Migration of small ligands in globins: Xe diffusion in truncated hemoglobin N.
In heme proteins, the efficient transport of ligands such as NO or O2 to the binding site is achieved via ligand migration networks. A quantitative assessment of ligand diffusion in these networks is thus essential for a better understanding of the function of these proteins. For this, Xe migration in truncated hemoglobin N (trHbN) of Mycobacterium Tuberculosis was studied using molecular dynamics simulations. Transitions between pockets of the migration network and intra-pocket relaxation occur on similar time scales (10 ps and 20 ps), consistent with low free energy barriers (1-2 kcal/mol). Depending on the pocket from where Xe enters a particular transition, the conformation of the side chains lining the transition region differs which highlights the coupling between ligand and protein degrees of freedom. Furthermore, comparison of transition probabilities shows that Xe migration in trHbN is a non-Markovian process. Memory effects arise due to protein rearrangements and coupled dynamics as Xe moves through it.


[Pigmented villonodular synovitis of the wrists invading bone--case report].
24-years-old patent suffers from wrist pain for 5 years and reduced wrist movement, especially on left wrist . Inflammatory lab tests were negative. Radiologic examination reveals bone destruction. Histopathological examination confirmed diagnosis: pigmented villonodular synovitis.


Marker-derived phylogeny of European cattle supports demic expansion of agriculture.
Using frequencies of 86 genes from 23 loci of blood group systems, blood and milk proteins, the genetic relationships among 14 cattle breeds including four native Balkan and four synthetic Balkan-Alp breeds were studied. The dendrogram and nonlinear map construction shows a consensus 'Balkan breed cluster', an 'Alp breed cluster', an unstable position of synthetic breeds and well-separated American breeds. Positive partial correlations between genetic distance and time elapsed since introduction of farming while keeping geographical distances constant, and regular patterns over thousands of kilometers indicate that large-scale cattle population movements together with human migration (in the Neolithic age) from the Near East into Europe across the Balkans are the most likely explanation for the genetic distances observed in our data. More recent breed differentiation and selection do not yet blur this initial pattern of European cattle populations.


The impact of interventions by a community-based organization on inner-city vaccination coverage: Fulton County, Georgia, 1992-1993.
To evaluate the impact of interventions by a community-based organization on immunization rates. Controlled community intervention trial. Children aged 3 to 59 months in Fulton County, Georgia, who were patients of 1 of 4 public clinics (clinic based), or residents of 1 of 9 inner-city communities (residence based). (1) Clinic-based intervention included monthly review of clinic vaccination records to identify undervaccinated children followed by contact with family (reminder-recall strategy); (2) residence-based intervention included door-to-door assessment and education campaigns followed by mobile van vaccinations, temporary on-site vaccination stations, free child care and transportation to providers, incentives of food and baby products, focus groups, and coalitions with local organizations (community saturation with vaccination messages and opportunities). Change in vaccination rates after 1 year based on clinic record reviews and population surveys. For clinic-based intervention, series completion rates improved from 43% (87/204) to 58% (99/170) in intervention clinics (P=.003), while rates in control clinics did not change from the baseline of 52% (81/157 to 78/150), for a net difference between intervention and control arms of +15 percentage points (P=.046). For residence-based intervention, age-appropriate vaccination rates improved from 44% (154/347) to 61% (260/429) in intervention communities (+17 percentage points; P<.001) compared with improvement of 44% (78/178) to 58% (129/221) for control communities (+14 percentage points; P=.004), but the difference between arms was not significant (+3 percentage points, P=.78). Reminder-recall activities by the community-based organization improved vaccination rates in intervention clinics compared with control clinics. A statistically significant impact on vaccination rates could not be detected for residence-based interventions by the community-based organization.


Towards Topology-Free Programming for Cyber-Physical Systems with Process-Oriented Paradigm.
The paper proposes a topology-free specification of distributed control systems by means of a process-oriented programming paradigm. The proposed approach was characterized, on the one hand, by a topologically independent specification of the control algorithm and, on the other hand, by the possibility of using existing formal verification methods by preserving the semantics of a centralized process-oriented program. The paper discusses the advantages of a topologically independent specification of distributed control systems, outlines the features of control software, argues why the use of a process-oriented approach to the development of the automation of cyber-physical systems is suitable for solving these problems, describes a general scheme for implementing a distributed control system according to a process-oriented specification, and proposes a formal heuristic algorithm for partitioning a sequential process-oriented program into independent clusters. We illustrate our algorithm with bottle-filling and sluice case studies.


Survey of dental prophylaxes rendered by pediatric dentists in New England.
The objective of this anonymous postal survey was to assess the provision of dental prophylaxis by pediatric dentists in New England. The questionnaire survey was sent by first class mail in September, 2001 to all 217 American Academy of Pediatric Dentistry (AAPD) members in active private practice in the six New England States of Connecticut, Massachusetts, Maine, New Hampshire, Rhode Island, and Vermont. A self-addressed, stamped envelope was provided to facilitate the returned response. The survey had a response rate of 70%. Most practitioners (93%) routinely recommended dental prophylaxis for their recall patients. The proportion of practitioners who considered the following indications for recommending dental prophylaxis was: plaque, stain, and/or calculus removal - 99%; caries prevention - 75%; prior to topical fluoride application - 82%; prior to sealant application - 58%; and for behavioral modification - 68%. Almost two thirds of the practitioners (62%) defined dental prophylaxis as referring to both rubber cup pumice prophylaxis as well as to toothbrush prophylaxis. However, only one in four practitioners (26%) had modified her/his clinical practice to substitute toothbrush prophylaxis in lieu of rubber cup pumice prophylaxis. Pediatric dentists in New England routinely provide dental prophylaxis to their recall patients.


[Anti-VEGF treatment for retinal angiomatous proliferation].
Retinal angiomatous proliferation (RAP) is a subform of neovascular age-related macular degeneration (AMD), which is characterized by a particularly poor prognosis. The aim of this study is to describe the loading phase and maintenance phase for RAP during intravitreal anti-VEGF treatment. A total of 82 eyes in 82 patients with RAP stages 1-3 were treated during upload therapy with repeated intravitreal injections of 1.25 mg bevacizumab at intervals of 4 weeks until the retinal edema resolved. Baseline examination included measurement of the best corrected distance visual acuity (ETDRS chart), central retinal thickness using optical coherence tomography (OCT), and fluorescein angiography (FLA). During maintenance therapy, the patients' distance visual acuity was monitored at 4- to 12-week intervals and OCT or FLA performed if needed. The average follow-up was 7.4 months (SD 4.2). Treatment with intravitreal anti-VEGF therapy was repeated if there was evidence of sub- or intraretinal fluid with a decrease in visual acuity of 5 points or more, increase of the central retinal thickness of 100 microm or more on OCT, or subjective deterioration with verifiable sub- or intraretinal fluid. During upload therapy an improvement in visual acuity of an average of +5.1 letters (mean, n=82 eyes) was observed. During maintenance therapy it was initially possible to sustain this treatment effect. However, 5 months after loading therapy was concluded, a deterioration of -5.5 letters (mean, n=31) was evident in comparison with the end of loading therapy. During the further course deterioration continued (12-month follow-up: -8.6 letters, n=7). Recurrence occurred in 60% of the cases, on average 8 weeks after termination of loading therapy. During an observation period of 6 months (n=66) a total of 3.6 injections were necessary. Therapy with intravitreal anti-VEGF medications represents a treatment option for RAP, but in the long term the disease continues to progress accompanied by functional deterioration. We thus recommend that patients with RAP be monitored at 4-week intervals to permit early treatment of recurrence.


[Classification of maxillary dental arches by correlation and principal component analyses].
To evaluate the morphology of dental arches. 62 (male: 36, female: 26) paired casts having normal dentitions and occlusion were selected from 396 (age: 18 to 26 years old; male: 257, female: 139) sets of dental study models. The maxillary dentitions were preliminarily classified as square, round-square, round and round V-shaped arches based on the conventional morphological descriptions. Midpoints of the incisor edge (I1(R), I1(L), I2(R), &I2(L)), summits of the cuspids (C(R) & C(L)), buccal cusps of the premolars (P1(R), P1(L), P2(R), & P2(L)), mesial buccal cusps of the first and second molars (M1(R), M1(L), M2(R), & M2(L)), and the midpoint (A) of line I1(R)-I1(L) were designated as reference points. From A, let a vertical line intersected line M2(R)-M2(L) at reference point B. The line A-B intersected CR-CL at reference point E. The following items were evaluated: (1) the protrusion of the cuspids by 1. angle I2(R)-C(R)-P1(R) (angle R) + angle I2(L)-C(L)-P1(L) (angle L); (2) the curvature of the anterior teeth by 2. A-B/C(R)-C(L), 3. 180 degrees-angle C(R)-A-C(L), and 4. A-E/C(R)-C(L); (3) the length to width ratio of the dental arch by 5. A-B/M2(R)-M2(L); (4) the degree of roundness of the maxillary arch by estimation of 6. (rtheta(5)-rtheta(4))(R)+(rtheta(5)-rtheta(4))(L); and (5) an item 7. for the differentiation of type I and type II round-square arches by relating the bilateral contour and position of break line P1-P2-M1-M2 (i) to line P1-M2 (ii). The data of items 1, 2, 3, 4, 5, and 6 were further standardized and summarized into three essential principal components: (1) the curvature of the anterior teeth, (2) the curvilinear contour of the dental arch, and (3) the length-to-width ratio of the dental arch. (1) 60% of the maxillary dentitions were round-square arches which showed no prominent principal component; (2) square maxillary arches distinctly showed a small 1. angle R+angle L; (3) round arches were characteristic by small 6 (rtheta(5)-rtheta(4))(R)+(rtheta(5)-rtheta(4))(L) values; and (4) round V-shaped arches had large 2, 3 and 4 values. We concluded that parameters 1, 2, 3, 4, 5, 6 and 7 were summarized into three principal components (first principal component, second principal component and third principal component). Through three principal component analysis, we can quickly evaluate the morphology of the dental arches clinically. This methods is simple and of validity. And we can also obtain the characteristics of maxillary dental arches.


Woofer-tweeter adaptive optics scanning laser ophthalmoscopic imaging based on Lagrange-multiplier damped least-squares algorithm.
We implemented a Lagrange-multiplier (LM)-based damped least-squares (DLS) control algorithm in a woofer-tweeter dual deformable-mirror (DM) adaptive optics scanning laser ophthalmoscope (AOSLO). The algorithm uses data from a single Shack-Hartmann wavefront sensor to simultaneously correct large-amplitude low-order aberrations by a woofer DM and small-amplitude higher-order aberrations by a tweeter DM. We measured the in vivo performance of high resolution retinal imaging with the dual DM AOSLO. We compared the simultaneous LM-based DLS dual DM controller with both single DM controller, and a successive dual DM controller. We evaluated performance using both wavefront (RMS) and image quality metrics including brightness and power spectrum. The simultaneous LM-based dual DM AO can consistently provide near diffraction-limited in vivo routine imaging of human retina.


Indoor Skydiving: An Emerging Cause of Anterior Shoulder Dislocations.
Background: The risks of indoor skydiving have not been extensively studied. Indoor skydiving facilities are often used for corporate events and parties and by relatively inexperienced participants who may not appreciate the risks involved. The abducted and externally rotated shoulder position, combined with nearby walls, tight spaces, and the strong airstream, has resulted in a pattern of shoulder dislocation injuries. Case Report: A 26-year-old male presented with recurrent left shoulder instability after developing an engaging Hill-Sachs lesion following traumatic anterior shoulder dislocation while indoor skydiving. He entered the wind tunnel with his arms abducted and externally rotated. The wind created an upward force that held his arms in this position. As he reached with his left arm for the side of the tunnel to exit, his arm was forced into further external rotation, dislocating the shoulder. The patient was treated arthroscopically with a remplissage procedure and repair of the glenoid labrum. Postoperatively, he resumed his active lifestyle and sports without further dislocations or instability. Conclusion: Indoor skydiving may pose a high risk of anterior dislocation because the shoulder is forced into abduction and external rotation in the free-fall position. We advise caution before participation in indoor skydiving by any individual, but especially those with a history of shoulder instability.


GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases.
CRISPR RNA-guided nucleases (RGNs) are widely used genome-editing reagents, but methods to delineate their genome-wide, off-target cleavage activities have been lacking. Here we describe an approach for global detection of DNA double-stranded breaks (DSBs) introduced by RGNs and potentially other nucleases. This method, called genome-wide, unbiased identification of DSBs enabled by sequencing (GUIDE-seq), relies on capture of double-stranded oligodeoxynucleotides into DSBs. Application of GUIDE-seq to 13 RGNs in two human cell lines revealed wide variability in RGN off-target activities and unappreciated characteristics of off-target sequences. The majority of identified sites were not detected by existing computational methods or chromatin immunoprecipitation sequencing (ChIP-seq). GUIDE-seq also identified RGN-independent genomic breakpoint 'hotspots'. Finally, GUIDE-seq revealed that truncated guide RNAs exhibit substantially reduced RGN-induced, off-target DSBs. Our experiments define the most rigorous framework for genome-wide identification of RGN off-target effects to date and provide a method for evaluating the safety of these nucleases before clinical use.


Development of a simulation-based sub-module in undergraduate medical education.
It is well documented that medical students struggle to bridge the theory-practice gap. Competently integrating classroom-taught skills into the clinical setting is a challenge. A survey of final year medical students identified a lack of confidence in managing common clinical emergencies that are experienced in the Intern role. The researchers devised a new sub-module to include high-fidelity simulation in the undergraduate medical curriculum. This development was underpinned by an educational curriculum model. Students attended small group teaching using high-fidelity simulation training (Sim-Man 3G ™) and practiced managing common emergencies. Training sessions were facilitated by clinical tutors and anaesthetic lecturers. Cases were supplied by clinical staff. An evidence-based format was utilised as an integrative teaching method. A simulation-based sub-module based on Finks Taxonomy was planned, introduced and evaluated. Utilisation of a high-fidelity simulation-based approach to teaching had a marked effect on student's confidence in their ability to manage clinical emergencies. Students reported very positive attitudes to this new method of teaching, specifically related to the integration of previously acquired knowledge and skills. A hands-on practical approach in a safe learning environment was valued by students. The module descriptor devised can be used as a template for further curricular reform. Simulation-based teaching is feasible in the undergraduate setting and may ease the transition to postgraduate teaching modalities.


A fossil "Social Stomach": Micro-CT imaging unearths first fossil record of ant proventriculi and comparative analysis with recent species.
Eusociality is an extreme form of social behavior found in some invertebrates, especially in ants, which allows them to thrive and dominate the environment through their cooperative and organized behavior. Their complex colony structure and social behavior is provided, among other things, by morphological adaptations, such as the unique structure and a huge variety of proventriculi relative to other insect groups. Here, we report the first discovery of proventriculus by X-ray microcomputed tomography (µCT) in the fossil ants †Oecophylla brischkei Mayr, 1868 and †Oecophylla crassinoda Wheeler, 1922. This is a remarkable fossil find, as it opens up new perspectives for research on phylogeny, the evolution of ant sociality and feeding behavior. We present a 3D model of the proventriculus in Oecophylla smaragdina (Fabricius, 1775) and a 3D model of the proventriculus in †Oe. brischkei and compare them. Fossil proventriculi are not fundamentally different in structure from the recent. Apparently, already in the late Eocene, the lifestyle and sociality of ants of the genus Oecophylla were similar to the one we are currently observing.


In vitro fertilization in inbred BALB/c mice I: isotonic osmolarity and increased calcium-enhanced sperm penetration through the zona pellucida and male pronuclear formation.
To optimize IVF conditions for BALB/c mice, which are known to have poor in vitro fertilizability, the requirements for sperm-ova interaction were studied by use of modified simplex optimization medium (mKSOM) as a basic medium. Modified human tubal fluid (mHTF) was used for sperm preincubation and acted as a positive control. When the two media were compared, neither capacitation nor fertilization was supported in mKSOM. Increasing the calcium concentration in mKSOM to 5 mM or more during sperm: ova coincubation improved zona penetration but not male pronuclear (MPN) formation to the same level as those cells incubated in mHTF. When medium osmolarity was varied from 230-305 mOsmol by NaCl at 5 mM CaCl2, MPN formation improved at 280 mOsmol or higher osmolarity to the same level as that found when using mHTF. When NaCl equivalent to 25-75 mOsmol was substituted with trehalose, no significant reduction in fertilization was observed. Substitution of NaCl equivalent to 75 mOsmol with other osmotic reagents (sucrose, choline chloride and sorbitol) resulted in similar levels of fertilization as found with mHTF, except for sorbitol, which reduced fertilization significantly caused by its detrimental effect on sperm viability. At isotonic osmolarity (305 mOsmol), maximum fertilization was observed at 5 mM CaCl2; lower or higher concentrations of CaCl2 resulted in reduced fertilization. Calcium and osmolarity, therefore, are important for sperm : ova interaction in BALB/c mice and the increases in calcium to 5 mM and osmolarity to 305 mOsmol are optimal for BALB/c sperm to penetrate through the zona and to form MPN.


Miniaturized thermocycler based on thermoelectric heating for diagnosis of sexually transmitted disease by DNA amplification.
Sexually transmitted disease (STD) is among the most common infectious diseases; therefore, it is necessary to develop sensitive early diagnostic techniques. As the gold standard, polymerase chain reaction (PCR) has been most widely employed for STD diagnosis; however, PCR requires large and expensive instruments. In this study, miniaturized thermal cycler using Peltier modules was developed for the PCR analysis. In comparison with the conventional PCR instrument, the Peltier-based micro-PCR (P-mPCR) device developed in this study enables one to amplify and successfully distinguish between DNA of different sizes. Furthermore, by using the clinical vaginal sample collected with the vaginal swab and tampon, different kinds of STD bacteria could be detected with high accuracy (∼94.19%) and high sensitivity (∼95.6%). Therefore, the P-mPCR device will be applicable in STD diagnosis as well as the detection of other bacteria/viruses using DNA amplification in regions including those with limited resources.


Using correspondence analysis in pharmacy practice.
Correspondence analysis (CA) and some of its uses in pharmacy practice are described. CA is a multivariate and graphic form of exploratory data analysis that reduces a multidimensional contingency table to a two-dimensional plot with minimal loss of information. An exploratory association between medication use and the number and severity of falls among the elderly living at home is used to illustrate the process of CA. Row profiles are constructed by dividing the count data in each cell by the total of the corresponding row, converting frequency data into relative frequencies. A graph of the data can be made by recalculating the frequencies to illustrate how the data might appear in a three-dimensional space with the axes defined by the three categories of falling (minimum, moderate, and major). For example, the number of patients using central nervous system (CNS) agents is plotted using the relative frequencies as vectors to locate a position in the three-dimensional space. All drug profiles and the average profile will lie exactly in a triangular plane defined by the terminal points of each axis. In this manner, data with three dimensions can be projected onto a two-dimensional space. Transferring the vectors named by pharmacologic class onto the plot requires creating two axes that cross at a common point or the origin. This point locates the average profile and defines an important reference for making comparisons. CA revealed that CNS agents may be associated with moderate to major falls, psychotherapeutic agents with moderate falls, and anticoagulants with moderate to minimum falls. CA has widespread uses in pharmacy practice. It can identify patients at risk for serious but preventable drug-related complications, enabling pharmacists to allocate pharmacy resources to the areas in most need and suggest hypotheses for future research. Clinicians can also use CA to analyze vast amounts of patient-related data to uncover hard-to-detect associations. The use of CA in pharmacy practice will allow new strategies for improved patient care to be more readily appreciated and implemented.


Utility of Talus Osteochondral Allograft Augmentation for Varying Hill-Sachs Lesion Sizes: A Cadaveric Study.
Humeral head reconstruction with fresh osteochondral allografts (OCA) serves as a potential treatment option for anatomic reconstruction. More specifically, talus OCA is a promising graft source because of its high congruency with a dense cartilaginous surface. To analyze the surface geometry of the talus OCA plug augmentation for the management of shoulder instability with varying sizes of Hill-Sachs lesions (HSLs). Controlled laboratory study. Seven fresh-frozen cadaveric shoulders were tested in this study. The humeral heads were analyzed using actual patients' computed tomography scans. Surface laser scan analysis was performed on 7 testing states: (1) native state; (2) small HSL; (3) talus OCA augmentation for small HSL; (4) medium HSL; (5) talus OCA augmentation for medium HSL; (6) large HSL; and (7) talus OCA augmentation for large HSL. OCA plugs were harvested from the talus allograft and placed in the most medial and superior aspect of each HSL lesion. Surface congruency was calculated as the mean absolute error and the root mean squared error in the distance. A 1-way repeated-measures analysis of variance was performed to evaluate the effects of the difference in the HSL size and associated talus OCA plugs on surface congruency and the HSL surface area. The surface area analysis of the humeral head with the large (1469 ± 75 mm2), medium (1391 ± 81 mm2), and small (1230 ± 54 mm2) HSLs exhibited significantly higher surface areas than the native state (1007 ± 88 mm2; P < .001 for all sizes). The native state exhibited significantly lower surface areas as compared with after talus OCA augmentation for large HSLs (1235 ± 63 mm2; P < .001) but not for small or medium HSLs. Talus OCA augmentation yielded improved surface areas and congruency after treatment in small, medium, and large HSLs (P < .001). Talus OCA plug augmentation restored surface area and congruency across all tested HSLs, and the surface area was best improved with the most common HSLs-small and medium. Talus OCA plugs may provide a viable option for restoring congruity of the shoulder in patients with recurrent anterior glenohumeral instability and an HSL.


Issues in management of pancreatic pseudocysts.
Pancreatic pseudocysts (PPs) comprise more than 80% of the cystic lesions of the pancreas and cause complications in 7-25% of patients with pancreatitis or pancreatic trauma. The first step in the management of PPs is to exclude a cystic tumor. A history of pancreatitis, no septation, solid components or mural calcification on CT scan and high amylase content at aspiration favor a diagnosis of PP. Endoscopic ultrasound (EUS)-guided FNAC is a valuable diagnostic aid. Intervention is indicated for PPs which are symptomatic, in a phase of growth, complicated (infected, hemorrhage, biliary or bowel obstruction) or in those occurring together with chronic pancreatitis and when malignancy cannot be unequivocally excluded. The current options include percutaneous catheter drainage, endoscopy and surgery. The choice depends on the mode of presentation, the cystic morphology and available technical expertise. Percutaneous catheter drainage is recommended as a temporizing measure in poor surgical candidates with immature, complicated or infected PPs. The limitations include secondary infection and pancreatic fistula in 10-20% of patients which increase complications following eventual definitive surgery. Endoscopic therapy for PPs including cystic-enteric drainage (and transpapillary drainage), is an option for PPs which bulge into the enteric lumen which have a wall thickness of less than 1 cm and the absence of major vascular structures on EUS in the proposed tract or those which communicate with the pancreatic duct above a stricture. Surgical internal drainage remains the gold standard and is the procedure of choice for cysts which are symptomatic or complicated or those having a mature wall,. Being more versatile, a cystojejunostomy is preferred for giant pseudocysts (>15 cm) which are predominantly inframesocolic or are in an unusual location. In PPs with coexisting chronic pancreatitis and a dilated pancreatic duct, duct drainage procedures (such as longitudinal pancreaticojejunostomy) should be preferred to a cyst drainage procedure.


An experimental and theoretical study of the mixing characteristics of a periodically reoriented irrotational flow.
The minimum-energy method to generate chaotic advection should be to use an irrotational flow. However, irrotational flows have no saddle connections to perturb in order to generate chaotic orbits. To the early work of Jones & Aref (Jones & Aref 1988 Phys. Fluids 31, 469-485 (doi:10.1063/1.866828)) on potential flow chaos, we add periodic reorientation to generate chaotic advection with irrotational experimental flows. Our experimental irrotational flow is a dipole potential flow in a disc-shaped Hele-Shaw cell called the rotated potential mixing flow; it leads to chaotic advection and transport in the disc. We derive an analytical map for the flow. This is a partially open flow, in which parts of the flow remain in the cell forever, and parts of it pass through with residence-time and exit-time distributions that have self-similar features in the control parameter space of the stirring. The theory compares well with the experiment.


Enhanced Hydrothermal Stability and Catalytic Performance of HKUST-1 by Incorporating Carboxyl-Functionalized Attapulgite.
Much attention has been paid to metal-organic frameworks (MOFs) due to their large surface areas, tunable functionality, and diverse structure. Nevertheless, most reported MOFs show poor hydrothermal stability, which seriously hinders their applications. Here a strategy is adopted to tailor the properties of MOFs by means of incorporating carboxyl-functionalized natural clay attapulgite (ATP) into HKUST-1, a well-known MOF. A new type of hybrid material was thus fabricated from the hybridization of HKUST-1 and ATP. Our results indicated that the hydrothermal stability of the MOFs as well as the catalytic performance was apparently improved. The frameworks of HKUST-1 were severely destroyed after hydrothermal treatment (hot water vapor, 60 °C), while that of the hybrid materials was maintained. For the hybrid materials containing 8.4 wt % of ATP, the surface area reached 1302 m(2)·g(-1) and was even higher than that of pristine HKUST-1 (1245 m(2)·g(-1)). In the ring-opening of styrene oxide, the conversion reached 98.9% at only 20 min under catalysis from the hybrid material, which was obviously higher than that over pristine HKUST-1 (80.9%). Moreover, the hybrid materials showed excellent reusability and the catalytic activity was recoverable without loss after six cycles. Our materials provide promising candidates for heterogeneous catalysis owing to the good catalytic activity and reusability.


Production of antibodies to sheep red blood cells and Brucella abortus in Mongolian gerbils (Meriones unguiculatus).
The levels of total and 2-mercaptoethanol (2-ME)-resistant antibodies in male Mongolian gerbils (Meriones unguiculatus) to sheep red blood cells (SRBC) were higher than those in male C57BL/6 mice after the first and second immunizations. On the other hand, the total antibody level to Brucella abortus (BA) in gerbils was comparable to that in mice, whereas 2-ME-resistant antibody titers were lower after the first and second immunizations than in mice. After injection of 8 x 10(4) SRBC, male gerbils did not produce either total or 2-ME-resistant antibodies after the first immunization, but they produced total and 2-ME-resistant antibodies after the first to the fourth immunizations with different dilutions of BA, and both types of antibody titers significantly increased with the repeated immunizations. There were no sex differences in total and 2-ME-resistant antibody production to SRBC.


Methyldopa hypersensitivity syndrome.
A 74-year-old man developed an extensive erythematous skin eruption, fever, lymphadenopathy and eosinophilia probably due to methyldopa. The symptoms reappeared shortly after unintentional reexposure to the drug. The positive results of two in vitro tests for the detection of drug allergy not only give further support to the causal relationship between the drug and the symptoms, but also indicate the involvement of an immunologic, allergic mechanism. Although every individual symptom represented in our case has been reported in previous cases, the association of the symptoms together in one patient has never before been reported. An awareness that adverse reactions to methyldopa can mimic a variety of acute infectious diseases and lymphoproliferative disorders is the key to diagnosis and might prevent unnecessary hospitalizations, procedures, and use of potentially toxic antibiotics and chemotherapy.


Headache Rounds: Sudden, Transient Neurologic Symptoms in a Woman With Migraine.
The following article is a Graham Headache Center headache rounds presentation at the Brigham and Women's Faulkner Hospital by fellows Roni Sharon and Melissa Rayhill with special guest Tobias Kurth. It summarizes the case of a 36-year-old female with no history of migraine in the past, presenting with several transient neurological episodes associated with headache. Her history, symptoms, imaging workup is reviewed. Following the case is a discussion of the differential diagnosis for the patient's symptoms along with a review of the association of migraine with and without aura with vascular neurological insult such as stroke. The article also discusses the clinical implications of migraine as a risk factor for stroke along with possible treatment recommendations.


An extensive review: How starch and gluten impact dough machinability and resultant bread qualities.
Wheat flour can form dough with a three-dimensional viscoelastic structure that is responsible for gas holding during fermentation and oven-rise, creating a typical fixed, open-cell foam structure of bread after baking. As the major components of dough, the continuous reticular skeleton formed by gluten proteins and the concentrated starch granules entrapped in gluten matrix predominantly determine dough rheological behaviors and bread qualities. This review surveys the latest literatures and draws out a conclusion from a plethora of information related to the filling effects of starch granules on gluten matrix and the cross-linking mechanisms between gluten proteins and starch granules, which is of great significance to provide sufficient scientific knowledge for development of bread with satisfactory attributes and quality control of end products.


Beneficial effects of inhaled nitric oxide with intravenous steroid in an ischemia-reperfusion model involving aortic clamping.
This study evaluated the effects of inhaled nitric oxide (iNO) therapy combined with intravenous (IV) corticosteroids on hemodynamics, selected cytokines, and kidney messenger RNA toll-like receptor 4 (mRNA TLR4) expression in ischemia-reperfusion injury animal model. The primary endpoint was the evaluation of circulatory, respiratory, and renal function over time. We also investigated the profile of selected cytokines and high-mobility group box 1 (HMGB1) protein, as well as renal mRNA TLR4 activation determined by quantitative real-time polymerase chain reaction analysis. Pigs (n = 19) under sevoflurane AnaConDa anesthesia/sedation were randomized and subjected to abdominal laparotomy and alternatively suprarenal aortic cross-clamping (SRACC) for 90 min or sham surgery: Group 1 (n = 8) iNO (80 ppm) + IV corticosteroids (25 mg ×3) started 30 min before SRACC and continued 2 h after SRACC release, followed with decreased iNO (30 ppm) until the end of observation, Group 2 (n = 8) 90 min SRACC, Group 3 (n = 3)-sham surgery. Renal biopsies were sampled 1 hr before SRACC and at 3 and 20 h after SRACC release. Aortic clamping increased TLR4 mRNA expression in ischemic kidneys, but significant changes were recorded only in the control group ( P = 0.016). Treatment with iNO and hydrocortisone reduced TLR4 mRNA expression to pre-ischemic conditions, and the difference observed in mRNA expression was significant between control and treatment group after 3 h ( P = 0.042). Moreover, animals subjected to treatment with iNO and hydrocortisone displayed an attenuated systemic inflammatory response and lowered pulmonary vascular resistance plus increased oxygen delivery. The results indicated that iNO therapy combined with IV corticosteroids improved central and systemic hemodynamics, oxygen delivery, and diminished the systemic inflammatory response and renal mRNA TLR4 expression.


Molecular pathogenesis of Kallmann's syndrome.
Hypogonadotrophic hypogonadism (HH) is characterized by delayed or absent pubertal development secondary to gonadotrophin deficiency. HH can result from mutations of the gonadotrophin-releasing hormone receptor 1, the gonadotrophin beta-subunits, or various transcription factors involved in pituitary gland development. HH occurs in DAX1 mutations when associated with adrenal insufficiency (adrenal hypoplasia congenita), and is also linked with obesity in patients with mutations of leptin and its receptor, as well as mutations in prohormone convertase 1. Rarely, HH has resulted from kisspeptin receptor (GPR54) mutations, a gene implicated in the regulation of pubertal onset. When occurring with anosmia (a lack of sense of smell), HH is referred to as Kallmann's syndrome (KS). Two KS-related loci are currently known: KAL1, encoding anosmin-1, responsible for X-linked KS, and KAL2, encoding the fibroblast growth factor receptor 1 (FGFR1), mutated in autosomal dominant KS. Anosmin-1 is an extracellular glycoprotein with some unique structural characteristics; it interacts with both urokinase-type plasminogen activator and FGFR1. It has previously been shown that anosmin-1 enhances FGFR1 signalling in a heparan sulphate-dependent manner, and proposed that anosmin-1 fine-tunes FGFR1 signalling during olfactory and GnRH neuronal development. Here, we review the known normosmic causes of HH, and discuss novel developmental and molecular mechanisms underlying KS; finally, we introduce three novel genes (NELF, PKR2, and CHD7) that may be associated with some phenotypic features of KS.


Rupture of a liposomal vesicle.
We discuss pore dynamics in osmotically stressed vesicles. A set of equations which govern the liposomal size, internal solute concentration, and pore diameter is solved numerically. We find that dependent on the internal solute concentration and vesicle size, liposomes can stay pore free, nucleate a short-lived pore, or nucleate a long-lived pore. The phase diagram of pore stability is constructed, and the different scaling regimes are deduced analytically.


Eugenol, a plant-derived phenolic nutraceutical, protects thiol (SH) group in myocardium from ROS-mediated oxidation under chemotherapeutic stress induced by arsenic trioxide - a in vivo model study.
Arsenic trioxide is an effective chemotherapeutic agent for the treatment of acute promyelocytic leukemia. The clinical usefulness of arsenic trioxide is narrow due to different organ toxicities. It is hypothesized that the generation of reactive oxygen species by arsenic trioxide leads to thiol-based oxidative damage in rat myocardium. In this study, the defensive effect of eugenol on thiol-based oxidative stress was investigated in arsenic trioxide-treated rats. Rats were orally administered with arsenic trioxide (4 mg/kg per day) alone and in combination with eugenol (5 mg/kg per day) for 30 days. Reduction in relative organ weight, total thiol level, protein thiol content, acid-soluble thiol content, thioredoxin activity, and protein content was witnessed in arsenic trioxide-treated rats. Additionally, the total antioxidant activity, tissue GSH level, and GSH/GSSG ratio were considerably diminished. However, the co-treatment of eugenol noticeably sheltered the arsenic trioxide-mediated cardiotoxicity. In conclusion, eugenol is a prospective phenolic compound, of natural origin, for protecting the thiol group in myocardium from oxidative stress by chemotherapeutic compounds.


Group effort in resuscitation teams.
Baumeister and colleagues underline that individual identification and differentiation of selves are important characteristics for group performance. They name specialization, moral responsibility, and efficiency as vital components of well-functioning groups. In my commentary, I transfer their framework to the group effort within resuscitation teams to discuss for the first time how these components determine teamwork during resuscitation.


DKK4 enhances resistance to chemotherapeutics 5-Fu and YN968D1 in colorectal cancer cells.
Dickkopf-related protein 4 (DKK4) is a target of the β-catenin/transcription factor 4 complex in colorectal cancer. Previous research has demonstrated that its expression level may vary and has indicated that it may have a role in the development of resistance to chemotherapy in colorectal cancer cells. In the present study, DKK4 was over expressed in several colorectal cancer cell lines. The DKK4 over-expressing cell lines were screened using reverse transcription quantitative polymerase chain reaction analysis and western blotting. Analysis of cell viability in the control and DKK4 over-expressing cell lines, following treatment with 5-fluorouracil (5-Fu), YN968D1 or both, indicated that DKK4 over-expressing cells exhibit increased drug resistance. The results of Transwell chamber assays suggested that DKK4 had an effect on cell migration. Furthermore, the results from flow cytometric analysis showed that the percentage of apoptotic cells was reduced in the DKK4 over-expressing cell lines, following drug treatment, compared with the control. The present data suggested that DKK4 may enhance the resistance of colorectal cancer cells to 5-Fu and YN968D1 treatment, when used alone or in combination.


Multiple Neural Networks Malfunction in Primary Blepharospasm: An Independent Components Analysis.
Primary blepharospasm (BPS) is a focal dystonia characterized by involuntary blinking and eyelid spasms. The pathophysiology of BPS remains unclear. Several neuroimaging studies have suggested dysfunction of sensory processing and sensorimotor integration, but the results have been inconsistent. This study aimed to determine whether patients with BPS exhibit altered functional brain connectivity and to explore possible correlations between these networks and clinical variables. Twenty-five patients with BPS and 25 healthy controls were enrolled. We found that the patient group exhibited decreased connectivity within the sensory-motor network (SMN), which involved regions of the bilateral primary sensorimotor cortex, supplementary motor area (SMA), right premotor cortex, bilateral precuneus and left superior parietal cortex. Within the right fronto-parietal network, decreased connections were observed in the middle frontal gyrus, dorsal lateral prefrontal cortex and inferior frontal gyrus. Regarding the salience network (SN), increased connectivity was observed in the left superior frontal gyrus and middle frontal gyrus. These findings suggest the involvement of multiple neural networks in primary BPS.


Palladium-catalyzed ring-opening thiocarbonylation of vinylcyclopropanes with thiols and carbon monoxide.
Palladium-catalyzed ring-opening thiocarbonylation of vinylcyclopropanes (VCPs) with thiols and carbon monoxide affords the corresponding unsaturated thioesters in moderate to excellent yields. This reaction provides a general method for the ring-opening thiocarbonylation of VCPs. It further demonstrates the utility of transition metal catalysts for the synthesis of organosulfur compounds.


Functions of preschool children's questions in coping with hospitalization.
This study was designed to determine the functions of hospitalized preschool children's questions. Subjects were 24 hospitalzied preschool children ranging in age from 3 years to 5 years, 11 months. Subjects' spontaneous questions, addressed to adults in a hospital setting, were recorded and categorized using Piaget's functional classification. The categories were: (a) causal explanation, (b) reality and history, (c) actions and intentions, (d) classification and evaluation, (e) rules, and (f) calculation. The frequency distribution of questions was determined by categories. The largest perentage of questions (53%) was about actions and intentions of persons, primarily hospital personnel. Small percentages dealt with classification and evaluation (13%) and reality and history (12%) Causal explanation questions (8%) were uncommon. A larger percentage of actions and intentions questions was found in the hospital than in nonstressful school settings. The findings suggest that hospitalized preschool children need orientation to the actions and intentions of personnel in regard to therapeutic activities, routine care, and play and social activity.


Gene expression changes in the retina following optic nerve transection.
To obtain and analyze the gene expression profiles of the retina following optic nerve transection (ONT). An axotomy animal model was generated by taking a cross section of the optic nerve with care not to damage the adjacent blood supply. The extent of cell loss was evaluated by counting cells in the ganglion cell layer (GCL) of flat-mounted retinas stained with cresyl violet. Gene expression profiles of control and ONT retinas were analyzed with Agilent's rat oligo microarray slides. Differentially expressed genes were identified from three independent experiments and clustered based on their functions with expression analysis systematic explorer software. Real-time quantitative and semiquantitative RT-PCR were used to validate changes in gene expression levels. Gene expression changes in axotomized retinas were analyzed with rat oligo microarray slides containing 22,575 oligonucleotide probes that represent over 20,000 genes and expressed sequence tags (ESTs). The expression of 493 genes was increased more than 1.5 fold, including 85 that were upregulated more than 2 fold, and the expression of 113 genes was decreased 1.5 fold or more, including 21 that were downregulated more than 2 fold. Differentially expressed genes were clustered based on their functions. Several novel genes expressed in the GCL have been identified and their expression patterns in different tissues were analyzed. Among the genes differentially regulated in retinas with induced retinal ganglion cell (RGC) death, we have identified 13 genes that are mapped to known glaucoma loci and can be considered for mutation screening in patients with inherited forms of glaucoma. The gene expression profiles of the ONT retinas can be used to identify RGC-expressed genes, which may be essential for the morphological and physiological characteristics of RGCs. The results of this study can also be used to evaluate the molecular changes taking place in the retina in response to RGC loss.


[Brain death, physiopathology, optimal care and hormonal therapy for cardiac donation].
Heart transplantation goes on leading the standard therapy for the terminal heart failure. The success of this procedure and the post-transplantation survival, meaning during the perioperative time, is variable because of the good performance of the donor graft, due to this situation, the pre-harvesting evaluation is very important. The brain death brought many changes: 1) first of all the "adrenergic storm" with its hemodynamic and cardiac dangerous events; 2) Thyroidal hormones, cortisol, vasopressin, and seric insulin depletion with its metabolic consequences respectively. Since 2002 during the Crystal Consensus started the using of the hormonal therapeutic with triyodothironine, vasopressin and metilprednisolone for improving, meaning those limitrophes ones, the performance donor hearts with brain death, and the goal has been to harvest more donated organs en quality and numbers. This hormonal therapeutic has had great acceptation and is successful; having more harvesting of solids organs: hearts, lungs and kidneys; with more performance and surviving at 1 year post-transplantation and less mortality. Our Hospital began the hormonal therapeutic since January 2011with excellent results. On this basis we can concluded that, hormonal rescue therapy, improves the availability and performance of organs for transplantation.


Complete occlusion after blunt injury to the abdominal aorta.
Injury to the abdominal aorta after blunt trauma is uncommon. When this injury results in complete vessel occlusion, the presentation is dramatic. Timely intervention is essential. After a case report, we examined all reported cases of complete occlusion after blunt injury to the abdominal aorta and reviewed the cause, presentation, and management of this injury. Complete vessel occlusion arises from intimal injury. The most frequent mechanism is compression from a seat belt or steering wheel during a motor vehicle crash. Patients present with absent femoral and distal pulses in association with lower extremity neuropathy. Intervention commonly involves bypass grafting of the abdominal aorta. Complete occlusion after blunt trauma to the abdominal aorta is rare. Neurologic deficits most commonly arise from peripheral nerve ischemia. Reperfusion within 6 hours confers a greater chance of limb salvage and neurologic recovery.


The Spatial Extent of Epiretinal Electrical Stimulation in the Healthy Mouse Retina.
Retinal prostheses use electrical stimulation to restore functional vision to patients blinded by retinitis pigmentosa. A key detail is the spatial pattern of ganglion cells activated by stimulation. Therefore, we characterized the spatial extent of network-mediated electrical activation of retinal ganglion cells (RGCs) in the epiretinal monopolar electrode configuration. Healthy mouse RGC activities were recorded with a micro-electrode array (MEA). The stimuli consisted of monophasic rectangular cathodic voltage pulses and cycling full-field light flashes. Voltage tuning curves exhibited significant hysteresis, reflecting adaptation to electrical stimulation on the time scale of seconds. Responses decreased from 0 to 300 µm, and were also dependent on the strength of stimulation. Applying the Rayleigh criterion to the half-width at half-maximum of the electrical point spread function suggests a visual acuity limit of no better than 20/946. Threshold voltage showed only a modest increase across these distances. The existence of significant hysteresis requires that future investigations of electrical retinal stimulation control for such long-memory adaptation. The spread of electrical activation beyond 200 µm suggests that neighbouring electrodes in epiretinal implants based on indirect stimulation of RGCs may be indiscriminable at interelectrode spacings as large as 400 µm.


Parent Management Training-Oregon Model: Adapting Intervention with Rigorous Research.
Parent Management Training-Oregon Model (PMTO(®) ) is a set of theory-based parenting programs with status as evidence-based treatments. PMTO has been rigorously tested in efficacy and effectiveness trials in different contexts, cultures, and formats. Parents, the presumed agents of change, learn core parenting practices, specifically skill encouragement, limit setting, monitoring/supervision, interpersonal problem solving, and positive involvement. The intervention effectively prevents and ameliorates children's behavior problems by replacing coercive interactions with positive parenting practices. Delivery format includes sessions with individual families in agencies or families' homes, parent groups, and web-based and telehealth communication. Mediational models have tested parenting practices as mechanisms of change for children's behavior and found support for the theory underlying PMTO programs. Moderating effects include children's age, maternal depression, and social disadvantage. The Norwegian PMTO implementation is presented as an example of how PMTO has been tailored to reach diverse populations as delivered by multiple systems of care throughout the nation. An implementation and research center in Oslo provides infrastructure and promotes collaboration between practitioners and researchers to conduct rigorous intervention research. Although evidence-based and tested within a wide array of contexts and populations, PMTO must continue to adapt to an ever-changing world.


[High and intensive care three years later; an evaluation of the experiences of patients and employees and the effect on coercive measures in psychiatry].
As part of the national campaign against the use of coercive measures in psychiatry, the Mental Health Service in Eindhoven set up the first modern high and intensive care centre (HIC) in Eindhoven in 2012. To study the progress of the numbers on coercive measures, and to evaluate the experiences of patients and treatment team members after working for three years according to the HIC model. We analysed the registration data, carried out a short survey and conducted interviews. We found that the number of coercive measures used between 2012 and 2015 had declined by 42%. Patients generally had a positive attitude to the treatment they had received. They appreciated the role played by the team and were pleased to have had access to modern technology. Team members had a positive attitude to working with the HIC model. The main goals of working according to the new HIC model have been achieved. However, it should be possible to increase cooperation with mobile teams, develop more links with patients' next-of-kin and make wider use of modern technology.


The diagnostic dilemma: why we need to change how we diagnose bipolar disorder in children.
Bipolar disorder diagnosis has been rising dramatically in children for the past decade. In response to this increase, writes Daniel Dickstein, M.D., of Bradley Hospital and Brown University, researchers at the National Institute of Mental Health and elsewhere are reviewing the diagnostic criteria. In coming years, Dickstein argues, recognizing and diagnosing bipolar disorder in children should be based more on biological markers, such as brain structure and the use of neural circuits, than on the inconsistent diagnostic categories laid out in the Diagnostic and Statistical Manual of Mental Disorders.


Inhibition by an alkyl-lysophospholipid of the uptake of epidermal growth factor in human breast cancer cell lines in relation to epidermal growth factor internalization.
The effects of 1-O-octadecyl-2-O-methyl-sn-glycerol-3-phosphocholine (ET-18-OCH3), an alkyl-lysophospholipid derivative, on the binding and uptake of labeled epidermal growth factor (EGF) in hormone-dependent (MCF-7 and ZR-75-1) and hormone-independent (BT-20) breast cancer cell lines were investigated at 4 degrees C and 37 degrees C. The total (bound and intracellular) EGF associated with breast cancer cells tested were largely temperature dependent. By pretreatment of the cells with ET-18-OCH3 (10 micrograms/ml) for 12 h, the EGF uptake at 37 degrees C was greatly reduced in both MCF-7 and ZR-75-1 (ET-18-OCH3-susceptible) but not in BT-20 (ET-18-OCH3-resistant) cell lines. The ET-18-OCH3 pretreatment slightly decreased the EGF uptake at 4 degrees C in MCF-7 and ZR-75-1 and had little effect on that in BT-20. The EGF binding at 37 degrees C was unaffected by ET-18-OCH3 in MCF-7 and BT-20 and slightly decreased in ZR-75-1. The EGF binding at 4 degrees C was not changed by ET-18-OCH3 in all cell lines tested. These results suggest that labeled EGF is taken up by the cells in a temperature-dependent manner and ET-18-OCH3 may inhibit this internalization process only in ET-18-OCH3-sensitive human breast cancer cell lines. It is inferrable that the inhibition of the internalization process for EGF may be one of the modes of antitumoral action of ET-18-OCH3.


Reduction of nitrate leaching with haying or grazing and omission of nitrogen fertilizer.
In some high-fertility, high-stocking-density grazing systems, nitrate (NO(3)) leaching can be great, and ground water NO(3)-N concentrations can exceed maximum contaminant levels. To reduce high N leaching losses and concentrations, alternative management practices need to be used. At the North Appalachian Experimental Watershed near Coshocton, OH, two management practices were studied with regard to reducing NO(3)-N concentrations in ground water. This was following a fertilized, rotational grazing management practice from which ground water NO(3)-N concentrations exceeded maximum contaminant levels. Using four small watersheds (each approximately 1 ha), rotational grazing of a grass forage without N fertilizer being applied and unfertilized grass forage removed as hay were used as alternative management practices to the previous fertilized pastures. Ground water was sampled at spring developments, which drained the watershed areas, over a 7-yr period. Peak ground water NO(3)-N concentrations before the 7-yr study period ranged from 13 to 25.5 mg L(-1). Ground water NO(3)-N concentrations progressively decreased under each watershed and both management practices. Following five years of the alternative management practices, ground water NO(3)-N concentrations ranged from 2.1 to 3.9 mg L(-1). Both grazing and haying, without N fertilizer being applied to the forage, were similarly effective in reducing the NO(3)-N levels in ground water. This research shows two management practices that can be effective in reducing high NO(3)-N concentrations resulting from high-fertility, high-stocking-density grazing systems, including an option to continue grazing.


Two efficient ribosomal frameshifting events are required for synthesis of mouse mammary tumor virus gag-related polyproteins.
The primary translation products of retroviral pol genes are polyproteins initiated in an upstream gene (gag). To investigate the manner in which the gag-initiated polyproteins of the mouse mammary tumor virus are produced, we determined the nucleotide sequence of a 1.8-kilobase DNA fragment that spans the region between gag and pol in the C3H strain of mouse mammary tumor virus. The sequence reveals three overlapping open reading frames: the first encodes products of gag (p27gag and p14gag); the second encodes a protein domain of unknown function (termed X) that is highly related to a similarly positioned sequence in simian type D retroviruses and the viral protease (pro); and the third encodes the reverse transcriptase. The reading frames are organized to permit uninterrupted readthrough from gag to pol if ribosomal frameshifts occur in the -1 direction within each of the two overlapping regions, one of which is 16 nucleotides in length and the other 13 nucleotides. Cell-free translation of RNA containing these overlap regions shows that fusion of the reading frames by ribosomal frameshifting occurs efficiently: about one-fourth of the ribosomes traversing the gag-X/pro overlap and one-tenth traversing the X/pro-pol overlap shift frames, generating gag-related polyproteins in ratios similar to those observed in vivo. Synthetic oligonucleotides containing either of the overlap regions inserted into novel contexts do not induce frameshifting; hence the overlapping portions of the reading frames are not sufficient to induce a frameshift event, and a larger sequence context or secondary structure may be implicated.


Derivation and Internal Validation of a Score to Predict Dehydration Severity in Patients over 5 Years with Acute Diarrhea.
Diarrheal disease accounts for more than one million deaths annually in patients over 5 years of age. Although most patients can be managed with oral rehydration solution, patients with severe dehydration require resuscitation with intravenous fluids. Scoring systems to assess dehydration have been empirically derived and validated in children under 5 years, but none have been validated for patients over 5 years. In this study, a prospective cohort of 2,172 patients over 5 years presenting with acute diarrhea to International Centre for Diarrhoeal Disease Research, Dhaka Hospital, Bangladesh, were assessed for clinical signs of dehydration. The percent difference between presentation and posthydration stable weight determined severe (≥ 9%), some (3-9%), or no (< 3%) dehydration. An ordinal regression model was derived using clinical signs and demographics and was then converted to a 13-point score to predict none (score of 0-3), some (4-6), or severe (7-13) dehydration. The Novel, Innovative Research for Understanding Dehydration in Adults and Kids (NIRUDAK) Score developed by our team included age, sex, sunken eyes, radial pulse, respiration depth, skin turgor, and vomiting episodes in 24 hours. Accuracy of the NIRUDAK Score for predicting severe dehydration, as measured by the area under the receiver operating characteristic curve, was 0.76 (95% confidence interval = 0.73-0.78), with a sensitivity of 0.78 and a specificity of 0.61. Reliability was also robust, with an Inter-Class Correlation Coefficient of 0.88 (95% confidence interval = 0.84-0.91). This study represents the first empirically derived and internally validated scoring system for assessing dehydration in children ≥ 5 years and adults with acute diarrhea in a resource-limited setting.


Quantifying spatial variability of selected soil trace elements and their scaling relationships using multifractal techniques.
Multifractal techniques were utilized to quantify the spatial variability of selected soil trace elements and their scaling relationships in a 10.24-ha agricultural field in northeast China. 1024 soil samples were collected from the field and available Fe, Mn, Cu and Zn were measured in each sample. Descriptive results showed that Mn deficiencies were widespread throughout the field while Fe and Zn deficiencies tended to occur in patches. By estimating single multifractal spectra, we found that available Fe, Cu and Zn in the study soils exhibited high spatial variability and the existence of anomalies ([α(q)max-α(q)min]≥0.54), whereas available Mn had a relatively uniform distribution ([α(q)max-α(q)min]≈0.10). The joint multifractal spectra revealed that the strong positive relationships (r≥0.86, P<0.001) among available Fe, Cu and Zn were all valid across a wider range of scales and over the full range of data values, whereas available Mn was weakly related to available Fe and Zn (r≥0.18, P<0.01) but not related to available Cu (r = -0.03, P = 0.40). These results show that the variability and singularities of selected soil trace elements as well as their scaling relationships can be characterized by single and joint multifractal parameters. The findings presented in this study could be extended to predict selected soil trace elements at larger regional scales with the aid of geographic information systems.


Avian model to mitigate gut-derived immune response and oxidative stress during heat.
The tissue injury at the early stages of the heat stress response triggers release of inflammatory and oxidative agents from intestinal content into the milieu of the body. Intestinal homeostasis (i.e., eubiosis) improves the barrier function and mitigates the gut-derived influx of endotoxins. In this study we have analyzed the mitigating role of embryonic stimulation of the gut homeostasis in chickens on immune and oxidative responses to heat. The animal trial was conducted on broiler chickens. The treatment included a single in ovo injection of the galactooligosaccharides (GOS) prebiotic into the air cell of the egg on day 12 of incubation. Control eggs were in ovo injected with the same volume of sterile physiological saline. After hatching, birds were raised in group pens (6 pens/group, 25 birds/pen). Short-term, mild heat stress was induced on day 32 post-hatching by increase in the ambient temperature above the thermal comfort (30 °C for 8.5 h). The spleen was harvested from randomly selected individuals. The relative gene expression study was conducted with RT-qPCR. The two gene panels were analyzed: (1) immune response genes (IL-1β, IL-2, IL-4, IL-6, IL-12p40 and IL-17) and (2) stress response genes (HSP25, HSP70, HSP90, BAG3, CAT and SOD). Data were evaluated by the analysis of variance in a 2 × 2 factorial design that included in ovo treatment and ambient temperature as factors. We have found that the immune-related and stress-related gene expression signatures were triggered in animals subjected to heat but with unbalanced intestinal flora (i.e., dysbiotic, without in ovo stimulation with GOS). These animals had increased expression of the genes involved in the immune responses (IL-4 and IL17) and stress responses (HSP25, HSP70, HSP90, CAT and SOD) to short-term heat stress that indicated presence of inflammatory and oxidative mediators (P < 0.05). The individuals that were in ovo stimulated with GOS did not mount the anti-inflammatory or antioxidative responses. Heat shock proteins (HSP25 and HSP70) were increased in both groups challenged with heat, which indicated their role in adaptation to heat.


Comparative Evaluation of Human Papillomavirus Type 16 L1 Protein Expressed in Plasmid- and Baculovirus-Based Systems in Insect Cells.
Human papillomavirus (HPV) has been associated with specific types of papillomas, lesions at particular anatomic sites, and malignancies. The HPV16 and HPV18 have been shown to play a role in a variety of carcinomas. The most documented HPV-associated cancer is cervical carcinoma. Suitable antigens are needed to be identified for the diagnostic tests and vaccines and the expression of L1 recombinant protein should be accelerated in papillomaviruses. Therefore, in this study, the expression of the L1 protein of HPV16 was evaluated and compared in insect cells using a plasmid and a baculovirus system. The expression of the L1 protein of HPV16 in insect cells was investigated using a plasmid (InsectDirect) and a baculovirus system (BacMagic). The expressed recombinant proteins were purified from the Sf9 lysate using Ni-NTA resin columns. The characterization of recombinant L1 protein expressed in both systems (BacMagic and InsectDirect) was performed using immunofluorescence, SDS-PAGE, western blot, and dot blot. The yields of the purified proteins from the plasmid- and baculovirus-based systems (10 ml culture; 107 cells) had the ranges of 455-495 µg/ml and 1.44-1.6 mg/ml as analyzed by spectrophotometer, respectively. The SDS-PAGE analysis of purified proteins revealed that the recombinant proteins with the expected size of 58 KDa were produced in both InsectDirect and baculovirus systems. A high degree (95%) of purification was achieved using this system as observed in SDS-PAGE. The purified L1 protein in the baculovirus system was clearly more efficient than the InsectDirect system. The results of this study indicate that the BacMagic system is an appropriate tool for large scale protein production and provides an alternative to the traditional baculovirus system. In addition, the InsectDirect system might provide a rapid and dependable pointer of whether a protein can be successfully produced in a baculovirus system. Both InsectDirect and BacMagic systems present remarkable savings in cost and time.


Role of imaging in the applicability of irreversible electroporation for the management of pancreatic adenocarcinoma.
Pancreatic ductal adenocarcinoma is one of the most aggressive malignant neoplasms, with a one-year survival rate below 20%. Axial methods (computed tomography and magnetic resonance imaging) play a fundamental role in the diagnosis and staging of the disease, because they provide adequate anatomical resolution in the assessment of key structures, mainly vascular structures. Pancreatic ductal adenocarcinoma is most often discovered in advanced stages, when surgical resection is no longer feasible. In that scenario, minimally invasive treatment alternatives have been developed in attempts to change the natural history of the disease. Irreversible electroporation, an interventional procedure that minimizes deleterious effects on adjacent tissues, has proven useful for the treatment of tumors traditionally considered unresectable. Despite the growing acknowledgment of this technique as a tool for the management of pancreatic ductal adenocarcinoma, it is still relatively unknown among radiologists. In this study, we sought to provide an overview of the main characteristics and eligibility criteria that must be considered for the indication of irreversible electroporation in cases of pancreatic ductal adenocarcinoma.


[Influence of the depth of the sperm counting chamber on sperm motility].
To investigate the influence of the depth of the sperm counting chamber on sperm motility. We measured the depths of sperm counting chambers using the Filmetrics F20 Spectral Reflectance Thin-Film Measurement System. Then, according to the WHO5 manual, we analyzed 36 semen samples for the percentages of progressively motile sperm (PR) and non-progressively motile sperm (NP) and sperm motility (PR + NP) with the Ruiqi CFT-9201 computer-aided sperm analysis system, and compared the results of analysis. The depths of the 4 sperm counting chambers were 9.8, 12.7, 15.7 and 19.9 microm, respectively, and the obtained PR were (44.00 +/- 11.63), (41.96 +/- 12.62), (40.86 +/- 11.71) and (37.78 +/- 11.38)%, NP (13.54 +/- 3.01), (14.13 +/- 2.94), (14.91 +/- 3.02) and (16.53 +/- 2.77)%, and sperm motility (57.53 +/- 11.06), (56.08 +/- 11.97), (55.78 +/- 11.55) and (54.31 +/- 12.11)% from the 4 chambers, respectively. The depth of the sperm counting chamber was correlated negatively with PR (r = -0.993, P < 0.05) and sperm motility (r = -0.978, P < 0.05), but positively with NP (r = 0.989, P < 0.05). There were statistically significant differences between the 9.8 microm and 19.9 microm deep chambers in PR and NP (P < 0.05) though not in sperm motility among the 4 chambers of different depths. The impact of the depth of the sperm counting chamber on sperm motility should not be ignored, for the deviation of the results from the chambers of different depths may lead clinicians to incorrect diagnosis and consequently inappropriate therapeutic approaches. Different reference ranges of sperm motility need to be normalized in correspondence to the depths of sperm counting chambers.


PD-L1 up-regulation restrains Th17 cell differentiation in STAT3 loss- and STAT1 gain-of-function patients.
Patients with hypomorphic mutations in STAT3 and patients with hypermorphic mutations in STAT1 share several clinical and cellular phenotypes suggesting overlapping pathophysiologic mechanisms. We, therefore, examined cytokine signaling and CD4+ T cell differentiation in these cohorts to characterize common pathways. As expected, differentiation of Th17 cells was impaired in both cohorts. We found that STAT1 was hyperphosphorylated in response to cytokine stimulation in both cohorts and that STAT1-dependent PD-L1 up-regulation-known to inhibit Th17 differentiation in mouse models-was markedly enhanced as well. Overexpression of SOCS3 strongly inhibited phosphorylation of STAT1 and PD-L1 up-regulation, suggesting that diminished SOCS3 expression may lead to the observed effects. Defects in Th17 differentiation could be partially overcome in vitro via PD-L1 inhibition and in a mouse model of STAT3 loss-of-function by crossing them with PD-1 knockout mice. PD-L1 may be a potential therapeutic target in several genetic diseases of immune deficiency affecting cytokine signaling.


Time to remission of proteinuria and incidence of relapse in patients with steroid-sensitive minimal change disease and focal segmental glomerulosclerosis: the Japan Nephrotic Syndrome Cohort Study.
Minimal change disease (MCD) is characterized by a nephrotic syndrome usually steroid-sensitive and a high incidence of relapse of proteinuria. Previous cohort studies have reported conflicting results regarding the association between the time to remission and incidence of relapse. This multicenter prospective cohort study included 102 adult patients with steroid-sensitive MCD or focal segmental glomerulosclerosis from a 5-year cohort study of primary nephrotic syndrome, the Japan Nephrotic Syndrome Cohort Study, who achieved remission of proteinuria within 2 months of immunosuppressive therapy (IST). The association between the time to remission of proteinuria after immunosuppressive therapy and incidence of relapse was assessed using Cox proportional hazards models adjusted for clinically relevant factors. Remission was observed at 3-7, 8-14, 15-21, 22-28, and 30-56 days after initiation of immunosuppressive therapy in 17 (16.7%), 37 (36.3%), 21 (20.6%), 13 (12.7%), and 14 (13.7%) patients, respectively. During a median observation period of 2.3 years after the end of the 2nd month after initiation of immunosuppressive therapy, 46 (45.1%) patients relapsed. The time to remission was associated with the incidence of relapse in an inverse U-shaped pattern (multivariable-adjusted hazard ratios [95% confidence intervals] of the time to remission of 3-7, 8-14, 15-21, 22-28, 30-56 days: 1.00 [reference], 1.76 [0.56, 5.51], 6.06 [1.85, 19.80], 5.46 [1.44, 20.64], and 2.19 [0.52, 9.30], respectively). The time to remission was identified as a significant predictor of relapse in steroid-sensitive patients.


Establishment of a cell line from a human fetal liver and its xenotransplantation to nude mice.
An epithelial cell line, NuE15-1, was established from the liver of a 24-week-old human fetus. The cells had round clear nuclei with clear (agranular) cytoplasm of polygonal shape and grew in a pavement-like manner. The karyotype was human and heteroploid. alpha-Fetoprotein and alpha 1-antitrypsin were detected in the concentrated culture medium but albumin was not. This may indicate the hepatocytic origin of the cell line. Malignant transformation was confirmed by xenotransplantability to nude mice. When inoculated intraperitoneally, the implanted cells caused lethal peritonitis carcinomatosa in 9 out of 10 mice within 3 weeks and liver metastasis was observed in one case. A possible role of alpha-fetoprotein-producing cells as precursors of tumor cells in hepatocarcinogenesis is discussed.


The removal index for evaluation of dialysis.
To find an index of adequacy that takes into consideration the effect of the decreasing concentration of urea nitrogen in hemodialysis (HD) and can be used before treatments to quantitate the prescriptions with the same criterion for both HD and continuous ambulatory peritoneal dialysis (CAPD). The removal index was obtained through mathematical theories and then compared with the urea index (KT/V) values of the sample patients. Thirty-two HD and 21 CAPD patients were included. All patients were dialyzed with optimal urea index values and had been stable for at least one year. The removal index in HD (xi HD) for each dialysis was O.62 +/- 0.07, and the normalized removal index in CAPD (xi CAPD) was 0.59 +/- 0.11. There was no statistical significance. This result is consistent with the fact that no difference of morbidity or mortality exists between these two modalities. After mathematical manipulation, the removal index in HD can be presented in the form of the urea reduction ratio, which is a retrospective measure to estimate the performance of hemodialysis. This study implies that the removal index is able to facilitate the prescriptions for adequate dialysis. The removal index can also be used to explain the reason why the urea index values are always larger in HD than in CAPD.


Activated monocytes capture platelets for heterotypic association in patients with severe carotid artery stenosis.
Inflammation and thrombosis, two processes influencing each other, are involved in the pathogenesis of cerebrovascular disease. We showed that in patients with acute ischaemic stroke circulating platelets are activated and exhausted. To identify whether activated haemostasis might be cause or effect, we investigated the role of leukocyte and platelet activation in patients with severe asymptomatic and symptomatic carotid artery disease. Flow cytometry analysis demonstrated that monocytes from symptomatic (acute stroke aetiology) and asymptomatic patients were highly activated, shown by significantly enhanced presentation of inflammatory markers CD11b and thrombospondin-1 (TSP-1) on the surface. Both correlated positively with monocyte-platelet association rate. However, increased monocyte activation and elevated levels of monocyte-platelet associates in asymptomatic patients were restricted to patients with echo-lucent plaques, providing a close link between monocyte activation and plaque morphology. Circulating single as well as monocyte-bound platelets from symptomatic patients showed significantly enhanced surface expression of P-selectin and TSP-1, whereas platelets from asymptomatic patients were not significantly activated. These results indicate that monocytes activated by inflammation rather than platelets might be the candidates to initiate platelet-monocyte rosetting during the pathogenesis of atherothrombotic cerebral ischaemia and that haemostasis might be activated secondarily by the first occurring inflammation.


Septate Junction Proteins Play Essential Roles in Morphogenesis Throughout Embryonic Development in Drosophila.
The septate junction (SJ) is the occluding junction found in the ectodermal epithelia of invertebrate organisms, and is essential to maintain chemically distinct compartments in epithelial organs, to provide the blood-brain barrier in the nervous system, and to provide an important line of defense against invading pathogens. More than 20 genes have been identified to function in the establishment or maintenance of SJs in Drosophila melanogaster Numerous studies have demonstrated the cell biological function of these proteins in establishing the occluding junction, whereas very few studies have examined further developmental roles for them. Here we examined embryos with mutations in nine different core SJ genes and found that all nine result in defects in embryonic development as early as germ band retraction, with the most penetrant defect observed in head involution. SJ genes are also required for cell shape changes and cell rearrangements that drive the elongation of the salivary gland during midembryogenesis. Interestingly, these developmental events occur at a time prior to the formation of the occluding junction, when SJ proteins localize along the lateral membrane and have not yet coalesced into the region of the SJ. Together, these observations reveal an underappreciated role for a large group of SJ genes in essential developmental events during embryogenesis, and suggest that the function of these proteins in facilitating cell shape changes and rearrangements is independent of their role in the occluding junction.


Software for dynamic analysis of tracer-based metabolomic data: estimation of metabolic fluxes and their statistical analysis.
Metabolic flux analysis of biochemical reaction networks using isotope tracers requires software tools that can analyze the dynamics of isotopic isomer (isotopomer) accumulation in metabolites and reveal the underlying kinetic mechanisms of metabolism regulation. Since existing tools are restricted by the isotopic steady state and remain disconnected from the underlying kinetic mechanisms, we have recently developed a novel approach for the analysis of tracer-based metabolomic data that meets these requirements. The present contribution describes the last step of this development: implementation of (i) the algorithms for the determination of the kinetic parameters and respective metabolic fluxes consistent with the experimental data and (ii) statistical analysis of both fluxes and parameters, thereby lending it a practical application. The C++ applications package for dynamic isotopomer distribution data analysis was supplemented by (i) five distinct methods for resolving a large system of differential equations; (ii) the 'simulated annealing' algorithm adopted to estimate the set of parameters and metabolic fluxes, which corresponds to the global minimum of the difference between the computed and measured isotopomer distributions; and (iii) the algorithms for statistical analysis of the estimated parameters and fluxes, which use the covariance matrix evaluation, as well as Monte Carlo simulations. An example of using this tool for the analysis of (13)C distribution in the metabolites of glucose degradation pathways has demonstrated the evaluation of optimal set of parameters and fluxes consistent with the experimental pattern, their range and statistical significance, and also the advantages of using dynamic rather than the usual steady-state method of analysis. Software is available free from http://www.bq.ub.es/bioqint/selivanov.htm


To a beginning psychotherapist: how to conduct individual psychotherapy.
The paper presents a realistic orientation to the practice of primarily insight psychotherapy. The principles, basic assumptions, and practical considerations are nevertheless applicable to all serious psychotherapy. Therapy is here understood not as a science or a matter of technique but as a collaborative and uniquely personal exploration of the patient's difficulties in living. Psychotherapy is thus seen as a search for meaning rather than for cure, with the hope that knowing what they are doing with their lives will enhance the ability of patients to live with more effectiveness and fulfillment. The limitations of therapy and therapists are fully recognized. Specific guidelines for initiating and conducting therapy are designed to promote genuine progress and reduce the risk of errors that are so easily made even by the experienced. As the medium of therapy, the patient-therapist interplay is presented from the perspective of both patient and therapist. The intelligent use of empathy and interpretation and the manifold forms of resistance all receive their share of attention. The paper concludes with a consideration of values, morality, and the importance of recognizing and respecting the spiritual dimension.


Intramolecular charge-assisted hydrogen bond strength in pseudochair carboxyphosphate.
Carboxyphosphate, a suspected intermediate in ATP-dependent carboxylases, has not been isolated nor observed directly by experiment. Consequently, little is known concerning its structure, stability, and ionization state. Recently, carboxyphosphate as either a monoanion or dianion has been shown computationally to adopt a novel pseudochair conformation featuring an intramolecular charge-assisted hydrogen bond (CAHB). In this work, additive and subtractive correction schemes to the commonly employed open-closed method are used to estimate the strength of the CAHB. Truhlar's Minnesota M06-2X functional with Dunning's aug-cc-pVTZ basis set has been used for geometry optimization, energy evaluation, and frequency analysis. The CHARMM force field has been used to approximate the Pauli repulsive terms in the closed and open forms of carboxyphosphate. From our additive correction scheme, differential Pauli repulsion contributions between the pseudochair (closed) and open conformations of carboxyphosphate are found to be significant in determining the CAHB strength. The additive correction modifies the CAHB prediction (ΔEclosed-open) of -14 kcal/mol for the monoanion and -12 kcal/mol for the dianion to -22.9 and -18.4 kcal/mol, respectively. Results from the subtractive technique reinforce those from our additive procedure, where the predicted CAHB strength ranges from -17.8 to -25.4 kcal/mol for the monoanion and from -15.7 to -20.9 kcal/mol for the dianion. Ultimately, we find that the CAHB in carboxyphosphate meets the criteria for short-strong hydrogen bonds. However, carboxyphosphate has a unique energy profile that does not result in the symmetric double-well behavior of low-barrier hydrogen bonds. These findings provide deeper insight into the pseudochair conformation of carboxyphosphate, and lead to an improved mechanistic understanding of this intermediate in ATP-dependent carboxylases.


Analysis of soil bacteria susceptibility to manufactured nanoparticles via data visualization.
The impact of ZnO and TiO2 manufactured nanoparticles (MNPs) on soil bacterial communities for different exposure periods and MNP doses was explored via data visualization techniques. Interrelationships between MNP treatments and responses of bacterial taxa were illustrated by bipartite graphs, allowing fast identification of important soil bacterial taxa that are susceptible to MNPs. Contribution biplots with subcompositional coherence property were generated via log-ratio analysis (LRA), which jointly display the treatment distribution and the variance (contribution) of bacterial taxa. The LRA contribution biplots and nonmetric multi-dimensional scaling (NMDS) of the dataset, along with hierarchical clustering, demonstrated that high doses of ZnO and TiO2 MNPs caused significant compositional changes in soil bacterial communities. The suitability of family level for MNP taxonomic impact assessment was demonstrated by both the LRA biplots and simplified NMDSs with quantification provided by the distance correlation between MNP impacts summarized at different taxonomic levels. The present study demonstrates that visual exploration could potentially assist in knowledge discovery and interpretation of data on soil bacterial communities exposed to MNPs and thus evaluate the potential for environmental impacts.


Systematic review with meta-analysis: the efficacy of faecal microbiota transplantation for the treatment of recurrent and refractory Clostridium difficile infection.
Clostridium difficile infection (CDI) is the commonest nosocomial cause of diarrhoea. Faecal microbiota transplantation (FMT) is an approved treatment for recurrent or refractory CDI but there is uncertainty about its use. To evaluate the efficacy of FMT in treating recurrent and refractory CDI and investigate outcomes from modes of delivery and preparation. A systematic review and meta-analysis was performed. MEDLINE, EMBASE, CINAHL, Cochrane Library, trial registers and conference proceedings were searched. Studies on FMT in recurrent and refractory CDI were included. The primary outcome was clinical resolution with subgroup analyses of modes of delivery and preparation. Random effects meta-analyses were used to combine data. Thirty seven studies were included; seven randomised controlled trials and 30 case series. FMT was more effective than vancomycin (RR: 0.23 95%CI 0.07-0.80) in resolving recurrent and refractory CDI. Clinical resolution across all studies was 92% (95%CI 89%-94%). A significant difference was observed between lower GI and upper GI delivery of FMT 95% (95%CI 92%-97%) vs 88% (95%CI 82%-94%) respectively (P=.02). There was no difference between fresh and frozen FMT 92% (95%CI 89%-95%) vs 93% (95%CI 87%-97%) respectively (P=.84). Administering consecutive courses of FMT following failure of first FMT resulted in an incremental effect. Donor screening was consistent but variability existed in recipient preparation and volume of FMT. Serious adverse events were uncommon. Faecal microbiota transplantation is an effective treatment for recurrent and refractory Clostridium difficile infection, independent of preparation and route of delivery.


Luteinizing hormone increases the abundance of various transcripts, independently of the androgens, in the rat prostate.
Differential display analysis was carried out to find, in the rat prostate, genes that could be regulated by Luteinizing Hormone (LH), independently of the androgens. Hypophysectomized and castrated adult rats were treated with either LH, testosterone or saline. Regulated discrete bands have been eluted and reamplified. After Northern blotting, the levels of mRNA corresponding to 8 PCR fragments were significantly increased by LH treatment. None of these inserts were found to be induced by testosterone. One insert was subcloned, sequenced and identified as the ribosomial protein S 23. A competitive RT-PCR assay was carried out on the full length S 23 cDNA and confirmed that its mRNA levels were stimulated by LH but not by testosterone. These results strongly suggest that the LH membrane receptor, previously shown to be expressed in the rat prostate, has a physiological significance in this organ. Moreover, it appears that the effect of LH on the rat prostate are independent of the androgens.


First Report of Anthracnose Crown Rot on Strawberry (Fragaria × ananassa Duch.) Caused by Colletotrichum pandanicola in Yunnan Provence, China.
Strawberry (Fragaria × ananassa Duch.), a widely grown octoploid species, is one of the most important economic fruit crops and has been widely cultivated in the world, including China. In December 2021, a serious crown rot disease (approximately 50% incidence) was observed in strawberry (cultivar Miaoxiang) plantations in Qujing City, Yunnan Province, China. Symptoms observed on aboveground part withered rapidly, reddish-brown marbled necrosis on crown. The roots were healthy and strong, but the plants finally died. To isolate the causal agent of this disease, crown tissues from five strawberry plants showing typical symptoms were cut into pieces of 5×5 mm, and the pieces were surface-sterilized with 75% ethanol for 45 s followed by 2.5% NaClO for 3 min and rinsed thrice with sterile water, and then placed onto potato dextrose agar (PDA) for 7 days at 25 ºC. After 3 to 4 days, extended single hyphal tips from the tissues were transferred to PDA and incubated for 7 days at 25 ºC. The colonies were initially white, later became somewhat zonate, velvety, cyan gray on the upper side and cyan ink pigment ring on the reverse side of plates, with concentric rings of salmon sporodochia. Many yellowish or orange creamy conidial droplets formed on PDA after 14 days at 25 ºC. Fifty-nine isolates were obtained, and three isolates QLYRR1, QLMCR9, and QLMCR39 were selected for further experiments. Conidia were hyaline, cylindrical with rounded ends, 12.17-19.35×3.71-6.30 μm (average±SD, 15.24±1.37×5.09±0.45 μm, n=150), L/W ratio = 2.99. The three isolates were molecularly identified using the genomic regions of internal transcribed spacer (ITS), actin (ACT), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-tubulin (TUB2) genes, and the sequences were deposited in GenBank (accession nos. QLYRR1, QLMCR9, QLMCR39: ON668272, ON668256, ON668257[ITS], ON684302, ON684300, ON684301[ACT], ON684316, ON684314, ON684315[CHS-1], ON684292, ON684290, ON684291[GAPDH], ON684286, ON684284, ON684285[TUB2]). The phylogenetic analysis of experimental strains was performed by Maximum-likelihood (ML) tree and Bayesian inference (BI) method. Nucleotide sequences exhibited three isolates were clustered with the ex-type strain C. pandanicola strain MFLUCC 170571T found in Thailand, C. pandanicola strains (SAUCC201152, SAUCC200204) found in Shandong Province, and the holotype stain C. parvisporum YMF 1.06942T found in Guangxi Province, China. Morphologically, isolates were easily distinguished from C. parvisporum by the colony on PDA and the size of conidia (Yu et al. 2022). Morphological characteristics and phylogenetic analyses revealed that QLYRR1, QLMCR9, and QLMCR39 belong to C. pandanicola, the members of the C. gloeosporioides species complex (Tibpromma et al. 2018; Mu et al. 2021). Koch's postulates were tested by strawberry plants (two cultivars, Akihime and Miaoxiang) in vivo, strawberry plants were tested for the three isolates by spraying 1×106 conidia/mL suspension on three seedlings. Three seedlings sprayed with sterile distilled water were served as control. All of the plants were transferred to a glasshouse with a 28/20 °C day/night temperature range and natural sunlight. After 6 weeks, QLYRR1-, QLMCR9-, and QLMCR39-sprayed seedlings were stunted and developed typical wilt symptoms similar to those observed in the field with the incidence for 3, 3, and 3 seedlings, respectively. The negative control remained asymptomatic. The fungi were reisolated again from lesions of diseased plants and leaves with 100% frequency, and morphological characteristics and tested gene sequences were identical to the original isolates in this note, thus fulfilling Koch's postulates. C. pandanicola was described from the healthy leaves of Pandanus sp. and the lesion fruits of Juglans regia. To our knowledge, this is the first report confirming C. pandanicola causes anthracnose crown rot on strawberries in China. C. pandanicola has the potential for causing serious losses to the strawberry industry, and research is needed on management strategies to minimize losses.


Diaphragmatic pulmonary sequestration.
Two cases of diaphragmatic extralobar pulmonary sequestrations are described. One of them simulated a suprarenal tumour and the other was associated with a diaphragmatic hernia. Both of them were firmly attached to the left hemidiaphragm and were partly located within it. They had an arterial supply from the abdominal aorta. Both sequestrations were treated surgically by sequesterectomy. The patients had an uneventful recovery.


Regulatory role of transcription factor SutR (YdcN) in sulfur utilization in Escherichia coli.
Sulfur makes up 1 % of the dry mass of bacteria, and it is an abundant element (0.1 %) on earth. Sulfur in the environment is, however, mostly in oxidized forms and inaccessible to living organisms. At present, the entire assimilation pathway of external sulfur to sulfur-containing biomolecules and its regulation in Escherichia coli remain poorly understood, except for the metabolic pathway of cysteine synthesis, the first-step metabolite of sulfur assembly. During the search for regulation targets of uncharacterized transcription factors by Genomic SELEX screening, we found that the hitherto uncharacterized YdcN regulates a set of genes involved in the utilization of sulfur, including the generation of sulfate and its reduction, the synthesis of cysteine, the synthesis of enzymes containing Fe-S as cofactors, and the modification of tRNA with use of sulfur-containing substrates. Taking these findings together, we propose renaming YdcN as SutR (regulator of sulfur utilization).


Systemic thrombolysis in ischaemic stroke patients with COVID-19.
Intravenous thrombolysis (IVT) with recombinant tissue plasminogen activator is the core medical therapy of acute ischaemic stroke (AIS). COVID-19 infection negatively modifies acute stroke procedures and, due to its pro-coagulative effect, may potentially impact on IVT outcome. Thus, short-term efficacy and safety of IVT were compared in patients with and without evidence of SARS-CoV-2. An observational, retrospective study included 70 patients with AIS, including 22 subjects (31%) with evidence of acute COVID-19 infection, consecutively treated with IVT in 4 stroke centres between 15 September and 30 November 2020. Patients infected with COVID-19 were characterized by higher median of National Institute of Health Stroke Scale (NIHSS) score (11.0 vs. 6.5; p < .01) and D-dimers (870 vs. 570; p = .03) on admission, higher presence of pneumonia (47.8% vs. 12%; p < .01) and lower percentage of 'minor stroke symptoms' (NIHSS 1-5 pts.) (2% vs., 18%; p < .01). Hospitalizations were longer in patients with COVID-19 than in those without it (17 vs. 9 days, p < .01), but impact of COVID-19 infection on patients' in-hospital mortality or functional status on dismission has been confirmed neither in uni- or multivariate analysis. SARS-CoV-2 infection prolongs length of stay in hospital after IVT, but does not influence in-hospital outcome.


Oncogenes in cellular immortalisation and differentiation (review).
Oncogenes have been shown to be able to overcome the limited proliferative capacity of normal mammalian cells in culture enforcing them to an immortalised phenotype, which in turn may act as a primary step in tumourigenesis. The oncogenes which display such immortalisation activity have the common feature of nuclear localisation, while the oncogenes which are capable of transforming cells are mainly cytoplasmic. Oncogenes from both families have been shown to interfere with the differentiation processes of several cell types. There is evidence that some of these proto-oncogenes may function as regulators of normal cell differentiation, and when immortalisation occurs this is a process of blocking the cell from achieving a differentiated state. This article focuses on the ability of some oncogenes with different functions, such as myc and ras, to override the limited cellular proliferative capacity and their effects on differentiation; finally it examines the recent implications that some onco-suppressor genes are actively participating in cellular differentiation processes.


Anti-HLA specificity determined by serum absorption with antigen-masked lymphocytes.
Lymphocyte or platelet absorption of multispecific sera is frequently used to establish HLA antibody specificity. Absorption of such sera with cells containing the desired antigenic combinations is often limited by the availability and the HLA characteristics of fresh or frozen cell panels. The method described permits improved absorption of specific HLA antibody with a preparation of cells containing a predetermined single HLA antigen. The procedure involves the screening of the patient's active sera against a panel of frozen cells. Analysis of the positive and negative reactions allows tentative assessment of recipient serum specificities. Cells with a single available HLA antigen type are prepared for serum absorption by suspending the cells in selected antisera to block (mask) HLA surface antigens while leaving just one type of antigen free for monospecific absorption. Following absorption of the test serum with the masked cells, the absorbed serum is then retested against the original panel of frozen cells. Should the results of this screen show a loss of a single HLA activity, the suspected specificity is assigned to the recipient's serum. The use of HLA antigen-masked lymphocytes has proven useful in the definition of HLA antibody specificity.