC) with or without distant metastases. To identify independent predictors of survival, evaluate the correlation between the amount of sarcomatoid differentiation and cancer-specific survival (CSS), and to design a multivariate prognostic model for patients with sarcomatoid RCC. We used the Mayo Clinic Nephrectomy Registry to identify 204 post-nephrectomy patients with sarcomatoid-variant RCC, as well as 207 patients with unilateral grade 4 RCC without sarcomatoid features for comparison. All slides were reviewed by one pathologist. CSS was estimated using the Kaplan-Meier method. The associations of clinical and pathological features with death from RCC were evaluated using Cox proportional hazards regression models. For all patients with grade 4 RCC, the presence of sarcomatoid differentiation was associated with a 58% increased risk of death from RCC (P < 0.001). For patients with grade 4 non-metastatic (M0) RCC, the presence of sarcomatoid differentiation was associated with an 82% increased risk of death from RCC (P < 0.001). For patients with sarcomatoid RCC, the 2009 primary tumour classifications, presence of regional lymph nodes and distant metastases, coagulative tumour necrosis, and the amount of sarcomatoid differentiation were each significantly associated with death from RCC in a multivariate setting. After adjusting for other prognostic variables, each 10% increase in the amount of sarcomatoid differentiation was associated with a 6% increased risk of death from RCC (P = 0.028). Patients whose tumours contained ≥30% (median amount) sarcomatoid differentiation were 52% more likely to die from RCC compared with patients whose tumours contained <30% sarcomatoid differentiation (hazard ratio 1.52; P = 0.018). Among patients with grade 4 RCC, either with or without distant metastases at surgery, sarcomatoid differentiation is significantly associated with adverse survival in a multivariate setting. We also suggest for the first time that the percentage of sarcomatoid differentiation is an independent prognostic feature in a multivariate setting. The 2009 primary tumour classifications, regional lymph node status, the presence of distant metastases classifications, coagulative tumour necrosis, and the amount of sarcomatoid differentiation are independent predictors of survival for patients with sarcomatoid RCC.


Clinical target volume and organs at risk segmentation for rectal cancer radiotherapy using the Flex U-Net network.
The aim of this study was to improve the accuracy of the clinical target volume (CTV) and organs at risk (OARs) segmentation for rectal cancer preoperative radiotherapy. Computed tomography (CT) scans from 265 rectal cancer patients treated at our institution were collected to train and validate automatic contouring models. The regions of CTV and OARs were delineated by experienced radiologists as the ground truth. We improved the conventional U-Net and proposed Flex U-Net, which used a register model to correct the noise caused by manual annotation, thus refining the performance of the automatic segmentation model. Then, we compared its performance with that of U-Net and V-Net. The Dice similarity coefficient (DSC), Hausdorff distance (HD), and average symmetric surface distance (ASSD) were calculated for quantitative evaluation purposes. With a Wilcoxon signed-rank test, we found that the differences between our method and the baseline were statistically significant (P< 0.05). Our proposed framework achieved DSC values of 0.817 ± 0.071, 0.930 ± 0.076, 0.927 ± 0.03, and 0.925 ± 0.03 for CTV, the bladder, Femur head-L and Femur head-R, respectively. Conversely, the baseline results were 0.803 ± 0.082, 0.917 ± 0.105, 0.923 ± 0.03 and 0.917 ± 0.03, respectively. In conclusion, our proposed Flex U-Net can enable satisfactory CTV and OAR segmentation for rectal cancer and yield superior performance compared to conventional methods. This method provides an automatic, fast and consistent solution for CTV and OAR segmentation and exhibits potential to be widely applied for radiation therapy planning for a variety of cancers.