Anatomic and magnetic resonance imaging bases for the naso-maxillo-cheek flap technique.
A transfacial approach to the deep cranio-maxillo-facial areas by the naso-maxillo-cheek flap technique (NMCF) is indicated for the treatment of some bulky tumors of the naso-pharynx. The procedure requires precise preoperative imaging. This study presents the morphologic bases of this surgical access and the reasonable limits of the excision preoperatively determined by magnetic resonance imaging (MRI). 18 facial and skull specimens were submitted to surgical facial dismantling by the NMCF technique according to Curioni's method. The clinical application in a 66-year-old patient suffering from a neuroblastoma of the olfactory nerve extended into the naso-pharynx is presented. Pre- and postoperative MRI correlations were made in transverse, sagittal and frontal acquisitions. Several structures were preserved in the procedure: facial reliefs, inferior orbital rim and orbital floor, posterior wall of the maxillary sinus covering the pterygopalatine fossa, lateral and medial pterygoid plates and pterygopalatine ganglion with its branches, lateral facial neurovascular pedicle, teeth and soft palate. Other structures were sacrificed: arteries and nerves located at the sites of skin and mucosal incision, and at the sites of osteotomies, ie the infraorbital nerve, the distal part of the greater palatine nerve, the nerves supplying the naso-pharynx, the nasal septum and the nasal conchae, nasolacrimal groove and lacrimal canal. The NMCF technique gives wide access to the deep nasal and nasopharyngeal areas. It is essential to preserve the lateral facial neurovascular pedicle to prevent necrosis of the midface structures. Preservation of the bony architecture surrounding the osteotomy sites is of great importance to allow precise final bone reassembly. Preoperative MRI appears of paramount importance to determine the borders of the lesion and the possibility of block resection.


Identification and characterization of peptides that bind the PPIase domain of Parvulin17.
Peptidyl-prolyl cis-trans isomerases (PPIases) are the enzymes that increase the rate of isomerization of the peptide bond N-terminal to the proline substrate. Par14 and its isoform Par17 belong to the Parvulin family of PPIases. Par14 can bind AT-rich double-stranded DNA and was shown to be part of the pre-ribosomal ribonucleoprotein (pre-rRNP) complexes, where it functions as an RNA processing factor that is involved in ribosome biogenesis. Its longer isoform Par17 is expressed only in cells of hominids, where it is targeted to the mitochondria. To find binding partners (peptides or proteins) for Par17, we applied the phage display technology. We panned 7-mer and 12-mer peptide libraries against Par17. The consensus sequence XHSXVHØ, where X can be any amino acid and Ø is a hydrophobic amino acid, was enriched from both libraries. We demonstrate the binding of this motif to the PPIase domain of Par17 using phage ELISA and NMR spectroscopy. We propose that residues Met90, Val91, Phe94, Gln95, Glu96, and Ala98 of Par17 are involved in substrate recognition, and that the phage display-selected motif XHSXVHØ can be recognized by Par17 PPIase domain in vivo.


Effect of Post-Infiltration Soil Aeration at Different Growth Stages on Growth and Fruit Quality of Drip-Irrigated Potted Tomato Plants (Solanum lycopersicum).
Soil hydraulic principles suggest that post-infiltration hypoxic conditions would be induced in the plant root-zone for drip-irrigated tomato production in small pots filled with natural soil. No previous study specifically examined the response of tomato plants (Solanum lycopersicum) at different growth stages to low soil aeration under these conditions. A 2 × 6 factorial experiment was conducted to quantify effects of no post-infiltration soil aeration versus aeration during 5 different periods (namely 27-33, 34-57, 58-85, 86-99, and 27-99 days after sowing), on growth and fruit quality of potted single tomato plants that were sub-surface trickle-irrigated every 2 days at 2 levels. Soil was aerated by injecting 2.5 liters of air into each pot through the drip tubing immediately after irrigation. Results showed that post-infiltration aeration, especially during the fruit setting (34-57 DAS) and enlargement (58-85 DAS) growth stages, can positively influence the yield, root dry weight and activity, and the nutritional (soluble solids and vitamin C content), taste (titratable acidity), and market quality (shape and firmness) of the tomato fruits. Interactions between irrigation level and post-infiltration aeration on some of these fruit quality parameters indicated a need for further study on the dynamic interplay of air and water in the root zone of the plants under the conditions of this experiment.


CRISPR adaptation from a structural perspective.
Bacterial CRISPR-Cas systems provide adaptive immunity against viruses and other mobile genome elements. During the adaptation step cells become immunized by insertion of short fragments of foreign DNA, termed spacers, into the genomic region called a CRISPR array. Selection, processing and insertion of new spacers is an elaborate and precisely orchestrated reaction, which relies on the Cas1-Cas2 integrase complex and accessory proteins that vary among different types of CRISPR-Cas systems. This review focuses on CRISPR adaptation from the structural perspective, with the spotlight on adaptation proteins employed by type I and type II CRISPR-Cas systems.


GO/PEDOT:NaPSS modified cathode as heterogeneous electro-Fenton pretreatment and subsequently aerobic granular sludge biological degradation for dye wastewater treatment.
Heterogeneous electro-Fenton (EF) technology has been wildly applied for the treatment of wastewater containing dyes and other organic pollutants. However, biologically treatment should be further applied after heterogeneous electro-Fenton process in order get better effluent quality. In the present study, a simple electropolymerization method using poly (3,4-ethylenedioxythiophene) (PEDOT) and graphene oxide (GO) was applied for graphite felt (GF) electrode modification as cathode in EF system, and coupling subsequently aerobic granular sludge (AGS) biological treatment for dye wastewater treatment. The modified electrode was characterized by scanning electron microscopy (SEM), Raman spectrum, and cyclic voltammetry (CV). Data implied that much higher H2O2 productivity, current response and coulomb efficiency (CE) were achieved by using GO/PEDOT:NaPSS modified GF. The results could be ascribed to the synergistic effect between PEDOT and GO that accelerated the electron transfer rate. Moreover, the H2O2 production capacity remained over 84.2% after 10-times reuses for GO/PEDOT:NaPSS modified GF, indicating that GO significantly improved the stability and life of electrode. Compared with the single system, the total organic carbon (TOC) and chemical oxygen demand (COD) removal efficiencies of the combined system degradation methylene blue (MB) wastewater were significantly improved. Therefore, this modified GF could be used as a potentially useful cathode in heterogeneous EF technology for actual wastewater treatment and the combined system have a promising engineering application value in MB wastewater degradation field.


Systematic Review of Management and Survival Outcome of Parotid Cancers with Lateral Skull Base Invasion.
Lateral skull base involvement from parotid cancers is a rare condition and is considered a poor prognostic indicator. The aim of this study was to systematically review the literature of parotid tumors with temporal bone invasion to analyze the survival outcome. A systematic literature review was performed in August 2022, without time limits, and 289 patients affected by parotid gland cancers and lateral skull base involvement were included. The most common symptoms in parotid tumors at the onset were indolent mass, facial weakness, pain, and hearing loss; the chi-square value is 23.1063, with a statistically significance (p = < 0.000121). The five most common histologies were adenoid cystic carcinoma, acinic cell carcinoma, mucoepidermoid carcinoma, adenocarcinoma, and squamous cell carcinoma. The facial nerve function after surgery showed statistically significance (functional vs. non-functional; chi-square was 91.7698, p = < 0.00001). Mean follow-up was 36.2 months (range 0.3-192). At the last follow-up, more patients died of disease (DOD; 60/289, 21%) than other causes (DOOC; 5/289, 2%). There is a statistically significant correlation between patients died for tumor (DOD) and patients died for other causes (DOOC) (p = < 0.0001), suggesting that the lateral skull base invasion negatively impacts on survival. Basing on the results of our systematic review, lateral skull base involvement from parotid recurrent/advance tumors should be considered a poor prognostic factor, as the majority of patients die due to this condition. It also would be necessary to have "clear"works, with full data (demographic, clinical, surgical data), and with a longer follow up, in order to assess the best treatment modality of these patients.


Comparison of the Effect of Adipocyte-derived Stem Cells and Curcumin Nanoliposomes with Phenytoin on Open Cutaneous Wound Healing in Rats.
Reducing the healing time of wounds can decrease the patient's immobility time and their medical costs, leading a faster return of the patients to daily work. The aim of the present study is to compare the effect of adipose-derived stem cells and curcumin- containing liposomal nanoparticles with phenytoin on wound healing. After anesthesia of the rats, open skin ulcers were made by a bistoury blade. Subsequently, stem cells were removed from the adipose tissue of the upper border of the epididymis. The originality of stem cells was then confirmed by the flow cytometry. The fusion method was used to prepare the liposome; and also, nanoliposomal particles were confirmed by using the DLS microscope. The percentage of recovery and the cell count was measured with IMAGEJ. The expression of genes was assessed by PCR. The number of fibroblasts was counted by immunohistochemistry techniques. The amount of collagen was determined by Tri-chromosome staining, and the number of capillaries was enumerated by H & E staining. The expression of the TGF-β1 gene, vascular number, wound healing rate and the number of fibroblasts increased significantly in adipose tissue-derived stem cells and curcumin nanoliposome groups (p<0.05); the wound surface was also decreased significantly (p<0.05). Based on the results of our research, adipose tissue-derived stem cells and curcumin nanoliposomes can heal wounds efficiently.


Carotenoid production and gene expression in an astaxanthin-overproducing Xanthophyllomyces dendrorhous mutant strain.
The primary carotenoid synthesized by Xanthophyllomyces dendrorhous is astaxanthin, which is used as a feed additive in aquaculture. Cell growth kinetics and carotenoid production were correlated with the mRNA levels of the idi, crtE, crtYB, crtI, crtS and crtR genes, and the changes in gene sequence between the wild-type and a carotenoid overproducer XR4 mutant strain were identified. At the late stationary phase, the total carotenoid content in XR4 was fivefold higher than that of the wild-type strain. Additionally, the mRNA levels of crtE and crtS increased during the XR4 growth and were three times higher than the wild-type strain in the late stationary phase. Moreover, the nucleotide sequences of crtYB, crtI and crtR exhibited differences between the strains. Both the higher crtE and crtS transcript levels and the crtYB, crtI and crtR mutations can, at least in part, act to up-regulate the carotenoid biosynthesis pathway in the XR4 strain.


Quantitative and qualitative analyses of urinary L-FABP for predicting acute kidney injury after emergency laparotomy.
The aim of this study was to explore the clinical utility of urinary L-FABP for earlier prediction of acute kidney injury (AKI) after emergency laparotomy, and to assess the clinical utility of a point-of-care (POC) kit for urinary L-FABP. Forty-eight patients undergoing emergency laparotomy were divided into AKI and non-AKI groups by the kidney diseases: improving global outcome (KDIGO) criteria. Ten patients were included in the AKI group. Urinary L-FABP, albumin, N-acetyl-β-D-glucosaminidase (NAG), TIMP-2, IGFBP7, serum creatinine (SCr), and blood presepsin were measured perioperatively and compared between groups. Perioperative urinary L-FABP was also evaluated qualitatively using a POC kit. L-FABP and albumin levels were significantly higher in the AKI group at all measurement points. NAG was significantly higher only postoperatively in the AKI group. There were no inter-group differences in [TIMP-2] × [IGFBP7] at any measuring point. The area under the receiver operating characteristic curve of urinary L-FABP was greater than 0.8 perioperatively, which was larger than that of other biomarkers throughout the study period. The correlation coefficient at 2 h after entering the operating room between quantitative and qualitative tests for urinary L-FABP was 0.714, which was the maximum. The sensitivity, specificity, and negative predictive value of the urinary L-FABP POC kit at 2 h after entry were 55.6%, 91.9%, and 89.5%, respectively. Quantitative L-FABP analyses is suitable for predicting postoperative AKI earlier in the perioperative period of emergency laparotomy. Conversely, the higher specificity of qualitative L-FABP analysis suggests that it may be useful for excluding the risk of AKI but its overall clinical validity should be further investigated.


Assessment of cardiac performance with quantitative radionuclide angiocardiography: right ventricular ejection fraction with reference to findings in chronic obstructive pulmonary disease.
A reproducible noninvasive technique for measuring righ ventricular ejection fraction was developed using first pass quantitative radionuclide angiocardiography. Studies were obtained in the anterior position with a computerized multicrystal scintillation camera with high count rate capabilities. Right ventricular ejection fraction was calculated on a beat to beat basis from the high frequency components of the background-corrected right ventricular time-activity curve. In 50 normal adults, right ventricular ejection fraction averaged 55 percent (range of 45 to 65 percent). This radionuclide measure of right ventricular function was reproducible, with minimal inter- and intraobserver variability, and was sensitive to changes in inotropic state induced with isoproterenol. In 36 patients with chronic obstructive pulmonary disease, right ventricular ejection fraction ranged from 19 to 71 percent. All 10 patients with corpulmonale, as well as 9 additional patients, had an abnormal right ventricular ejection fraction. Arterial oxygen tension and forced expiratory volume were depressed significantly more in patients with abnormal right ventricular ejection fraction than in subjects with normal right ventricular function. There was no relation between abnormalities in right and left ventricular ejection fraction.


Two new bacteria isolated from vagina of a patient with vaginosis: Atopobium massiliense sp. nov. and Butyricimonas vaginalis sp. nov.
Two new bacterial strains, Marseille-P4126 (=CSURP4126) and Marseille-P4593 (=CSURP4593), were isolated from the vaginal sample of a French woman with vaginosis. These strains were identified and characterized using the taxonogenomics method. The findings from phylogenetic tree interpretation, phenotypic criteria and genomic analysis provided here distinctly display that Atopobium massiliense sp. nov. and Butyricimonas vaginalis sp. nov. are new members of the genus Atopobium and Butyricimonas, respectively.


Brain-stiffness-mimicking tilapia collagen gel promotes the induction of dorsal cortical neurons from human pluripotent stem cells.
The mechanical properties of the extracellular microenvironment, including its stiffness, play a crucial role in stem cell fate determination. Although previous studies have demonstrated that the developing brain exhibits spatiotemporal diversity in stiffness, it remains unclear how stiffness regulates stem cell fate towards specific neural lineages. Here, we established a culture substrate that reproduces the stiffness of brain tissue using tilapia collagen for in vitro reconstitution assays. By adding crosslinkers, we obtained gels that are similar in stiffness to living brain tissue (150-1500 Pa). We further examined the capability of the gels serving as a substrate for stem cell culture and the effect of stiffness on neural lineage differentiation using human iPS cells. Surprisingly, exposure to gels with a stiffness of approximately 1500 Pa during the early period of neural induction promoted the production of dorsal cortical neurons. These findings suggest that brain-stiffness-mimicking gel has the potential to determine the terminal neural subtype. Taken together, the crosslinked tilapia collagen gel is expected to be useful in various reconstitution assays that can be used to explore the role of stiffness in neurogenesis and neural functions. The enhanced production of dorsal cortical neurons may also provide considerable advantages for neural regenerative applications.


Integron-encoded GES-type extended-spectrum beta-lactamase with increased activity toward aztreonam in Pseudomonas aeruginosa.
A Pseudomonas aeruginosa strain expresses an extended-spectrum beta-lactamase, GES-9, which differs from GES-1 by a Gly243Ser substitution, is inhibited by clavulanic acid and imipenem, and hydrolyzes aztreonam. The bla(GES-9) gene was located inside a class 1 integron structure containing two copies of a novel insertion sequence belonging to the IS1111 family.


Restraint use and injury patterns among children involved in motor vehicle collisions.
Motor vehicle collisions are the leading cause of death among children older than 1 year. Use of appropriate restraint systems is associated with reductions in morbidity and mortality in this age group. No studies have evaluated the association between specific injury patterns and restraint use among children. The purpose of this study was to evaluate differences in risks of injuries in different body regions according to restraint use among children 0 to 11 years of age. The 1995 to 1999 National Automotive Sampling System data files were used. The National Automotive Sampling System is a national probability sample of passenger vehicles involved in police-reported tow-away collisions. Information on occupant (seating position, restraint use), collision (change in velocity, vehicle intrusion), and outcome characteristics was evaluated. Risks of injuries in different body regions (Abbreviated Injury Scale, 1990 Revision score > or = 2) were calculated and compared according to restraint use. Between 1995 and 1999, there were approximately 1.5 million children 0 to 11 years of age involved in police-reported tow-away MVCs who met the inclusion criteria for this study. Compared with unrestrained children, properly restrained children had significantly lower overall injury risk (risk ratio [RR], 0.37); significant risk reductions were also observed for injuries to the head (RR, 0.18), thorax (RR, 0.35), and lower extremities (RR, 0.26), and mortality (RR, 0.26). Significant risk reductions were not noted when comparing improperly restrained children with unrestrained children. Proper restraint use among children is associated with lower risk of injury. Educational initiatives should focus not only on encouraging restraint use but also on ensuring that parents know the appropriate age-dependent restraint method and how to use it properly.


Reliability of COVID-19 data: An evaluation and reflection.
The rapid proliferation of COVID-19 has left governments scrambling, and several data aggregators are now assisting in the reporting of county cases and deaths. The different variables affecting reporting (e.g., time delays in reporting) necessitates a well-documented reliability study examining the data methods and discussion of possible causes of differences between aggregators. To statistically evaluate the reliability of COVID-19 data across aggregators using case fatality rate (CFR) estimates and reliability statistics. Cases and deaths were collected daily by volunteers via state and local health departments, as primary sources and newspaper reports, as secondary sources. In an effort to begin comparison for reliability statistical analysis, BroadStreet collected data from other COVID-19 aggregator sources, including USAFacts, Johns Hopkins University, New York Times, The COVID Tracking Project. COVID-19 cases and death counts at the county and state levels. Lower levels of inter-rater agreement were observed across aggregators associated with the number of deaths, which manifested itself in state level Bayesian estimates of COVID-19 fatality rates. A national, publicly available data set is needed for current and future disease outbreaks and improved reliability in reporting.


CART peptide and opioid addiction: Expression changes in male rat brain.
Previous studies have shown the prominence of cocaine- and amphetamine-regulated transcript (CART) peptide in rewarding and reinforcing effects of drugs of abuse specially psychostimulants. The data regarding the effects of different stages of opioid addiction on CART expression and the interconnection between CART and opioids are not much available. Here we have studied the changes in the expression level of CART mRNA and protein in various parts of the brain reward pathway in different stages of opioid addiction. Groups of male rats received acute low-dose (10mg/kg), acute high-dose (80mg/kg) and chronic escalating doses of morphine. In addition, withdrawal and abstinence states were evaluated after injection of naloxone (1mg/kg) and long-term maintenance of addicted animals, respectively. Expression of CART mRNA in the brain was measured by real-time PCR method. Western blotting was used to quantify the protein level. CART mRNA and protein were both up-regulated in high-dose morphine-administered animals and also in the withdrawal group in the nucleus accumbens (NAc), striatum and prefrontal cortex (PFC). In the addicted group, CART mRNA and protein were both down-regulated in NAc and striatum. In the abstinent group, CART mRNA was down-regulated in NAc. In the hippocampus, the only observed change was the up-regulation of CART mRNA in the withdrawal group. We suggest that the modulatory role of CART peptide in rewarding and reinforcing effects of opioids weakens when opioids are used for a long time and is stimulated when acute stress such as naloxone-induced withdrawal syndrome or acute high-dose administration of morphine occurs to the animal.


Pulmonary cryptococcosis mimicking lung cancer: 3 case report.
Pulmonary cryptococcosis is an opportunistic pathogen. However, it is difficult to differentiate it from other diseases by imaging. Therefore, the clinical presentation and imaging features of patients with pulmonary cryptococcosis are analyzed and summarized to improve clinicians' early recognition of the disease. We present a case where preoperative imaging was difficult to differentiate between pulmonary cryptococcosis and non-small cell lung cancer. A pathological biopsy was taken by surgical treatment to clarify the diagnosis. Three cases of Cryptococcus pulmonary patients diagnosed by pathology in our department were analyzed. We believe that the diagnosis should be confirmed as soon as possible through surgery, relevant laboratory tests, and pathological examinations, and antifungal treatment should be carried out in time. We believe that an earlier and clearer diagnosis of pulmonary cryptococcosis requires an earlier interplay of imaging, pathology, testing, and individual patient differences.


[Popper's critical rationalism and the biomedical sciences].
Popper's rationalism makes an important contribution of the 20th century philosophy to the methodology of natural sciences. Through its criterion of falsification, it enabled the scientists to take a critical but constructive view on hypotheses, conjectures and theories. This attitude found its application also in medicine.


A voluntary summer program to expose nursing students to the perioperative specialty.
Perioperative courses often are not available in undergraduate nursing programs, even though students are interested in the specialty. A voluntary summer program can provide opportunities that other clinical experiences cannot and can be a rewarding experience for faculty members and students. One such program has been established at a college in the southeastern United States.


AP4 directly downregulates p16 and p21 to suppress senescence and mediate transformation.
Here we analyzed the function of the c-MYC-inducible basic helix-loop-helix leucine-zipper transcription factor AP4 in AP4-deficient mouse embryo fibroblasts (MEFs). Loss of AP4 resulted in premature senescence and resistance towards immortalization. Senescence was accompanied by induction of the cyclin-dependent kinase inhibitor-encoding genes p16, a known tumor suppressor, and p21, a previously described target for repression by AP4. Notably, AP4 directly repressed p16 expression via conserved E-box motifs in MEFs and human diploid fibroblasts. Senescence caused by AP4-deficiency was prevented by depletion of p16 and/or p21, demonstrating that these factors mediate senescence caused by AP4 loss. As senescence induced by the loss of AP4 was rescued by ectopic AP4, secondary lesions were not involved in causing premature senescence. Activation of c-MYC resulted in repression of p21 and p16 in AP4(+/+), but not in AP4(-/-) MEFs. Furthermore, after combined expression of c-MYC and mutant RAS in MEFs, AP4 was required for colony formation, anchorage-independent growth and tumor formation in mice. In addition, combined ectopic expression of AP4 and mutant RAS in MEFs resulted in colony formation. However, additional loss of the p53 tumor suppressor was necessary for anchorage-independent growth and tumor formation of MEFs by combined AP4 and mutant RAS expression. In conclusion, this study identified AP4 as an oncogenic antagonist of cellular senescence. AP4 achieves this effect by direct repression of p16 and p21, and may thereby critically contribute to c-MYC function and tumor progression.


Measurement of anti-factor VIII IgG, IgG4 and IgM alloantibodies in previously untreated hemophilia A patients treated with recombinant factor VIII. Kogenate Japanese Clinical Study Group.
Using enzyme-linked immunosorbent assay (ELISA), we measured anti-factor VIII immunoglobulin G (IgG), IgG4 and IgM in serum samples obtained from hemophilia A patients treated with a recombinant factor VIII (rFVIII) preparation (Kogenate). Twelve pre- and post-treatment serum samples from 3 previously untreated patients who developed an inhibitor (alloantibody) were evaluated. Both rFVIII and plasma-derived factor VIII (pdFVIII) were used as antigen. Serum samples were diluted 37.5-fold. For the measurement of IgM antibody, protein A Sephadex suspension was added to the patients' serum samples and the supernatant was assayed. Anti-human IgG, IgG4, and IgM monoclonal antibodies labelled with peroxidase were added and absorbance at 450nm was determined. The reactivity of the IgG antibody with rFVIII and pdFVIII was extremely low. Samples containing the IgG4 inhibitor with a neutralizing activity of approximately 7.5 Bethesda units (BU)/ml resulted in absorbance values of 0.451-0.551, thus demonstrating a considerably high degree of sensitivity. The correlation between the neutralizing activity and reactions of the IgG4 antibody with rFVIII and pdFVIII antigens was high, with correlation coefficients of 0.912 and 0.966, respectively. Furthermore, the correlation coefficient between the measured absorbance values for the antibody reacted with pdFVIII and rFVIII was 0.961. No correlation was found between the reactivity of IgM to rFVIII and pdFVIII.


The influence of cylindrospermopsin on oxidative DNA damage and apoptosis induction in HepG2 cells.
Cylindrospermopsin (CYN) a potent cyanobacterial cytotoxin and protein synthesis inhibitor is increasingly being found in surface freshwaters worldwide. Due to its genotoxic activity and potential carcinogenicity it was recognized as a potential threat to humans. However, the mechanisms of CYN genotoxicity are not well understood. We explored whether CYN at non-cytotoxic exposure conditions causes DNA damage through induction of oxidative stress and whether it induces apoptosis in HepG2 cells. With the DCFH-DA probe a significant increase in the intracellular formation of reactive oxygen species (ROS) was observed, which steadily increased with incubation. Induction of oxidative DNA damage was determined with the modified comet assay with formamidopyrimidine glycosylase (Fpg) digestion. No DNA damage was observed after 4h exposure to CYN. After 12 and 24 h exposure, CYN (at 0.25 and 0.5 μg mL(-1)) induced significant increase of DNA strand breaks, but not oxidative DNA damage, suggesting minor role of oxidative stress in CYN mediated genotoxicity. CYN also significantly increased the mitochondrial membrane potential (MMP), determined with the JC-1 probe, while no induction of caspase 3 and 7 activity and no increase in the number of apoptotic cells, measured with Annexin V/PI staining, could be determined. These results show that at non-cytotoxic concentrations CYN induced DNA damage was not the consequence of oxidative stress and that CYN did not induce apoptosis, which may add to the hazard of this toxin, as cells with damaged DNA are not removed from the population, enhancing the risk of mutations and consequently carcinogenesis.


[Detection of hyperthermophilic archaea of the genus Desulforococcus by hybridization with oligonucleotide probes].
Based on the analysis of nucleotide sequences of 16S rRNA, oligonucleotide probes were designed for the detection and identification of representatives of the genus Desulfurococcus (kingdom Crenarchaeota of the domain Archaea). The detection procedure included obtaining of PCR products on DNA isolated from pure cultures, enrichments, or natural samples with a Crenarchaeota-specific primer pair designed: Cren 7F (5'-TTCCGGTTGATCCYGCCGGACC-3') and Cren 518R (5'-GCTGGTWTTACCGCGGCGGCTGA-3'). The PCR products were hybridized with Dig-11-dUTP-labeled oligonucleotide probes targeting the genus Desulfurococcus (Dco 198, 5'-CGTTAACYCCYGCCACACC-3) and its species D. mobilis (Dco_mob 198, 5'-CGTTAACCCCTGCCACACC-3') and D. amylolyticus (Dco_amy 198, 5'-CGTTAACCCCCGCCACACC-3'). With the use of these primers and probes, four new strains isolated from hydrotherms of Kamchatka and Kunashir Island were identified as members of the species Desulfurococcus amylolyticus. Desulfurococcus representatives were detected in several natural samples, including a sample taken from a marine hydrotherm at the Kunashir Island; this demonstrates that representatives of this genus occur not only in terrestrial but also in marine environments.


Effects of extensive splenomegaly in patients with myelofibrosis undergoing a reduced intensity allogeneic stem cell transplantation.
Changes in spleen size postallogeneic haematopoietic stem cell transplantation (HSCT) in patients with primary myelofibrosis have been poorly characterized. We analysed 10 patients with myelofibrosis and splenomegaly following a reduced-intensity allogeneic HSCT. All patients fully engrafted donor cells including five patients with extensive splenomegaly. Extensive splenomegaly was associated with a prolonged time to neutrophil and platelet recovery. In all 10 patients, a progressive reduction of splenomegaly was documented within 12 months post-transplant and paralleled the reduction of marrow fibrosis. These findings suggest that myelofibrosis patients with extensive splenomegaly may proceed with allogeneic HSCT without prior splenectomy.


Catalysis by Nature's photoenzymes.
Photoenzymes use light to initiate biochemical reactions. Although rarely found in nature, their study has advanced understanding of how light energy can be harnessed to facilitate enzyme catalysis, which is also of importance to the design and engineering of man-made photocatalysts. Natural photoenzymes can be assigned to one of two families, based broadly on the nature of the light-sensing chromophores used, those being chlorophyll-like tetrapyrroles or flavins. In all cases, light absorption leads to excited state electron transfer, which in turn initiates photocatalysis. Reviewed here are recent findings relating to the structures and mechanisms of known photoenzymes. We highlight recent advances that have deepened understanding of mechanisms in biological photocatalysis.


Male sex is an independent risk factor for patellar osteochondral fractures following acute patellar dislocation in pediatric patients.
Although most patellar dislocations are associated with medial patellofemoral ligament (MPFL) injury, many patients also sustain concomitant patellar osteochondral fractures following a patellar dislocation. Few prior studies have described or evaluated risk factors for patellar osteochondral fractures in pediatric patients. The purpose of the present study was to describe the incidenceand location of patellar osteochondral fractures following acute patellar dislocation in pediatric patients. In addition, we described risk factors for patellar osteochondral fractures in this population. We hypothesized that most fractures would occur at the inferomedial quadrant of the patella following a traumatic injury mechanism. Following Institutional Review Board approval, the electronic medical record was queried to identify pediatric patients ≤ 18 years old who underwent MPFL reconstruction (MPFLR) or non-operative treatment for patellar instability between July 2016 and February 2020. Osteochondral fractures were defined as full-thickness chondral injuries with attached subchondral bone or purely osseous injuries measuring ≥ 3 mm (mm) in at least two magnetic resonance imaging (MRI) planes. Patients were included in the study if they had complete preoperative x-ray and MRI studies with minimum 6-month follow-up. Patients were excluded if they had incomplete imaging, isolated chondral fractures, or less than 6 months follow-up. Univariate analysis was used to identify patient factors associated with osteochondral fractures. Multivariate regression analysis was used to identify risk factors for osteochondral fractures. Sixty patients (63 knees) were included in the study, 15 (23.8%) of whom had a patellar osteochondral fracture. The majority of osteochondral fractures (87%) involved the inferomedial quadrant of the patella. Univariate analysis showed an association between male sex (p = 0.041), skeletal immaturity (p = 0.028), and decreased patellar tilt (p = 0.021) and patellar osteochondral fractures. Multivariate regression analysis identified male sex as an independent risk factor for osteochondral fractures (relative risk: 4.8, 95% confidence interval [CI]: 1.08-20.9, p = 0.039). No patients had recurrent dislocation at minimum 6-month follow-up. All patients with osteochondral fractures returned to sports at most recent follow up. In this study, 23% of pediatric patients with acute patellar dislocations have a concomitant patellar osteochondral fracture. The majority of patellar osteochondral fractures involve the inferomedial quadrant of the patella. Male sex is an independent risk factor for patellar osteochondral fractures, and skeletal immaturity is associated with patellar osteochondral fractures in this population. Level III.


Cell-Free NADPH Oxidase Activation Assays: A Triumph of Reductionism.
The superoxide (O2·-)-generating NADPH oxidase complex of phagocytes comprises a membrane-associated heterodimeric flavocytochrome, known as cytochrome b 558 (consisting of NOX2 and p22phox) and four cytosolic regulatory proteins, p47phox, p67phox, p40phox, and the small GTPase Rac. Under physiological conditions, in the resting phagocyte, O2·- generation is initiated by engagement of membrane receptors by a variety of stimuli, followed by signal transduction sequences leading to the translocation of the cytosolic components to the membrane and their association with the cytochrome, a process known as NADPH oxidase assembly. A consequent conformational change in NOX2 initiates the electron flow along a redox gradient, from NADPH to molecular oxygen (O2), leading to the one-electron reduction of O2 to O2·-. Historically, methodological difficulties in the study of the assembled complex derived from stimulated cells, due to its lack of stability, led to the design of "cell-free" systems (also known as "broken cells" or in vitro systems). In a major paradigm shift, the cell-free systems have as their starting point NADPH oxidase components derived from resting (unstimulated) phagocytes, or as in the predominant method at present, recombinant proteins representing the components of the NADPH oxidase complex. In cell-free systems, membrane receptor stimulation and the signal transduction sequence are absent, the accent being placed on the actual process of assembly, all of which takes place in vitro. Thus, a mixture of the individual components of the NADPH oxidase is exposed in vitro to an activating agent, the most common being anionic amphiphiles, resulting in the formation of a complex between cytochrome b 558 and the cytosolic components and O2·- generation in the presence of NADPH. Alternative activating pathways require posttranslational modification of oxidase components or modifying the phospholipid milieu surrounding cytochrome b 558. Activation is commonly quantified by measuring the primary product of the reaction, O2·-, trapped immediately after its generation by an appropriate acceptor in a kinetic assay, permitting the calculation of rates of O2·- production, but numerous variations exist, based on the assessment of reaction products or the consumption of substrates. Cell-free assays played a paramount role in the identification and characterization of the components of the NADPH oxidase complex, the performance of structure-function studies, the deciphering of the mechanisms of assembly, the search for inhibitory drugs, and the diagnosis of various forms of chronic granulomatous disease (CGD).


Trends in mortality from cerebrovascular disease in France from 1968 to 1978. With reference to cardiovascular and all causes of death.
Cerebrovascular disease mortality in France during the period 1968-1978 was compared with cardiovascular and all other causes of death. Our study demonstrated a 25% decline in the age-adjusted cerebrovascular disease mortality rates in both sexes and particularly in the middle-aged groups. This decline is greater than that of the general causes of mortality, which was on the order of 20%. It parallels the decline in congestive heart failure mortality but differs from that of ischemic heart disease. Nevertheless, despite the increase in the proportion of elderly people in the population, the total number of deaths from cerebrovascular diseases has remained almost unchanged, although the deaths occur at a higher age than previously. If this trend is confirmed, cerebrovascular diseases will remain one of the more frequent causes of death in the elderly and thus a social problem of crucial importance.


Diarylidene-N-Methyl-4-Piperidones and Spirobibenzopyrans as Antioxidant and Anti-Inflammatory Agents.
Curcumin has antioxidant properties resulting from its radical scavenging ability and inhibition of inflammation-associated factors. However, its lack of solubility, instability, and poor bioavailability are impediments to its therapeutic use. As potential alternatives, we synthesized and performed chemical analysis of thirty diarylidene-N-methyl-4-piperidone (DANMP), diheteroarylidene-N-methyl-4-piperidone (DHANMP), and spirobibenzopyran (SBP) derivatives, one of which was also characterized by single crystal X-ray diffraction. All compounds were evaluated for antioxidant activity via 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and for drug-like properties in silico. A subset of five compounds was investigated in terms of aqueous solubilities, which were significantly improved compared to that of curcumin. In vitro assessments of cellular and anti-inflammatory effects were conducted via real time polymerase chain reaction (RT-PCR) and Griess assays to evaluate the presence of inflammatory/activated (M1) markers and production of nitric oxide (NO) species, which are associated with inflammation. The five compounds reduced levels of markers and NO to extents similar to or better than curcumin in inflamed cells, and showed no adverse effects on cell viability. We show that these compounds possess anti-inflammatory properties and may be used as curcumin-substitutes with improved characteristics.


The reliability of microleakage studies using dog and bovine primary teeth instead of human primary teeth.
This was to investigate if dental restorations on human, dog and bovine primary teeth are equitable for microleakage analysis, and the respective marginal adaptation deficiency in in vitro conditions. In order to determine the level of microleakage in Class V poly acid-modified composite restorations of human, dog and bovine primary teeth, samples were evaluated by dye-leakage method in sections of the inner area of the restorations. We found no leakage in 6 out of 10 (60%) human restorations, in 7 out of 10 (70%) of dog and in 7 out of 10 (70%) of bovine teeth. The levels of dye-microleakage were tested with Kruskal-Wallis one-way variant analysis method. The relative leakage differences were not statistically significant among all species (p>0.05). We concluded that dog and bovine primary teeth might be suitable for in vitro studies instead of human ones.


Durability of effects from short-term economic incentives for clinic attendance among HIV positive adults in Tanzania: long-term follow-up of a randomised controlled trial.
Conditional economic incentives are shown to promote medication adherence across a range of health conditions and settings; however, any long-term harms or benefits from these time-limited interventions remain largely unevaluated. We assessed 2-3 years outcomes from a 6-month incentive programme in Tanzania that originally improved short-term retention in HIV care and medication possession. We traced former participants in a 2013-2016 trial, which randomised 800 food-insecure adults starting HIV treatment at three clinics to receive either usual care (control) or up to 6 months of cash or food transfers (~US$11/month) contingent on timely attendance at monthly clinic appointments. The primary intention-to-treat analysis estimated 24-month and 36-month marginal risk differences (RD) between incentive and control groups for retention in care and all-cause mortality, using multiple imputation for a minority of missing outcomes. We also estimated mortality HRs from time-stratified Cox regression. From 3 March 2018 to 19 September 2019, we determined 36-month retention and mortality statuses for 737 (92%) and 700 (88%) participants, respectively. Overall, approximately 660 (83%) participants were in care at 36 months while 43 (5%) had died. There were no differences between groups in retention at 24 months (86.5% intervention vs 84.4% control, RD 2.1, 95% CI -5.2 to 9.3) or 36 months (83.3% vs 77.8%, RD 5.6, -2.7 to 13.8), nor in mortality at either time point. The intervention group had a lower rate of death during the first 18 months (HR 0.27, 95% CI 0.10 to 0.74); mortality was similar thereafter (HR 1.13, 95% CI 0.33 to 3.79). These findings confirm that incentives are a safe and effective tool to promote short-term adherence and potentially avert early deaths at the critical time of HIV treatment initiation. Complementary strategies are recommended to sustain lifelong retention in HIV care. NCT01957917.


Tehran cohort study (TeCS) on cardiovascular diseases, injury, and mental health: Design, methods, and recruitment data.
Cardiovascular disease, mental health, and injury are among the top health issues globally. In Tehran Cohort Study, we aimed to determine the prevalence, incidence, and trend of cardiovascular diseases, psychiatric symptoms, injury, and risk factors in Tehran households. We enrolled 4215 households in the recruitment phase from March 2016 to March 2019. Demographic characteristics, past medical history, medications, and familial history of the participants were collected. Rose angina pectoris, general health Questionnaire-28 (GHQ-28), and injury questionnaires were completed. Fasting blood samples were collected to measure routine biochemistry and store samples in the biobank. Anthropometric and physiological measurements and electrocardiograms were performed. The participants are followed every three years for up to 12 years. In total, 8296 individuals participated in the cardiovascular section, 10247 completed the GHQ-28, and 4167 households completed the injury questionnaire. The mean age of the participants was 48.2 (16.41), and 46.5% were male. 64.3% of recruited individuals had no symptoms of psychiatric disorders, and 3729 (89.5%) households did not have any severe injury requiring treatment. The participants' diversity and their invaluable data will help us provide a general picture of the current prevalence and incidence of the main study objectives.


Thrombin inhibitors built on an azaphenylalanine scaffold.
A series of azaphenylalanine derivatives were investigated as novel thrombin inhibitors based on the prodrug principle. By systematic structural modifications we have identified optimal groups for this series that led us to potent inhibitors of thrombin incorporating the benzamidine fragment at the P1 position, and their potentially orally active benzamidoxime prodrugs. The binding modes in the thrombin active site of two representative compounds were identified by X-ray crystallographic analysis.


Alternative splicing results in differential expression, activity, and localization of the two forms of arginyl-tRNA-protein transferase, a component of the N-end rule pathway.
The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. The underlying ubiquitin-dependent proteolytic system, called the N-end rule pathway, is organized hierarchically: N-terminal aspartate and glutamate (and also cysteine in metazoans) are secondary destabilizing residues, in that they function through their conjugation, by arginyl-tRNA-protein transferase (R-transferase), to arginine, a primary destabilizing residue. We isolated cDNA encoding the 516-residue mouse R-transferase, ATE1p, and found two species, termed Ate1-1 and Ate1-2. The Ate1 mRNAs are produced through a most unusual alternative splicing that retains one or the other of the two homologous 129-bp exons, which are adjacent in the mouse Ate1 gene. Human ATE1 also contains the alternative 129-bp exons, whereas the plant (Arabidopsis thaliana) and fly (Drosophila melanogaster) Ate1 genes encode a single form of ATE1p. A fusion of ATE1-1p with green fluorescent protein (GFP) is present in both the nucleus and the cytosol, whereas ATE1-2p-GFP is exclusively cytosolic. Mouse ATE1-1p and ATE1-2p were examined by expressing them in ate1Delta Saccharomyces cerevisiae in the presence of test substrates that included Asp-betagal (beta-galactosidase) and Cys-betagal. Both forms of the mouse R-transferase conferred instability on Asp-betagal (but not on Cys-betagal) through the arginylation of its N-terminal Asp, the ATE1-1p enzyme being more active than ATE1-2p. The ratio of Ate1-1 to Ate1-2 mRNA varies greatly among the mouse tissues; it is approximately 0.1 in the skeletal muscle, approximately 0.25 in the spleen, approximately 3.3 in the liver and brain, and approximately 10 in the testis, suggesting that the two R-transferases are functionally distinct.


Electrogenerated chemiluminescence detector based on Ru(bpy)3(2+) immobilized in cation exchange resin for high-performance liquid chromatography: An approach to stable detection.
In this work, an electrogenerated chemiluminescence (ECL) detector with improved stability was developed for high-performance liquid chromatography (HPLC) detection of hydrochlorothiazide (HCTZ). The detector was prepared by packing cation exchanged resin particles in a glass tube, followed by inserting Pt wires (working electrode) in this tube and sealing. The leakage of Ru(bpy)3(2+) from the resin was compensated by adding a small amount of Ru(bpy)3(2+) in the mobile phase. Factors affected the performance of the proposed ECL detector were investigated. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of HCTZ in the range of 5.0 × 10(-8) g mL(-1)-2.5 × 10(-5) g mL(-1) and the detection limit was 2.0 × 10(-8) g mL(-1) (S/N=3). Application of the detector to the analysis of HCTZ in human serum proved feasible.


Prime and probability: causal knowledge affects inferential and predictive effects on self-agency experiences.
Experiences of having caused a certain outcome may arise from motor predictions based on action-outcome probabilities and causal inferences based on pre-activated outcome representations. However, when and how both indicators combine to affect such self-agency experiences is still unclear. Based on previous research on prediction and inference effects on self-agency, we propose that their (combined) contribution crucially depends on whether people have knowledge about the causal relation between actions and outcomes that is relevant to subsequent self-agency experiences. Therefore, we manipulated causal knowledge that was either relevant or irrelevant by varying the probability of co-occurrence (50% or 80%) of specific actions and outcomes. Afterwards, we measured self-agency experiences in an action-outcome task where outcomes were primed or not. Results showed that motor prediction only affected self-agency when relevant actions and outcomes were learned to be causally related. Interestingly, however, inference effects also occurred when no relevant causal knowledge was acquired.


Recreational Runners With a History of Injury Are Twice as Likely to Sustain a Running-Related Injury as Runners With No History of Injury: A 1-Year Prospective Cohort Study.
To investigate (1) the cumulative incidence proportion and the most common anatomical locations of running-related injuries in recreational runners, and (2) the associations between running-related injuries and previous injury, running experience, weekly running distance, age, sex, and body mass index. A 1-year prospective cohort study. Two hundred twenty-four recreational runners (average weekly running distance for the past 12 months, 15 km) were included (89 women, 135 men). Pain information was reported weekly for 1 year, and all running-related injuries that resulted in time loss or medical consultation were recorded. We accounted for censoring when calculating cumulative incidence proportion, and used crude Cox proportional hazards regression to evaluate whether the variables of interest were associated with running-related injuries. The 1-year cumulative incidence proportion of running-related injuries was 45.9% (95% confidence interval [CI]: 38.4%, 54.2%). The most common anatomical locations were the knee (20/75, 27%) and Achilles tendon/calf (19/75, 25%). Previous injury was associated with a higher injury rate (hazard rate ratio = 1.9; 95% CI: 1.2, 3.2), while the other variables had no statistically significant association with injury. There were 75 running-related injuries during the 1-year surveillance period, for a cumulative incidence proportion of 46%. The most common injuries were to the knee and Achilles tendon/calf. Recreational runners with a previous injury were twice as likely to sustain a running-related injury as runners with no previous injury. J Orthop Sports Phys Ther 2021;51(3):144-150. Epub 25 Dec 2020. doi:10.2519/jospt.2021.9673.


Redefining CD56 as a Biomarker and Therapeutic Target in Multiple Myeloma.
Multiple myeloma cells aberrantly express surface antigens compared with normal plasma cells. Among others, CD56 is present at variable levels in approximately 70% of patients with multiple myeloma; however, very little is known about CD56 role in multiple myeloma. We demonstrated that patients with multiple myeloma with more than 10% of CD56-expressing clonal multiple myeloma cells have inferior clinical outcomes. By gain-of and loss-of function models, we revealed that CD56 promotes multiple myeloma cell growth, survival, and adhesion to stromal cells. These protumoral effects are induced by the activation of the RSK2/CREB1 signaling pathway, with increased mRNA and protein levels of the anti-apoptotic genes BCL2 and MCL1. Consequently, the genomic and pharmacological inhibition of RSK2 or CREB1 specifically induced multiple myeloma cell death in CD56-expressing multiple myeloma cells. Finally, we observed that CD56 signaling decreases CRBN expression, reducing responses to lenalidomide. RSK2 or CREB1 inhibition increased CRBN levels and were synergic with lenalidomide in inducing cell death, especially in CD56-expressing multiple myeloma cells. In conclusion, our findings demonstrate that CD56 promotes multiple myeloma cell growth, and pave the way to novel therapies based on targeting CD56, along with the use of CD56 as a predictive biomarker for multiple myeloma therapies. Multiple myeloma is an incurable, genetically heterogeneous disease, without available tailored therapeutic approaches. CD56 signaling promotes multiple myeloma growth and adhesion, by activating CREB1 target genes, MCL1 and BCL2. Inhibition of CREB1 alone or in combination with lenalidomide is an unexplored synthetic lethal approach in CD56-expressing patients with multiple myeloma.


Biological injection therapy with leukocyte-poor platelet-rich plasma induces cellular alterations, enhancement of lubricin, and inflammatory downregulation in vivo in human knees: A controlled, prospective human clinical trial based on mass spectrometry imaging analysis.
To investigate the in vivo biological effects of leukocyte-poor platelet-rich plasma (LpPRP) treatment in human synovial layer to establish the cellular basis for a prolonged clinical improvement. Synovial tissues (n = 367) were prospectively collected from patients undergoing arthroscopic surgery. Autologous-conditioned plasma, LpPRP, was injected into the knees of 163 patients 1-7 days before surgery to reduce operative trauma and inflammation, and to induce the onset of regeneration. A total of 204 patients did not receive any injection. All samples were analyzed by mass spectrometry imaging. Data analysis was evaluated by clustering, classification, and investigation of predictive peptides. Peptide identification was done by tandem mass spectrometry and database matching. Data analysis revealed two major clusters belonging to LpPRP-treated (LpPRP-1) and untreated (LpPRP-0) patients. Classification analysis showed a discrimination accuracy of 82%-90%. We identified discriminating peptides for CD45 and CD29 receptors (receptor-type tyrosine-protein phosphatase C and integrin beta 1), indicating an enhancement of musculoskeletal stem cells, as well as an enhancement of lubricin, collagen alpha-1-(I) chain, and interleukin-receptor-17-E, dampening the inflammatory reaction in the LpPRP-1 group following LpPRP injection. We could demonstrate for the first time that injection therapy using "autologic-conditioned biologics" may lead to cellular changes in the synovial membrane that might explain the reported prolonged beneficial clinical effects. Here, we show in vivo cellular changes, possibly based on muscular skeletal stem cell alterations, in the synovial layer. The gliding capacities of joints might be improved by enhancing of lubricin, anti-inflammation by activation of interleukin-17 receptor E, and reduction of the inflammatory process by blocking interleukin-17.


International palliative care: Middle East experience as a model for global palliative care.
Care for elderly people with life-limiting illness cannot be delivered primarily by geriatricians or palliative care practitioners. The role of these clinicians is to help carers become adept in palliative care medicine. In a culture in which family ties run deep, the offer of palliative care from an outsider may be met with suspicion. The family bond in the Middle East is strong, but the emotional response to terminal illness may push families to request futile treatments, and physicians to comply. When palliative care is well developed and well understood, it provides a viable alternative to such extreme terminal measures.


The Added Value of Family Material in the Discovery of Multiple Sclerosis Genes.
A mutation in a nuclear receptor (NR1H3) gene was detected in a familial, progressive form of multiple sclerosis (PPMS). Further analyses showed a significant association between a common NR1H3 variant in PPMS patients and loss of function for the encoded protein.


Analysis of the Reproductive Outcome of Patients with Cesarean Scar Pregnancy Treated by High-Intensity Focused Ultrasound and Uterine Artery Embolization: A Retrospective Cohort Study.
To investigate risk factors for infertility and recurrent cesarean scar pregnancy (CSP) after previous CSP. A retrospective cohort study (Canadian Task Force classification II-1). University hospital. Between January 2007 and April 2016, a total of 650 patients were included, all diagnosed with CSP and treated by high-intensity focused ultrasound (HIFU) and uterine artery embolization (UAE), followed by suction curettage under hysteroscopic guidance. None. Follow-up of the reproductive outcomes ended in June 2017. A total of 135 of the 650 patients with CSP were involved in the study, among whom 32 became infertile, 78 had an intrauterine pregnancy, and 25 had recurrent CSP after previous CSP. Age (≥35 years; odds ratio [OR], 4.252; p = .002), β-human chorionic gonadotropin (≤5000 mIU/mL; OR, 3.778; p = .011), and longer duration of amenorrhea (>56 days; OR, 2.507; p = .05) were risk factors for infertility. Treatment with UAE (OR, 5.796; p = .003), more abortions (≥4; OR, 2.851; p = .022), and being asymptomatic (OR, 4.175; p = .039) were risk factors for recurrent CSP. There was no statistically significant difference in the subsequent outcomes of pregnant women in the HIFU and UAE groups (p >.05). More attention should be given to subsequent reproductive outcomes after CSP, not only for intrauterine pregnancy, but also for infertility and recurrent CSP. Early diagnosis and treatment of CSP could reduce the risk of infertility and recurrent CSP. HIFU seemed to be superior to UAE in reducing the risk of recurrent CSP. Patients with CSP should adhere to strict contraception if they do not desire more children.


Vulvar manifestations of human papillomavirus infection.
As technology continues to improve, we are increasingly able to detect the presence of HPV in both clinically diseased and histologically and clinically normal vulvar tissue. Unfortunately, the technology for effective treatment of these conditions has not advanced as rapidly. Still, fundamental information about the interaction of HPV with the host cell continues to be collected and will provide us with an increased understanding of the importance of latent infection and its progression to clinical vulvar disease. There are a host of therapeutic modalities available for the treatment of HPV-associated conditions of the vulva, ranging from simple topical medication like podophyllin to sophisticated combination therapies involving carbon dioxide laser destruction in conjunction with interferon. Regrettably, there is no "magic bullet" that is totally effective for any of the HPV-associated vulvar diseases. This makes it more important to weigh the benefits and morbidity of treatment regimens before they are used. Until the success rate of available therapies improves, it is important to have definitive goals for therapy, because it is currently impossible to completely eradicate HPV from tissues. Examples of therapeutic goals might range from removal of visible lesions or treatment of symptomatic areas to treatment of premalignant or malignant lesions. At present, perhaps the most important service we can provide for patients is close follow-up of HPV-associated vulvar diseases in an attempt to prevent the development of invasive malignancy. Regardless of the choice of treatment, the primary objective of the clinician should be to help the patient but to do no harm.


Senior house officers' experience of a six month post in a hospice.
Hospices provide care for patients with a terminal prognosis--a very small number of hospices employ Senior House Officers. A survey was carried out to determine the experiences of SHOs working in hospices and their perceptions of the learning opportunities available. Perceived stress was measured by a Visual Analogue scale and psychological distress by the GHQ 12. Of the 38 posts identified 23 SHOs (60%) responded and the majority had qualified during the last five years. Formal teaching was limited, but experiential teaching was very positively described. Many SHOs described their posts as stressful and cited staff conflict and caring for young patients as particularly stressful. Median stress scores as measured on the VAS was 55 mm (range 0-98 mm). Five respondents 22% scored for identifiable psychological distress on the GHQ 12. Conclusions include the need to acknowledge the important learning opportunities provided within hospices but also the need for consultant staff to be aware of the sources of stress for SHOs and their need for support.


Giant insulinoma in a 15-year-old man: A case report.
Giant insulinomas are extremely rare pancreatic neuroendocrine tumor. A 15-year-old man presenting with acute onset of lightheadedness was found to have serum glucose of 1.5mmol/L. The blood collected from the hypoglycemic episode showed an inappropriately high insulin and C-peptide level. Abdominal computerized tomography showed a 12.5cm well-defined, lobulated hypervascular mass at pancreatic tail, without any evidence of metastasis. En bloc resection with distal pancreatectomy, and splenectomy was successfully performed. The pathological examination confirmed insulinoma, with benign characteristics. Follow-up after the procedure revealed neither hypoglycemic, nor hyperglycemia. We report the youngest case of a giant insulinoma. Despite the size of the tumor, the pathological report confirmed the benign characteristics. However, long-term follow-up is still essential to detect recurrence in the future.


[Total Peroxidase and Catalase Activity of Luminous Basidiomycetes Armillaria borealis and Neonothopanus nambi in Comparison with the Level of Light Emission].
The peroxidase and catalase activities in the mycelium of luminous basidiomycetes Armillaria borealis and Neonothopanus nambi in normal conditions and under stress were compared. An increase in the luminescence level was observed under stress, as well as an increase in peroxidase and catalase activities. Moreover, the peroxidase activity in extracts of A. borealis mycelium was found to be almost one and a half orders of magnitude higher, and the catalase activity more than two orders of magnitude higher in comparison with the N. nambi mycelium. It can be suggested that the difference between the brightly luminescent and dimly luminescent mycelium of N. nambi is due to the content of H2O2 or other peroxide compounds.


Infection of surgical wounds in the maxillofacial region and free flap donor sites with methicillin-resistant Staphylococcus aureus.
We studied the acquisition of methicillin-resistant Staphylococcus aureus (MRSA) in two groups of patients who had free flap reconstructions of defects after excision of lesions in the head and neck. The first group (n=31) was given a 5-day perioperative course of antibiotic prophylaxis (long-term) and the second (n=33) a 24-h course (short-term). MRSA was the main infecting organism. Seventeen of the 64 patients (27%) developed an infection with MRSA, including seven at donor sites of free flaps. All infections were acquired after operation, and delayed healing or discharge in five patients with MRSA and four with infections with other organisms. The median length of hospital admission was greater for patients that acquired MRSA (p=0.005). There were significantly fewer patients with wounds infected by MRSA in the short-term group (4/33 compared with 13/31, p=0.01). We recommend the short-term use of an antibiotic for surgical prophylaxis and the application of cross-infection control measures on the ward.


Pulmonary function and bronchial reactivity in asthmatics during low-level formaldehyde exposure.
This study evaluated whether formaldehyde, at concentrations similar to those found in the indoor environment, could produce adverse effects on the lower airway of 15 asthmatic persons with documented bronchial hyperresponsiveness who were exposed for 90 min in a climate chamber to clean air containing formaldehyde vapor at levels of 0.85 mg/m3, 0.12 mg/m3, and 0.008 mg/m3. No significant changes in forced expiratory volume in 1 sec (FEV1), airway resistance (Raw), specific airway resistance (SRaw), and flow-volume curves could be detected during formaldehyde exposure. Furthermore, histamine challenge tests performed immediately after formaldehyde exposure showed no evidence of changes in bronchial reactivity. No late reactions were registered during the first 14-16 hr after exposure. The results suggest that residential levels of formaldehyde are of minor importance in the emergence of pulmonary symptoms. Discrepancies between the present study and previous data may be due to differences in environmental conditions.


5-day storage of single-donor platelets obtained using a blood cell separator.
Platelets were collected from normal donors via a blood cell separator (Fenwal CS-3000). Platelets were stored initially in two separate 1000-ml bags (average count per bag, 2.3 +/- 0.5 x 10(11)) in 100 ml of autologous plasma for 5 days. Little change in platelet count was noted after 5 days of storage; however, the white cell count fell from 5.1 +/- 1.7 x 10(9) at Time 0 to 3.4 +/- 2.3 x 10(9) per l at Day five. The initial lactate values were 32 +/- 11 mg per dl and rose to 165 +/- 28 mg per/dl by 5 days. Platelet aggregation was impaired both by the collection procedure and during storage: whereas the response to ADP of the donors' platelets before the procedure was 100 percent, samples taken from the product immediately after collection had only a 45 percent response, which fell to 12 percent by Day 5. Aggregation using epinephrine was similarly affected, with a 75 percent response after collection and 0 percent response by Day 5. The plasma beta-thromboglobulin (beta-TG) level was high, both after collection (5.0 micrograms/ml at Time 0) and after storage (11.0 micrograms/ml), indicating a considerable effect of collection on platelet alpha granule release. In vivo recovery of these platelets was very good at 67 +/- 6 percent, with an average survival of 7.3 +/- 1.4 days (multiple hit; n = 4).(ABSTRACT TRUNCATED AT 250 WORDS)


The combined ratios of L-arginine and asymmetric and symmetric dimethylarginine as biomarkers in spontaneously hypertensive rats.
Hypertension and hypertensive end-organ damage have been associated with decreased nitric oxide (NO) bioavailability. Asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) both can inhibit NO availability by competition with L-arginine (L-Arg). However, whether a combined analysis of these 3 parameters can serve as an ideal biomarker of hypertension remains unclear. We measured the plasma and renal levels of L-Arg, ADMA, and SDMA in spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) control rats at 3 stages: 4 weeks old (prehypertensive), 12 weeks old (hypertensive), and 24 weeks old (end-organ damage). The plasma and renal L-Arg/ADMA ratio (AAR) and the ADMA/SDMA ratio (ASR) were computed for all 3 age stages. Our results revealed an ADMA level increase, and an AAR decrease in plasma and kidneys may develop early on, even before the onset of hypertension in 4-week-old SHRs. The renal ADMA level and AAR were restored in SHRs at 24 weeks of age, which might protect SHRs against kidney injury. We found that the plasma AAR is superior to the levels of L-Arg and ADMA in plasma, and it predicted blood pressure and urinary NOx levels. Renal AAR is a strong independent marker of renal dimethylarginine dimethylaminohydrolase (DDAH) activity. The plasma ASR was correlated strongly to blood pressure. However, renal DDAH activity was related to the renal ASR but not the plasma ASR. In conclusion, the AAR and ASR may serve as better markers for disease activity and progression than each individual parameter. Clinical use of these ratios to elucidate the role of ADMA in hypertension awaits further validation.


Day Hospital Treatment as a Missing Link for Single Patients with Posttraumatic Stress Disorder (PTSD): a Preliminary Study.
The aim of this study was to examine the day hospital treatment outcome on severity of clinical manifestations, general neuroticism and coping mechanisms in patients suffering from chronic combat-related PTSD. The sample consisted of 38 consecutive patients admitted to the Day Hospital treatment of PTSD during one year observation period. The average age of the sample was 46.03 years. The patients completed 3 self-report measures upon admission to the hospital and upon discharge: The Mississippi scale for combat-related PTSD (M-PTSD), The Crown-Crisp experiential index (CCEI), and The COPE inventory. There was no significant change in the severity of clinical manifestations of PTSD, general neuroticism and coping mechanisms among the whole sample. However, compared to married participants and participants with children, single participants and those without children reported higher levels of anxiety when admitted to the hospital, but lower levels at discharge. In addition, patients without children reduced their avoidance behavior during the treatment. This preliminary study showed that single patients and those without children may benefit more from the day hospital treatment program. Our findings emphasize the importance of social support in the recovery process of severely traumatized persons, and may assist with the development of more effective therapeutic approaches.


Effect of laser groove treatment on shear bond strength of resin-based luting agent to polyetheretherketone (PEEK).
The mechanical properties of polyetheretherketone (PEEK) are ideally suited for fixed dental prostheses. However, PEEK typically has low adhesion strength to resin-based luting agent. This study assessed the shear bond strength between laser groove treated PEEK and resin-based luting agent. A total of 230 specimens were randomly divided into five groups (n=46): no-treatment, air abrasion treatment, 100μm-deep, 150μm-deep, and 200μm-deep laser groove treatments. The surface roughness was measured, scanning electron microscopy was used to observe the specimen surfaces, and X-ray photoelectron spectroscopy (XPS) was used to analyze the surfaces. Each group was divided into four resin-based luting agent subgroups: Panavia V5, RelyX Ultimate Resin Cement, G-CEM Link Force, and Super-Bond C&B. After the resin-based luting agent was bonded to the specimens, the bond strength was measured using shear tests and the failure modes were assessed by stereomicroscopy. The surfaces were also observed by scanning electron microscopy (SEM) after the shear bond strength measurements. The data were statistically analyzed using a two-way analysis of variance and Tukey's honest significant difference test (α=0.05). The PEEK surface after laser groove treatment groups exhibited the highest mean Ra values. In the XPS analysis, the laser treated PEEK surface exhibited an effective surface composition for bonding with resin-based luting agent. The shear bond strengths for the laser groove treated samples were significantly higher (p<0.05) than those of the no-treatment and air abrasion treatment groups. The shear bond strength between PEEK and resin-based luting agent was substantially improved by laser groove treatment.


Successful treatment of severe ABO antibody-mediated rejection using bortezomib: a case report.
Our program for ABO-incompatible renal transplantation includes antigen-specific immunoadsorption (extracorporeal columns with the A or B trisaccharides), rituximab, and standard maintenance immunosuppression. Anti-A or -B titers ≤ 8 in the indirect antiglobulin test (IAT) against panel A1 or B RBC are acceptable for transplantation. A previously healthy, 15-month-old girl was diagnosed with Wilms' tumor and proteinuria. Denys-Drash syndrome was confirmed. Bilateral nephrectomy was performed. At 3.5 years of age she received an ABO-incompatible renal transplant from her father (A1 to O). The anti-A titers before transplantation were low. She was treated preoperatively with rituximab, immunoadsorption, immunoglobulin and mycophenolate mofetil (MMF). The maintenance immunosuppression protocol included basiliximab, tacrolimus, MMF, and prednisolone. The initial postoperative course was uncomplicated with rapid normalization of serum creatinine. The anti-A titers started to increase on postoperative day 5 (8 NaCl/16 IAT). Despite daily immunoadsorptions the titers rose to 1024 NaCl/1024 IAT on day 9. Renal function deteriorated and hemodialysis was started. A renal biopsy on day 9 showed acute severe antibody-mediated rejection. Additional treatment with bortezomib was given and after 2 doses the titers started to decline, renal allograft function improved and hemodialysis was stopped. On day 21 posttransplant the titers went down, creatinine was 28 μmol/L, and no more immunoadsorptions were performed. By using bortezomib, we were able to successfully reverse a severe ABO antibody-mediated rejection.


Direct access to the cooperative substructure of proteins and the protein ensemble via cold denaturation.
The modern view of protein thermodynamics predicts that proteins undergo cold-induced unfolding. Unfortunately, the properties of proteins and water conspire to prevent the detailed observation of this fundamental process. Here we use protein encapsulation to allow cold denaturation of the protein ubiquitin to be monitored by high-resolution NMR at temperatures approaching -35 degrees C. The cold-induced unfolding of ubiquitin is found to be highly noncooperative, in distinct contrast to the thermal melting of this and other proteins. These results demonstrate the potential of cold denaturation as a means to dissect the cooperative substructures of proteins and to provide a rigorous framework for testing statistical thermodynamic treatments of protein stability, dynamics and function.


Saccade-confounded image statistics explain visual crowding.
Processing of shape information in human peripheral visual fields is impeded beyond what can be expected by poor spatial resolution. Visual crowding, the inability to identify objects in clutter, has been shown to be the primary factor limiting shape perception in peripheral vision. Despite the well-documented effects of crowding, its underlying causes remain poorly understood. Given that spatial attention both facilitates learning of image statistics and directs saccadic eye movements, we propose that the acquisition of image statistics in peripheral visual fields is confounded by eye-movement artifacts. Specifically, the image statistics acquired under a peripherally deployed spotlight of attention are systematically biased by saccade-induced image displacements. These erroneously represented image statistics lead to inappropriate contextual interactions in the periphery and cause crowding.


Autotaxin inhibitor IOA-289 reduces gastrointestinal cancer progression in preclinical models.
Autotaxin (ATX) is a secreted enzyme that converts lysophosphatidylcholine to lysophosphatidic acid (LPA). LPA stimulates cell proliferation and migration and promotes wound repair following tissue damage. ATX levels are directly correlated with stage and grade in several human cancers. Several small molecule ATX inhibitors have been developed in recent years. IOA-289 is a potent ATX inhibitor, developed to treat cancers containing fibrosis. In this study, we tested IOA-289 treatment on different gastrointestinal tract tumor cell lines, in order to evaluate its effects on viability and motility. To determine the effects on cell viability and proliferation of treatment with increasing concentrations of IOA-289, we used the crystal violet assay, a clonogenic assay in matrigel, and we evaluated the inhibitor's effect on formation of 3D spheroids in an in vitro model. The effect of IOA-289 on cell cycle phases was analysed with a redox dye reagent. Cell migration capacity was evaluated by wound healing assay and transwell migration assay. To evaluate the pro-apoptotic effect of the inhibitor, cells were stained with Annexin V and immunofluorescence and flow cytometry analysis were performed. An antibody array was also used, to discriminate, in various samples, the differential expression of 43 proteins involved in the apoptosis pathway. We found that IOA-289 is able to inhibit both growth and migration of gastrointestinal tract tumor cell lines, both in 2D (crystal violet assay) and 3D in vitro models (spheroid formation and clonogenic assay in matrigel). This effect is dose-dependent, and the drug is most effective when administered in FBS-free culture medium. The inhibitory effect on cell growth is due to a pro-apoptotic effect of IOA-289. Staining with FITC-conjugated Annexin V showed that IOA-289 induced a dose-dependent increase in fluorescence following incubation for 24 h, and apoptotic cells were also distinguished in flow cytometry using Annexin/PI staining. The antibody array shows that treatment with IOA-289 causes the increased expression of several pro-apoptotic proteins in all tested cell lines. These results indicate that IOA-289 may be an effective drug for the treatment of tumors of the gastrointestinal tract, particularly those characterized by a high degree of fibrosis.


Anterior transperitoneal approach for treatment of urolithiasis.
An anterior transperitoneal incision was used to remove stones from 20 kidneys in 18 patients (2 bilateral). Our experience with 35 kidneys operated upon by this method (15 previously reported) has been favorable. Use of percutaneous nephropyelostomy tubes to drain obstructed kidneys in septic patients has permitted all procedures to be done under non-emergency conditions. Careful alignment of x-ray equipment and adjustment of exposure technique in the operating room before induction of anesthesia result in consistently good quality preoperative and intraoperative roentgenograms. The transperitoneal approach has afforded good access to the anterior portion of the renal pelvis even in those patients who have had previous operations on the kidney by the flank approach. Additional intra-abdominal procedures done at the same operation have included urinary diversion by ileal loop, revision of previous ileal loop, ureterolithotomy, pyeloureteroplasty, partial nephrectomy, cholecystectomy and appendectomy. The procedures done with the patients in the supine position have been well tolerated even in patients with serious coincident medical diseases.


Lymph node metastasis in bladder cancer.
We reviewed the literature on nodal staging in patients with bladder cancer treated with radical cystectomy and lymphadenectomy. Fractionating the lymph node specimen significantly increases the node count, whereas results are contradictory as to whether that increase improves detection of positive nodes. Pathoanatomic data indicate that extending lymph node dissection to the aortic bifurcation improves nodal staging. That approach might be beneficial, especially in cases of T3/T4a tumours, which more often have lymph node metastases above the iliac bifurcation as compared to less advanced tumours. In node-negative patients, extended lymph node dissection probably removes undetected micrometastases and thereby increases disease-free survival. Four studies suggested that a minimum of 8, 10, 10-14, and 16 nodes must be removed, to improve survival, and in another investigation aortic bifurcation was proposed as the upper limit for dissection. Some patients with positive nodes can be cured by surgery alone, even those with gross adenopathy. There is no evidence that extended lymphadenectomy increases surgery-related morbidity. The TNM classification is apparently insufficient for stratifying node-positive patients because several larger cystectomy series could not verify differences in survival between N groups. Fractionating the lymphadenectomy specimen increases the lymph node count. In node-negative patients, more meticulous and extended lymph node dissection (8-16 nodes or to the aortic bifurcation) probably improves disease-free survival by removing undetected micrometastases. Patients with positive lymph nodes should also be offered radical cystectomy.


In situ hybridization of oxytocin messenger RNA: macroscopic distribution and quantitation in rat hypothalamic cell groups.
Oxytocin mRNA was detected in the rat hypothalamus by in situ hybridization to a single stranded 35S-labelled DNA probe and the distribution of oxytocin mRNA-containing cell groups was studied at the macroscopic level. Specificity of hybridization was confirmed by comparison to vasopressin mRNA hybridization in parallel tissue sections. Cell groups containing oxytocin mRNA were confined to a set of hypothalamic cell groups, i.c. the supraoptic, paraventricular, anterior commissural nuclei, nucleus circularis and scattered hypothalamic islets. These cell groups displayed similar densities of autoradiographic signals indicating that the oxytocin gene is expressed at approximately the same average level at these various sites.


Ubiquitin ligases and a processive proteasome facilitate protein clearance during the oocyte-to-embryo transition in Caenorhabditis elegans.
The ubiquitin-mediated degradation of oocyte translational regulatory proteins is a conserved feature of the oocyte-to-embryo transition. In the nematode Caenorhabditis elegans, multiple translational regulatory proteins, including the TRIM-NHL RNA-binding protein LIN-41/Trim71 and the Pumilio-family RNA-binding proteins PUF-3 and PUF-11, are degraded during the oocyte-to-embryo transition. Degradation of each protein requires activation of the M-phase cyclin-dependent kinase CDK-1, is largely complete by the end of the first meiotic division and does not require the anaphase-promoting complex. However, only LIN-41 degradation requires the F-box protein SEL-10/FBW7/Cdc4p, the substrate recognition subunit of an SCF-type E3 ubiquitin ligase. This finding suggests that PUF-3 and PUF-11, which localize to LIN-41-containing ribonucleoprotein particles, are independently degraded through the action of other factors and that the oocyte ribonucleoprotein particles are disassembled in a concerted fashion during the oocyte-to-embryo transition. We develop and test the hypothesis that PUF-3 and PUF-11 are targeted for degradation by the proteasome-associated HECT-type ubiquitin ligase ETC-1/UBE3C/Hul5, which is broadly expressed in C. elegans. We find that several GFP-tagged fusion proteins that are degraded during the oocyte-to-embryo transition, including fusions with PUF-3, PUF-11, LIN-41, IFY-1/Securin, and CYB-1/Cyclin B, are incompletely degraded when ETC-1 function is compromised. However, it is the fused GFP moiety that appears to be the critical determinant of this proteolysis defect. These findings are consistent with a conserved role for ETC-1 in promoting proteasome processivity and suggest that proteasomal processivity is an important element of the oocyte-to-embryo transition during which many key oocyte regulatory proteins are rapidly targeted for degradation.


γ-Oryzanol ameliorates fine dust-induced premature endothelial senescence and dysfunction via attenuating oxidative stress.
Various cardiovascular diseases are associated with endothelial senescence, and a recent study showed that fine dust (FD)-induced premature endothelial senescence and dysfunction is associated with increased oxidative stress. The aim of the present study was to investigate protective effect of rice bran extract (RBE) and its major component of γ-Oryzanol (γ-Ory) against FD-induced premature endothelial senescence. Porcine coronary artery endothelial cells (PCAECs) were treated with FD alone or with RBE or γ-Ory. Senescence-associated β-galactosidase (SA-β-gal) activity, expression of cell cycle regulatory proteins, and oxidative stress levels were evaluated. The results indicated that SA-β-gal activity in the FD-treated PCAECs was attenuated by RBE and γ-Ory. Additionally, γ-Ory inhibited FD-induced cell cycle arrest, restored cell proliferation, and reduced the expression of cell cycle regulatory proteins. γ-Ory also inhibited oxidative stress and prevented senescence-associated NADPH oxidase and LAS activity in FD-exposed ECs suggesting that γ-Ory could protect against FD-induced ECs senescence and dysfunction.


Clinical Variables as Indicative Factors for Endoscopy in Adolescents with Esophageal Atresia.
Gastro-esophageal reflux disease (GERD) occurs frequently in patients operated for esophageal atresia (EA). Longstanding esophagitis may lead to dysphagia, strictures, columnar metaplasia, and dysplasia with an increased risk of adenocarcinoma. Are clinical factors and non-invasive assessments reliable indicators for follow-up with endoscopy? A follow-up study with inclusion of EA adolescents in Norway born between 1996 and 2002 was conducted. Clinical assessment with pH monitoring, endoscopy with biopsies, along with interviews and questionnaires regarding gastroesophageal reflux disease (GERD) and dysphagia were performed. We examined 68 EA adolescents. 62% reported GERD by interview, 22% by questionnaire. 85% reported dysphagia by interview, 71% by questionnaire. 24-hour pH monitoring detected pathological reflux index (RI) (>7%) in 7/59 (12%). By endoscopy with biopsy 62 (92%) had histologic esophagitis, of whom 3 (4%) had severe esophagitis. Gastric metaplasia was diagnosed in twelve (18%) adolescents, intestinal metaplasia in only one (1.5%). None had dysplasia or carcinoma. Dysphagia and GERD were statistically correlated to esophagitis and metaplasia, but none of the questionnaires or interviews alone were good screening instruments with high combined sensitivity and specificity. A compound variable made by simply taking the mean of rescaled RI and dysphagia by interview showed to be the best predictor of metaplasia (85% sensitivity, 67% specificity). The questionnaires and interviews used in the present study were not good screening instruments alone. However, combining dysphagia score by interview and RI may be helpful in assessing which patients need endoscopy with biopsy at each individual follow-up examination. Level II prognostic study.


Amyotrophic Lateral Sclerosis After Exposure to Manganese from Traditional Medicine Procedures in Kenya.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by motor neuron loss and widespread muscular atrophy. Despite intensive investigations on genetic and environmental factors, the cause of ALS remains unknown. Recent data suggest a role for metal exposures in ALS causation. In this study we present a patient who developed ALS after a traditional medical procedure in Kenya. The procedure involved insertion of a black metal powder into several subcutaneous cuts in the lower back. Four months later, general muscle weakness developed. Clinical and electrophysiological examinations detected widespread denervation consistent with ALS. The patient died from respiratory failure less than a year after the procedure. Scanning electron microscopy and X-ray diffraction analyses identified the black powder as potassium permanganate (KMnO4). A causative relationship between the systemic exposure to KMnO4 and ALS development can be suspected, especially as manganese is a well-known neurotoxicant previously found to be elevated in cerebrospinal fluid from ALS patients. Manganese neurotoxicity and exposure routes conveying this toxicity deserve further attention.


Feed intake of the sow and playful creep feeding of piglets influence piglet behaviour and performance before and after weaning.
Creep feed intake is variable and may be partly homeostatically and exploratory driven. We studied effects of maternal feed restriction and a 'play-feeder' on piglet behaviour and performance. 37 Litters received creep feed in a conventional (CON) or play-feeder (PL) and their sows were full-fed (FF) or restrictedly-fed (RES). Eaters were determined via rectal swabs. At weaning (d24) four piglets from the same treatment were grouped (n = 36 pens). RES hindered piglet growth by 41 g/d and enhanced time eating, creep feed intake and percentage of eaters at weaning versus FF. RES-PL had the largest proportion of moderate and good eaters. PL stimulated feeder exploration and attracted more piglets to the feeder than CON. Post-weaning, RES increased exploratory behaviours, feed intake between d0-5, and growth between d0-2, and reduced body lesions between d0-2 (within CON), drinking and ear biting. PL increased ingestive behaviours, feed intake and growth between d0-15, and BW at d15 post-weaning by 5%. PL also lowered the prevalence of watery diarrhoea, number of body lesions and piglets with ear (within FF) and tail (within RES) damage at d15 post-weaning. Treatments did not affect FCR. To conclude, RES and particularly PL (broader and for longer) result in less weaning-associated-problems.


Efficacy and safety of pharmacotherapy for refractory or unexplained chronic cough: a systematic review and network meta-analysis.
Refractory chronic cough (RCC) has a significant impact on patient's health-related quality of life and represents a challenge in clinical management. However, the optimal treatment for RCC remains controversial. This study aimed to investigate and compare the efficacy and safety of the current pharmacological therapeutic options for RCC. A systematic review was performed by searching PubMed, Web of Science, Embase, and Ovid databases from January 1, 2008 to March 1, 2023. All randomised control trials (RCTs) reporting outcomes of efficacy or/and safety were included in the Bayesian network meta-analysis. Here, we compared the effects on Leicester Cough Questionnaire (LCQ), Visual Analogue Scale (VAS), and objective cough frequency of patients with RCC. Besides, we also compared the incidence of adverse events (AEs) for analysis of safety. PROSPERO registration: CRD42022345940. 19 eligible RCTs included 3326 patients and 7 medication categories: P2X3 antagonist, GABA modulator, Transient Receptor Potential (TRP) modulator, NK-1 agonist, opioid analgesic, macrolide, and sodium cromoglicate. Compared with placebo, mean difference (MD) of LCQ and 24 h cough frequency for P2X3 antagonist relief were 1.637 (95% CI: 0.887-2.387) and -11.042 (P = 0.035). Compared with placebo, effect sizes (MD for LCQ and cough severity VAS) for GABA modulator were 1.347 (P = 0.003) and -7.843 (P = 0.003). In the network meta-analysis, gefapixant is the most effective treatment for patients with RCC (The Surface Under the Cumulative Ranking Curves (SUCRA) is 0.711 in LCQ, 0.983 in 24 h cough frequency, and 0.786 in cough severity VAS). Lesogaberan had better efficacy than placebo (SUCRA: 0.632 vs. 0.472) in 24 h cough frequency. Eliapixant and lesogaberan had better efficacy than placebo in cough severity VAS. However, TRP modulator had worse efficacy than placebo. In the meta-analysis of AEs, the present study found P2X3 antagonist had a significant correlation to AEs (RR: 1.129, 95% CI: 1.012-1.259), especially taste-related AEs (RR: 6.216, P < 0.05). In this network meta-analysis, P2X3 antagonist showing advantages in terms of efficacy is currently the most promising medication for treatment of RCC. GABA modulator also showed potential efficacy for RCC but with AEs of the central system. Nevertheless, the role of TRP modulator needed to be revisited. Further research is needed to determine the potential beneficiary population for optimizing the pharmacological management of chronic cough. National Natural Science Foundation of China (81870079), Guangdong Science and Technology Project (2021A050520012), Incubation Program of National Science Foundation for Distinguished Young Scholars (GMU2020-207).


Establishment, characterization, and response to cytotoxic and radiation treatment of three human melanoma cell lines.
Three human melanoma cell lines were derived from tumor specimens and established in culture. CAL 1 originated from a bone marrow metastasis and CAL 4 and CAL 7 were derived from solid tumor fragments. CAL 1 and CAL 7 were cloned before establishment. Ultrastructural and chromosome analysis were carried out along with the response to nine chemotherapeutic agents at various concentrations. Survival curves after irradiation were also plotted. The uncloned cell line, CAL 4, displayed some differences from the other two cell lines as regards ploidy and response to chemotherapy. Greater spread of chromosome numbers were observed with this cell line, which contained both hypoploid and a hyperploid modal numbers. All three cell lines showed a relatively high extrapolation number after irradiation, suggesting that inherent cellular properties may be partly responsible for the clinical radioresistance of malignant melanomas.


Environmental impacts, human health, and energy consumption of nitrogen management for maize production in subtropical region.
Over-application of fertilizers could not improve crop yield and agronomic efficiency, but result in increasing nitrogen (N) surplus and adverse effects on the ecosystem sustainability. Although some previous studies have addressed one or a few environmental aspects in crop production, an integrated assessment for the effects of N fertilizer on multiple environmental impacts, and the optional steps of normalization and weighting is required. A consecutive 2-year plot-based field experiment was conducted with five N fertilizer levels (0, 90, 180, 270, and 360 kg N ha-1) in maize production at three sites in Southwest China, to evaluate the environmental performance and sustainability through joint use of life cycle assessment (LCA) and energy consumption analysis. Results demonstrated that the optimal N rate (180 kg N ha-1) showed greater potential for maintaining high yield (achieved 86% of the yield potential) and reducing the global warming (- 31%), acidification (- 47%), eutrophication (- 44%) compared to farmers' practice, and energy depletion potentials, by reducing pollutants emission during the production and transportation of N fertilizer and Nr losses at farm stage. Optimal N treatment indirectly reduced the land use, life-cycle human toxicity, aquatic eco-toxicity, and terrestrial eco-toxicity potentials by improving grain yield and agronomic efficiency. In addition, the optimal N treatment reduced the energy consumption by enhancing the energy use efficiency (EUE) (+ 74%) and reducing non-renewable energy form (- 45%) than the farmer's practice. This study will provide comprehensive information for both scientists and farmers involved in maize production and N management in subtropical region.


Trichothecene resistance in wheat: Development of molecular markers for PDR-type ABC transporter genes.
Infection withFusarium graminearum andF. culmorum not only causes severe yield and quality losses, the most relevant concern is the contamination of cereal foods and feeds with trichothecenes (e.g. deoxynivalenol, DON). The ability to synthesize trichothecenes has been shown to be a virulence factor ofF. graminearum on wheat and, on the other hand, toxin resistance is most likely an important component of field resistance. Our hypothesis is that pleiotropic drug resistance mediated by PDR-type ABC transporter proteins (acting as membrane located drug efflux pumps) is a relevant mechanism of DON resistance not only in yeast but also in wheat. Goal of this project is the development of molecular markers for this gene family for use in marker-assisted plant breeding programs. The technical difficulties caused by the large size of the PDR-family are discussed.


Intracellular calcium waves generated by Ins(1,4,5)P3-dependent mechanisms.
Cellular oscillations of cytosolic free Ca2+ ([Ca2+]i) have been observed in many cell types in response to cell surface receptor agonists acting through inositol 1,4,5-trisphosphate (InsP3). In a number of cases where appropriate spatial and temporal resolution have been used to examine these [Ca2+]i oscillations, they have been found to be organized as repetitive waves of Ca2+ increase that propagate through the cytosol of individual cells. In some cases Ca2+ waves also occur as a single pass through stimulated cells. This review discusses the factors underlying the spatial organization of [Ca2+]i signals in the form of Ca2+ waves. In addition, potential mechanisms for the initiation and subsequent propagation of these Ca2+ waves are described.


An interesting case of whitish pleural effusion: think beyond the obvious.
A whitish pleural fluid calls for further biochemical and microbiological investigations beyond routine pleural fluid analysis as it decides the aetiology and management. https://bit.ly/3GwGpzS.