Indolicidin action on membrane permeability: carrier mechanism versus pore formation.
Indolicidin, a 13-residue cationic peptide with extremely high tryptophan content, exhibits broad-spectrum antimicrobial as well as hemolytic activity. To gain insight into the mechanism of indolicidin action on membrane permeability, liposome leakage induced by this peptide was studied by using various probes with vesicles of different lipid compositions. In liposomes containing negatively charged lipids, indolicidin induced rather unselective permeabilization. By contrast, the peptide appeared to be selective in provoking leakage of neutral, egg phosphatidylcholine (PC) liposomes: it effectively induced the release of negatively charged fluorescent dyes, carboxyfluorescein (CF), calcein and sulforhodamine B, but was unable to induce the leakage of a neutral compound, glucose, and that of positively charged doxorubicin. Moreover, organic anions, such as fatty acids, were found to suppress the indolicidin-induced CF leakage of egg PC liposomes. Based on these results, we concluded that indolicidin facilitates the dye release from uncharged lipid vesicles not by formation of membrane pores as it is generally accepted for the majority of antimicrobial peptides but rather via translocation of dye molecules across the membrane in the form of dye-peptide complexes, i.e. indolicidin operates as an organic anion carrier. This conclusion was supported by observing the formation of complexes between indolicidin and pyrenebutyrate in solution. The indolicidin analog having only one arginine was ineffective in pyrenebutyrate binding and CF transport. The mode of action proposed here for indolicidin can be related to that previously postulated for oligoarginine derivatives which are able to carry organic anions across liposomal and bulk phase membranes [Sakai N. & Matile S. J. Am. Chem. Soc. 2003, 125:14348-14356]. The newly identified mechanism of peptide ionophoric activity in uncharged lipid membranes may be involved in hemolytic action of indolicidin via induction of plasma membrane permeability for important anionic metabolites which disturbs regulation of osmotic balance ultimately leading to erythrocyte membrane rupture.


Proteome changes of fibroblasts and endothelial cells upon incubation with human cytomegalovirus subviral Dense Bodies.
Human cytomegalovirus (HCMV) is a pathogen of high medical relevance. Subviral Dense Bodies (DB) were developed as a vaccine candidate to ameliorate the severe consequences of HCMV infection. Development of such a candidate vaccine for human application requires detailed knowledge of its interaction with the host. A comprehensive mass spectrometry (MS)- based analysis was performed regarding the changes in the proteome of cell culture cells, exposed to DB.


Prediction of Technical Failure of Inframalleolar Angioplasty in Patients with Chronic Limb Threatening Ischaemia.
This study sought to: determine anatomically evaluated predictors of the technical failure of inframalleolar angioplasty (IMA), develop a predictive model for unsuccessful IMA, and investigate the effect of IMA on clinical outcomes in patients with chronic limb threatening ischaemia (CLTI). This single centre retrospective observational study enrolled 159 patients with CLTI who underwent IMA for de novo occluded lesions between November 2017 and May 2021. These patients were divided into two groups: the Failed IMA group (n = 62) and the Successful IMA group (n = 97). In multivariable analysis, no target vessel outflow (OR 39.8, 95% CI 10.7 - 148, p < .001), medial artery calcification (MAC) grade (OR 4.91, 95% CI 1.40 - 17.3, p = .010), and occluded pedal arch (OR 5.2, 95% CI 1.2 - 22.7, p = .030) were identified as independent predictors of IMA technical failure. The risk prediction model had an area under the receiver operating characteristic curve (AUC) of 0.93; after bootstrapping adjustment for optimism, this value represented a corrected AUC of 0.95. The patients in the Successful IMA group had a significantly higher proportion of wound healing at 12 months than those in the Failed IMA group (log rank p = .030). IMA technical failure was associated with a significant change in the proportion of wound healing (HR 0.59, 95% CI 0.37- 0.94, p = .030). No target vessel outflow, MAC grade, and occluded pedal arch were independent predictors of IMA technical failure. Additionally, successful IMA was associated with better wound healing outcomes at 12 months. Furthermore, a model incorporating these three predictors precisely predicted IMA technical failure.


Types of nerve terminals in fetal and neonatal rabbit myocardium.
With the use of electron microscopy 4 types of axonal profiles were observed in the developing myocardium of rabbits: 1) adrenergic axons which contained mainly small dense-core vesicles and which presumably can store 5-hydroxy-dopamine; 2) cholinergic axons which contained small clear synaptic vesicles and which were acetylcholinesterase-positive; 3) axons which contained large vesicles filled with moderately electron-dense material and which resembled purinergic axons; and 4) profiles filled with mitochondria, vesicles of various sizes, lysosome-like bodies, and microtubules and which resembled sensory terminals.


Single-agent docetaxel in patients with platinum-refractory metastatic or recurrent squamous cell carcinoma of the head and neck (SCCHN).
The objective of this retrospective study was to investigate the efficacy and tolerability of single-agent docetaxel in patients with platinum-refractory squamous cell carcinoma of the head and neck (SCCHN). Platinum-refractory disease was defined as cancer with documented tumor progression during platinum-based treatment or recurrence within 6 months after platinum-based chemoradiotherapy. Patients fulfilling the following criteria were enrolled: histologically confirmed SCCHN, excluding nasopharyngeal cancer; measurable metastatic lesions as assessed by the Response Evaluation Criteria in Solid Tumors (RECIST); and platinum-refractory disease. Docetaxel (60 mg/m2) was administered every 3-4 weeks and continued unless there was evidence of disease progression or unacceptable toxicity. Twenty patients were recruited. Overall response rate was 10% (2/20) and tumor control rate was 25% (5/20). Median progression-free and median overall survival times were 1.7 and 4.6 months, respectively. The most common hematological toxicities were leucopenia (grade 4: 35%) and neutropenia (grade 4: 30%). Grade 3 febrile neutropenia and grade 3 mucositis (functional/symptomatic) each occurred in two patients (10%). One fatal bleeding occurred during this treatment, however, the relation between this event and docetaxel was unlikely. Median inpatient period during treatment was 5.4 days (range, 0-50 days). A single-agent docetaxel regimen appeared to offer an acceptable clinical profile in patients with platinum-refractory SCCHN.


Effects of some commonly used Saudi folk herbal medications on the metabolic activity of CYP2C9 in human liver microsomes.
To investigate the potential effects of eleven of the most commonly used Saudi folk herbal medications on the metabolic activity of CYP2C9 in human liver microsomes. CYP2C9-mediated 4'-hydroxylation of tolbutamide (TB) to 4'-hydroxytolbutamide (4-OH-TB) was utilized to assess the metabolic activity of CYP2C9. In the present study, an initial screening of the eleven herbs was carried out by incubating TB with microsomes and NADPH in absence or presence of a fixed concentration (25 μg/ml) of alcoholic extracts of different herbs and the metabolite formed was measured by HPLC. Herbs that showed significant effects were further investigated at a lower range of concentration. Among the investigated herbal extracts, only aniseed and curcuma showed statistically significant effects on the formation of 4-OH-TB in human liver microsomes. Curcuma produced a potent inhibition on the metabolite formation and its maximum (about 45% inhibition) was observed at the highest extract concentrations (10 and 25 μg/ml). On the other hand, aniseed significantly activates the formation of 4-OH-TB and the maximum activation (about 55%) was observed at 2.5 μg/ml of aniseed extract. The results of this study have shown that alcoholic extracts of curcuma and aniseed were capable of inhibiting and activating; respectively, the CYP2C9-mediated 4-OH-TB formation in human liver microsomes, suggesting that these herbs have the potential to interact with CYP2C9 drug substrates. None of the other nine investigated herbs was able to produce any statistically significant effect.


Retinopathy of prematurity and pulse oximetry: a national survey of recent practices.
To determine if practices related to the use of pulse oximetry in the first 2 weeks following birth and after 2 weeks of age have a relationship to the rate of retinopathy of prematurity (ROP) and retinal ablation surgery in infants < or =1500 g. A questionnaire was mailed in July 2001 to 318 neonatal intensive care units (NICUs) in the United States and information was collected regarding SpO2 guidelines and the rate of both severe ROP and retinal ablation surgery. A total of 142 surveys were returned (45%). In all, 87% of the NICUs had SpO2 guidelines, and 60% of these centers maintained a different range of SpO2 for infants < or = or >2 weeks of age. The range of SpO2 was 82 to 100% with an average minimum (min) and maximum (max) of 89 and 95%, respectively. In the NICUs with an SpO2 max of >98% in the first 2 weeks following birth, the rate of retinal ablation surgery was 5.5 vs 3% in those units with a max SpO2 >98% (p<0.05). After 2 weeks of age, the rate of retinal ablation surgery was 3.3% when max SpO2 was >92 vs 1.3% when the max SpO2 was < or =92% (p<0.00001). The rate of > or =stage 3 ROP after 2 weeks of age was 5.5% when max SpO2 was >92 vs 2.4% when max SpO2 was < or =92% (p<0.0005). NICUs in the US today have a wide range of SpO2 guidelines. The results of this survey show a "gradient of risk" towards less retinal ablation surgery when the max SpO2 is <98% in the first 2 weeks following birth (p<0.05). There was a statistically significant lower rate of > or =stage 3 ROP and retinal ablation surgery when the max SpO2 was < or =92% after the first 2 weeks of age. A randomized, controlled trial is needed to establish a safe upper limit of SpO2 in the premature infant at risk for developing ROP.


UPARSE: highly accurate OTU sequences from microbial amplicon reads.
Amplified marker-gene sequences can be used to understand microbial community structure, but they suffer from a high level of sequencing and amplification artifacts. The UPARSE pipeline reports operational taxonomic unit (OTU) sequences with ≤1% incorrect bases in artificial microbial community tests, compared with >3% incorrect bases commonly reported by other methods. The improved accuracy results in far fewer OTUs, consistently closer to the expected number of species in a community.


A matched and controlled longitudinal cohort study of dehydrated human amniotic membrane allograft sheet used as a wraparound nerve bundles in robotic-assisted laparoscopic radical prostatectomy: a puissant adjunct for enhanced potency outcomes.
We sought to explore the potency outcomes in two systematically controlled, non-randomized, matched, homogenous patient cohorts, which either underwent intervention (INT) with placement of dehydrated human amniotic membrane (dHAM) around nerve bundles (NVB) during robotic-assisted laparoscopic radical prostatectomy (RALP) or did not (CON). It is hypothesized that dHAM use would lead to better potency outcomes. 1400 eligible informed, consented patients underwent full bilateral nerve-sparing RALP by a single surgeon, wherein 700 patients had dHAM allograft wrapped around the NVB and 700 did not. Groups were matched demographically, clinically, and biochemically. Potency was defined as the ability to have satisfactory penetrative intercourse > 50% of time with SHIM score of ≥ 17 with or without of phosphodiesterase-5 inhibitors. A retrospective matched longitudinal cohort study was performed at 1 year. The first noticeable erection sufficient enough for a satisfactory penetrative intercourse was significantly earlier (p < 0.01; 34.6 ± 3.6 days), whereas the decrease in SHIM score was lower (4.27 ± 0.14 days) in INT. Binary logistic regression demonstrated that INT was an independent significant (p < 0.001) predictor of achieving potency at 1 year, such that INT was 3.86 times (95% CI 2.43-6.13) more likely to achieve potency in the same period when compared with CON. Chi square analysis demonstrated that recovery of potency in INT was better (p < 0.05) in every quarter compared to CON. A higher (p < 0.005) percentage (93.1%) of INT regained potency versus CON (87.1%) at 1 year.


Photosynthetic pathways and the ecological distribution of the chenopodiaceae in Isreal.
Fifty-four species of the Chenopodiaceae in Israel were examined for their anatomical features, δ13C values, habitat and phytogeographical distribution. 17 species have δ13C values between -20‰ and -30‰and non-Kranz anatomy (NK) and are therefore considered as C3 plants. 37 species have δ13C values between -10‰ and -18‰ and Kranz or C4-Suaeda type anatomy and are therefore considered as C4 plants. Some C4 plants have leaf structure which seems to be intermediate between the Kranz and the C4-Suaeda type of leaf anatomy.The segregation of the species into photosynthetic groups shows tribal and phytogeographical grouping. Most of the C3 Chenopods are either mesoruderal plants or coastal halophytes, with a distribution area which covers the Euro-Siberian as well as the Mediterranean phytogeographical regions. The C4 Chenopods are mainly desert or steppe xerohalophytes with a distribution area which includes the Saharo-Arabian and/or Irano-Turanian phytogeographical regions.


Gastroesophageal reflux and atrial fibrillation: is there any correlation?
Atrial fibrillation (AF), gastroesophageal reflux disease (GERD) and hiatal hernias are commonly seen in clinical practice. GERD and hiatal hernias have been proposed to be a possible cause of AF. In this paper, we will briefly review GERD, AF and hiatal hernias, consider the available literature covering the association between these diseases and provide further insight into the topic in general.


Termitomyces pakistanensis, a new mushroom species from Pakistan based on scanning electron microscopy and ITS-rDNA barcoding.
Termitomyces pakistanensis sp. nov. is a member of an edible genus generally distributed in Asia and Europe. This species has been described as new species based on its different morphology, and scanning electron microscopy (SEM) of basidiospores. The novelty and degree of endemism is confirmed by analyzing the genetic variation of the internal transcribed spacer regions (ITS1-5.8 S-ITS2) of the ribosomal DNA gene, a universal fungal marker. The evolutionary affinities of new species is also evaluated with Asian and European species by phylogenetic analysis based on ITS sequences. In our phylogenetic analysis, this genus is found monophyletic comprising of two monophyletic sub clades: Clade I, Microcarpus, with small sized fruiting bodies generally less than 5 cm without pseudorrhiza and Clade II, Macrocarpus, with large sized fruiting bodies generally more than 5 cm having pseudorrhiza. All collections of Pakistani species clustered independently in Microcarpus clade showing their endemic genetic makeup as it is clustering independently. A comprehensive description, photographs of the basidiocarps and Scanning electron microscopy (SEM) micrographs of spores are provided. RESEARCH HIGHLIGHTS: It has a new species from Pakistan to world based on the scanning electron microscopy and further confirmed by DNA barcoding. The exact shape and size of basidiospores of this novel species is first time introduced by using SEM analysis. This genus is rarely described from Pakistan. This paper has introduced a two clade, Microcarpus and macrocarpus, in the world for this genus.


Third or more repeat hepatectomy for recurrent hepatocellular carcinoma.
We sought to evaluate the surgical results of third or more repeat hepatectomy for recurrent hepatocellular carcinoma (HCC). The role of repeat hepatectomy for recurrent HCC, especially in cases with third or more repeat hepatectomy, is controversial. We performed A retrospective, cohort study to analyze the surgical results of repeat hepatectomy performed at a single medical center from 1989 to 2011. A total of 1,000 hepatectomies for HCC were divided into 3 groups: A first hepatectomy group (n = 791), second hepatectomy group (n = 163), and third or more hepatectomy group (n = 46). Operative results and patient prognoses were compared among the 3 groups. There were no differences in early surgical results such as mortality and morbidity among the 3 groups. The 5-year survival rates after the first, second, and third or more hepatectomy were 67%, 60%, and 43%, respectively (P = .1913). There was a significant difference in disease-free survival among the 3 groups, and the 5-year disease-free survival rates after first, second, and third or more hepatectomy were 37%, 29%, and 18%, respectively (P = .0169). Third or more repeat hepatectomy for recurrent HCC was performed safely and associated with relatively long-term survival. Third or more repeat hepatectomy for recurrent HCC seems justified, but high rate of HCC recurrence remains a problem.


miR-145 suppresses cell invasion in hepatocellular carcinoma cells: miR-145 targets ADAM17.
miR-145 is a candidate tumor suppressor miRNA. However, it is unknown whether miR-145 is involved in the invasion of hepatocellular carcinoma (HCC). Therefore, we aimed to explore the effect and mechanism of miR-145 in the control of HCC cell invasion. HCC cell invasion was evaluated by transwell assays after transfection with pre-miR-145 or anti-miR-145. A luciferase reporter assay was used to determine whether a disintegrin and metalloprotease 17 (ADAM17) were a target of miR-145. The levels of miR-145 and ADAM17 mRNA were detected by a real-time polymerase chain reaction assay, and the level of ADAM17 protein was measured by western blot analysis. Pearson's correlation test was used to assess the correlation between ADAM17 mRNA expression and miR-145 expression in 20 HCC tissue samples. miR-145 was significantly downregulated in HCC tissues and cell lines. The loss of miR-145 expression was associated with the tumor-node-metastasis stage, vascular invasion and intrahepatic metastasis. The overexpression of miR-145 was able to suppress tumor MHCC-97H cell invasion, whereas the knockdown of miR-145 expression induced SMMC-7721 cell invasion. We demonstrated that miR-145 bound directly to the 3'-untranslated region of ADAM17 and inhibited the expression of ADAM17. The knockdown of ADAM17 in SMMC-7721 cells could partially reverse the effects of anti-miR-145. miR-145 expression was inversely associated with ADAM17 expression in 20 HCC tissue specimens. Our findings indicate that miR-145 could inhibit HCC cell invasion by regulating the expression of ADAM17.


Association of PON1 genotype and haplotype with susceptibility to coronary artery disease and clinical outcomes in dual antiplatelet-treated Han Chinese patients.
The aim of this study was to evaluate the association of PON1 genetic variants with the susceptibility to coronary artery disease (CAD) and with the clinical endpoints in aspirin and clopidogrel (dual antiplatelet therapy)-treated Han Chinese patients with CAD after percutaneous coronary intervention (PCI). A total of 538 Han Chinese patients undergoing PCI and receiving dual-antiplatelet therapy were sequentially recruited to the study and followed for up to 1 year. Healthy controls (n = 539) were enrolled during the same period. All study participants were genotyped for five genetic variants in PON1 and the cytochrome P450 2C19*2 mutation (CYP2C19*2). The effect of genetic variants on disease risk and clinical outcome of major adverse cardiac events (MACE) within 1 year or bleeding within 6 months was assessed. CYP2C19*2 was associated with a higher risk of MACE (adjusted P = 0.0098), but a lower risk of bleeding events (adjusted P = 0.0016). The PON1 Q192R polymorphism was significantly associated with a lower risk of bleeding events [odds ratio (OR) 0.61, 95% confidence interval (CI) 0.43-0.87, adjusted P = 0.0066). The haplotype bearing the PON1 -126C allele was associated with a higher risk to CAD (OR 1.48, 95% CI 1.04-2.09, P = 0.029) and a higher risk of bleeding events (OR 1.68, 95% CI 1.10-2.56, P = 0.017) compared to the most frequent haplotype. The transcription activity of haplotype p-162A-126C-108C in the PON1 promoter was 2.6-fold higher than that of the most frequent haplotype (p-162G-126G-108T). Based on these results, we suggest that the haplotype-bearing PON1 -126C allele contributes to the disease risk and the risk of bleeding events in dual antiplatelet-treated CAD patients after PCI.


Bulbar encephalitis complicating trigeminal zoster in the acquired immune deficiency syndrome.
A 30-year-old homosexual man with the acquired immune deficiency syndrome and a recent history of zoster involving the mandibular division of the right trigeminal nerve was found at autopsy to have a predominantly demyelinating lesion restricted to the ipsilateral spinal trigeminal tract and nucleus. Cowdry A inclusions were readily apparent in the nuclei of numerous glia and isolated neurons, and inclusion-bearing cells were immunoreactive with an antiserum to the varicella zoster virus (VZV). This represents the first demonstration that zoster-associated trigeminal encephalitis is the result of brainstem invasion by VZV. The restricted topography of this lesion implicates axonal transport in its pathogenesis.


Unrecognized small vesicovaginal fistula as a cause of persistent urinary incontinence.
Three cases of persistent urinary incontinence from unrecognized small vesicovaginal fistula after abdominal hysterectomy for benign gynecologic disease are presented. Visualization of urine in the posterior fornix was suggestive; diagnosis was confirmed by methylene blue test and cystoscopy. Two patients had undergone interval Marshall-Marchetti-Krantz operations for suspected stress-related urinary incontinence. An extensive literature survey confirms that the vesicovaginal fistula were an unlikely complication of the Marshall-Marchetti-Krantz procedure.


Measuring mitochondrial function in intact cardiac myocytes.
Mitochondria are involved in cellular functions that go beyond the traditional role of these organelles as the power plants of the cell. Mitochondria have been implicated in several human diseases, including cardiac dysfunction, and play a role in the aging process. Many aspects of our knowledge of mitochondria stem from studies performed on the isolated organelle. Their relative inaccessibility imposes experimental difficulties to study mitochondria in their natural environment-the cytosol of intact cells-and has hampered a comprehensive understanding of the plethora of mitochondrial functions. Here we review currently available methods to study mitochondrial function in intact cardiomyocytes. These methods primarily use different flavors of fluorescent dyes and genetically encoded fluorescent proteins in conjunction with high-resolution imaging techniques. We review methods to study mitochondrial morphology, mitochondrial membrane potential, Ca(2+) and Na(+) signaling, mitochondrial pH regulation, redox state and ROS production, NO signaling, oxygen consumption, ATP generation and the activity of the mitochondrial permeability transition pore. Where appropriate we complement this review on intact myocytes with seminal studies that were performed on isolated mitochondria, permeabilized cells, and in whole hearts.


Objective quantification of arm rigidity in MPTP-treated primates.
Rigidity is a cardinal symptom of Parkinson's disease and is frequently used as an outcome measure in clinical and non-human primate studies examining the effects of medication or surgical intervention. A limitation of current rigidity assessment methods is that they are inherently subjective. To better understand the physiological mechanisms of rigidity and how various therapeutic approaches work, a more objective and quantitative method is needed. In this study, an automated arm rigidity testing (ART) system was developed to objectively quantify rigidity while the primate's limb was moved between two user-specified angles. Recordings of normal force versus elbow-angle were categorized according to area and slope. These quantitative measures of rigidity were investigated in three rhesus macaque monkeys treated with 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine and compared with clinical assessment methods. The ART system incorporates electromyographical recordings that can detect and differentiate active from actual resistance. The ART system detected significant changes in rigidity measures following administration of apomorphine or deep brain stimulation of the globus pallidus internus. The most sensitive measures were total area, extension slope, and flexion slope. The ART system provides precise and reliable measures of rigidity that are objective and quantitative.


Cavernous brain malformations and their relation to black blood MRI in respect to vessel wall contrast enhancement.
Inflammatory responses are implicated as crucial patho-mechanisms of vascular brain malformations. Inflammation is suggested to be a key contributor to aneurysm rupture; however it is unclear whether inflammation contributes similarly to bleeding of cerebral cavernous malformations (CCMs). Black blood MRI is a sequence which identifies inflammation in blood vessel walls and in the present study is used to detect inflammatory response in CCMs. Fifteen patients with 17 CCMs treated in our department in 2017 were retrospectively analysed. All patients received black blood MRIs and the results were analysed in correlation with, size and bleeding of CCMs. Size and bleeding status of CCMs did not correlate with contrast enhancement in the CCM wall. One of 3 patients with bleeding displayed contrast enhancement in black blood MRI, whereas the others had non enhancing lesions. Because of the small number of cases a statistical analysis was not performed. In this limited cohort, inflammatory reactions in CCMs could not be detected by black blood MRI suggesting that the level of inflammation is minimal in these lesions and those different patho-mechanisms play a more important role in the rupture of CCMs.


Unilateral fixed dilation of the pupil as a false-localizing sign with intracranial hemorrhage: case report and literature review.
Although other focal signs may prove "false localizing," it is a neurosurgical axiom that unilateral fixed dilation of the pupil occurs ipsilateral to a supratentorial mass. A 25-year-old man collapsed with a dense right hemiplegia and a Glasgow Coma Scale score of 6 (eye opening, 1; motor, 4; verbal, 1) after rupture of a left middle cerebral artery aneurysm associated with an intrasylvian hematoma. Initially, both pupils had remained equal-sized and reactive: however, within hours, the right (contralateral) pupil became fixed and dilated (i.e., false localizing). For some time, the left (ipsilateral) pupil remained small and reactive; at emergency craniotomy, this also became fixed and equally dilated. After evacuation of the clot and wrapping of the aneurysm, both pupils rapidly became equal-sized and reactive. Twenty-four hours later, concurrent with massive left hemispheric swelling and a midline shift, the left (ipsilateral) pupil became unilaterally fixed and dilated (i.e., false localizing). Eventually, the right (contralateral) pupil also became fixed and dilated, concurrent with cardiovascular collapse. Death occurred within 10 hours. Unilateral fixed dilation of the pupil in patients with hemispheric mass lesions may be false localizing. Furthermore, disparate "herniating mechanisms" can arise despite mass effect emanating from the same side. Because such mechanisms cannot be witnessed, their nature remains speculative. An extensive review is contained in this article.


Bactericidal and virucidal activity of ethanol and povidone-iodine.
Ethanol and povidone-iodine (PVP-I) are important microbicides that inactivate bacteria and viruses. The present study provides a review of literature data on the concentration-dependent bactericidal and virucidal activity of ethanol and PVP-I in vitro. A systematic search was performed using the meta-database for biomedicine PubMed. Eventually, 74 studies with original data on the reduction of bacterial and viral infectivity using in vitro tests were analyzed. A safe bactericidal effect of ethanol can be expected at concentrations between 60% and 85%, and the exposure times vary between ≤0.5 and ≥5 min. Within an exposure of up to 5 min, 80%-90% ethanol also exerts virucidal/low-level activity, which includes its action against enveloped viruses plus adeno-, noro-, and rotaviruses. For PVP-I, the best bactericidal and virucidal/high-level effect is present at a concentration range of approx. 0.08%-0.9% depending on the free iodine concentration. The maximum exposure times are 5 min for bacteria and 60 min for viruses. The available data may help optimize the significant inactivation of bacteria and viruses in various areas. However, as the conditions in application practice can vary, concrete recommendations for the application can only be derived to a limited extent.


Sodium entry in rat diaphragm induced by depolarizing drugs.
1. An increased uptake of labelled sodium was found in the end-plate region of rat diaphragm following brief exposure to solution containing (24)Na plus carbachol (100 muM), with a wash in inactive saline. Tetrodotoxin (0.1 muM) was also present. Comparable results were obtained with decamethonium and suberyldicholine.2. With carbachol (100 muM) the influx of labelled sodium at the end-plate region was increased by a factor of at least three as compared with that at the end of the fibres.3. After entry the labelled sodium spread along the fibres with an apparent diffusion coefficient which was half that expected in the external solution.4. The dose-response curve for the effect of carbachol gave a half-maximal value of 72 muM.5. In muscles depolarized by potassium methyl sulphate the effect of carbachol on the entry of sodium was reduced although demonstrable.6. The entry of labelled sodium at the end-plate was maintained during prolonged exposure to carbachol (100 muM) or decamethonium (100 muM).7. The rate of entry of (24)Na obtained with carbachol, after corrections for the wash, was estimated as 1.5 x 10(3) ions channel(-1) sec(-1), measured over a period of 15 sec.8. Labelled decamethonium and labelled carbachol also accumulated at the end-plate region. After extrapolation to allow for the effects of the wash the entry of decamethonium when expressed as a clearance (pl. mg(-1) sec(-1)) was comparable to that of sodium, as expected if decamethonium and sodium enter through the same channels.


Grounded theory: issues for research in nursing.
The grounded theory approach to research appears to be well suited to providing nurses with understanding of social behaviours so that they can enhance patient care Here, the author examines the principles of grounded theory and describes how the approach can influence the way that nurses practise.


Multimodal investigation reveals the neuroprotective mechanism of Angong Niuhuang pill for intracerebral hemorrhage: Converging bioinformatics, network pharmacology, and experimental validation.
Angong Niuhuang Pill (ANP) is a traditional Chinese medicine formula that has been used clinically for many years in the treatment of cerebral hemorrhage. It is composed of ingredients such as calculus bovis, moschus, and others. Ancient texts have documented that ANP's multiple components possess properties such as heat-clearing, detoxification, and sedation, which can be effective in treating conditions such as coma and stroke. However, the underlying mechanisms of ANP's potential actions are still under investigation. ANP is a Chinese medicine widely utilized for the treatment of intracerebral hemorrhage (ICH). However, the precise mechanism underlying the therapeutic effects remains largely elusive. The present study aims to unravel the effects and pharmacological molecular mechanisms of ANP in combatting ICH, employing a comprehensive network pharmacology approach and experimental validation. The molecular targets of ANP and ICH were obtained from various databases, followed by the construction of protein-protein interaction (PPI) networks using the STRING database. Further, gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) analyses were conducted using the Metascape database and Cytoscape, respectively. Finally, molecular docking was performed. We performed a series of behavioral tests, immunohistochemical staining, TUNEL staining, and Western Blot to verify the effects of ANP. IL-6, JUN, MMP9, IL-1β, VEGFA were the main candidate targets and were associated with fluid shear stress and atherosclerosis, TNF signaling pathway, etc. It is suggested that the potential mechanism of ANP against ICH may be mainly related to pyroptosis, inflammation. In vivo validation showed that ANP treatment significantly reduced the number of TUNEL-positive cells and ANP inhibited the activation of Iba-1 positive neurons, and suppressed the expression of inflammatory factors and pyroptosis indicators. In addition, ANP improved the cognitive level and motor ability of ICH mice. The results of the study combined with virtual screening and experimental validation showed that ANP has an important contribution in protecting the brain from neuronal damage by regulating the pathways of inflammation and pyroptosis, laying the foundation and innovative ideas for future studies.


Effect of dexamethasone on albendazole cysticidal activity in experimental cysticercosis by Taenia crassiceps in BALB/c mice: In vitro and in vivo evaluation.
Taenia solium is a parasite whose larvae (cysticerci) can locate in the central nervous system of humans and cause neurocysticercosis (NC). The introduction of cysticidal drugs such as albendazole (ABZ) for the treatment of NC has significantly improved its prognosis. However, treatment is not always effective, and the high levels of corticosteroids used to prevent inflammatory complications in this disease could be, partly, the cause of this observation. In this context, this study investigated, using the experimental mouse model of intraperitoneal infection with Taenia crassiceps, the influence of corticosteroid administration on the therapeutic efficacy of ABZ. We evaluated and compared the effects of ABZ, dexamethasone (DXM) and their combination (ABZ + DXM) on cyst viability, both in vitro and in vivo. Serum levels of IL-4, IFN-gamma, IL-6 and IL-10 were evaluated in the in vivo study. Results showed that the treatment with ABZ, in vitro and in vivo, was associated with a high number of parasites deaths. Concomitant treatment with DXM did not alter ABZ in vitro cysticidal activity but reduced its effectiveness significantly in the in vivo experimental model. Cytokine serum levels did not change significantly in treated mice compared to the controls. The results of this study are relevant as they indicate a negative effect of corticosteroids on the efficacy of cysticidal therapy. In human neurocysticercosis, control of inflammation is of great importance to most patients in order to avoid complications. Corticosteroids are generally used for this purpose and the results of this study demonstrate the need to find other therapeutic strategies. Further studies are needed to better understand the mechanisms involved.


Effects of melophlins on colony formation of Chinese hamster V79 cells and IL-8 production in PMA-stimulated HL-60 cells.
We have recently isolated four new melophlins P (1), Q (2), R (3), and S (4) together with seven known melophlins A (5), D (6), E (7), G (8), H (9), I (10), and O (11) from two marine sponges of the genus Melophlus collected in Palau. In this study, the influence of these compounds on the colony formation of Chinese hamster V79 cells and the production of IL-8 in PMA-stimulated HL-60 cells were examined. These 11 compounds did not show any effect on IL-8 production. The EC50 values of compounds 2, 3, 4, 5, 7, 9, 10, and 11 against V79 cells were 44.0, 13.3, 16.7, 27.2, 19.8, 8.5, 23.1, and 9.6 microM, respectively. The linear-chain-type compounds (1, 6, and 8) were not active against V79 cells at 50 microM. Although the growth inhibitory activity of these melophlins was not remarkable, some structure-activity relationships of these compounds against V79 and murine leukemia L1210 cells were observed.


Hepatotoxicity of di-(2-ethylhexyl)phthalate is attributed to calcium aggravation, ROS-mediated mitochondrial depolarization, and ERK/NF-κB pathway activation.
Di-(2-ethylhexyl)phthalate (DEHP) is a widely used plasticizer found in a variety of polyvinyl chloride medical products. Although DEHP-induced cytotoxicity and apoptosis are well studied in various cell types, the precise mechanisms are not well understood so far. This study, aimed at going beyond the toxicology approach, focuses on the molecular mechanisms through which DEHP causes hepatotoxicity. We show that DEHP induces apoptotic cell death in a dose-dependent manner, as proven by an increase in annexin V-positively stained cells, DAPI/PI staining, and immunofluorescence studies. The DEHP-induced decrease in cell viability was significantly inhibited by adding catalase (CAT), but CAT treatment did not suppress the DEHP-stimulated calcium flux in the hepatocytes, whereas BAPTA-AM significantly reduced the DEHP-stimulated DCF intensity. These results demonstrate that DEHP increases the intracellular calcium level, which mediates the generation of H(2)O(2) in hepatocytes. Investigating cell-signaling mechanisms, we found that DEHP induced apoptotic cell death by mitochondrial-dependent caspase-3 activation and PARP cleavage. These changes due to DEHP exposure were associated with increased IKK and NF-κB phosphorylation. Preexposure of hepatocytes to an IKK inhibitor (PS-1145) prevented DEHP-induced caspase-3 and PARP cleavage. DEHP also markedly increased the activity of ERK1/2 MAPK. Pretreatment with the ERK inhibitor PD98059 attenuated NF-κB and IKK phosphorylation, indicating that ERK MAPK is mainly involved in DEHP-induced NF-κB activation. These results, for the first time, reveal that DEHP induces apoptosis in hepatocytes via the activation of the ERK/NF-κB signaling pathway, in which calcium ions and hydrogen peroxide act as the pivotal mediators of the apoptotic signaling.


Alloactivation of Naïve CD4+CD8-CD25+T Regulatory Cells: Expression of CD8α Identifies Potent Suppressor Cells That Can Promote Transplant Tolerance Induction.
Therapy with alloantigen-specific CD4+CD25+ T regulatory cells (Treg) for induction of transplant tolerance is desirable, as naïve thymic Treg (tTreg) are not alloantigen-specific and are weak suppressor cells. Naïve tTreg from DA rats cultured with fully allogeneic PVG stimulator cells in the presence of rIL-2 express IFN-gamma receptor (IFNGR) and IL-12 receptor beta2 (IL-12Rβ2) and are more potent alloantigen-specific regulators that we call Ts1 cells. This study examined additional markers that could identify the activated alloantigen-specific Treg as a subpopulation within the CD4+CD25+Foxp3+Treg. After culture of naïve DA CD4+CD8-CD25+T cells with rIL-2 and PVG alloantigen, or rIL-2 without alloantigen, CD8α was expressed on 10-20% and CD8β on <5% of these cells. These cells expressed ifngr and Il12rb2. CD8α+ cells had increased Ifngr that characterizes Ts1 cells as well was Irf4, a transcription factor induced by TCR activation. Proliferation induced by re-culture with rIL-12 and alloantigen was greater with CD4+CD8α+CD25+Treg consistent with the CD8α+ cells expressing IL-12R. In MLC, the CD8α+ fraction suppressed responses against allogeneic stimulators more than the mixed Ts1 population, whereas the CD4+CD8-CD25+T cells were less potent. In an adoptive transfer assay, rIL-2 and alloantigen activated Treg suppress rejection at a ratio of 1:10 with naïve effector cells, whereas alloantigen and rIL-2 activated tTreg depleted of the CD8α+ cells were much less effective. This study demonstrated that expression of CD8α by rIL-2 and alloantigen activation of CD4+CD8-CD25+Foxp3+T cells was a marker of activated and potent Treg that included alloantigen-specific Treg.


High-throughput analysis of algal crude oils using high resolution mass spectrometry.
Lipid analysis often needs to be specifically optimized for each class of compounds due to its wide variety of chemical and physical properties. It becomes a serious bottleneck in the development of algae-based next generation biofuels when high-throughput analysis becomes essential for the optimization of various process conditions. We propose a high-resolution mass spectrometry-based high-throughput assay as a 'quick-and-dirty' protocol to monitor various lipid classes in algal crude oils. Atmospheric pressure chemical ionization was determined to be most effective for this purpose to cover a wide range of lipid classes. With an autosampler-LC pump set-up, we could analyze algal crude samples every one and half minutes, monitoring several lipid species such as TAG, DAG, squalene, sterols, and chlorophyll a. High-mass resolution and high-mass accuracy of the orbitrap mass analyzer provides confidence in the identification of these lipid compounds. MS/MS and MS3 analysis could be performed in parallel for further structural information, as demonstrated for TAG and DAG. This high-throughput method was successfully demonstrated for semi-quantitative analysis of algal oils after treatment with various nanoparticles.


Vacuum erection devices to treat erectile dysfunction and early penile rehabilitation following radical prostatectomy.
Vacuum erection devices (VED) are becoming first-line therapies for erectile dysfunction and preservation (rehabilitation) of erectile function following treatment for prostate cancer. Currently, phosphodiesterase-5 inhibitors have limited efficacy in elderly patients or patients with moderate to severe diabetes, hypertension, and coronary artery disease. Alternative therapies, such as VED, have emerged as a primary option for patients refractory to oral therapy. VED has also been successfully used in combination treatment with oral therapy and penile injections. More recently, there has been interest in the use of VED in early intervention protocols to encourage corporeal rehabilitation and prevention of post-radical prostatectomy venoocclusive dysfunction. This is evident by the preservation of penile length and girth seen with the early use of the VED following radical prostatectomy. There are ongoing studies to help preserve penile length and girth with early use of VED following prostate brachytherapy and external beam radiation for prostate cancer. Recently, there has also been interest in VED to help maintain penile length following surgical correction of Peyronie's disease and to increase penile size before implantation of the penile prosthesis.


Allelic and genotypic frequencies of platelet glycoprotein polymorphisms in a Portuguese population.
We studied genotypic and allelic frequencies of polymorphisms that can affect platelet function, namely the Kozak, VNTR and HPA-2 polymorphisms of glycoprotein Ibα, the Pl(A) polymorphism of glycoprotein IIIa and the C807T polymorphism of glycoprotein Ia, in a Portuguese population composed of 227 donors. PCR-RFLP was used to assess the Kozak, HPA-2, Pl(A) and C807T polymorphisms. The VNTR polymorphism was discriminated by different weight bands on electrophoresis. All genotypic frequencies were in Hardy-Weinberg equilibrium and do not differ from other Caucasian populations. Genotypic frequencies were 68.3%, 26.9% and 4.8% for Pl(A1/A1), Pl(A1/A2) and Pl(A2/A2) genotypes of the Pl(A) polymorphism, 79.3%, 20.3% and 0.4% for TT, TC and CC genotypes of the Kozak polymorphism, 81.1%, 18.9% and 0.0% for aa, ab and bb genotypes of the HPA-2 polymorphism, 15.4%, 0.9%, 70.5%, 11.5%, 1.3% and 0.4% for BC, BD, CC, CD, DD and CE genotypes of the VNTR polymorphism, and 39.7%, 50.2% and 10.1% for CC, CT and TT genotypes of the C807T polymorphism. The Portuguese population has now been characterized in terms of major platelet glycoprotein polymorphisms, which will be an important tool for further studies to assess the role of platelet glycoproteins in individual predisposition to prothrombotic conditions and response to antithrombotic therapy.


Developmentally informed evaluation of depression: evidence-based instruments.
This article has two primary aims: (1) to describe how to incorporate evidence-based assessment procedures into diagnostic practice and (2) to present a review of the more commonly used interview methods and clinical measures of depression among preschoolers, school-age children, and adolescents.


Assessing organic material on single-use vessel sealing devices: a comparative study of reprocessed and new LigaSure™ devices.
Reprocessed devices must be thoroughly cleaned prior to sterilization to ensure efficacy of sterilization agents. Many single-use devices are not designed to be thoroughly cleaned. Interlocking design features inherent to LigaSure™ vessel sealing devices may prevent thorough cleaning and promote accumulation of human tissue that cannot be removed. Thus, the aim of this study was to compare industry reprocessed and new LigaSure™ vessel sealing devices for organic material. A total of 168, 84 new and 84 reprocessed, vessel sealing devices were disassembled and inspected for the presence of residual organic matter using visual, microscopic, and chemical analysis. Devices were randomized and test conductors blinded to group membership. Devices were aseptically disassembled and sent through visual inspection. Next, devices were either examined using light microscopy, scanning electron microscopy (SEM) or exposed to a solution that luminesces in the presence of hemoglobin. Additionally, 165 reprocessed devices were sent to a 3rd party lab for sterility testing via direct immersion culture for 14 days. Significant amounts of remnant organic material (C, N, O, S, Na, P) were observed with 81/84 reprocessed and 0/84 new devices failing inspection protocols. When tested for the presence of hemoglobin, only 1/12 reprocessed devices passed inspection. SEM of reprocessed devices revealed residues with liquid patterns and diffuse soiling with foreign material. Sterility testing of reprocessed devices revealed a sterility level < 6-3. The abundance of material resembling human tissue observed on reprocessed VSDs suggests inadequate cleaning prior to sterilization. Atomic and morphological analyses of the remnant materials suggest that bacterial biofilms could also be present. Additionally, surface degradation and release of reinforcing glass fibers from the device were observed. Devices designed for single use can harbor significant amounts of remnant material that likely interfere with the sterilization process.


Versatile peroxidase as a valuable tool for generating new biomolecules by homogeneous and heterogeneous cross-linking.
The modification and generation of new biomolecules intended to give higher molecular-mass species for biotechnological purposes, can be achieved by enzymatic cross-linking. The versatile peroxidase (VP) from Pleurotus eryngii is a high redox-potential enzyme with oxidative activity on a wide variety of substrates. In this study, VP was successfully used to catalyze the polymerization of low molecular mass compounds, such as lignans and peptides, as well as larger macromolecules, such as protein and complex polysaccharides. Different analytical, spectroscopic, and rheological techniques were used to determine structural changes and/or variations of the physicochemical properties of the reaction products. The lignans secoisolariciresinol and hydroxymatairesinol were condensed by VP forming up to 8 unit polymers in the presence of organic co-solvents and Mn(2+). Moreover, 11 unit of the peptides YIGSR and VYV were homogeneously cross-linked. The heterogeneous cross-linking of one unit of the peptide YIGSR and several lignan units was also achieved. VP could also induce gelation of feruloylated arabinoxylan and the polymerization of β-casein. These results demonstrate the efficacy of VP to catalyze homo- and hetero-condensation reactions, and reveal its potential exploitation for polymerizing different types of compounds.


Potential role of left atrial strain in estimation of left atrial pressure in patients with chronic heart failure.
In a large proportion of heart failure with reduced ejection fraction (HFrEF) patients, echocardiographic estimation of left atrial pressure (LAP) is not possible when the ratio of the peak early left ventricular filling velocity over the late filling velocity (E/A ratio) is not available, which may occur due to several potential causes. Left atrial reservoir strain (LASr) is correlated with LV filling pressures and may serve as an alternative parameter in these patients. The aim of this study was to determine whether LASr can be used to estimate LAP in HFrEF patients in whom E/A ratio is not available. Echocardiograms of chronic HFrEF patients were analysed and LASr was assessed with speckle tracking echocardiography. LAP was estimated using the current ASE/EACVI algorithm. Patients were divided into those in whom LAP could be estimated using this algorithm (LAPe) and into those in whom this was not possible because E/A ratio was not available (LAPne). We assessed the prognostic value of LASr on the primary endpoint (PEP), which comprised the composite of hospitalization for the management of acute or worsened HF, left ventricular assist device implantation, cardiac transplantation, and cardiovascular death, whichever occurred first in time. We studied 153 patients with a mean age of 58 years of whom 76% men and 82% who were in NYHA class I-II. A total of 86 were in the LAPe group and 67 in the LAPne group. LASr was significantly lower in the LAPne group as compared with the LAPe group (15.8% vs. 23.8%, P < 0.001). PEP-free survival at a median follow-up of 2.5 years was 78% in LAPe versus 51% in LAPne patients. An increase in LASr was significantly associated with a reduced risk of the PEP in LAPne patients (adjusted hazard ratio: 0.91 per %, 95% confidence interval 0.84-0.98). An abnormal LASr (<18%) was associated with a five-fold increase in reaching the PEP. In HFrEF patients in whom echocardiographic estimation of LAP is not possible due to due to unavailability of E/A ratio, assessing LASr potentially carries added clinical and prognostic value.


IKKβ regulates antimicrobial innate immune responses in the yellow mealworm, Tenebrio molitor.
Toll and IMD pathways regulate antimicrobial innate immune responses in insect model systems. The transcriptional activation of antimicrobial peptides (AMPs) confers humoral immunity in the host against invaded pathogens. The IKK kinase complex (IKKα, IKKβ, and the regulatory subunit IKKγ/NEMO) centrally regulates the NF-κB response to various stimuli. It triggers an appropriate antimicrobial immune response in the host. In this study, a TmIKKβ (or TmIrd5) homolog was screened from the RNA-seq database of the coleopteran beetle, Tenebrio molitor. A single exon characterizes the TmIKKβ gene, and the open reading frame (ORF) comprises of 2112 bp that putatively encodes a polypeptide of 703 amino acid residues. TmIKKβ contains a serine/threonine kinase domain and is phylogenetically close to Tribolium castaneum IKKβ homolog (TcIKKβ). TmIKKβ transcripts were highly expressed in the early pupal (P1) and adult (A5) stages. Among the tissues, TmIKKβ showed higher expression in the integument of the last instar larvae and the fat body and hemocytes of 5-day-old adults. TmIKKβ mRNA was upregulated post-E. coli challenge to the host. Moreover, RNAi-based TmIKKβ mRNA silencing increased host larvae' susceptibility against E. coli, S. aureus and C. albicans. TmIKKβ RNAi in the fat body led to a downregulation in mRNA expression of ten out of fourteen AMP genes, including TmTenecin1, -2, and -4; TmDefensin, and -like; TmColeoptericinA, and -B; and TmAttacin1a, -1b, and -2, suggesting the requirement of the gene in antimicrobial innate immune responses. Further, a decrease in the mRNA expression of NF-κB factors such as TmRelish, TmDorsal1, and TmDorsal2 in the fat body of T. molitor larvae was observed post-microorganisms challenge. Thus, TmIKKβ regulates antimicrobial innate immune responses in T. molitor.


Development of Coriolis mass flowmeter with digital drive and signal processing technology.
Coriolis mass flowmeter (CMF) often suffers from two-phase flowrate which may cause flowtube stalling. To solve this problem, a digital drive method and a digital signal processing method of CMF is studied and implemented in this paper. A positive-negative step signal is used to initiate the flowtube oscillation without knowing the natural frequency of the flowtube. A digital zero-crossing detection method based on Lagrange interpolation is adopted to calculate the frequency and phase difference of the sensor output signals in order to synthesize the digital drive signal. The digital drive approach is implemented by a multiplying digital to analog converter (MDAC) and a direct digital synthesizer (DDS). A digital Coriolis mass flow transmitter is developed with a digital signal processor (DSP) to control the digital drive, and realize the signal processing. Water flow calibrations and gas-liquid two-phase flowrate experiments are conducted to examine the performance of the transmitter. The experimental results show that the transmitter shortens the start-up time and can maintain the oscillation of flowtube in two-phase flowrate condition.


Toward Progress in Quantitative Translational Medicine: A Call to Action.
Quantitative translational medicine (QTM) is envisioned as a multifaceted discipline that will galvanize the path from idea to medicine through quantitative translation across the discovery, development, regulatory, and utilization spectrum. Here, we summarize results of an American Society for Clinical Pharmacology and Therapeutics (ASCPT) survey on barriers relevant to the advancement of QTM and propose opportunities for its deployment. Importantly, we offer a call to action to break down these barriers through patient-centered stewardship, effective communication, cross-sector collaboration, and a modernized educational curriculum.


Falling through the cracks: the hidden economic burden of chronic illness and disability on Australian households.
Major reform plus targeted strategies have the potential to provide relief.


Local experience with the use of nasal bubble CPAP in infants with bronchiolitis admitted to a combined adult/paediatric intensive care unit.
Bronchiolitis is an acute inflammatory disease of the lower small airways predominantly occurring in infants younger than 1 year of age. As a result of the respiratory distress associated with bronchiolitis, infants frequently require admission to an intensive care unit for respiratory support. Thirty-five infants diagnosed with bronchiolitis were admitted to a combined adult/paediatric tertiary intensive care unit over a 2-year period for nasal bubble continuous positive airway pressure (CPAP). Following this therapy, 20 (57.14%) of these infants could be transferred to the medical ward of the onsite paediatric hospital. The remainder required transfer to the national paediatric intensive care unit (PICU) for ongoing observation and/or positive pressure ventilation. Nasal bubble CPAP is a simple therapy that can be easily set up at the bedside. The use of nasal bubble CPAP enabled infants to remain in their geographical area, thus improving family visiting access and reducing the demand for paediatric beds in the national PICU.


A general method to determine ionization constants of responsive polymer thin films.
A general method has been developed to determine the ionization constants of polymer thin films based on the stimuli-responsiveness of the polymer. Robust polymer films were fabricated on silicon wafers and gold slides using perfluorophenyl azide (PFPA) as the coupling agent. The ionization constants were measured by a number of techniques including ellipsometry, dynamic contact angle goniometry, and surface plasmon resonance imaging (SPRi). Using poly(4-vinylpyridine) (P4VP) as the model system, P4VP thin films were fabricated and the ionization constants of the films were measured taking advantage of the pH responsive property of the polymer. The pK(a) determined by ellipsometry, ~4.0, reflects the swelling of the polymer film in response to pH. The pK(a) value calculated from the dynamic contact angle measurements, ~5.0, relies on the change in hydrophilicity/hydrophobicity of the films as the polymer undergoes protonation/deprotonation. The pK(a) value measured by SPRi, ~4.9, monitors in situ the change of refractive index of the polymer thin film as it swells upon protonation. This was the first example where SPRi was used to measure the ionization constants of polymers.


Keeping kids in school: modelling school-based testing and quarantine strategies during the COVID-19 pandemic in Australia.
In 2021, the Australian Government Department of Health commissioned a consortium of modelling groups to generate evidence assisting the transition from a goal of no community COVID-19 transmission to 'living with COVID-19', with adverse health and social consequences limited by vaccination and other measures. Due to the extended school closures over 2020-21, maximizing face-to-face teaching was a major objective during this transition. The consortium was tasked with informing school surveillance and contact management strategies to minimize infections and support this goal. Outcomes considered were infections and days of face-to-face teaching lost in the 45 days following an outbreak within an otherwise COVID-naïve school setting. A stochastic agent-based model of COVID-19 transmission was used to evaluate a 'test-to-stay' strategy using daily rapid antigen tests (RATs) for close contacts of a case for 7 days compared with home quarantine; and an asymptomatic surveillance strategy involving twice-weekly screening of all students and/or teachers using RATs. Test-to-stay had similar effectiveness for reducing school infections as extended home quarantine, without the associated days of face-to-face teaching lost. Asymptomatic screening was beneficial in reducing both infections and days of face-to-face teaching lost and was most beneficial when community prevalence was high. Use of RATs in school settings for surveillance and contact management can help to maximize face-to-face teaching and minimize outbreaks. This evidence supported the implementation of surveillance testing in schools in several Australian jurisdictions from January 2022.


Phenotype switching and mutations in random environments.
Cell populations can benefit from changing phenotype when the environment changes. One mechanism for generating these changes is stochastic phenotype switching, whereby cells switch stochastically from one phenotype to another according to genetically determined rates, irrespective of the current environment, with the matching of phenotype to environment then determined by selective pressure. This mechanism has been observed in numerous contexts, but identifying the precise connection between switching rates and environmental changes remains an open problem. Here, we introduce a simple model to study the evolution of phenotype switching in a finite population subject to random environmental shocks. We compare the successes of competing genotypes with different switching rates, and analyze how the optimal switching rates depend on the frequency of environmental changes. If environmental changes are as rare as mutations, then the optimal switching rates mimic the rates of environmental changes. If the environment changes more frequently, then the optimal genotype either maximally favors fitness in the more common environment or has the maximal switching rate to each phenotype. Our results also explain why the optimum is relatively insensitive to fitness in each environment.


Production of fructooligosaccharides by crude enzyme preparations of beta-fructofuranosidase from Aureobasidium pullulans.
Fructooligosaccharides (FOS) were produced from sucrose by using crude enzyme preparations of beta-fructofuranosidases (FFases) obtained from sucrose-cultured cells of Aureobasidium pullulans DSM 2404. When the preparation mainly consisted of FFase I, that has high transfructosylating activity, the FOS yield was 62%. When the reaction was carried out with additional commercial glucose isomerase (GI) at an activity ratio of FFase and GI of 1:2, the maximum FOS yield reached 69%. This value was higher than those obtained previously using other Aureobasidium spp. (53-59%).


Chiral dendrimer encapsulated Pd and Rh nanoparticles.
The synthesis of a series of chiral PAMAM dendrimers and the formation of chiral dendrimer encapsulated metal nanoparticles are described.


Cultivation of different strains of king oyster mushroom (Pleurotus eryngii) on saw dust and rice straw in Bangladesh.
Pleurotus eryngii is a popular mushroom due to its excellent consistency of cap and stem, culinary qualities and longer shelf life. In Bangladesh, where Pleurotus mushrooms are very popular, P. eryngii may take position among the consumers, but currently this mushroom is not cultivated in large scale there. In this study, 3 strains of P. eryngii such as Pe-1 (native to Bangladesh), Pe-2 (germplasm collected from China) and Pe-3 (germplasm collected from Japan) were cultivated on saw dust and rice straw and their growth and yield parameters were investigated. Pe-1 on saw dust showed the highest biological yield and efficiency (73.5%) than other strains. Also, the mycelium run rate and number of fruiting bodies were higher in Pe-1 than other two strains. The quality of mushroom strains was near about similar. On saw dust, the yield and efficiency were better than those cultivated on rice straw, however, on straw; the mushroom fruiting bodies were larger in size. This study shows the prospects of P. eryngii cultivation in Bangladesh and suggests further study in controlled environment for higher yield and production.


The endohyphal microbiome: current progress and challenges for scaling down integrative multi-omic microbiome research.
As microbiome research has progressed, it has become clear that most, if not all, eukaryotic organisms are hosts to microbiomes composed of prokaryotes, other eukaryotes, and viruses. Fungi have only recently been considered holobionts with their own microbiomes, as filamentous fungi have been found to harbor bacteria (including cyanobacteria), mycoviruses, other fungi, and whole algal cells within their hyphae. Constituents of this complex endohyphal microbiome have been interrogated using multi-omic approaches. However, a lack of tools, techniques, and standardization for integrative multi-omics for small-scale microbiomes (e.g., intracellular microbiomes) has limited progress towards investigating and understanding the total diversity of the endohyphal microbiome and its functional impacts on fungal hosts. Understanding microbiome impacts on fungal hosts will advance explorations of how "microbiomes within microbiomes" affect broader microbial community dynamics and ecological functions. Progress to date as well as ongoing challenges of performing integrative multi-omics on the endohyphal microbiome is discussed herein. Addressing the challenges associated with the sample extraction, sample preparation, multi-omic data generation, and multi-omic data analysis and integration will help advance current knowledge of the endohyphal microbiome and provide a road map for shrinking microbiome investigations to smaller scales. Video Abstract.


The effect of the Ontario Fertility Program on IVF utilization and outcomes in women of advanced reproductive age.
Are the demographics and clinical outcomes similar for patients aged ≥40 but <43 years seeking IVF in Ontario, Canada, before and after implementation of the Ontario Fertility Program (OFP), which supports public funding of IVF up to age 43? Retrospective database review using the Canadian Assisted Reproductive Technologies Registry Plus (CARTR Plus) and Better Outcomes Registry & Network (BORN) Ontario databases. Cycles from women who underwent autologous IVF and who were aged ≥40 and <43 years were analysed during a 2-year period prior to (2014-2015) and after (2016-2017) introduction of publicly funded IVF through the OFP. There was an almost doubling of treatment cycles in women aged 40-42 in Ontario after the OFP launch. Clinical pregnancy rate per cycle start (17.0% versus 13.3%, P < 0.001) and cumulative clinical pregnancy rate per stimulation cycle (20.5% versus 16.8%, P < 0.001) were statistically higher in women before OFP implementation. While cumulative live birth rate per cycle start was statistically lower after funding was introduced (12.5% versus 10.5%, P = 0.027), the clinical importance of this difference appears small. Outcomes were above the 10% live birth per cycle threshold recommended by the Advisory Process for Infertility Services panel, commissioned by the Ministry of Health, to determine access to publicly funded IVF. Use of IVF in women over age 40 doubled with access to OFP funding; however, eligibility criteria based on age still meet the target of achieving a cumulative live birth rate of at least 10%.


Effect of nifedipine on the sick sinus syndrome.
The effect of nifedipine (N) on sinus node (SN) function was studied in 15 patients (9 males, 6 females) sixty-two to seventy-six (mean 68.1 +/- 11) years old, with sick sinus syndrome (SSS). SSS was characterized electrophysiologically by a prolonged corrected sinus node recovery time (CSNRT greater than 535 msec) and/or prolonged sinoatrial conduction time (SACT greater than 125 msec), assessed by applying premature atrial stimulation. Ten mg N was given sublingually, and CSNRT and SACT were again evaluated sixty minutes after N administration, and again ten minutes after 1.5 mg atropine (A) was given IV. Heart rate increased significantly after N (p less than 0.005), systolic blood pressure (SBP) diminished significantly (p less than 0.005), and CSNRT and SACT shortened significantly (p less than 0.005, p less than 0.005) and became normal in 7 and 5 patients respectively. After A administration, a further significant increase of heart rate (p less than 0.005) and decrease of CSNRT (p less than 0.005) and SACT (p less than 0.005) were observed. CSNRT and SACT became normal in 8 and 7 patients respectively. SBP remained stable.


Hyperlipidemia modifies innate immune responses to lipopolysaccharide via the TLR-NF-κB signaling pathway.
The objective of this study was to investigate the effect of hyperlipidemia (HLP) on innate immune responses to lipopolysaccharide (LPS). Male New Zealand white rabbits were fed a normal diet (ND) or a high-fat diet (HFD) for 8 weeks. In vivo, the rabbits were injected intravenously with LPS for 24 h. In vitro, peripheral mononuclear cells were collected and stimulated (or unstimulated) with LPS for 24 h. Assay results were analyzed with one-way ANOVA or an equivalent non-parametric test. A P value of 0.05 was considered statistically significant. Despite having no influence in body weight, the HFD intake significantly increased serum lipids, C-reactive protein (CRP), nuclear factor (NF)-κB subunit p65, Toll-like receptor (TLR)-4, SR-A and FAS. Although we found increased circulating tumor necrosis factor (TNF)-α, interleukin (IL)-6, CRP, IL-1β, and IL-10 in the ND-fed rabbits, no significant difference was found in the LPS-stimulated production of TNF-α, IL-6, and CRP in the HFD-fed rabbits. The macrophages harvested from the HFD-fed rabbits developed a blunted inflammatory response, with lower mRNA expression of TNF-α, IL-6, CRP, TLR-4, SR-A, FAS, and Bcl-2 than that expressed by the ND group. In the HFD-fed animals, LPS incubation decreased NF-κB subunit p65 expression, whereas the cytoplasmic phosphorylation of the inhibitor of NF-κB protein was enhanced. These data indicate that HLP displayed a form of innate immune paralysis, including reduced pro- and anti-inflammatory cytokine release to external stimulus, which was related to the altered TLR-NF-κB signaling pathway and altered pro- and anti-apoptotic processes in macrophages.


Protecting Human Health from Climate Change: Legal Obligations and Avenues of Redress under International Law.
In this contribution, we explore how human health can be protected from climate change and its adverse effects by reliance on States' obligations under international law. We achieved this by reviewing the principal legal instruments that establish the right to health, as well as those that recognize that climate change has an adverse impact on health (Part II). We then examine the means of redress that may be available to those whose human right to health has been interfered with or violated because of climate change (Part III). Finally, we draw some conclusions as to the current effectiveness and future direction of these developments.


Draft Genome Sequences of Two Heat-Resistant Mutant Strains (A52 and B41) of the Photosynthetic Hydrogen-Producing Bacterium Rhodobacter capsulatus.
The draft genome sequences of two heat-resistant mutant strains, A52 and B41, derived from Rhodobacter capsulatus DSM 1710, and with different hydrogen production levels, are reported here. These sequences may help understand the molecular basis of heat resistance and hydrogen production in R. capsulatus.


Effect of kainate-induced experimental epilepsy on NADPH-diaphorase and calcium-binding proteins in rat hippocampal neurons.
Experimental epilepsy induced in rats by infusion of kainic acid into the lateral cerebral ventricles decreased the number of NADPH-diaphorase-positive neurons in the hippocampal formation by 55-79% and increased activity of this enzyme in CA1 and CA3 pyramidal neurons. All parvalbumin-immunoreactive cells were highly resistant to the cytotoxic effects of kainate in contrast to calbindin- and calretinin-positive interneurons, whose amount decreased by 50%.


Regression, kinetics and isotherm models for biosorption of organic pollutants, suspended and dissolved solids by environmentally friendly and economical dried Phragmites australis.
Low cost adsorbents such as P. australis have received considerable interest owing to their low cost and easy availability. The aim of the present study was the evaluation of the removal of chemical oxygen demand (COD), biological oxygen demand (BOD), total suspended and dissolved solids (TSS and TDS) using dried P. australis in influent wastewater to a wastewater treatment plant. The results of the COD and BOD concentration reduction with P. australis at optimum operating conditions were determined for maximum reduction and adsorption isotherms. The maximum reduction of COD, BOD, TSS and TDS concentrations under the optimum operating conditions was 92.27%, 93.89%, 94.38% and 91.61%, respectively. The results demonstrate that the new dried biosorbent is able to adsorb all the aforementioned contamination. It achieved an adsorption capacity for COD of 72.5 mg g-1 and an adsorption capacity for BOD of 43.93 mg g-1. The results were well fitted by the pseudo-second order model with R 2 = 0.984.


Quality control of astrocyte-directed Cre transgenic mice: the benefits of a direct link between loss of gene expression and reporter activation.
Cre recombinase activity for cell-type restricted deletion of floxed target genes (i.e., flanked by Cre recognition loxP-sites) is often measured by separate matings with recombination-activated reporter gene mice. Using a floxed Gja1 (Cx43) allele, we demonstrate the benefits of a direct link between reporter gene expression and target gene deletion to overcome critical limitations of the Cre/loxP system. The widely used human glial fibrillary acidic protein (hGFAP)-Cre transgene exhibits variable recombination activity and requires postexperimental validation. Such quality control is essential to correlate the extent of Cre-mediated Gja1 ablation with phenotypical alterations and to maintain the activity status of hGFAP-Cre in transgenic mouse colonies. We present several strategies to control for the fidelity of hGFAP-Cre mediated recombination. (c) 2008 Wiley-Liss, Inc.


Identification of the major sites of enzymic glycosylation of myelin basic protein.
In the presence of porcine submaxillary N-acetylgalactosaminyltransferase and uridine diphospho-N-acetyl-D-galactosamine, approx. 1.2-1.5 mol of N-acetylgalactosamine were transfered per mol of myelin basic protein. Tritium-labelled N-acetylgalactosamine-labelled basic protein was digested with trypsin and the peptides were separated by HPLC and the radioactivity measured. Most of the radioactivity was associated with three peptide peaks (I, II and III) containing 17, 69 and 6% of the total radioactivity, respectively. The remaining radioactivity was distributed amongst several peptides, each containing less than 2.5% of the total radioactivity. Glycosylation of the basic proteins isolated from human, bovine and guinea pig myelins showed that they were all equally good acceptors. In spite of differences in the peptide profiles of the basic proteins from different species, the distribution of radioactivity between the three peptide peaks was similar for all the species studied. The transfer of N-acetylgalactosamine to peptide II was much faster than to peptides I and III. The apparent Km values of the three peptides were within a narrow range of 0.52-0.63 mM, whereas the Vmax values were considerably different. The glycosylated peptide peaks (I, II and III) were separated by electrophoresis, the radioactivity measured, and amino acid compositions determined after hydrolysis. The major radioactive peptides of the human basic protein were identified with tryptic peptides containing the following sequences: (formula; see text)


Interleukin-1beta reduces transcellular monocyte diapedesis and compromises endothelial adherens junction integrity.
Diapedesis occurs through endothelial cell-cell junctions (paracellular) or through individual endothelial cells without disrupting junctions (transcellular). While in vitro studies have provided considerable insight into mechanisms controlling paracellular diapedesis, little is known about what regulates transcellular diapedesis. The authors investigated whether transcellular diapedesis is susceptible to IL-1beta exposure of the endothelium. Laser scanning confocal microscopy and biochemical analysis were used to determine the effect of IL-1beta pretreatment of the endothelium on adherens junctional morphology and monocyte transcellular diapedesis in cocultures of human peripheral blood monocytes and coronary artery endothelial cells. IL-1beta pretreatment caused a 40% decrease in the number of migrating monocytes that used a transcellular route of diapedesis, and resulted in elongate endothelial cell morphology, a reorganization of the F-actin cytoskeleton, and a significant decrease in transendothelial electrical resistance. In IL-1beta treated monolayers, VE-cadherin and its associated catenins were distributed in a punctate pattern in comparison to the lacy pattern seen in control monolayers. Coimmunoprecipitation of VE-cadherin molecular assemblies revealed that IL-1beta-mediated changes in distribution were associated with a decrease in the presence of cadherin/catenin complexes in the detergent insoluble fraction. IL-1beta-induced rearrangement of interendothelial adherens junctions facilitates paracellular diapedesis at the expense of transcellular diapedesis.


Increased cortical thickness and caudate volume precede atrophy in PSEN1 mutation carriers.
Neuroimaging studies of familial Alzheimer's disease allow investigation of the disease process before clinical onset. We performed semi-automated MRI analysis to evaluate cortical thickness (CTh), grey matter (GM) volumes, and GM diffusivity indexes in PSEN1 mutation carriers (MC). We recruited 11 MC from 4 families with PSEN1 mutations (L286P, M139T, K239N) and 6 familial and 12 non-familial healthy controls. MC were classified as either asymptomatic (n=6) or symptomatic (n=5). Subjects underwent structural and diffusion-weighted 3-Tesla MRI scanning. CTh and GM volumes of subcortical structures and diffusivity indexes were calculated and group comparisons were performed. Structural images were reanalyzed with voxel-based morphometry methodology. Cerebrospinal fluid amyloid-β1-42 levels (Aβ) were measured. We found that symptomatic MC presented widespread cortical thinning, especially in precuneus and parietotemporal areas (p<0.01) and increased mean diffusivity (MD) in these areas compared to controls. Unexpectedly, asymptomatic MC, 9.9 years prior to the predicted age of disease onset, presented increased CTh in the precuneus and parietotemporal areas (p<0.01), increased caudate volumes (p<0.01), and decreased MD (p<0.05) in these areas compared to HC. In MC, CTh correlated with adjusted age. Aβ values were within normal limits in AMC. In conclusion, at early preclinical stages, CTh in the precuneus and parietotemporal regions and caudate volume increase in PSEN1 MC and decrease thereafter with disease progression. The different trends in MD in asymptomatic and symptomatic MC suggest that different microstructural changes underlie the contrasting morphometric findings. Reactive neuronal hypertrophy or/and inflammation may account for increased CTh and decreased MD in asymptomatic MC.


Divalent cations influence the dimerization mode of murine S100A9 protein by modulating its disulfide bond pattern.
S100A9, with its congener S100A8, belongs to the S100 family of calcium-binding proteins found exclusively in vertebrates. These two proteins are major constituents of neutrophils. In response to a pathological condition, they can be released extracellularly and become alarmins that induce both pro- and anti-inflammatory signals, through specific cell surface receptors. They also act as antimicrobial agents, mainly as a S100A8/A9 heterocomplex, through metal sequestration. The mechanisms whereby divalent cations modulate the extracellular functions of S100A8 and S100A9 are still unclear. Importantly, it has been proposed that these ions may affect both the ternary and quaternary structure of these proteins, thereby influencing their physiological properties. In the present study, we report the crystal structures of WT and C80A murine S100A9 (mS100A9), determined at 1.45 and 2.35 Å resolution, respectively, in the presence of calcium and zinc. These structures reveal a canonical homodimeric form for the protein. They also unravel an intramolecular disulfide bridge that stabilizes the C-terminal tail in a rigid conformation, thus shaping a second Zn-binding site per S100A9 protomer. In solution, mS100A9 apparently binds only two zinc ions per homodimer, with an affinity in the micromolar range, and aggregates in the presence of excess zinc. Using mass spectrometry, we demonstrate that mS100A9 can form both non-covalent and covalent homodimers with distinct disulfide bond patterns. Interestingly, calcium and zinc seem to affect differentially the relative proportion of these forms. We discuss how the metal-dependent interconversion between mS100A9 homodimers may explain the versatility of physiological functions attributed to the protein.


[Stem cell-based treatment of neurologic diseases].
Therapeutic strategies based on stem cells are being increasingly used to treat a wide range of neurological diseases. Although these strategies were initially designed to replace dead cells in injured tissue, the potential of stem cells to migrate, secrete trophic factors, and immunomodulate allows their therapeutic use as a vehicle for gene therapy, as in Parkinson's disease, or as immunomodulators and neuroprotectors in diseases such as multiple sclerosis. This review will focus on current clinical and experimental evidence on the treatment of neurological disorders with strategies based on stem cells.


A systematic analysis of predicted MiR-31-targets identifies a diagnostic and prognostic signature for lung cancer.
Recent studies have shown that miR-31 could play a potential role as diagnostic and prognostic biomarkers of several cancers including lung cancer. The aim of this study is to globally summarize the predicting targets of miR-31 and their potential function, pathways and networks, which are involved in the biological behavior of lung cancer. We have conducted the natural language processing (NLP) analysis to identify lung cancer-related molecules in our previous work. In this study, miR-31 targets predicted by combinational computational methods. All target genes were characterized by gene ontology (GO), pathway and network analysis. In addition, miR-31 targets analysis were integrated with the results from NLP analysis, followed by hub genes interaction analysis. We identified 27 hub genes by the final integrative analysis and suggested that miR-31 may be involved in the initiation, progression and treatment response of lung cancer through cell cycle, cytochrome P450 pathway, metabolic pathways, apoptosis, chemokine signaling pathway, MAPK signaling pathway, as well as others. Our data may help researchers to predict the molecular mechanisms of miR-31 in the molecular mechanism of lung cancer comprehensively. Moreover, the present data indicate that the interaction of miR-31 targets may be promising candidates as biomarkers for the diagnosis, prognosis and personalized therapy of lung cancer.


A role for neural cell adhesion molecule in the formation of the avian inner ear.
The inner ear forms by a series of folds within an ectodermal placode. Previous work has shown that changes in surrounding tissues play a more prominent role in invagination than changes in the cytoskeleton of the primordium. Interference with the integrity of the extracellular matrix causes abnormalities in the folding process, primarily related to abnormalities in the paraxial mesoderm which lies ventral to the placode. In this study, the role of the neural cell adhesion molecule (N-CAM) was investigated, based on the expression of this component of the plasmalemma at the time the otic placode begins to fold. Microinjection of blocking antibodies to N-CAM into the paraxial mesoderm adjacent to the otic placode resulted in two major classes of defects, detachment of the primordium from the neural tube and interference with formation of the folds. Microinjection of saline, control immunoglobulin, or antibody against cytoplasmic domain had no effect. These defects correlate with the pattern of N-CAM expression at the time of injection, along the neural ectoderm and otic epithelium and the mesenchyme cells ventral to the primordium. It seems likely that N-CAM is playing a role in heterophilic associations rather than through the homophilic binding domain during formation of the otic vesicle.


Microextraction based on liquid-solid phase transition of benzoic acid: Extraction of statins from human urine followed by chromatographic analysis.
Herein, a microextraction method was reported based on the liquid-solid phase transition of benzoic acid to quantify two statins, namely lovastatin and simvastatin in authentic human urine. The principle of the method is based on the phase transition of benzoic acid by altering the pH of the sample solution enabling efficient dispersion and phase separation in one step. Due to the moderate melting point of benzoic acid, its solidification is performed at ambient temperature without the need for sample cooling. Various experimental parameters that affect the performance of the analytes (i.e. extractant type and its concentration, acid type and concentration, and sample volume) have been examined and optimized. The method was validated based on the total error concept. For this purpose, accuracy profiles were constructed in the concentration range of 100-5000 ng mL-1 while β-expectation tolerance intervals fell within ±15% demonstrating that 95% of future results will not exceed the defined bias limits. The intra-day and inter-day method precision was less than 4.7% and 4.3% for both analytes, while the limit of detection was 15 ng mL-1 for both analytes. It was also proved that the usage of benzoic acid is advantageous in minimizing the potential inter-conversion of the analytes during the acidification step of the extraction procedure. The green potential of the proposed analytical scheme was examined based on Green Analytical Procedure index. The proposed sample pretreatment technique proved to be a valuable tool offering selectivity and rapidness. The developed method was used for the analysis of real human urine obtained after the administration of statin-based pharmaceutical formulations.


[Fertility preservation in boys: spermatogonial stem cell transplantation and testicular grafting].
Spermatogonial stem cells (SSC) are the founder cells of spermatogenesis and are responsible for the lifelong production of spermatozoa. The cryopreservation and transplantation of these cells has been proposed as a fertility preservation strategy for young boys at risk for stem cell loss, i.e. patients undergoing chemotherapy for cancer or as a conditioning treatment for bone marrow transplantation. To prevent lifelong sterility in boys, two fertility restoration strategies are being developed: the injection of SSC and the grafting of testicular tissue containing SSC. Depending on the disease of the patient one of these two approaches will be applicable. Grafting has the advantage that SSC can reside within their natural niche, preserving the interactions between germ cells and their supporting cells and may therefore be regarded as the first choice strategy. However, in cases where the risk for malignant contamination of the testicular tissue is real, e.g. leukemia, transplantation of SSC by injection is preferable over grafting.


[Diversity research about facial morphology of unilateral complete cleft lip and palate affecting by different repair periods of palate repair].
To evaluate facial morphology characteristics of patients with unilateral complete cleft lip and palate (UCCLP) after cleft palate repair in different periods. 46 nonsyndromic UCCLP patients were chosen as test group and divided into three kinds (under 4-year-old, 4-7-year-old, over 7-year-old) according to periods of palate repair. 38 age and gender matched non-clefts children were involved as control group. 26 cephalometric measurements were measured to evaluate facial morphology of four groups. Compared with patients who received palate repair before 4-year-old, patients who received repair after 4-year-old had more protrusive ANS point (Ba-N-ANS, Ba-ANS) and less Y-axis angle. Patients who received palate repair after 7-yesr-old had greater A-PMP and ANS-PMP than patients who received palate repair before 4-year-old, patients who received palate repair before 7-year-old had smaller ANS-Me but after had no significant difference when compared with non-clefts. Whenever palate repair is undertaken in the period of maxillary growth and development, patients' maxillary growth would be restrained, the delayed repair sample have better lower face height, maxillary protrusion, maxillary sagittal length and mandible growing direction when compared with the early repair sample, that is to say, the delayed repair sample have preferable head-face morphology.


[Congenital "cardio-esophageal" syndromes. Congenital cardiopathies and multiple abnormalities syndromes in the child with esophageal abnormalities].
A case of esophageal stenosis and tracheo-esophageal fistula associated with dextrocardia due to dextrorotation, ventricular septal defect and complete double arotic arch is reported. In 116 cases of esophageal atresia and tracheo-esophageal fistula 9 had an associated cardiac malformation (4 right-sided aortic arch and 4 ventricular septal defects). The possible individualization of multiple abnormality syndromes in which these two types of malformations are associated with others such as vertebral arch defects, limb anomalies, imperforate anus, etc. is discussed. These are so-called "Vater" or "Vactel" syndromes.


Reassessment of the Widal test in the diagnosis of typhoid.
The Widal test was done in 100 nontyphoidal febrile pients, 53 patients with proven typhoid, and in 61 patients with immunological disturbances in an are where typhoid is endemic. Salmonella typhi H or O agglutinin titers of 1:160 or more were seen in only 1% of nontyphoidal fevers. Using these criteria, the Widal test was of diagnostic value in 50 of 53 (94.3%) of patients with typhoid. The Widal test was positive in 85.7% of patients even in the first week of the illness, possibly owing to the hyperimmune state of the patients. A false-positive test was found in 11.5% of patients with major immunological disturbances in whom the Widal test must therefore be interpreted with caution.


Assays for quantification of male and female gametocytes in human blood by qRT-PCR in the absence of pure sex-specific gametocyte standards.
Malaria transmission from humans to Anopheles mosquitoes requires the presence of gametocytes in human peripheral circulation, and the dynamics of transmission are determined largely by the density and sex ratio of the gametocytes. Molecular methods are thus employed to measure gametocyte densities, particularly when assessing transmission epidemiology and the efficacy of transmission-blocking interventions. However, accurate quantification of male and female gametocytes with molecular methods requires pure male and female gametocytes as reference standards, which are not widely available. qRT-PCR assays were used to quantify levels of sex-specific mRNA transcripts in Plasmodium falciparum female and male gametocytes (pfs25 and pfMGET, respectively) using synthetic complimentary RNA standards and in vitro cultured gametocytes. Assays were validated and assay performance was investigated in blood samples of clinical trial participants using these standards and compared to absolute quantification by droplet digital PCR (ddPCR). The number of transcript copies per gametocyte were determined to be 279.3 (95% CI 253.5-307.6) for the female-specific transcript pfs25, and 12.5 (95% CI 10.6-14.9) for the male-specific transcript pfMGET. These numbers can be used to convert from transcript copies/mL to gametocyte/mL. The reportable range was determined to be 5.71 × 106 to 5.71 female gametocytes/mL for pfs25, and 1.73 × 107 to 1.73 × 101 male gametocytes/mL for pfMGET. The limit of detection was 3.9 (95% CI 2.5-8.2) female gametocytes/mL for pfs25, and 26.9 (95% CI 19.3-51.7) male gametocytes/mL for PfMGET. Both assays showed minimal intra-assay and inter-assay variability with coefficient of variation < 3%. No cross-reactivity was observed in both assays in uninfected human blood samples. Comparison of results from ddPCR to qRT-PCR assays on clinical blood samples indicated a high-level agreement (ICC = 0.998 for pfs25 and 0.995 for pfMGET). This study reports the validation of qRT-PCR assays that are able to accurately quantify female and male P. falciparum gametocytes at sub-microscopic densities. The assays showed excellent reproducibility, sensitivity, precision, specificity, and accuracy. The methodology will enable the estimation of gametocyte density in the absence of pure female and male gametocyte standards, and will facilitate clinical trials and epidemiological studies.


Sexual activities and attitudes of women with vulvar dysesthesia.
To assess the relationship between sexual activities and attitudes of women with and without vulvar dysesthesia. Women with vulvar dysesthesia, 18-60 years old, and ethnically matched women without this disorder were enrolled in this cross-sectional study, completed a 27-page questionnaire, and had a physical examination. We compared sexual activities and attitudes between patients and controls using univariate and multivariable analyses. Between January 21, 2001, and December 12, 2002, we enrolled 63 women with vulvar dysesthesia and 62 controls who reported having a current sexual partner. Women with vulvar dysesthesia were less likely than controls to have had, during the previous month, intercourse (66.7% versus 83.9%, P =.03) and orgasms (57.6% versus 78.7%, P =.02), although the association with intercourse was no longer statistically significant after controlling for age, education, and smoking status (P =.07). Of those having had intercourse in the past month, the frequency of this activity was less among patients (3.0 +/- 2.7 versus 6.2 +/- 4.5 episodes, P <.001). Frequencies of orgasms, fellatio, cunnilingus, and masturbation did not significantly differ between patients and controls. Patients rated sex less important and rated themselves more negatively as sexual people than did controls (P <.001). Women with vulvar dysesthesia are similar to those without in many sexual activities but are less likely to have had an orgasm in the past month or to have had intercourse at the same frequency as controls. Although some attitudes were similar, sexual interest and self-concept were decreased among those with this disorder.


Amyloid-β and tau pathologies relate to distinctive brain dysconnectomics in preclinical autosomal-dominant Alzheimer's disease.
The human brain is composed of functional networks that have a modular topology, where brain regions are organized into communities that form internally dense (segregated) and externally sparse (integrated) subnetworks that underlie higher-order cognitive functioning. It is hypothesized that amyloid-β and tau pathology in preclinical Alzheimer's disease (AD) spread through functional networks, disrupting neural communication that results in cognitive dysfunction. We used high-resolution (voxel-level) graph-based network analyses to test whether in vivo amyloid-β and tau burden was associated with the segregation and integration of brain functional connections, and episodic memory, in cognitively unimpaired Presenilin-1 E280A carriers who are expected to develop early-onset AD dementia in ∼13 y on average. Compared to noncarriers, mutation carriers exhibited less functional segregation and integration in posterior default-mode network (DMN) regions, particularly the precuneus, and in the retrospenial cortex, which has been shown to link medial temporal regions and cortical regions of the DMN. Mutation carriers also showed greater functional segregation and integration in regions connected to the salience network, including the striatum and thalamus. Greater tau burden was associated with lower segregated and integrated functional connectivity of DMN regions, particularly the precuneus and medial prefrontal cortex. In turn, greater tau pathology was related to higher segregated and integrated functional connectivity in the retrospenial cortex and the anterior cingulate cortex, a hub of the salience network. These findings enlighten our understanding of how AD-related pathology distinctly alters the brain's functional architecture in the preclinical stage, possibly contributing to pathology propagation and ultimately resulting in dementia.


Association between aspirin use during pregnancy and cardiovascular risk factors 2-7 years after delivery: The nuMoM2b Heart Health Study.
To evaluate the association between aspirin use during first pregnancy and later maternal cardiovascular risk. In this secondary analysis of a prospective cohort, we included participants who carried their first pregnancy to 20 + weeks, had data regarding aspirin use, and attended a study visit 2-7 years following delivery. The exposure was aspirin use during the first pregnancy. We calculated aspirin use propensity scores from logistic regression models including baseline variables associated with aspirin use in pregnancy and cardiovascular risk. Outcomes of interest were incident cardiovascular-related diagnoses 2-7 years following delivery. Robust Poisson regression calculated the risk of outcomes by aspirin exposure, adjusting for the aspirin use propensity score. The primary outcome was a composite of incident cardiovascular diagnoses at the time of the study visit: cardiovascular events, chronic hypertension, metabolic syndrome, prediabetes or type 2 diabetes, dyslipidemia, and chronic kidney disease. Of 4,480 women included, 84 (1.9%) reported taking aspirin during their first pregnancy. 52.6% of participants in the aspirin-exposed group and 43.0% in the unexposed group had the primary outcome. After adjusting for the aspirin use propensity scores, aspirin use during the first pregnancy was not associated with any of the outcomes. We did not detect an association between aspirin use during the first pregnancy and cardiovascular-related diagnoses 2-7 years later. Our study was only powered to detect a large difference in relative risk, so we cannot rule out a smaller difference that may be clinically meaningful.