SemiMAR: Semi-Supervised Learning for CT Metal Artifact Reduction.
Metal artifacts lead to CT imaging quality degradation. With the success of deep learning (DL) in medical imaging, a number of DL-based supervised methods have been developed for metal artifact reduction (MAR). Nonetheless, fully-supervised MAR methods based on simulated data do not perform well on clinical data due to the domain gap. Although this problem can be avoided in an unsupervised way to a certain degree, severe artifacts cannot be well suppressed in clinical practice. Recently, semi-supervised metal artifact reduction (MAR) methods have gained wide attention due to their ability in narrowing the domain gap and improving MAR performance in clinical data. However, these methods typically require large model sizes, posing challenges for optimization. To address this issue, we propose a novel semi-supervised MAR framework. In our framework, only the artifact-free parts are learned, and the artifacts are inferred by subtracting these clean parts from the metal-corrupted CT images. Our approach leverages a single generator to execute all complex transformations, thereby reducing the model's scale and preventing overlap between clean part and artifacts. To recover more tissue details, we distill the knowledge from the advanced dual-domain MAR network into our model in both image domain and latent feature space. The latent space constraint is achieved via contrastive learning. We also evaluate the impact of different generator architectures by investigating several mainstream deep learning-based MAR backbones. Our experiments demonstrate that the proposed method competes favorably with several state-of-the-art semi-supervised MAR techniques in both qualitative and quantitative aspects.


Fluorescence based primer extension technique to determine transcriptional starting points and cleavage sites of RNases in vivo.
Fluorescence based primer extension (FPE) is a molecular method to determine transcriptional starting points or processing sites of RNA molecules. This is achieved by reverse transcription of the RNA of interest using specific fluorescently labeled primers and subsequent analysis of the resulting cDNA fragments by denaturing polyacrylamide gel electrophoresis. Simultaneously, a traditional Sanger sequencing reaction is run on the gel to map the ends of the cDNA fragments to their exact corresponding bases. In contrast to 5'-RACE (Rapid Amplification of cDNA Ends), where the product must be cloned and multiple candidates sequenced, the bulk of cDNA fragments generated by primer extension can be simultaneously detected in one gel run. In addition, the whole procedure (from reverse transcription to final analysis of the results) can be completed in one working day. By using fluorescently labeled primers, the use of hazardous radioactive isotope labeled reagents can be avoided and processing times are reduced as products can be detected during the electrophoresis procedure. In the following protocol, we describe an in vivo fluorescent primer extension method to reliably and rapidly detect the 5' ends of RNAs to deduce transcriptional starting points and RNA processing sites (e.g., by toxin-antitoxin system components) in S. aureus, E. coli and other bacteria.


Scoping Review of Pediatric Early Warning Systems (PEWS) in Resource-Limited and Humanitarian Settings.
Pediatric Early Warning Systems (PEWS) aim to identify hospitalized children at increased risk of deterioration by assigning a score based on vital signs and clinical status and guiding interventions using a response algorithm to improve outcomes. When implemented with quality improvement methodology, these systems have been shown to be effective in high-resource settings and have the potential to improve the care of children in humanitarian and resource-limited settings (RLS). The purpose of this review is to summarize the current evidence for use of PEWS in RLS and identify areas for further research. A review of the current PEWS literature in RLS was performed using Web of Science, PubMed, Scopus, Cumulative Index of Nursing and Allied Health Literature (CINAHL), EMBASE, Portal Regional da BVS, and TRIP Database. While there is limited research available on this topic, eight studies on the use of PEWS, or a PEWS score in a pediatric population in low- or middle-income countries were identified. Two studies assessed the clinical effect of implementation of PEWS; one reported a reduction in clinical deterioration events and the other a reduction in mortality. The remaining studies assessed the association of a PEWS score with signs of clinical deterioration or mortality without a response algorithm. Further research on the impact of PEWS implementation on inpatient care and outcomes in RLS is needed.


Social stigma and the dilemmas of providing care to substance users in a safety-net emergency department.
There has been increasing interest in how social stigma affects health care delivery to vulnerable patients but few examples of the dilemmas that may arise for providers who care for stigma-vulnerable populations. Using qualitative data, the authors examine care delivery in a safety-net emergency department where many patients were socially disadvantaged and vulnerable to stigma because of substance use problems. Analysis of our data revealed five themes related to the dilemmas of providing care for this patient population: 1) providers valued assisting vulnerable and underserved patients; 2) providers' interactions with patients could be challenging; 3) providers did not know if substance involved patients provided accurate medical histories; 4) providers were concerned about drug-seeking behavior; and 5) providers had to balance substance-involved patients' needs with the necessity of managing limited resources. We discuss how these themes relate to care dynamics and social stigma in the safety-net setting.


Stroke risk in anticoagulated patients with atrial fibrillation undergoing endoscopy.
To determine the risk of stroke in anticoagulated patients with atrial fibrillation (AF) when anticoagulation is adjusted for bronchoscopy, gastroscopy, or colonoscopy, and to identify factors that could modify this risk. The authors reviewed patients with AF undergoing endoscopies between 1995 and 2001, with specific analysis of patients with AF in whom anticoagulation was adjusted for the procedures. The authors calculated the stroke rate within 30 days of the procedures. Twelve strokes occurred in 987 patients undergoing 1,137 procedures (1.06%/procedure). The risk ranged from 0.31% for patients with nonvalvular AF undergoing routine procedures to 2.93% for complex patients undergoing endoscopies combined with other procedures or with comorbid illnesses. Patients with stroke were more likely to be complex (7/12 vs 219/975, p = 0.04); to be older than 80 years (6/12 vs 187/975, p = 0.017); to have a history of stroke (7/12 vs 194/975, p = 0.004), hypertension (10/12 vs 508/975, p = 0.04), or hyperlipidemia (9/12 vs 334/975, p = 0.005); or to have a family history of vascular disease (10/12 vs 502/975, p = 0.039). The risk of stroke in patients with AF whose anticoagulation is adjusted for endoscopies is low, but almost tenfold higher in patients with complex clinical situations. Age, history of stroke, hypertension, hyperlipidemia, and family history of vascular disease may increase the risk of stroke.


Expression changes of cation chloride cotransporters in the rat spinal cord following intraplantar formalin.
Cation chloride cotransporters, K(+)-Cl(-) cotransporter 2 (KCC2) and Na(+)-K(+)-Cl(-) cotransporter 1 (NKCC1) are reported to be expressed in the neurons in the spinal cord and regulate intracellular Cl(-) concentration. Evidence has been accumulating that the expression of cation chloride cotransporters changes in inflammatory or neuropathic pain, and such changes take a part in pathophysiology of the persistent pain states. However, it is largely unknown how these cotransporters contribute to hyperalgesia in the acute pain state. We, therefore, investigated expression changes of KCC2 and NKCC1 in the spinal dorsal horn of the rat after the intraplantar injection of formalin as an acute nociceptive stimulus. The rats showed two phases (phases 1 and 2) of increase in pain-related behavior in response to formalin. We found that expression of KCC2-like immunoreactivity (IR) was reduced in lamina I and II in the lumbar spinal cord on the stimulated side in phase 1, and then recovered gradually. In contrast, the number of NKCC1-like IR-positive cells was unchanged over the period examined. These results suggest that KCC2, rather than NKCC1, mainly contributes to modulating excitability of the dorsal spinal cord neurons in the initial stage of formalin-evoked hyperalgesia.


Hepatic artery pseudoaneurysm and hemobilia following laparoscopic cholecystectomy.
Iatrogenic or accidental injury to the right hepatic artery or its branches can result in formation of pseudoaneurysm, which may rupture into the biliary system, leading to life-threatening hemobilia. We describe one such case following laparoscopic cholecystectomy and discuss its management.


[The morbidity of radical prostatectomy using a retropubic approach: a series of 150 cases].
The authors studied the morbidity of retropubic radical prostatectomy in 150 patients operated between December 1983 and March 1993. The perioperative mortality was zero. A rectal injury occurred in 3 cases (2%) and was repaired immediately without colostomy. The median operative blood loss was 1,500 ml. 36.6% of cases developed perioperative complications. 8% of early postoperative complications (< 1 month) required reoperation. Major complications (thromboembolic and septic) were observed in 3.2% of cases. The late postoperative complications included fibrous stenosis of the urethrovesical anastomosis in 12% of cases with a median time to onset of 4 months: 3.6% of patients developed an anastomotic stenosis due to local recurrence. 12.6% of cases developed persistent urinary incontinence (6.6% of minor incontinence, 5.3% of moderate incontinence and 0.7% of severe incontinence). Incontinence was more frequent (p < 0.05) among the first 75 patients of the series. Erectile function was preserved in 19.8% of patients. The incidence of post-operative impotence was significantly lower (p < 0.01) in patients under the age of 60 years (65% vs 94.8%). Radical prostatectomy, considered to be the most appropriate treatment for localised prostatic cancer, can now be performed with acceptable morbidity.


A two-dimensional silver-iodide organic network constructed from a unique [Ag6I6] hexagonal prism-based one-dimensional column motif.
A novel two-dimensional coordination polymer, poly[[μ2-1,3-bis(2-methyl-1H-imidazol-1-yl)propane]di-μ4-iodido-di-μ3-iodido-silver(I)], [Ag4I4(C11H16N4)]n, (I), has been synthesized by solvothermal reaction of AgNO3, KI and 1,3-bis(2-methyl-1H-imidazol-1-yl)propane (bmimp). In (I), the two unique Ag(I) cations have AgNI3 and AgI4 four-coordinated tetrahedral geometries. The bmimp ligand has imposed twofold symmetry. The Ag(I) cations and iodide anions form a unique one-dimensional polymeric column motif incorporating [Ag6I6] hexagonal prisms, which are then connected by bmimp ligands to form two-dimensional organic-inorganic layers. The layers are arranged in parallel in an ABAB fashion and are packed into the resultant three-dimensional supramolecular framework by van der Waals interactions.


Effects of bisphenol A on antioxidant system and lipid profile in rats.
We investigated the effects of bisphenol A (BPA) on antioxidant system enzymes, blood lipid profile and histologic structure of liver and pancreas in rats. We used 40 8-week-old male Wistar albino rats. The animals were divided into five groups of eight: control, vehicle, BPA-5, BPA-50 and BPA-500. BPA was dissolved in ethanol, then mixed with corn oil. The control group was untreated. The vehicle group was given the ethanol-corn oil mixture. The BPA 5, BPA 50 and BPA 500 groups were given 5, 50, and 500 µg/kg body weights/day, respectively. After 8 weeks, blood and tissue samples were obtained from the animals and plasma GSH, TBARS, SOD, GPx, CAT, NO, total cholesterol, triglyceride, HDL, LDL, insulin and glucose were measured. The sections were stained using histochemical and immunohistochemical methods. BPA significantly decreased the levels of GSH, SOD, GPx and CAT, and increased the levels of TBARS and NO in plasma. There was no significant difference among the groups in plasma insulin and glucose levels. The percentage of insulin immunoreactive cells in islets increased significantly in the BPA-500 group. The H-score of the BPA-5 and BPA-50 groups decreased significantly compared to controls. We found that BPA caused oxidative stress and disruption of pancreatic β-cell function. Therefore, BPA is a risk factor for animal and human health.


Foam solar sea power plant.
In the accompanying report Beck suggests a new type of open-cycle system for obtaining power from the ocean's thermal gradient. A modification of this open-cycle plant which will ensure a high efficiency, and also a low capital cost per unit power output, is described here.


Bone marrow mastocytosis associated with an undifferentiated extramedullary tumor of hemopoietic origin.
The case of a 62-year-old man who presented with acute abdominal pain and a widespread tumor involving the retroperitoneum is described. Three weeks after initial presentation, the patient died suddenly of acute cardiac failure with signs of arrhythmia. Autopsy revealed a disseminated tumor with infiltration of the retroperitoneal fat, as well as nodules in the left testis and the right atrium. The tumor cells were reactive for CD45, vimentin, and chloroacetate esterase, but were unreactive with a broad spectrum of antibodies against myelomonocytic and lymphocytic antigens and with antibodies against tryptase and c-kit (CD117), which are characteristic markers for mast cells. However, the bone marrow exhibited the typical picture of mastocytosis, with disseminated clusters of differentiated spindle-shaped cells that stained strongly for tryptase, c-kit, and chloroacetate esterase. No infiltrates of well-differentiated mastocytosis could be detected in any of the extramedullary tissues investigated. A diagnosis of bone marrow mastocytosis with an associated undifferentiated extramedullary tumor of hemopoietic origin was established. By definition, the extramedullary tumor could not be diagnosed as a granulocytic sarcoma or (differentiated) mastocytoma, but the possibility that a mast cell progenitor could be involved in the evolution of both tumors cannot be ruled out.


Roy Schafer: a beginning.
The author provides a biographical overview of Schafer's life, culled from his published work and focused primarily on his professional development. This biography is used to demonstrate some of Schafer's central theoretical insights on narrativity and language, and reveals the consistency of his thinking over his long career. A brief discussion of his writing on King Lear provides a bridge between theoretical and biographical material.


Mass analysis of mobility-selected ion populations using dual gate, ion mobility, quadrupole ion trap mass spectrometry.
An electrospray ionization, dual gate, ion mobility, quadrupole ion trap mass spectrometer (ESI-DG-IM-QIT-MS) was constructed and evaluated for its ability to select mobility-filtered ions prior to mass analysis. While modification of the common signal-averaged ion mobility experiment was required, no modifications to the QIT were necessary. The dual gate scanning mode of operation was used to acquire mobility spectra, whereas the single mobility monitoring experiment selectively filtered ions for concentration and subsequent fragmentation within the QIT. Ion mobility separation of positively charged peptides and negatively charged carbohydrates, followed by MS fragmentation, was demonstrated. For a 1-min acquisition time, it was possible to obtain complete de novo sequence information for the examined peptides. Fragmentation of the negative carbohydrate chlorine adducts yielded ions characteristic of cross-ring and glycosidic bond cleavage. Previous unions of atmospheric pressure ion mobility and mass spectrometry have been limited in their ability to reproducibly obtain MSn data for mobility separation ions. The union of high-pressure ion mobility with quadrupole ion trap mass spectrometry presents the unique opportunity to obtain more detailed information regarding the chemistries of gas-phase ions.


Joint dysfunction associated with venous malformations of the limbs: Which patients are at high risk?
Objectives Joint dysfunction occurs frequently in patients with venous malformations of the extremities. This study aims to describe the risk factors of joint dysfunction in these patients. Method We retrospectively collected clinical and radiological aspects of 168 patients with extremity venous malformations from January 2013 to August 2015. Patients were categorized into "with joint dysfunction" or "without joint dysfunction" groups according to the symptoms. Results Forty-four (26%) patients were assigned into with joint dysfunction group. Univariate analysis showed that the significant variables were age (P = 0.008), location (P = 0.005), size (P < 0.001), involved tissue (P < 0.001), visual analogue scale scores of pain (P = 0.004), pain duration (P = 0.018), and therapy history (P < 0.001). Multivariate analysis showed that the factors associated with joint dysfunction were age below 10 years (adjusted odds ratio = 2.70, P = 0.004), lower limb (adjusted odds ratio = 2.72, P = 0.042), pain duration over 1 year (adjusted odds ratio = 3.47, P = 0.001) and therapy history (yes) (adjusted odds ratio = 3.71, P = 0.004). Conclusion This study provides preliminary evidence indicating which patients are at high risk for joint dysfunction associated with limb venous malformations.


Metabolomics analysis of anaphylactoid reaction reveals its mechanism in a rat model.
Anaphylactoid reactions, accounting for more than 77% of all immune-mediated immediate hypersensitivity reactions, have become a serious threat to public health, but their effect mechanism is not clear and diagnostic tests are limited. Comprehensive metabolite analysis may reveal the anaphylactoid effect mechanism systematically and provide reference for future diagnostic purposes. Plasma from Brown Norway rats given intravenous injection of saline, compound 48/80 (2.5 mL/kg) or ovalbumin (20 mL/kg) in 20 s for the first time was used to study the effect mechanism of anaphylactoid reactions through metabolomics (UPLC-qTOF-MS/MS). Metabolomics integrated with proteomics data were used to analyze the anaphylactoid pathways by MetaboAnalyst followed by integrated pathway analysis. Thirty metabolites were identified through the METLIN database by MS/MS and 18 of them were confirmed by authentic standards. The results showed that adenosine, histamine, N-acetylhistamine, N(α)-γ-glutamylhistamine, malate and xanthine are important indices for anaphylactoid reactions. It could be concluded that the effect mechanism is mainly composed of histidine metabolism, arachidonic acid metabolism, energy metabolism, purine metabolism and other small molecules through 30 metabolites. Multiple linear regression analysis indicated that not only histamine but also N(α)-γ-glutamylhistamine and arachidonic acid could be used to evaluate anaphylactoid symptoms of animals. Furthermore, the citrate cycle, histidine metabolism and arachidonic acid metabolism could be the main pathways of anaphylactoid reactions as determined by MetaboAnalyst. The results may provide a reference to improve diagnostic accuracy and predict and monitor treatment efficacy in anaphylactoid reactions in the clinical setting.


A pilot study: Exploring the influence of COVID-19 on cardiovascular physiology and retinal microcirculation.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) affects the cardiovascular system. The current study investigated changes in heart rate (HR), blood pressure (BP), pulse wave velocity (PWV), and microcirculation in patients recovering from Coronavirus disease 2019 (COVID-19) infection. Out of 43 initially contacted COVID-19 patients, 35 (30 males, 5 females; age: 60 ± 10 years; and body mass index (BMI): 31.8 ± 4.9) participated in this study. Participants were seen on two occasions after hospital discharge; the baseline measurements were collected, either on the day of hospital discharge if a negative PCR test was obtained, or on the 10th day after hospitalization if the PCR test was positive. The second measurements were done 60 days after hospitalization. The vascular measurements were performed using the VICORDER® device and a retinal blood vessel image analysis. A significant increase in systolic BP (SBP) (from 142 mmHg, SD: 15, to 150 mmHg, SD: 19, p = 0.041), reduction in HR (from 76 bpm, SD: 15, to 69 bpm, SD: 11, p = 0.001), and narrower central retinal vein equivalent (CRVE) (from 240.94 μm, SD: 16.05, to 198.05 μm, SD: 17.36, p = 0.013) were found. Furthermore, the trends of increasing PWV (from 11 m/s, SD: 3, to 12 m/s, SD: 3, p = 0.095) and decreasing CRAE (from 138.87 μm, SD: 12.19, to 136.77 μm, SD: 13.19, p = 0.068) were recorded. The present study investigated cardiovascular changes following COVID-19 infection at two-time points after hospital discharge (baseline measurements and 60 days post-hospitalization). Significant changes were found in systolic blood pressure, heart rate, and microvasculature indicating that vascular adaptations may be ongoing even weeks after hospitalization from COVID-19 infection. Future studies could involve conducting additional interim assessments during the active infection and post-infection periods.


The role of tuning curve variables and threshold measures in the estimation of sensory cell loss.
Auditory-evoked potential tuning curves were collected at six frequencies before and 30 days after various noise exposures in 363 chinchillas using a simultaneous masking paradigm. Traditional bivariate and multiple linear regression/correlation analyses were performed in an effort to determine the extent to which sensory cell damage could be estimated from a knowledge of audiometric and tuning curve variables. The results showed strong correlations between percent outer hair cell (%OHC) loss and permanent threshold shift (PTS) and between %OHC loss and the tuning curve variables Q10 dB and high- and low-frequency slopes (SHF, SLF). The correlations were strongest between PTS and %OHC loss. However, the proportion of variability (r2) in %OHC loss attributable to variability in the predictor variable(s) (i.e., PTS) could be increased significantly by adding the Q10 dB of the tuning curve whose probe frequency was centered in the octave band length of the cochlea corresponding to the frequency at which the PTS occurred. The r2 values could be further increased by including audiometric and tuning curve variables from frequencies adjacent to the octave band being evaluated.


Recollections on Sixty Years of NBS Ionizing Radiation Programs for Energetic X Rays and Electrons.
These recollections are on ionizing radiation programs at the National Bureau of Standards (NBS) that started in 1928 and ended in 1988 when NBS became the National Institute of Standards and Technology (NIST). The independent Council on Ionizing Radiation Measurements and Standards (CIRMS) was formed in 1992. This article focuses on how measurements and standards for x rays, gamma rays, and electrons with energies above 1 MeV began at NBS and how they progressed. It also suggests how the radiation processors of materials and foods, the medical radiographic and radiological industries, and the radiological protection interests of the government (including homeland security) represented in CIRMS can benefit from NIST programs.


Drivers and annual estimates of marine wildlife entanglement rates: A long-term case study with Australian fur seals.
Methods of calculating wildlife entanglement rates are not standardised between studies and often ignore the influence of observer effort, confounding comparisons. From 1997-2013 we identified 359 entangled Australian fur seals at Seal Rocks, south-eastern Australia. Most entanglement materials originated from commercial fisheries; most frequently entangling pups and juveniles. Using Generalized Additive Mixed Models, which incorporated observer effort and survey frequency, we identified that entanglements were observed more frequently amongst pups from July to October as they approached weaning. Neither the decline in regional fishing intensity nor changing seal population size influenced the incidence of entanglements. Using the models, we estimated that 302 (95% CI=182-510) entangled seals were at Seal Rocks each year, equivalent to 1.0% (CI=0.6-1.7%) of the site population. This study highlights the influence of observer effort and the value of long-term datasets for determining the drivers of marine debris entanglements.


NBI - plasma vaporization hybrid approach in bladder cancer endoscopic management.
A prospective study was performed aiming to evaluate the surgical efficacy, perioperative safety profile, diagnostic accuracy and medium term results of a multi-modal approach consisting in narrow band imaging (NBI) cystoscopy and bipolar plasma vaporization (BPV) when compared to the standard protocol represented by white light cystoscopy (WLC) and transurethral resection of bladder tumors (TURBT). A total of 260 patients with apparently at least one bladder tumor over 3 cm were included in the trial. In the first group, 130 patients underwent conventional and NBI cystoscopy followed by BPV, while in a similar number of cases of the second arm, classical WLC and TURBT were applied. In all non-muscle invasive bladder tumors' (NMIBT) pathologically confirmed cases, standard monopolar Re-TUR was performed at 4-6 weeks after the initial intervention, followed by one year' BCG immunotherapy. The follow-up protocol included abdominal ultrasound, urinary cytology and WLC, performed every 3 months for a period of 2 years. The obturator nerve stimulation, bladder wall perforation, mean hemoglobin level drop, postoperative bleeding, catheterization period and hospital stay were significantly reduced for the plasma vaporization technique by comparison to conventional resection. Concerning tumoral detection, the present data confirmed the NBI superiority when compared to standard WLC regardless of tumor stage (95.3% vs. 65.1% for CIS, 93.3% vs. 82.2% for pTa, 97.4% vs. 94% for pT1, 95% vs. 84.2% overall). During standard Re-TUR the overall (6.3% versus 17.4%) and primary site (3.6% versus 12.8%) residual tumors' rates were significantly lower for the NBI-BPV group. The 1 (7.2% versus 18.3%) and 2 (11.5% versus 25.8%) years' recurrence rates were substantially lower for the combined approach. NBI cystoscopy significantly improved diagnostic accuracy, while bipolar technology showed a higher surgical efficiency, lower morbidity and faster postoperative recovery. The combined technique offered a reduced rate of residual tumors at Re-TUR, both globally as well as for orthotopic tumors. Substantially lower recurrence rates were found at 1 and 2 years among the NBI-BPV cases.


Lignoren, a new sesquiterpenoid metabolite from Trichoderma lignorum HKI 0257.
The new compound lignoren (1) was isolated from Trichoderma lignorum HKI 0257 by chromatographic methods. This metabolite has a santalane-like structure, which was elucidated by mass spectrometric and NMR spectroscopic investigations. Lignoren (1) shows a moderate narrow-spectrum of antibacterial and antifungal activity.


Neurotrophic interactions in the development of spinal cord motoneurons.
The final number of spinal cord motoneurons is attained by a two-step process involving the proliferation of precursor cells and the loss by cell death of a proportion (approximately 50%) of the post-mitotic neurons. Although the mechanisms responsible for the proliferation of stereotyped numbers of motoneurons are not understood, considerable evidence from in vitro as well as in vivo studies indicates that the second step in attaining population size (cell death) is controlled by the interaction of motoneurons with both their efferent targets and their afferent inputs. Target influences on motoneuron survival are thought to be regulated by muscular activity and by competition for limited amounts of neurotrophic factors derived from striated skeletal muscles. However, evidence that such putative neurotrophic factors actually modulate motoneuron survival in vivo has been lacking. Using crude and partially purified extracts from embryonic hindlimbs (Days 8-9) we have found that the treatment of chick embryos in ovo with these agents during the normal cell death period (Days 5-10) rescues a significant number of motoneurons from degeneration. Kidney or lung extracts and heat-inactivated hindlimb extracts were ineffective. The survival-inducing activity of partially purified extract was dose dependent and developmentally regulated. The survival of sensory, sympathetic and a population of cholinergic sympathetic preganglionic neurons was unaffected by treatment with hindlimb extract. The massive motoneuron death that occurs after early target (hindlimb) removal was partially ameliorated by daily treatment with the hindlimb extract. Survival-inducing activity of the extract is lost after trypsin treatment. Taken collectively these results indicate that a target-derived protein or polypeptide neurotrophic factor is involved in the regulation of motoneuron survival in vivo.


Does neural blockade and other neurosurgical modalities offer a durable pain relief for spinal disorders?
Various minimally invasive techniques have been performed to fight pain originating at bony and soft tissue spine elements. Of these, denervation of the nerve supply to the facet joints is the only technique for which three multidisciplinary clinical studies have been conducted, according to state-of-the-art study criteria, to produce evidence-based efficacy data. Of all of the morphopathological factors contributing to chronic spinal pain, facet joints play a major role in generating spinal pain due to hypermobility or joint degeneration. This is commonly referred to as facet joint syndrome (FJS). Other important non-morphological co-morbidity factors (mostly psychiatric, social or occupational in origin) also play an important role in mediating spinal pain. From the existing evidence-based data, there seems to be no durable pain relief in FJS using standard lesioning techniques (thermal and/or cryo). Efficacy compared to placebo lasts for only 4 weeks. Only those patients with spinal canal stenosis and concomitant chronic FJS appear to benefit after microsurgical decompression of the narrowed spinal canal. However, the database for this claim is low since low back pain is not the major focus in the clinical symptoms reported by the patients.


VAB-3 combination chemotherapy in primary mediastinal germ cell tumors.
Ten patients with primary mediastinal nonseminomatous germ cell tumors were treated with the VAB-3 protocol. All patients with measurable disease showed tumor regression with chemotherapy, but only two had sustained partial response allowing thoracotomy and resection of residual tumor after the second induction. Both patients received additional induction therapy and one also received irradiation; they have been free of disease 41+ and 40+ months from the beginning of chemotherapy. All other patients died from the disease.


[Tocolysis with a nitroglycerin patch].
We would like to present a 22 year old nullipara where preterm labor was successfully tocolysed using a nitro-glycerine path without sequellae side effects. Potential advantages and disadvantages of a transdermal nitro-glycerine delivery system in comparison to the commonly used intravenous fenoterol tocolysis is discussed.


Ten-Year Follow-Up of a Fragment Reattachment to an Anterior Tooth: A Conservative Approach.
This report describes the 10-year follow-up data of a patient who underwent fragment reattachment to the maxillary central incisor after coronal fracture with pulp exposure as well as the procedures followed for functional and esthetic adjustments. A 9-year-old female patient presented at the clinic of dentistry at the State University of Rio de Janeiro with a coronal fracture and pulp exposure of the right maxillary central incisor that had occurred immediately after an accident. The intact tooth fragment was recovered at the accident site and stored in milk. The treatment plan followed was to perform direct pulp capping and tooth fragment reattachment. When the patient was 14 years old, adhesion between fragment and remaining tooth was lost, and fragment reattachment was performed. Five years later, the same tooth presented clinical discoloration and absence of sensitivity during pulp vitality tests. Subsequently, a new treatment plan was formulated, which included endodontic treatment, followed by nonvital tooth bleaching and light-cured composite resin restoration. An esthetic and natural-looking restoration was achieved. Tooth fragment reattachment is not a temporary restorative technique and requires functional and esthetic adjustments over time to maintain the biomimetic characteristics of traumatized anterior teeth and predictable outcomes.


Trichostatin A induces p53-dependent endoplasmic reticulum stress in human colon cancer cells.
Trichostatin A (TSA) has been demonstrated to exhibit various anticancer effects that influence cell cycle arrest, cell proliferation and apoptosis of cancer cells. A potential association between TSA and endoplasmic reticulum (ER) function has been suggested but its anticancer mechanism involving the induction of ER stress is unknown. p53 has previously been demonstrated to regulate ER function in response to stress but its role involving TSA and ER stress in cancer cells is poorly understood. The current study identified that TSA induced ER stress in wild type (WT) HCT116 human colon cancer cells. Following TSA treatment, the ER stress markers GRP78 and GRP94 significantly increased without hyperacetylation of their promoter regions. The inositol-requiring enzyme 1 α (IRE1α)/X-box binding protein 1 (XBP1) pathway was implicated due to an association of phosphorylated IRE1α and spliced XBP1 with ER stress. However, luciferase reporter assay indicated that splicing events were attenuated in HCT116 TP53(-/-) cells. Furthermore, cell viability and apoptosis were revealed to depend on p53 during TSA treatment. Cell viability increased and the apoptosis rate decreased in HCT116 TP53(-/-) cells compared with WT HCT116 cells undergoing TSA treatment. In conclusion, the current study revealed that TSA may induce ER stress via a p53-dependent mechanism in colon cancer cells. This provides information that may assist the development of treatments that exploit the anticancer function of TSA.


Atrial fibrillation and mitral prolapse in a subject affected by Refetoff syndrome.
Inappropriate secretion of TSH (IST) refers to a heterogeneous group of syndromes in which patients show unsuppressed TSH levels in spite of high serum free thyroid hormone concentrations. It has been recognised that IST can be due to both thyroid hormone resistance (RTH) and pituitary TSH-secreting tumours. The former can be generalised (GRTH) or pituitary (PRTH) if the resistance is more severe in the pituitary than in the remaining tissues. This case report describes a peculiar coexistence of atrial fibrillation and mitral valve prolapse in a patient affected by generalized resistance to thyroid hormone. This finding is suggestive for a major and almost physiological sensitivity of the myocardium to the thyroid hormones activity which in the course of years may determine the modifications responsible for the pathologies described.


Characterization of the suppressor cell activity in human cord blood lymphocytes.
Newborn mononuclear cells are known to have increased suppressor activity when compared with adult cells. However, the precise phenotypic description of the suppressor cell subpopulation has not yet been reported. This study was designed to examine the surface markers on human cord blood cells as defined by monoclonal antibodies and to characterize cord blood mononuclear cells that suppress in vitro pokeweed mitogen-driven immunoglobulin production. The pattern of fluorescence with the 9,6-monoclonal antibody suggested either a decreased density or a partial blocking of E-receptors on cord blood lymphocytes. Otherwise, human cord and adult cells had similar proportions of T cell subpopulations. Different subsets of newborn cells isolated by a monoclonal antibody rosetting technique were tested for suppressor activity. Cord blood lymphocytes recognized by the OKT8 monoclonal antibody constituted the major functional suppressor cell subpopulation. These cells were present in both E-rosetting and non-E-rosetting populations. Removal of OKT8+ mononuclear cord blood cells abrogated most of the suppressor activity. Thus the major suppressor cell present in cord blood is an OKT8 positive lymphocyte.


A critique of animal models in antibiotic research.
The interactions between microbes and antimicrobial agents in vitro are often markedly different from those in vivo, presumably because of pharmacological factors and host defenses. Although animal models have long been used to evaluate the efficacy of chemotherapeutic agents, there are few guidelines for the conduct of these studies. Therefore, the results of these investigations must be assessed critically in terms of the balance between the infectious challenge and the therapeutic response, and in terms of their relevance to human disease. It has been clearly demonstrated that the "freshness" of the infecting organism, the interval between infection and therapy, the order of administration of drug combinations, and the presence of foreign bodies, may have a crucial impact upon the outcome of animal trials. Less well studied have been the influence of the quantity and growth phase of the infecting inoculum, the use of single- as opposed to multiple-dose therapy, and the anatomical and pharmacological attributes of the particular animal species. Greater understanding of these factors would resolve some of the apparently contradictory results of various investigations, and would permit more rational design of these studies in the future.


Inflammatory markers are increased in youth with type 1 diabetes: the SEARCH Case-Control study.
Increased inflammation may contribute to type 1 diabetes (T1D) complications. The objective of the study was to investigate the association of inflammation with obesity, hyperglycemia and dyslipidemia in youth with T1D. This was a cross-sectional study of youth with and without T1D. The study was conducted in Colorado and South Carolina. SEARCH Case-Control participants with T1D [n = 553, mean age 15 yr (range 10-22), median duration 2.7 yr] and without diabetes [n = 215, mean age 15 yr (range 10-22)]. This was an observational study. IL-6, high-sensitivity C-reactive protein (hsCRP), fibrinogen, and leptin were measured. Inflammatory markers were evaluated by diabetes status, quartiles of glycated hemoglobin, and obesity using multiple linear regression analyses, adjusted for age, sex, study site, race/ethnicity, T1D duration, body mass index, and pubertal status. Compared with controls, youth with T1D had higher IL-6 and fibrinogen levels at all levels of glycemia and obesity, and hsCRP levels were significantly higher in youth with T1D in the top three quartiles of glycated hemoglobin (> or = 7.2%) and among normal-weight subjects. Leptin was lower in youth with poor glycemic control. Higher hsCRP and fibrinogen were correlated with higher total and LDL cholesterol, and apolipoprotein B in youth with T1D, whereas higher fibrinogen was correlated with higher LDL and apolipoprotein B in controls. T1D is characterized by excess inflammation, independent of adiposity and glycemic control. Even T1D youth in good glycemic control had higher levels of IL-6 and fibrinogen than controls. Elevated inflammatory markers were associated with an atherogenic lipid profile, which may contribute to accelerated atherosclerosis in youth with T1D.


Immunohistochemical studies of intrahepatic tumour necrosis factor alpha in chronic liver disease.
To determine the intrahepatic production of tumour necrosis factor alpha (TNF alpha) in chronic liver disease three monoclonal antibodies were used against TNF alpha in immunohistochemical studies of liver tissue sections from patients with chronic liver disease. All three monoclonal antibodies stained infiltrating mononuclear cells. Monoclonal antibody II 7C2 also stained the cytoplasm or nucleus, or both, of a varied number of hepatocytes from nine patients with chronic hepatitis B virus infection, suggesting that the antigenic epitope related to hepatitis B core antigen (HBcAg) crossreacted with II7C2. The other two monoclonal antibodies, III2F3 and IV3E5, stained significantly larger numbers of mononuclear cells in cases of chronic active hepatitis B than in chronic persistent hepatitis B, or hepatitis B related liver cirrhosis. III2F3 stained significantly larger numbers of mononuclear cells in non-A, non-B chronic active hepatitis than in chronic persistent hepatitis B or hepatitis B related liver cirrhosis. These results indicate that TNF alpha is produced and secreted by infiltrating mononuclear cells in focal inflammatory areas of the liver, and suggest that TNF alpha may have a role in the inflammatory activity of chronic liver disease.


Degradation and metabolism of tetrabromobisphenol A (TBBPA) in submerged soil and soil-plant systems.
Contamination by tetrabromobisphenol A (TBBPA), the most widely used brominated flame retardant, is a matter of environmental concern. Here, we investigated the fate and metabolites of (14)C-TBBPA in a submerged soil with an anoxic-oxic interface and planted or not with rice (Oryza sativa) and reed (Phragmites australis) seedlings. In unplanted soil, TBBPA dissipation (half-life 20.8 days) was accompanied by mineralization (11.5% of initial TBBPA) and the substantial formation (60.8%) of bound residues. Twelve metabolites (10 in unplanted soil and 7 in planted soil) were formed via four interconnected pathways: oxidative skeletal cleavage, O-methylation, type II ipso-substitution, and reductive debromination. The presence of the seedlings strongly reduced (14)C-TBBPA mineralization and bound-residue formation and stimulated debromination and O-methylation. Considerable radioactivity accumulated in rice (21.3%) and reed (33.1%) seedlings, mainly on or in the roots. While TBBPA dissipation was hardly affected by the rice seedlings, it was strongly enhanced by the reed seedlings, greatly reducing the half-life (11.4 days) and increasing monomethyl TBBPA formation (11.3%). The impact of the interconnected aerobic and anaerobic transformation of TBBPA and wetland plants on the profile and dynamics of the metabolites should be considered in phytoremediation strategies and environmental risk assessments of TBBPA in submerged soils.


Serial depletion of marine invertebrates leads to the decline of a strongly interacting grazer.
We investigated the relative roles of natural factors and shoreline harvest leading to recent declines of the black leather chiton (Katharina tunicata) on the outer Kenai Peninsula, Alaska (U.S.A.). This intertidal mollusk is a strongly interacting grazer and a culturally important subsistence fishery for Sugpiaq (Chugach Alutiiq) natives. We took multiple approaches to determine causes of decline. Field surveys examined the significant predictors of Katharina density and biomass across 11 sites varying in harvest pressure, and an integrated analysis of archaeological faunal remains, historical records, traditional ecological knowledge, and contemporary subsistence invertebrate landings examined changes in subsistence practices through time. Strong evidence suggests that current spatial variation in Katharina density and biomass is driven by both human exploitation and sea otter (Enhydra lutris) predation. Traditional knowledge, calibrated by subsistence harvest data, further revealed that several benthic marine invertebrates (sea urchin, crab, clams, and cockles) have declined serially beginning in the 1960s, with reduced densities and sizes of Katharina being the most recent. The timing of these declines was coincident with changes in human behavior (from semi-nomadic to increasingly permanent settlement patterns, improved extractive technologies, regional commercial crustacean exploitation, the erosion of culturally based season and size restrictions) and with the reestablishment of sea otters. We propose that a spatial concentration in shoreline collection pressure through time, increased harvest efficiency, and the serial depletion of alternative marine invertebrate prey have led to intensified per capita predator impacts on Katharina and thus its recent localized decline.


Significant Differences in Personality Styles of Securely and Insecurely Attached Psychotherapists: Data, Reflections and Implications.
This is a contribution to the research on the therapist variable aiming to improve effectiveness of psychotherapy. It is shown that attachment styles shape personality styles of psychotherapists in a favorable or unfavorable manner. Data on personality (PSDI) and attachment (RSQ) styles was collected from 430 psychological psychotherapists of the DACH countries using an online survey. The 88 insecurely attached psychotherapists differed significantly from their 342 securely attached colleagues in 9 of 14 personality styles: They were - even though well within normal range - more paranoid, borderline, schizoid, dependent, negativistic, self-sacrificing, avoidant, and depressive, as well as less optimistic. This corresponds to results of other researchers. Data regarding their effectiveness was not available. It is argued that a secure attachment style predispose to be a good psychotherapist. Yet, insecurely attached psychotherapists possibly compensate their adverse traits through self-therapy, continuous education, and supervision.


Ultrasound imaging in medical student education: Impact on learning anatomy and physical diagnosis.
Ultrasound use has expanded dramatically among the medical specialties for diagnostic and interventional purposes, due to its affordability, portability, and practicality. This imaging modality, which permits real-time visualization of anatomic structures and relationships in vivo, holds potential for pre-clinical instruction of students in anatomy and physical diagnosis, as well as providing a bridge to the eventual use of bedside ultrasound by clinicians to assess patients and guide invasive procedures. In many studies, but not all, improved understanding of anatomy has been demonstrated, and in others, improved accuracy in selected aspects of physical diagnosis is evident. Most students have expressed a highly favorable impression of this technology for anatomy education when surveyed. Logistic issues or obstacles to the integration of ultrasound imaging into anatomy teaching appear to be readily overcome. The enthusiasm of students and anatomists for teaching with ultrasound has led to widespread implementation of ultrasound-based teaching initiatives in medical schools the world over, including some with integration throughout the entire curriculum; a trend that likely will continue to grow. Anat Sci Educ 10: 176-189. © 2016 American Association of Anatomists.


Rectal myectomy in Hirschsprung disease: a decade of experience.
Short-segment aganglionosis is becoming more frequently recognized in view of more sophisticated diagnostic techniques, as well as the greater clinical awareness of the entity. A series of 37 patients who underwent rectal myectomy during a ten-year period was reviewed. Selected patients with short-segment aganglionosis, proved by muscle biopsy, responded well to a single transanal operation that required an overnight hospital stay. The complications were minimal, and the cure or marked improvement has persisted for as long as 11 years. When major surgery of an abdominoperineal pull-through type was required after the myectomy (four patients), no technical difficulties were encountered. Within the limits of its applicability, rectal myectomy has a definite place in the treatment of short-segment Hirschsprung disease.


Firing statistics of a neuron model driven by long-range correlated noise.
We study the statistics of the firing patterns of a perfect integrate and fire neuron model driven by additive long-range correlated Ornstein-Uhlenbeck noise. Using a quasistatic weak noise approximation we obtain expressions for the interspike interval (ISI) probability density, the power spectral density, and the spike count Fano factor. We find unimodal, long-tailed ISI densities, Lorenzian power spectra at low frequencies, and a minimum in the Fano factor as a function of counting time. The implications of these results for signal detection are discussed.


Pregnancy-associated plasma protein-A-induced inhibition of human leukocyte elastase: an artifact.
Pregnancy-associated plasma protein A (PAPP-A), was reported to be an inhibitor in many in vitro systems. Since it was shown that the inhibition of coagulation and complement activity attributed to PAPP-A was in fact due to a contamination by heparin occurring during the purification process, we undertook the present study to see whether the reported PAPP-A-induced inhibition of human leukocyte elastase (HLE) could also be attributed to heparin contamination. PAPP-A was purified from maternal pregnancy EDTA plasma by a method which was previously shown to eliminate contaminating heparin: this preparation was inactive in the HLE assay. But PAPP-A isolated by heparin-Sepharose chromatography, or a PAPP-A-free washing of the heparin-Sepharose column were both inhibitors of HLE. Furthermore the inactive PAPP-A preparation, when incubated with the PAPP-A-free washing of the heparin-Sepharose column, yielded a high molecular weight preparation which inhibited HLE. It is concluded that PAPP-A is not an inhibitor of HLE and that the inhibition of HLE previously attributed to PAPP-A was due to contaminating heparin.


Do the Side Effects of BRAF Inhibitors Mimic RASopathies?
Recent advances in targeted anticancer therapies have substantially improved the prognosis of several cancers. Such targeted therapies are not, however, free of side effects, and these side effects are clearly distinct from those induced by classical cytotoxic chemotherapies. This is likely so because targeted therapies are designed to interfere with specific oncogenic signaling pathways rather than to inhibit cell proliferation in general. In fact, interference with specific signaling pathways may lead to effects that mimic those associated with genetic disorders due to alterations in the corresponding signaling pathways. Here, we compare the clinical effects of treatment with BRAF inhibitors with those of genetic RASopathies and find a striking overlap between the inhibitor-induced, iatrogenic dermatoses with the genodermatoses seen in patients with corresponding congenital RASopathies. We hope that such comparisons lead to a better understanding of the side effects of targeted therapies.


DNTTIP2 Expression is Associated with Macrophage Infiltration and Malignant Characteristics in Low-Grade Glioma.
Low-grade gliomas (LGG) are primary brain tumors that often affect predominantly young adults, which usually have a painless course, and have a longer survival period compared to patients with high-grade gliomas. Relatively established treatment options include surgery, radiotherapy, chemotherapy or combination therapy, as well as individualized management based on tumor location, histology, molecular features and patient characteristics. Due to the rapid development of targeted therapies, the development of new molecular targets is now a very promising research direction. We explored the diagnostic value, clinical relevance, and molecular function of deoxynucleotidyl transferase terminal-interacting proteins 2 (DNTTIP2) in LGG using MethSurv, MEXPRESS, STRING, cBioPortal, Tumor Immunity Estimation Resource (TIMER) database, Gene Expression Profiling Interactive Analysis (GEPIA) databases. Besides, the "CIBERSORT" algorithm was conducted to estimate immune cells infiltration abundance, with "ggplot2" package visualizing the results. In vivo and vitro experiments were used to verify the speculations of bioinformatics analysis. In LGG patients, DNTTIP1/2 were over-expressed at mRNA levels and high DNTTIP1/2 levels correlated with poor survival in LGG patients. We confirmed that DNTTIP2 significantly promotes M2 macrophage activation and angiogenesis, which may be related to the IL6/JAK/STAT3 signaling pathway. In addition, we found that DNTTIP2 amplification was associated with an unfavorable prognosis in LGG patients. We demonstrated, finally, a correlation between DNTTIP2 gene hypermethylation and a poor prognosis in LGG. This study demonstrated that DNTTIP1/2 had diagnostic and prognostic value in LGG patients. The biological mechanisms of DNTTIP2 regarding angiogenesis and macrophage activation may provide new insights into the treatment of glioma.


Sunlight-Driven Photocatalytic Active Fabrics through Immobilization of Functionalized Doped Titania Nanoparticles.
Frequent washing of textiles poses a serious hazard to the ecosystem, owing to the discharge of harmful effluents and the release of microfibers. On one side, the harmful effluents from detergents are endangering marine biota, while on the other end, microplastics are observed even in breastfeeding milk. This work proposes the development of sunlight-driven cleaning and antibacterial comfort fabrics by immobilizing functionalized Zn-doped TiO2 nanoparticles. The research was implemented to limit the use of various detergents and chemicals for stain removal. A facile sol-gel method has opted for the fabrication of pristine and Zn-doped TiO2 nanoparticles at three different mole percentages of Zn. The nanoparticles were successfully functionalized and immobilized on cotton fabric using silane coupling agents via pad-dry-cure treatment. As-obtained fabrics were characterized by their surface morphologies, availability of chemical functionalities, and crystallinity. The sunlight-assisted degradation potential of as-functionalized fabrics was evaluated against selected pollutants (eight commercial dyes). The 95-98% degradation of dyes from the functionalized fabric surface was achieved within 3 h of sunlight exposure, estimated by color strength analysis with an equivalent exposition of bactericidal activities. The treated fabrics also preserved their comfort and mechanical properties. The radical trapping experiment was performed to confirm the key radicals responsible for dye degradation, and h+ ions were found to be the most influencing species. The reaction pathway followed the first order kinetic model with rate constant values of 0.0087 min-1 and 0.0131 min-1 for MB and MO dyes, respectively.


Cross calibration between XRF and ICP-MS for high spatial resolution analysis of ombrotrophic peat cores for palaeoclimatic studies.
Ombrotrophic peatlands are remarkable repositories of high-quality climatic signals because their only source of nutrients is precipitation. Although several analytical techniques are available for analysing inorganic components in peat samples, they generally provide only low-resolution data sets. Here we present a new analytical approach for producing high-resolution data on main and trace elements from ombrotrophic peat cores. Analyses were carried out on a 7-m-long peat core collected from Danta di Cadore, North-Eastern Italy (46° 34' 16″ N, 12° 29' 58″ E). Ca, Ti, Cr, Fe, Cu, Zn, Ga, Sr, Y, Cd, Ba and Pb were detected at a resolution of 2.5 mm with a non-destructive X-ray fluorescence core scanner (XRF-CS). Calibration and quantification of the XRF-CS intensities was obtained using collision reaction cell inductively coupled plasma quadruple mass spectrometry (CRC-ICP-QMS). CRC-ICP-QMS measurements were carried out on discrete samples at a resolution of 1 cm, after dissolution of 150-mg aliquots with 9 ml HNO3 and 1 ml HF at 220 °C in a microwave system. We compare qualitative XRF-CS and quantitative CRC-ICP-MS data and, however the several sources of variability of the data, develop a robust statistical approach to determine the R (2) and the coefficient of a simple regression model together with confidence intervals. Perfect positive correlations were estimated for Cd, Cr, Pb, Sr, Ti and Zn; high positive correlations for Ba (0.8954), Y (0.7378), Fe (0.7349) and Cu (0.7028); while moderate positive correlations for Ga (0.5951) and Ca (0.5435). With our results, we demonstrate that XRF scanning techniques can be used, together with other well-established geochemical techniques (such as ICP-MS), to produce high-resolution (up to 2.5 mm) quantitative data from ombrotrophic peat bog cores.


Using computer assisted image analysis to determine the optimal Ki67 threshold for predicting outcome of invasive breast cancer.
Ki67 positivity in invasive breast cancers has an inverse correlation with survival outcomes and serves as an immunohistochemical surrogate for molecular subtyping of breast cancer, particularly ER positive breast cancer. The optimal threshold of Ki67 in both settings, however, remains elusive. We use computer assisted image analysis (CAIA) to determine the optimal threshold for Ki67 in predicting survival outcomes and differentiating luminal B from luminal A breast cancers. Quantitative scoring of Ki67 on tissue microarray (TMA) sections of 440 invasive breast cancers was performed using Aperio ePathology ImmunoHistochemistry Nuclear Image Analysis algorithm, with TMA slides digitally scanned via Aperio ScanScope XT System. On multivariate analysis, tumours with Ki67 ≥14% had an increased likelihood of recurrence (HR 1.941, p=0.021) and shorter overall survival (HR 2.201, p=0.016). Similar findings were observed in the subset of 343 ER positive breast cancers (HR 2.409, p=0.012 and HR 2.787, p=0.012 respectively). The value of Ki67 associated with ER+HER2-PR<20% tumours (Luminal B subtype) was found to be <17%. Using CAIA, we found optimal thresholds for Ki67 that predict a poorer prognosis and an association with the Luminal B subtype of breast cancer. Further investigation and validation of these thresholds are recommended.


Advances in gene therapy technologies to treat retinitis pigmentosa.
Retinitis pigmentosa (RP) is a class of diseases that leads to progressive degeneration of the retina. Experimental approaches to gene therapy for the treatment of inherited retinal dystrophies have advanced in recent years, inclusive of the safe delivery of genes to the human retina. This review is focused on the development of gene therapy for RP using recombinant adenoassociated viral vectors, which show a positive safety record and have so far been successful in several clinical trials for congenital retinal disease. Gene therapy for RP is under development in a variety of animal models, and the results raise expectations of future clinical application. Nonetheless, the translation of such strategies to the bedside requires further understanding of the mutations and mechanisms that cause visual defects, as well as thorough examination of potential adverse effects.


Investigation of basic imaging properties in digital radiography. I. Modulation transfer function.
The effect of various digital parameters, such as the sampling aperture, sampling distance, and display aperture, on the modulation transfer function (MTF) of digital radiographic imaging systems was investigated by means of theoretical simulation studies. The MTFs were also determined experimentally to confirm the relationship used in the simulation studies. The results indicate that the overall MTF of a digital system cannot specify the resolution properties in the same way as can the MTFs of analog systems. The MTF of a digital system may include a "false" response due to aliasing, which could lead to an incorrect interpretation of the resolution properties. The magnitude of aliasing that will occur in a digitized signal depends on the sampling parameters chosen and on the frequency content of the radiologic object being imaged. Thus, the type of object to be detected as well as various digital parameters must be considered in the design and evaluation of digital imaging systems.


Adaptations to research within the intellectual disability population during the Covid-19 pandemic: Lessons learned from the CAREVIS study.
The COVID-19 pandemic resulted in changes in all areas of clinical practice, including clinical research and within the intellectual disability population. While there have been some benefits from this rapid adoption of change, those involved in research have had to overcome a number of additional challenges. These adaptive changes, which have included the use of technology, closure of social spaces, working with specific groups who are more vulnerable to COVID-19, and mask use impairing communication, have had both positive and negative impacts on research. As the pandemic and related restrictions evolve, it is important to examine the changes that have occurred. In the future, the adoption of a hybrid model in research is likely to be a common approach, establishing a balance between technology and in-person interaction.


Do young and older adults rely on different processes in source memory tasks? A neuropsychological study.
Source memory has consistently been associated with prefrontal function in both normal and clinical populations. Nevertheless, the exact contribution of this brain region to source memory remains uncertain, and evidence suggests that processes used by young and older adults may differ. The authors explored the extent to which scores on composite measures of neuropsychological tests of frontal and medial temporal function differentially predicted the performance of young and older adults on source memory tasks. Results indicated that a frontal composite measure, consistently associated with source memory performance in older adults, was unrelated to source memory in young adults, although it was sensitive to a demanding working memory task. The memory composite score, however, predicted performance in the young group. In addition, item and source memory were correlated in young but not older people. Findings are discussed in terms of age-related differences in working memory and executive functions, and differential binding processes necessary for item and source memory. The requirement to integrate item and source information at encoding appears to place greater demands on executive or working memory processes in older adults than in younger adults.


Utility of interphase FISH to stratify patients into cytogenetic risk categories at diagnosis of AML in an Eastern Cooperative Oncology Group (ECOG) clinical trial (E1900).
We evaluated the efficacy of FISH to detect chromosome anomalies in the evaluation of young (<60 years) patients with AML. Patients were enrolled in E1900, an ECOG clinical trial for AML. The protocol was designed to collect bone marrow or blood for both cytogenetic and FISH studies at study entry (diagnosis). FISH for each patient was performed and utilized eight probe sets to detect t(8;21), t(9;22), t(11;var), t(15;17), inv(16), +8, -5/5q, and -7/7q. We analyzed 237 specimens with complete cytogenetic and FISH results. Results for each specimen were classified by probe set into one of six categories. The concordance rate between cytogenetic and FISH results ranged from 98 to 100% for all probe sets and kappa analysis for concordance had a p-value of <0.0001. The high level of agreement between cytogenetic and FISH results demonstrate the accuracy of a panel of eight FISH probe sets for the detection of significant abnormalities in AML. Data from this investigation support the use of FISH as an adjunct method to increase the yield of useful cytogenetic results in large cooperative trials and demonstrate the potential of FISH as a follow-up study of minimal residual disease in ECOG trials.


Microbial amelioration of salinity stress in HD 2967 wheat cultivar by up-regulating antioxidant defense.
An experiment was conducted to investigate the potential of Piriformospora indica and plant growth-promoting bacteria (PGPB) to ameliorate salinity stress in HD 2967 wheat cultivar. Plants were treated with four different levels of salinity viz. 0, 50, 100 and 200 mM NaCl (electrical conductivity value 0.01, 5.84, 11.50 and 21.4 mS cm-1, respectively) under greenhouse conditions, using a completely randomized design experiment. Plants inoculated with PGPB and P. indica showed decrease in lipid peroxidation, relative membrane permeability and lipoxygenase enzyme (LOX) activity as compared to uninoculated plants. The result of this study showed that PGPB and P. indica inoculated HD 2967 wheat plants accumulated higher content of proline, α-tocopherol and carotenoid as compared to uninoculated plants. The HD 2967 wheat plants either inoculated with PGPB or P. indica showed significantly higher activities of antioxidant enzymes viz. superoxide dismutase, catalase and ascorbate peroxidase than that of the uninoculated plants. Moreover, PGPB inoculated plants showed greater activity of antioxidant enzymes than the plants inoculated with P. indica. Salinity stress tolerance was more pronounced in the PGPB inoculated than P. indica inoculated plants. This study revealed the potentiality of PGPB and P. indica as bio-ameliorator under salinity stress, and suggests that this plant microbial association could be a promising biotechnological tool to combat the deleterious effects of salinity stress.


Lipopolysaccharide-coated erythrocytes activate human neutrophils via CD14 while subsequent binding is through CD11b/CD18.
Interaction of LPS with monocytes and neutrophils is known to occur via CD14 and is strongly enhanced by LPS-binding protein (LBP). Integrins as well as CD14 play a role in the interaction of erythrocytes (E) coated with LPS or whole Gram-negative bacteria with phagocytes. We reasoned that the density of LPS on a particle is an important determinant in these interactions. Therefore, E were coated with different concentrations of LPS (ELPS). The binding of these ELPS to neutrophils was evaluated by flow cytometry. Simultaneously, we measured fMLP receptor expression to evaluate neutrophil activation. ELPS only bound to neutrophils in the presence of LBP. Blocking CD14 inhibited both activation and binding, whereas blocking complement (C) receptor 3 (CR3) inhibited binding but not activation. TNF activation restored ELPS binding in CD14-blocked cells but not in cells in which CR3 was blocked. Salmonella minnesota did bind to neutrophils independent of CR3 or CD14. The addition of LBP enhanced binding twofold, and this surplus was dependent upon CD14 but not on CR3. We conclude that ELPS interact with neutrophils via CD14, initially giving rise to cell activation; subsequently, binding is solely mediated by activated CR3.


Newer vascular targets: endosialin (review).
The identification of cell surface markers expressed selectively by tumor vasculature is challenging. To get as close to the human disease as possible, investigators have isolated endothelial cells from fresh human tumor specimens and subjected them to RNA-based gene expression analysis. The data indicate that there are few proteins that distinguish tumor vasculature from normal vasculature and re-enforce the notion that the endothelium is a tissue specialized cell-type. Endosialin was identified as a cell surface tumor endothelial marker. The selective expression of endosialin by tumor vasculature and stroma has been confirmed. Although the function of endosialin remains to be elucidated, the expression pattern for this protein may be favorable for cancer therapy.


Doxycycline and therapeutic targeting of the DNA damage response in cancer cells: old drug, new purpose.
There is a small proportion of cells within a tumour with self-renewing properties, which is resistant to conventional therapy, and is responsible for tumour initiation, maintenance and metastasis. These cells are known as cancer stem cells (CSCs) or tumour-initiating cells (TICs) [1]. Recent publications identify several antibiotics, such as salinomycin or doxycycline, as selective CSCs inhibitors [2-4]. However, the mechanisms of action of these antibiotics on CSCs are not fully understood.


[Evaluation of serum tissue polypeptide antigen (TPA) in patients with urogenital cancer].
The level of serum TPA was determined by radio-immunoassay in 19 healthy subjects and 90 patients with urogenital cancer. The normal level of serum TPA was 86 +/- 24 U/l, and the level of more than 134 U/l was determined positive. The positive rate of TPA was 38.9% in 90 patients, while that of CEA was 25.6%. In 19 patients with bladder tumor and 7 with testicular tumor, the positive rates of TPA were 52.6% and 71.4%, respectively, and the level of serum TPA was high in these positive patients. Considering the low positive rate of CEA, TPA may be a more useful marker than CEA in patients with bladder tumor and testicular tumor. Serial determinations of serum TPA and CEA showed the considerable variation of serum TPA compared with serum CEA and a temporary elevation of serum TPA following radical nephrectomy and retroperitoneal lymphadenectomy. However, the level of serum TPA fell significantly after the successful treatment in 8 patients (2 with renal cell cancer, 3 with bladder tumor, 1 with prostate cancer, 2 with testicular tumor) and rose sharply with recurrent or metastatic disease in 4 patients (2 with bladder tumor, 2 with testicular tumor). Although there was no correlation between the levels of serum TPA and serum PAP, the level of serum TPA tended to change in parallel with the level of serum AFP or HCG in 3 patients with testicular tumor.


Utilizing logical relationships in genomic data to decipher cellular processes.
The wealth of available genomic data has spawned a corresponding interest in computational methods that can impart biological meaning and context to these experiments. Traditional computational methods have drawn relationships between pairs of proteins or genes based on notions of equality or similarity between their patterns of occurrence or behavior. For example, two genes displaying similar variation in expression, over a number of experiments, may be predicted to be functionally related. We have introduced a natural extension of these approaches, instead identifying logical relationships involving triplets of proteins. Triplets provide for various discrete kinds of logic relationships, leading to detailed inferences about biological associations. For instance, a protein C might be encoded within an organism if, and only if, two other proteins A and B are also both encoded within the organism, thus suggesting that gene C is functionally related to genes A and B. The method has been applied fruitfully to both phylogenetic and microarray expression data, and has been used to associate logical combinations of protein activity with disease state phenotypes, revealing previously unknown ternary relationships among proteins, and illustrating the inherent complexities that arise in biological data.


Slater ice rules and H-bond dynamics in KDP-type systems.
We report on the first observation of the dynamics of correlated hydrogen switching among the six Slater D2PO4 configurations, induced by unpaired D3PO4 and DPO4 Takagi group diffusion in a deuteron glass Rb0.5(ND4)0.5D2PO4. The results obtained by two-dimensional (2D) 31P exchange NMR prove the validity of the Slater-Takagi ice rules and allow for the direct determination of the correlation time for the unpaired Takagi group visits to a given D2PO4 group.


F199E substitution reduced toxicity of Clostridium perfringens epsilon toxin by depriving the receptor binding capability.
Epsilon toxin (ETX), a potent toxin, is produced by types B and D strains of Clostridium perfringens, which could cause severe diseases in humans and domestic animals. Mutant rETXF199E was previously demonstrated to be a good vaccine candidate. However, the mechanism concerned remains unknown. To clarify how F199E substitution reduced ETX toxicity, we performed a series of experiments. The results showed that the cell-binding and pore-forming ability of rETXF199E was almost abolished. We speculated that F199E substitution reduced toxicity by depriving the receptor binding capability of ETX, which contributed to the hypothesis that domain I of ETX is responsible for cell binding. In addition, our data suggested that ETX could cause Ca2+ release from intracellular Ca2+ stores, which may underlie an alternate pathway leading to cell death. Furthermore, ETX induced crenation of the MDCK cells was observed, with sags and crests first appearing on the surface of condensed MDCK cells, according to scanning electron microscopy. The data also demonstrated the safety and potentiality of rETXF199E as a vaccine candidate for humans. In summary, findings of this work potentially contribute to a better understanding of the pathogenic mechanism of ETX and the development of vaccine against diseases caused by ETX, using mutant proteins.


Non-bacterial chronic recurrent osteomyelitis of the clavicle.
This report details the case of a 12-year-old girl with a painful, progressive swelling of the medial portion of the clavicle with no history of trauma or other constitutional symptoms. All laboratory investigations were normal except for an elevated erythrocyte sedimentation rate (ESR). Initial plain radiographs showed a destructive lesion with magnetic resonance imaging showing features of malignancy. Biopsies revealed osteomyelitis, but with negative bacterial cultures and no evidence of malignancy. Treatment with antibiotics did not result in a favourable response. Over time, the swelling increased in size with episodic exacerbations of pain. Follow-up radiographs showed sclerosis and hyperostosis. After five years, this was recognized as non-bacterial chronic recurrent osteomyelitis of the clavicle. Nonbacterial, chronic recurrent, osteomyelitis, clavicle.


Efficient and safe small-bowel adhesiolysis.
Small-intestine adhesiolysis can be very time consuming and may be associated with bowel wall damage. The risk for injuries to the small or large bowel resulting in increased morbidity and costs is considerable. Both efficient and gentle dissection of adhesions is important in order to avoid intraoperative perforation or, worse, postoperative intestinal leaks. We present a technique using drops of body-warm isotonic saline solution to create an edematous swelling of the adhesions. This procedure not only protects the bowel from cooling and drying, but also simplifies the dissection and, thus, lowers the risk of intestinal lesions.


Molecular analysis improves sensitivity of breast sentinel lymph node biopsy: results of a multi-institutional prospective cohort study.
Pathologic evaluation may lack the sensitivity required for accurate staging of the axilla in breast cancer patients. We have completed enrollment of a multi-institutional prospective cohort study designed to determine if molecular analyses can improve axillary staging. In subset analyses, we have attempted to address the following questions: (1) Does molecular analysis improve the sensitivity of sentinel lymph node biopsy (SLNB) and (2) is the sentinel lymph node (SLN) hypothesis valid at the molecular level? Four hundred eighty-nine subjects with T1, T2, or T3 breast cancer and no evidence of axillary lymph node (ALN) involvement were enrolled. ALNs were analyzed by routine pathology (hematoxylin-eosin staining), and by multimarker real-time reverse transcriptase-polymerase chain reaction analysis CRT-PCR to detect breast cancer metastases. Pathology and molecular data for both SLNs and nonsentinel ALNs were available for a subset of 207 subjects. The sensitivity of pathologic analysis of the SLN to predict the pathologic status of ALNs was 84.1%. The sensitivity of combining pathologic with molecular analysis of the SLN to predict the pathologic status of ALNs was 92.8%, a statistically significant increase in sensitivity (P = .031 by the McNemar test for correlated proportions). Finally, the sensitivity of combining pathologic with molecular analysis of the SLN to predict the pathologic or molecular status of ALNs was 85.4%. The combination of pathologic and molecular analysis of SLNs resulted in the highest sensitivity for prediction of the pathologic status of ALNs. The data also provide evidence that the SLN hypothesis remains valid at the molecular level.


Calcium determines the supramolecular organization of fibrillin-rich microfibrils.
Microfibrils are ubiquitous fibrillin-rich polymers that are thought to provide long-range elasticity to extracellular matrices, including the zonular filaments of mammalian eyes. X-ray diffraction of hydrated bovine zonular filaments demonstrated meridional diffraction peaks indexing on a fundamental axial periodicity (D) of approximately 56 nm. A Ca2+-induced reversible change in the intensities of the meridional Bragg peaks indicated that supramolecular rearrangements occurred in response to altered concentrations of free Ca2+. In the presence of Ca2+, the dominant diffracting subspecies were microfibrils aligned in an axial 0.33-D stagger. The removal of Ca2+ caused an enhanced regularity in molecular spacing of individual microfibrils, and the contribution from microfibrils not involved in staggered arrays became more dominant. Scanning transmission electron microscopy of isolated microfibrils revealed that Ca2+ removal or addition caused significant, reversible changes in microfibril mass distribution and periodicity. These results were consistent with evidence from x-ray diffraction. Simulated meridional x-ray diffraction profiles and analyses of isolated Ca2+-containing, staggered microfibrillar arrays were used to interpret the effects of Ca2+. These observations highlight the importance of Ca2+ to microfibrils and microfibrillar arrays in vivo.


Pan-cancer analysis of the oncogenic role of the core osteosarcoma gene VCAN in human tumors.
Versican (VCAN), a member of the multifunctional glycoprotein family, is involved in various aspects of cancer progression. However, the role of VCAN in diverse cancers remains poorly defined. This research aimed to investigate the correlation between VCAN expression and the oncogenic role, as well as visualize its prognostic landscape in pan-cancer. Raw data in regard to VCAN expression in cancer patients were acquired from GEO GeneChip public database in NCBI. Besides, we selected microarray data GSE16088 for analysis. We retrieved the genes associated with osteosarcoma (OS) from the OMIM database and identified their intersection with the core module. VCAN was suggested to be a potential marker gene for OS. Subsequently, we conducted Gene Set Enrichment Analysis (GSEA) to explore gene functional enrichment. Moreover, we performed pan-cancer analysis on VCAN to gain a comprehensive understanding of its implications across various cancer types. The VCAN expression in the tumor tissue was higher than that in normal tissue. Elevated expression of VCAN was associated with high the tumor stage and poor long-term survival. There was a significant positive correlation between VCAN and cancer fibroblasts in all pan cancers. Moreover, FBN1 was the intersection gene of VCAN-related genes and linker genes. ANTXR1, COL5A2, CSGALNACT2, and SPARC were the target genes of VCAN genes. GSEA analysis showed that VCAN was mainly enriched in the extracellular matrix (ECM) signaling pathway. VCAN can be used as a marker molecule for the early diagnosis of OS and holds significance as a molecule in cases of OS with distant metastasis. The ECM signaling pathway may be a core pathway in OS development and distant metastasis. These findings shed new light on therapeutics of cancers.


Genetic regulation of entry into meiosis in Caenorhabditis elegans.
The Caenorhabditis elegans germline is composed of mitotically dividing cells at the distal end that give rise to meiotic cells more proximally. Specification of the distal region as mitotic relies on induction by the somatic distal tip cell and the glp-1 signal transduction pathway. However, the genetic control over the transition from mitosis to meiosis is not understood. In this paper, we report the identification of a gene, gld-2, that has at least two functions in germline development. First, gld-2 is required for normal progression through meiotic prophase. Second, gld-2 promotes entry into meiosis from the mitotic cell cycle. With respect to this second function, gld-2 appears to be functionally redundant with a previously described gene, gld-1 (Francis, R., Barton, M. K., Kimble, J. and Schedl, T. (1995) Genetics 139, 579-606). Germ cells in gld-1(o) and gld-2 single mutants enter meiosis at the normal time, but germ cells in gld-2 gld-1(o) double mutants do not enter meiosis. Instead, the double mutant germline is mitotic throughout and forms a large tumor. We suggest that gld-1 and gld-2 define two independent regulatory pathways, each of which can be sufficient for entry into meiosis. Epistasis analyses show that gld-1 and gld-2 work downstream of the glp-1 signal transduction pathway. Therefore, we hypothesize that glp-1 promotes proliferation by inhibiting the meiosis-promoting functions of gld-1 and gld-2.


[What is appropriate in postmenopausal osteoporosis?].
The term of postmenopausal osteoporosis created 1940/1941 by Albright and his coworkers corresponds on our days no more to the original conception, because it is used now in a broader form. The resulting practical consequences, e.g. the postulates of a general medical prophylaxis, induce a new discussion which is adapted to the scientific state of to-day and the real importance.


Prevalence and associated survival of high-risk HPV-related adenoid cystic carcinoma of the salivary glands.
Adenoid cystic carcinoma (SACC) is a rare malignancy, but a frequent subtype in minor and major salivary glands. The molecular alterations or biomarkers that underlie its development and progression as well as therapy outcomes are poorly characterized. The main study goal was to investigate reliable biomarkers and patient-related factors that may have impact on recurrence and long-term survival of SACC. The prevalence of human papilloma virus (HPV) in SACC was determined by HPV-DNA genotyping and p16 immunostaining. Epithelial growth factor receptor (EGFR), p53 and Ki-67 expression were also evaluated. Twenty-eight (42%) of 67 patients were HPV-DNA positive. Kaplan-Meier analysis indicated that SACC patients with metastases (P=0.03) had a poor overall survival (OS) and a shorter recurrence-free survival (P<0.001). Positive resection margins significantly predicted shorter recurrence-free survival (P=0.01). In the multivariate analysis, non-metastatic disease (P=0.033) and p16 positivity (P=0.005) have shown their prediction value for OS while non-metastatic disease (P=0.002), HPV positivity (P=0.041) and negative resection margin predicted a better recurrence-free survival. The present study documents for the first time the positivity for HPV infection and overexpression of certain markers (p16, Ki-67, EGFR and p53) used in diagnostics in SACC as well as characterizes clinical entities. These factors might be exploited in the future as biomarkers for its prognostic value. Using the clinical and pathological basis for predicting different outcomes could significantly facilitate SACC stratification and potentially directing treatment.


Efficacy and safety of multitarget therapy with cyclophosphamide and tacrolimus for lupus nephritis: a prospective, single-arm, single-centre, open label pilot study in Japan.
Background Pulsed cyclophosphamide or mycophenolate mofetil for lupus nephritis has limited efficacy. We previously reported a case of mixed-class IV + V lupus nephritis successfully treated with cyclophosphamide and tacrolimus. This study assessed the efficacy and safety of multitarget therapy with cyclophosphamide and tacrolimus for the treatment of lupus nephritis. Methods In a prospective, single-arm, open label pilot study, we recruited 15 patients aged 18-64 years with active lupus nephritis who met the American College of Rheumatology criteria for a diagnosis of systemic lupus erythematosus (1997). The treatment protocol was a starting dose of prednisolone of 0.6-1.0 mg/kg/day for 2 weeks and then tapered to a maintenance dose, intravenous cyclophosphamide (500 mg biweekly for 3 months) and tacrolimus (3.0 mg/day). Tacrolimus was continued as maintenance therapy. Complete remission was defined as a spot urine protein/creatinine ratio of < 0.5 g/gCr with no active urine casts and a serum creatinine level that was either normal or within 30% of a previously abnormal baseline level. We retrospectively compared results for the study patients with those of 18 historical controls conventionally treated with cyclophosphamide and prednisolone. Results At baseline, the mean patient age was 41.5 ± 14.6 years (male:female ratio 2:13), urine protein/creatinine ratio 3.9 ± 2.3 g/gCr and serum creatinine 84.6 ± 34.6 µmol/L. Lupus nephritis classifications included classes IV ( n = 8), III + V ( n = 1), IV + V ( n = 5) and unclassified ( n = 1). Eleven patients completed the treatment protocol and four withdrew. At 6 months, 12 of 15 (80.0%) had achieved complete remission using intention-to-treat analysis, significantly more than historical controls (seven of 18 patients, 38.9%). A transient increase in serum creatinine and gastric symptoms occurred in three cases. One patient withdrew due to cytomegalovirus antigenemia and severe diabetes, and one patient died of thrombotic microangiopathy. Conclusions Multitarget therapy with cyclophosphamide and tacrolimus can be a therapeutic option for lupus nephritis. Clinical trials registration Combination therapy of tacrolimus and intravenous cyclophosphamide for remission induction of lupus nephritis, UMIN: 000004893, URL: https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr.cgi?function=brows&action=brows&type=summary&recptno=R000005830&language=E . Date of registration: 18 January 2011.


Sputum processing: a new method to improve cytospin quality.
Sputum examination is being increasingly used as a non-invasive method for studying airway inflammation. However, the application of sputum still presents some methodological problems and the results of sputum analysis may be substantially flawed by salivary contamination, cell and mucus debris. In addition, much work is needed to deepen the possibility of extensive application of cell and molecular biology techniques to sputum analysis. In an attempt to improve the technique of sputum processing, we investigated the effect of: (i) 20 and 11 microm filtration in addition to 40 microm on salivary contamination; (ii) Percoll density gradient centrifugation on sputum slides quality; (iii) a culture medium (Minimum Essential Medium containing HEPES 22 mm, pH 7.4: MEM) as washing and suspension solution compared to PBS on cell viability. Induced sputum samples were obtained in 37 asthmatics. 21 samples were processed as selected sputum and 16 samples as entire expectorates. After dithiotreitol (DTT) homogenization, each specimen was aliquoted in two parts of equal volume. One portion was processed with the usual method, the other using a modified method: cell pellet was suspended in sterile MEM, filtered through 40, 20 and 11 microm net filters and separated from the residual debris by Percoll gradient centrifugation. As compared to the current sputum processing this method resulted in: (i) no selective bronchial cellular loss; (ii) a significant decrease of salivary contamination, particularly in entire expectorates in which squamous cells were reduced from 47 (36) to 15.5% (20) as median values and interquartile range; (iii) a higher proportion of good quality cytospins; (iv) maintenance of cell viability over the time (88% vs. 81% in MEM and PBS, respectively) 1 h after sample collection. In the present study we demonstrated that the proposed method is feasible and makes it possible to overcome most of the technical limits met with the commonly used method, pointing to a potential extension of induced sputum application for more sophisticated techniques.


Transcription factor HNF1β regulates expression of the calcium-sensing receptor in the thick ascending limb of the kidney.
Mutations in hepatocyte nuclear factor 1β (HNF1β) cause autosomal dominant tubulointerstitial kidney disease (ADTKD-HNF1β), and patients tend to develop renal cysts, maturity-onset diabetes of the young (MODY), and suffer from electrolyte disturbances, including hypomagnesemia, hypokalemia, and hypocalciuria. Previous HNF1β research focused on the renal distal convoluted tubule (DCT) to elucidate the ADTKD-HNF1β electrolyte phenotype, although 70% of Mg2+ is reabsorbed in the thick ascending limb of Henle's loop (TAL). An important regulator of Mg2+ reabsorption in the TAL is the calcium-sensing receptor (CaSR). This study used several methods to elucidate the role of HNF1β in electrolyte reabsorption in the TAL. HNF1β ChIP-seq data revealed a conserved HNF1β binding site in the second intron of the CaSR gene. Luciferase-promoter assays displayed a 5.8-fold increase in CaSR expression when HNF1β was present. Expression of the HNF1β p.Lys156Glu mutant, which prevents DNA binding, abolished CaSR expression. Hnf1β knockdown in an immortalized mouse kidney TAL cell line (MKTAL) reduced expression of the CaSR and Cldn14 (claudin 14) by 56% and 48%, respectively, while Cldn10b expression was upregulated 5.0-fold. These results were confirmed in a kidney-specific HNF1β knockout mouse, which exhibited downregulation of the Casr by 81%. Cldn19 and Cldn10b expression levels were also decreased by 37% and 83%, respectively, whereas Cldn3 was upregulated by 4.6-fold. In conclusion, HNF1β is a transcriptional activator of the CaSR. Consequently, patients with HNF1β mutations may have reduced CaSR activity in the kidney, which could explain cyst progression and hyperabsorption of Ca2+ and Mg2+ in the TAL resulting in hypocalciuria.


Nucleotide sequence of rice dwarf phytoreovirus genome segment 2: completion of sequence analyses of rice dwarf virus.
The complete nucleotide sequence of rice dwarf phytoreovirus genome segment 2 (S2) was determined to be 3,512 nucleotides long with one open reading frame initiating at nucleotide 15 and terminating at nucleotide 3363. The encoded polypeptide was predicted to have 1,116 residues with a relative molecular weight of 123 kD. Comparison of S2 of two isolates showed they had identical lengths and 97 and 98.3% nucleotide and amino acid sequence identities, respectively. A search of the Swiss-Prot data base (R 22.0) failed to find any proteins with significant homology to the S2-encoded protein. Determination of the nucleotide sequence of the S2 has completed the sequence determination of the genome of rice dwarf virus. Homology searches made for proteins encoded by each of the genomic segments showed that the polypeptide encoded by S11 has similarity to histone H1 protein and VP6 of blue tongue virus, indicating it might possess nucleic acid binding properties.


Combinatorial Synthesis of Oxysulfides in the Lanthanum-Bismuth-Copper System.
Establishing synthesis methods for a target material constitutes a grand challenge in materials research, which is compounded with use-inspired specifications on the format of the material. Solar photochemistry using thin film materials is a promising technology for which many complex materials are being proposed, and the present work describes application of combinatorial methods to explore the synthesis of predicted La-Bi-Cu oxysulfide photocathodes, in particular alloys of LaCuOS and BiCuOS. The variation in concentration of three cations and two anions in thin film materials, and crystallization thereof, is achieved by a combination of reactive sputtering and thermal processes including reactive annealing and rapid thermal processing. Composition and structural characterization establish composition-processing-structure relationships that highlight the breadth of processing conditions required for synthesis of LaCuOS and BiCuOS. The relative irreducibility of La oxides and limited diffusion indicate the need for high temperature processing, which conflicts with the temperature limits for mitigating evaporation of Bi and S. Collectively the results indicate that alloys of these phases will require reactive annealing protocols that are uniquely tailored to each composition, motivating advancement of dynamic processing capabilities to further automate discovery of synthesis routes.


Quantitative Analysis of Milk-Derived microRNAs and Microbiota during the Manufacturing and Ripening of Soft Cheese.
MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per 200 mg/200 μl of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a timedependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR- 223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.


Investigations on the humidity-induced transformations of salbutamol sulphate particles coated with L-leucine.
The crystallization and structural integrity of micron-sized inhalable salbutamol sulphate particles coated with L-leucine by different methods are investigated at different humidities. The influence of the L-leucine coating on the crystallization of salbutamol sulphate beneath the coating layer is explored. The coated particles are prepared by an aerosol flow reactor method, the formation of the L-leucine coating being controlled by the saturation conditions of the L-leucine. The coating is formed by solute diffusion within a droplet and/or by vapour deposition of L-leucine. The powders are humidified at 0%, 44%, 65% and 75% of relative humidity and the changes in physical properties of the powders are investigated with dynamic vapour sorption analysis (DVS), a differential scanning calorimeter (DSC), and a scanning electron microscope (SEM). Visual observation show that all the coated particles preserve their structural integrity whereas uncoated salbutamol sulphate particles are unstable at 65% of relative humidity. The coating layer formed by diffusion performs best in terms of its physical stability against moisture and moisture-induced crystallization. The degree of crystallization of salbutamol in the as-prepared powders is within the range 24-35%. The maximum degree of crystallization after drying ranges from 55 to 73% when the salbutamol crystallizes with the aid of moisture. In addition to providing protection against moisture, the L-leucine coating also stabilizes the particle structure against heat at temperatures up to 250 degrees C. In order to preserve good flowability together with good physical stability, the best coating would contain two L-leucine layers, the inner layer being formed by diffusion (physical stability) and the outer layer by vapour deposition (flowability).


Proteomics Improves the New Understanding of Honeybee Biology.
The honeybee is one of the most valuable insect pollinators, playing a key role in pollinating wild vegetation and agricultural crops, with significant contribution to the world's food production. Although honeybees have long been studied as model for social evolution, honeybee biology at the molecular level remained poorly understood until the year 2006. With the availability of the honeybee genome sequence and technological advancements in protein separation, mass spectrometry, and bioinformatics, aspects of honeybee biology such as developmental biology, physiology, behavior, neurobiology, and immunology have been explored to new depths at molecular and biochemical levels. This Review comprehensively summarizes the recent progress in honeybee biology using proteomics to study developmental physiology, task transition, and physiological changes in some of the organs, tissues, and cells based on achievements from the authors' laboratory in this field. The research advances of honeybee proteomics provide new insights for understanding of honeybee biology and future research directions.