Effects of Skin Thickness on Cochlear Input Signal Using Transcutaneous Bone Conduction Implants.
Intracochlear sound pressures (PIC) and velocity measurements of the stapes, round window, and promontory (VStap/RW/Prom) will show frequency-dependent attenuation using magnet-based transcutaneous bone conduction implants (TCBCIs) in comparison with direct-connect skin-penetrating implants (DCBCIs). TCBCIs have recently been introduced as alternatives to DCBCIs. Clinical studies have demonstrated elevated high-frequency thresholds for TCBCIs as compared with DCBCIs; however, little data exist examining the direct effect of skin thickness on the cochlear input signal using TCBCIs. Using seven cadaveric heads, PIC was measured in the scala vestibuli and tympani with fiber-optic pressure sensors concurrently with VStap/RW/Prom via laser Doppler vibrometry. Ipsilateral titanium implant fixtures were placed and connected to either a DCBCI or a TCBCI. Soft tissue flaps with varying thicknesses (no flap and 3, 6, and 9 mm) were placed successively between the magnetic plate and sound processor magnet. A bone conduction transducer coupled to custom software provided pure-tone stimuli between 120 and 10,240 Hz. Stimulation via the DCBCI produced the largest response magnitudes. The TCBCI showed similar PSV/ST and VStap/RW/Prom with no intervening flap and a frequency-dependent nonlinear reduction of magnitude with increasing flap thickness. Phase shows a comparable dependence on transmission delay as the acoustic baseline, and the slope steepens at higher frequencies as flap thickness increases, suggesting a longer group delay. Proper soft tissue management is critical to optimize the cochlear input signal. The skin thickness-related effects on cochlear response magnitudes should be taken into account when selecting patients for a TCBCI.


[Acute febrile state].
A 34 year old airline pilot, who had spent nine days in Cameroon (Westafrica) presented for his yearly physical examination two weeks later. The physical examination and routine laboratory tests were within normal limits. The patient complained about mild pain of joints and extremities and about not feeling quite well. The same evening (a few hours after the physical examination) he experienced chills and fever (up to 39.5 degrees Celsius). He was seen subsequently by a tropical medicine specialist, who diagnosed Plasmodium falciparum on blood smears. The patient was immediately placed on Riamet, fever and symptoms disappeared completely within a few days.


Challenges for effective marker-assisted selection in plants.
The basic principle of Marker-Assisted Selection (MAS) is to exploit Linkage Disequilibrium (LD) between markers and QTLs. With strong enough LD, MAS should in theory be easier, faster, cheaper, or more efficient than classical (phenotypic) selection. I briefly review the major MAS methods, describing some 'success stories' where MAS was applied successfully in the context of plant breeding, and detailing other cases where efficiency was not as high as expected. I discuss the possible causes explaining the difference between theoretical expectations and practical observations. Finally, I review the principal challenges and issues that must be tackled to make marker-assisted selection in plants more effective in the future, namely: managing and controlling QTL stability to apply MAS to complex traits, and integrating MAS in traditional breeding practices to make it more economically attractive and applicable in developing countries.


Disc hemorrhages and treatment in the early manifest glaucoma trial.
To evaluate the effect of intraocular pressure (IOP)-reducing treatment on the development of disc hemorrhages in patients with glaucoma. Prospective cohort study of patients in the Early Manifest Glaucoma Trial, followed up to 11 years (median = 8 years). Patients with newly detected glaucoma randomized to argon laser trabeculoplasty plus betaxolol (n = 129) or no initial treatment (n = 126), followed with tonometry, perimetry, and ophthalmoscopy every 3 months, and fundus photography every 6 months. Logistic regression expressed as odds ratios (OR) and 95% confidence intervals (CIs), analysis of variance, and Cox time-dependent models, expressed as hazard ratios (HRs) and CIs. Presence (yes/no) and frequency of disc hemorrhages. Disc hemorrhages were identified in approximately 55% of all patients, whether by ophthalmoscopy or review of photographs. In analyses including data up to the time of progression, disc hemorrhages were equally common among treated and control patients: 51.2% versus 45.2%, respectively (P = 0.34), based on ophthalmoscopy, and 50.4% versus 44.4%, respectively (P = 0.34), based on photographs. Gender was the only factor related to the presence of disc hemorrhages detected by both ophthalmoscopy (OR = 0.48; CI, 0.26-0.88; P = 0.022) and photographs (OR = 0.64; CI, 0.38-1.09; P = 0.099) for male patients. The frequency of disc hemorrhages over time did not differ between treated and control patients: 8.4% versus 8.5%, respectively (P = 0.93), based on ophthalmoscopy, and 12.4% versus 11.2%, respectively (P = 0.36), based on photographs. Disc hemorrhages were significantly associated with time to progression (HR = 1.02; CI, 1.01-1.04), and there was no evidence of interaction between treatment group and disc hemorrhages. IOP-reducing treatment was unrelated to the presence or frequency of disc hemorrhages. The results may suggest that disc hemorrhages cannot be considered an indication of insufficient IOP-lowering treatment, and that glaucoma progression in eyes with disc hemorrhages cannot be totally halted by IOP reduction. The results also suggest that disc hemorrhages do not occur in all patients with glaucoma.


Pseudomonas aeruginosa Elastase Contributes to the Establishment of Chronic Lung Colonization and Modulates the Immune Response in a Murine Model.
Chronic infection by Pseudomonas aeruginosa in cystic fibrosis (CF) patients is a major contributor to progressive lung damage and is poorly treated by available antibiotic therapy. An alternative approach to the development of additional antibiotic treatments is to identify complementary therapies which target bacterial virulence factors necessary for the establishment and/or maintenance of the chronic infection. The P. aeruginosa elastase (LasB) has been suggested as an attractive anti-virulence target due to its extracellular location, its harmful degradative effects on host tissues and the immune system, and the potential to inhibit its activity using small molecule inhibitors. However, while the relevance of LasB in acute P. aeruginosa infection has been demonstrated, it is still unclear whether this elastase might also play a role in the early phase of chronic lung colonization. By analyzing clinical P. aeruginosa clonal isolates from a CF patient, we found that the isolate RP45, collected in the early phase of persistence, produces large amounts of active LasB, while its clonal variant RP73, collected after years of colonization, does not produce it. When a mouse model of persistent pneumonia was used, deletion of the lasB gene in RP45 resulted in a significant reduction in mean bacterial numbers and incidence of chronic lung colonization at Day 7 post-challenge compared to those mice infected with wild-type (wt) RP45. Furthermore, deletion of lasB in strain RP45 also resulted in an increase in immunomodulators associated with innate and adaptive immune responses in infected animals. In contrast, deletion of the lasB gene in RP73 did not affect the establishment of chronic infection. Overall, these results indicate that LasB contributes to the adaptation of P. aeruginosa to a persistent lifestyle. In addition, these findings support pharmacological inhibition of LasB as a potentially useful therapeutic intervention for P. aeruginosa-infected CF patients prior to the establishment of a chronic infection.


Clinical guidelines for the management of acute viral infections in patients with systemic lupus erythematosus.
In recent decades, many research groups have focused on the role of viral infections in the etiopathogenesis of systemic lupus erythematosus (SLE), the so-called "viral hypothesis". The main candidates are herpes viruses such as Epstein-Barr virus (EBV) and cytomegalovirus (CMV), which have a high seroprevalence in the general population. However, a viral causal agent of SLE has not yet been discovered, although many interesting clinical findings on the complex interactions between viruses and SLE have been made. This review analyzes 88 cases of acute viral infections in adult patients with SLE and identifies situations in which viral infections influenced the diagnosis, prognosis or treatment of SLE. We also propose clinical guidelines for the management of these infections in patients with SLE.


Retroperitoneal Sarcomas: An Update on the Diagnostic Pathology Approach.
Retroperitoneal sarcomas are a heterogenous group of rare tumors arising in the retroperitoneum. Retroperitoneal sarcomas comprise approximately 10% of all soft tissue sarcomas. Though any soft tissue sarcoma histologic types may arise in the retroperitoneal space, liposarcoma (especially well-differentiated and dedifferentiated types) and leiomyosarcoma do so most commonly. Retroperitoneal sarcomas are diagnostically challenging, owing to their diversity and morphological overlap with other tumors arising in the retroperitoneum. An accurate diagnosis is necessary for correct management and prognostication. Herein, we provide an update on the diagnostic approach to retroperitoneal sarcomas and review their key histologic findings and differential diagnoses.


[Micronutrients in oncology. Current data about vitamin D, selenium, L-carnitine and vitamin C].
Many patients receiving cancer treatment use micronutrient supplements, with the intention to complement their cancer treatment, or help them cope with the therapy- and disease-associated side-effects. Up to 90% of the cancer patients are adding antioxidants without the knowledge of the treating physician. There are many concerns that antioxidants might decrease the effectiveness of chemotherapy, but increasing evidence suggests a benefit when antioxidants and other micronutrients, such as selenium, L-carnitine and vitamin D are added to conventional cytotoxic therapies. It is imperative that physicians discuss the use ofantioxidant and other micronutrient supplements with their cancer patients and educate them about potentially negative, but also potentially beneficial effects.


[Epstein-Barr virus bilateral dacryoadenitis: case report].
The authors present a rare case of acute bilateral dacryoadenitis in association with infectious mononucleosis. A 27 year-old woman had acute bilateral lacrimal gland enlargement with inflammatory signs. The CT scan findings and laboratorial investigations confirmed the diagnosis of infectious mononucleosis. The systemic treatment with intravenous steroids was initiated leading to complete remission of the signs and symptoms. Dacryoadenitis is an uncommon manifestation of infectious mononucleosis and may minimize other signs of the disease. Mononucleosis has to be considered in acute dacryoadenitis cases. Systemic steroids contribute to the fast regression of inflammatory condition.


[Complementary addressed alkylation of 16S rRNA of Escherichia coli by 2',3'-O-[4-N-methyl-N-(2-chloroethyl)-amino]benzylidene derivatives of oligodeoxyribonucleotides. V. Study of the factors affecting selectivity of modification].
We studied the effect of different factors (reagent concentration, temperature, presence of oligonucleotide-effector (3',5'-diphenazinium derivative of oligodeoxyribonucleotide) stabilizing duplex RNA.reagent) on the selectivity of the site-directed modification of 16S rRNA with 2,3'-O-[4-N-methyl-N-(2-chloroethyl)-amino]-benzylidene derivative of oligonucleotide p(dTTTGCTCCCC)rA (reagent I) under conditions of secondary structure stability. The constant of cooperative binding of the reagent and oligonucleotide-effector with 16s rRNA was determined. The temperature rise from 20 to 40 degrees C brought about a 1.5-fold increase in the relative extent of modification at the target site 771-781. In the presence of oligonucleotide-effector, which is a full complementary copy of the 782-789 fragment of 16S rRNA (reagent concentration is 1 x 10(-6) M), the selectivity of the RNA modification at the target site is doubled and a high level of the modification is retained. When the reagent concentration in the reaction mixture was decreased down to 1 x 10(-7) M, the same level of selectivity was achieved without the oligonucleotide-effector. Under these conditions, however, a drastic (20-fold) drop of the level of the 16S rRNA alkylation was observed.


Decombinator V4: an improved AIRR compliant-software package for T-cell receptor sequence annotation?
Analysis of the T-cell receptor repertoire is rapidly entering the general toolbox used by researchers interested in cellular immunity. The annotation of T-cell receptors (TCRs) from raw sequence data poses specific challenges, which arise from the fact that TCRs are not germline encoded, and because of the stochastic nature of the generating process. In this study, we report the release of Decombinator V4, a tool for the accurate and fast annotation of large sets of TCR sequences. Decombinator was one of the early Python software packages released to analyse the rapidly increasing flow of T-cell receptor repertoire sequence data. The Decombinator package now provides Python 3 compatibility, incorporates improved sequencing error and PCR bias correction algorithms, and provides output which conforms to the international standards proposed by the Adaptive Immune Receptor Repertoire Community. The entire Decombinator suite is freely available at: https://github.com/innate2adaptive/Decombinator. Supplementary data are available at Bioinformatics online.


The impact of single embryo transfer policy on pregnancy outcomes after legislative change.
We evaluated the effect of mandatory single embryo transfer (SET) on live birth rates and pregnancy outcomes in Turkey. A retrospective study was conducted in Zekai Tahir Burak Women's Health Education and Research Hospital. Four hundred and four patients undergoing intracytoplasmic sperm injection (ICSI) cycles were included in the study. In Turkey, the number of embryos to be transferred in an assisted cycle was limited to three under normal circumstances until 6 March 2010. After that, new legislation was introduced to promote the application of SET. Outcomes were compared in periods of 1 year before and after the new law. We compared pregnancy outcomes of all assisted reproductive cycles in SET cycles (group 1: n = 281) with double embryo transfer (DET) cycles (group 2: n = 123). There were significant differences in oocyte number, multiple pregnancy, gestational age, birth weight and perinatal fetal morbidity between the groups (p = 0.023, 0.001, 0.001, 0.001, 0.001, respectively). But there were no differences in age, baseline FSH, infertility period, stimulation protocol, stimulation day, gonadotrophin dose, clinical pregnancy rate, abortion rate, live birth rate and cesarean rate (p > 0.05). These results suggest that under the new legislation multiple pregnancy rates and perinatal complications are significantly reduced without causing a significant decline in the pregnancy rates.


A challenging case of a chyle leak following axillary lymph node clearance.
Chyle leak following axillary lymph node clearance is a rarely reported complication. We present a case of chylous leakage following axillary lymph node clearance, which was diagnosed on clinical grounds. Surgical re-exploration was undertaken due to ongoing high output. However, the chylous leak recurred post-operatively, if at a lower rate. The patient was subsequently managed successfully with conservative measures, primarily utilizing regular aspiration and compression bandaging to the axilla.


Structural study of the low-temperature phase of TlH2PO4 at 180 K.
The low-temperature phase of TlH(2)PO(4) has been studied by X-ray diffraction. A structural model is proposed with space group P1. This model is compared with the structure of the antiferroelectric phase of TlD(2)PO(4) at room temperature to analyze the expected isomorphism at low temperature. Given the structural distortion of TlH(2)PO(4), such isomorphism present in the common high-temperature phase is not recovered in this phase. Through the analysis of the displacements of the PO(4) groups there is some evidence that the ordering of the H atoms responsible for the appearance of antiferroelectricity seems to be incomplete. An exhaustive study of the detected ferroelastic domains is also performed.


Production of antigen-specific human antibodies from mice engineered with human heavy and light chain YACs.
Our paper describes the introduction of large fragments of both the human heavy and light chain Ig genes into the mouse germline to create a mouse strain capable of producing a broad repertoire of antigen-specific, fully human antibodies. The human immunoglobulin gene sequences were functional in the context of the mouse machinery for antibody recombination and expression, either in the presence or absence of functional endogenous genes. This was demonstrated by their ability to undergo diverse rearrangement, to be expressed at significant levels, and to exclude expression of mouse immunoglobulins irrespective of their copy number or site of integration. The decrease in susceptibility to influence by adjacent genomic sequences may reflect the greater size, variable gene content, or structural integrity of the human Ig YACs and/or the presence of unidentified but important regulatory elements needed for optimal expression of the human immunoglobulin genes and their correct regulation. Our results show that mouse B cells coexpressing human heavy and kappa chains, upon immunization, can produce antigen-specific, fully human antibodies. Furthermore, the human heavy and kappa chain YACs induced differentiation and maturation of the growth-arrested B-cell lineage in mice with inactivated endogenous Ig genes, leading to the production of a diverse repertoire of fully human antibodies at levels approaching those in normal serum. These results suggest the potential value of these mice as a source of fully human antibodies for human therapy. Furthermore, it is expected that such mice would lack immunological tolerance to and thus readily yield antibodies to human proteins, which may constitute an important class of targets for monoclonal antibody therapy. Our findings suggest that the introduction of even larger portions of the human heavy and light chain loci, which should be achievable with the ES cell-yeast spheroplast fusion technology described, will result in strains of mice ultimately capable of recapitulating the full antibody repertoire characteristic of the human humoral response to infection and immunization. The present and future mouse strains may prove to be valuable tools for studying the molecular mechanisms and regulatory sequences influencing the programmed assembly and expression of human antibodies in the normal immune response, as well as the abnormal response characteristic of autoimmune disease and other disorders. The strategy we have described for the introduction of large segments of the human genome into mice in conjunction with the inactivation of the corresponding mouse loci may also have broad applicability to the investigation of other complex or uncharacterized loci.


Acetylation regulates DNA repair mechanisms in human cells.
The p300-mediated acetylation of enzymes involved in DNA repair and replication has been previously shown to stimulate or inhibit their activities in reconstituted systems. To explore the role of acetylation on DNA repair in cells we constructed plasmid substrates carrying inactivating damages in the EGFP reporter gene, which should be repaired in cells through DNA mismatch repair (MMR) or base excision repair (BER) mechanisms. We analyzed efficiency of repair within these plasmid substrates in cells exposed to deacetylase and acetyltransferase inhibitors, and also in cells deficient in p300 acetyltransferase. Our results indicate that protein acetylation improves DNA mismatch repair in MMR-proficient HeLa cells and also in MMR-deficient HCT116 cells. Moreover, results suggest that stimulated repair of mismatches in MMR-deficient HCT116 cells is done though a strand-displacement synthesis mechanism described previously for Okazaki fragments maturation and also for the EXOI-independent pathway of MMR. Loss of p300 reduced repair of mismatches in MMR-deficient cells, but did not have evident effects on BER mechanisms, including the long patch BER pathway. Hypoacetylation of the cells in the presence of acetyltransferase inhibitor, garcinol generally reduced efficiency of BER of 8-oxoG damage, indicating that some steps in the pathway are stimulated by acetylation.


[Finite element simulation of the dilatation of coronary vascular implants].
Intravascular stents are small tube-like structures expanded into stenotic arteries to restore blood flow. The stent is mounted on a balloon catheter and delivered to the site of blockage. First the balloon is inflated, so the stent expands and is pressed against the stenotic wall of the coronary artery. After balloon deflation the stent remains in the coronary artery and the balloon is removed. In this paper, the finite element method is used to simulate the expansion and recoil after ballOon deflation of the stent as well as the cyclic loading of the stent caused by the changing blood pressure.


Pineal attrition, loss of cognitive plasticity, and onset of puberty during the teen years: is it a modern maladaptation exposed by evolutionary displacement?
Cognitive plasticity, a developmental trait that promotes acquisition of complex skills such as language or playing musical instruments, diminishes substantially during puberty. The loss of plasticity has been attributed to surge of sex steroids during adolescence, but the phenomenon remains poorly understood. We hypothesize that pineal involution during puberty may contribute to plasticity decay. The pineal gland produces melatonin, the level of which declines dramatically during onset of puberty. Emerging evidence suggest that melatonin may modulate cognitive plasticity, independent of the effects of sex steroids, and low sex steroids and high melatonin may be simultaneously required to maintain cognitive plasticity. Potential mechanisms by which melatonin may modulate plasticity are examined within the sleep and hippocampal long-term potentiation frameworks. Implications for psychiatric conditions that involve sleep disorders and learning dysfunctions such as schizophrenia and autism are discussed, and the potential adaptive roles of postprandial and postcoital sleep are explored. From the Darwinian perspective, development and reproductive maturity may represent distinct phases that require tailored cognitive strategies to maximize fitness. While cognitive flexibility and susceptibility to new skills may be paramount during development, reduced cognitive flexibility and increased cognitive determinism may enable more efficient responses to stimuli during adulthood. Thus, cognitive plasticity and cognitive determinism may represent trade-off adaptations and different dimensions of intelligence. The decline of plasticity and emergence of puberty during the second decade may be relics of prehistoric times when the human lifespan was short and the environment was relatively simple and static. Today, when the environment is more complex and dynamic, and humans are living far longer, the early obsolescence of plasticity during puberty may represent a Darwinian inefficiency exposed by evolutionary displacement. Regulation of plasticity may be a systemic phenomenon, as exemplified by the association of learning disability with allergic conditions, a form of immune plasticity dysfunction. Ramifications for other plastic functions that decline during puberty such as wound healing and hyaline cartilage regeneration are explored. Like the plasticity of immunity and cognition, the plasticity of hyaline cartilage during youth may enable hosts to respond to ecologic opportunities and generate the optimally adapted adult phenotype. Pineal involution may represent a potential target for therapeutic extension or restoration of plasticity after puberty. Extending plasticity may have far-reaching consequences for human evolution.


Regulation of antiviral immune response by African swine fever virus (ASFV).
African swine fever (ASF) is a highly contagious and acute hemorrhagic viral disease with a high mortality approaching 100% in domestic pigs. ASF is an endemic in countries in sub-Saharan Africa. Now, it has been spreading to many countries, especially in Asia and Europe. Due to the fact that there is no commercial vaccine available for ASF to provide sustainable prevention, the disease has spread rapidly worldwide and caused great economic losses in swine industry. The knowledge gap of ASF virus (ASFV) pathogenesis and immune evasion is the main factor to limit the development of safe and effective ASF vaccines. Here, we will summarize the molecular mechanisms of how ASFV interferes with the host innate and adaptive immune responses. An in-depth understanding of ASFV immune evasion strategies will provide us with rational design of ASF vaccines.


Dialysis in adults in year 2000 in the Republic of Macedonia.
1,019 adult patients with terminal renal failure were treated with dialysis (D) in the first part of the year 2000 in the Republic of Macedonia. 1,010 patients (99%) were treated with chronic intermittent (maintenance) hemodialysis (HD) while nine patients (1%) were on continuous ambulatory peritoneal dialysis (CAPD). For the children, a special peritoneal dialysis program was developed; 509 patients per million of the population (PMP) were on dialysis. The Republic of Macedonia is, therefore, among those central and eastern European countries with a higher PMP number in the treatment of end-stage renal disease, following Croatia, the Czech Republic and Slovenia. The patients were treated at 18 Centers in a network of HD Centers at a distance of 30-50 km. from their place of residence in order to facilitate their access to treatment and to work. All patients who have had symptoms indicating need for treatment with D were accepted for treatment. The government payed all the expenses of the treatment and the salaries of the staff. 56% were male and 44% were female patients. The youngest patient was aged 9 and the oldest was 82 years old. There has been an increase in the age of the patients on D as well as an increase in their number. In 1993 we had 727 patients being treated with D, and now we have 1,019 with a constant increase in the number of patients with ESRD and a need for D and renal transplantation. Mortality per year at the different Centers ranged from 8-19% in 1999 and the average is 12%. Glomerulonephritis (GN)--both primary and secondary--is the main cause of renal failure (RF) in some Centers up to 45%. Tubulo-interstitial disease follows GN. ADPKD patients constitute 9.4% with a difference among the Centers of 3-29%, and diabetic nephropathy is found in 10%, 5-15% in different Centers. 11-61% of patients have an unknown etiology. 352 patients are on treatment with human recombinant erythropoietin (rhuEPO) - in some Centers up to 60%. The mode of application was subcutaneous and the initial dose is 20 U/kg body weight and the mean maintenance dose of EPO per patient weekly is 4,000 U. The Cimino-Brescia arteriovenous fistula is being applied as a standard vascular access. The survival rate of our patients treated with maintenance HD at 5 years was 58%. CAPD and particularly renal transplantation are to be further developed as alternative methods in treating terminal renal failure.


Stomatal Response and Leaf Injury of Pisum sativum L. with SO(2) and O(3) Exposures : I. INFLUENCE OF POLLUTANT LEVEL AND LEAF MATURITY.
Plants of Pisum sativum L. ;Alsweet' were grown under a controlled environment and exposed to SO(2) and O(3) to determine whether changes in stomatal aperture during exposure were related to subsequent leaf injury. Stomata consistently closed with injurious levels of SO(2) and O(3). Measurements with diffusion porometers demonstrated approximately 75 and 25% lower conductance with SO(2) and O(3) exposures, respectively, compared to the conductance of control plants. Stomata also showed a closing response with noninjurious levels of SO(2) but an opening response with noninjurious levels of O(3). Stomata closed to the same degree with combinations of SO(2) plus O(3) as with SO(2) alone. Stomata of expanding leaves closed more during pollutant exposures than stomata of expanded leaves. The abaxial and adaxial stomata both exhibited closure with SO(2) and combinations of SO(2) plus O(3), but abaxial stomata tended to close and adaxial stomata tended to open with exposure to O(3) alone.The changes in stomatal aperture were not closely correlated with the amount of leaf injury produced by different pollutant levels. Stomata closed, not only with exposure to pollutant levels that caused severe necrosis, but also with levels that caused only a trace of injury. There was no evidence of a reduced amount of closure or even stomatal opening with combinations of SO(2) and O(3) compared to plants exposed to the pollutants alone to explain the large amount of injury to plants exposed to pollutant combinations.


Alternative Routes for the Synthesis of 5-Aminolevulinic Acid in Maize Leaves : II. Formation from Glutamate.
Intact plastids from greening maize (Zea mays L.) leaves converted [(14)C]glutamate and [(14)C]2-ketoglutarate (KG) to [(14)C]5-aminolevulinic acid (ALA). Glutamate appeared to be the immediate precursor of ALA, while KG was first converted to glutamate, as shown by the effect of various inhibitors of amino acid metabolism. Plastids from greening leaves contained markedly higher activity as compared with etioplasts or chloroplasts. The synthesis of ALA by intact plastids was light dependent. The enzyme system resides in the stroma of plastids or may be lightly bound to membranes. The solubilized system showed maximal activity around pH 7.9 and required Mg(2+), ATP, and NADPH although dependence on the latter was not clear-cut. A relatively high level of activity could be extracted from etioplasts. Maximal activity was obtained from plastids of leaves which had been illuminated for 90 minutes, after which activity declined sharply. The enzyme system solubilized from plastids also catalyzed the conversion of putative glutamate 1-semialdehyde to ALA in a reaction which was not dependent on the addition of an amino donor.The system in maize greatly resembled the one which had been reported from barley. It is suggested that this system is the one responsible for the biosynthesis of ALA destined for chlorophyll formation.


Catalytic characteristics of MnO2 nanostructures for the O2 reduction process.
Nanorods with an α type MnO(2) structure and a diameter ranging from 25 to 40 nm, along with tipped needles with a β MnO(2) structure and a diameter of 100 nm were obtained. The 25 nm diameter α MnO(2) nanorods showed the best catalytic activity for dissociation of HO(2)(-) formed during oxygen reduction in a KOH solution. The MnO(2) nanostructures preferably followed a two-electron oxygen reduction mechanism in a LiOH solution. The size of the catalyst also affected the specific capacities of the non-aqueous Li/O(2) batteries fabricated using the MnO(2) based air electrode. The highest specific capacity of 1917 mA h g(-1) was obtained for an α MnO(2) nanorod catalyst having a diameter of 25 nm. The cation present in the MnO(2) nanostructures appears to determine the catalytic activity of MnO(2).


Inactivation of pea leaf phosphofructokinase by light and dithiothreitol.
When intact pea (Pisum sativum L.) plants are illuminated, the glycolytic enzyme phosphofructokinase is inactivated. In crude extracts the enzyme is inhibited by dithiothreitol. It would seem that this cytoplasmic enzyme, like glucose-6-P dehydrogenase, is light-inactivated when the enzymes of photosynthetic carbon metabolism are light-activated.


Evaluation of genetic differentiation in Bos indicus cattle breeds from Marathwada region of India using microsatellite polymorphism.
Elucidation of genetic variability and genetic relationship among breeds has direct relevance with the issues of sustainable use of domestic animal genetic resources. In the present study, genetic polymorphism was evaluated using 22 microsatellite loci in unrelated samples of Red Kandhari and Deoni cattle breeds inhabiting the same geographical area of Marathwada region in Maharashtra state (western India). This work was mainly aimed at assessing the current genetic diversity to understand whether the two zebu populations in question are genetically differentiated. A total of 164 alleles were detected with an average of 5.82 and 5.86 alleles per locus (MNA) in Red Kandhari and Deoni breeds, respectively. The estimated mean observed (Ho) and expected (He) heterozygosity were 0.47 and 0.64 in Red Kandhari vs. 0.57 and 0.69 in Deoni cattle, respectively, demonstrating considerable level of genetic variation in both the populations. Mean estimates of F statistics were: F (FIT) = 0.315 +/- 0.035, f(FIS) = 0.231 +/- 0.031, theta(FST) = 0.110 +/- 0.022, with both the breeds exhibiting significant deficit of heterozygotes (FIS = 0.179 in Deoni; 0.278 in Red Kandhari). The multilocus FST values implied that 11.0% of the total genetic variation corresponds to breed and were statistically greater than zero for the two populations, suggesting population division. The evaluation of exact test also indicated that allele frequencies across all the loci differed significantly (P < 0.001) between two zebu breeds, further supporting population differentiation. Different genetic distance measures showed considerable levels of distances between the two cattle breeds (0.318 = Nei's standard DS; 0.250 = Nei's DA; 0.416 = Cavalli-Sforza and Edwards's Dc; 0.164 = Reynold's, and 2.64 = Delta mu square (dmicro)2. Bayesian statistical approach to assign each individual to the population also supported considerable differentiation between the two cattle breeds, possibly reflecting the limited gene flow between the two Marthwada cattle populations. The existence of cohesive breeding structure of both the breeds was further substantiated by allele-sharing distance measures (DAS) among individual animals. The results of this study thus revealed that the two Bos indicus breeds sharing the common breeding tracts are genetically differentiated enough as separate breeds.


Can we really diagnose diabetes during pregnancy?
To identify the accuracy of diagnosing postpartum diabetes and glucose intolerance using antepartum glycosylated hemoglobin (HbA1c) and fasting glucose values. A retrospective Hawaiian cohort of women with gestational diabetes during 2004-2011 were evaluated. Antepartum HbA1c and postpartum 75-g glucose tolerance tests were obtained. An antepartum HbA1c value of ≥6.5% had a 45.7% sensitivity, a 96% specificity and a 40% positive predictive value (PPV) for predicting postpartum diabetes. An antepartum HbA1c value of ≥6.5% had a 6.6% sensitivity, a 94.2% specificity and a 27% PPV for predicting postpartum impaired glucose tolerance. An antepartum HbA1c value of ≥6.5% had a 10.3% sensitivity, a 95.7% specificity and a 33.3% PPV for predicting postpartum impaired fasting glucoses. We could not demonstrate a clinically useful PPV for diagnosing postpartum diabetes or glucose intolerance using an antepartum elevated HbA1c value of ≥6.5% or a fasting glucose level of ≥90 mg/dL.


Ester vs. amide on folding: a case study with a 2-residue synthetic peptide.
Although known for their inferiority as hydrogen-bonding acceptors when compared to amides, esters are often found at the C-terminus of peptides and synthetic oligomers (foldamers), presumably due to the synthetic readiness with which they are obtained using protected peptide coupling, deploying amino acid esters at the C-terminus. When the H-bonding interactions deviate from regularity at the termini, peptide chains tend to "fray apart". However, the individual contributions of C-terminal esters in causing peptide chain end-fraying goes often unnoticed, particularly due to diverse competing effects emanating from large peptide chains. Herein, we describe a striking case of a comparison of the individual contributions of C-terminal ester vs. amide carbonyl as a H-bonding acceptor in the folding of a peptide. A simple two-residue peptide fold has been used as a testing case to demonstrate that amide carbonyl is far superior to ester carbonyl in promoting peptide folding, alienating end-fraying. This finding would have a bearing on the fundamental understanding of the individual contributions of stabilizing/destabilizing non-covalent interactions in peptide folding.


Pentoxifylline for treatment of sepsis and necrotizing enterocolitis in neonates.
Mortality and morbidity due to neonatal sepsis and necrotizing enterocolitis (NEC) is high despite the use of potent antimicrobial agents. Agents that modulate inflammation may improve outcomes. Pentoxifylline, a phosphodiesterase inhibitor, is one such agent. The primary objectives were to assess the effect on mortality and the safety of intravenous pentoxifylline as an adjunct to antibiotic therapy in neonates with suspected or confirmed sepsis and NEC. The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 2, 2011), MEDLINE, EMBASE and CINAHL, Science Citation Index for articles referencing Lauterbach 1996, proceedings of the Pediatric Academic Societies (1990 to 2011), BIOSIS (1992 to 2011), conference proceedings (1992 to 2011), ongoing trials and reference lists of identified RCTs were searched in July 2011. Randomised or quasi-randomised trials assessing the efficacy of pentoxifylline as an adjunct to antibiotics for treatment of suspected or confirmed sepsis or NEC in neonates were eligible. Two review authors independently abstracted information for the outcomes of interest. Typical relative risk (RR) and risk difference (RD) with 95% confidence intervals (CI) using fixed effects model are reported for dichotomous outcomes and mean differences for continuous outcomes. NNT was calculated for outcomes for which there was a statistically significant reduction in RD. In four randomised controlled trials, 227 neonates with suspected or confirmed sepsis were randomised to pentoxifylline or placebo. Pentoxifylline therapy significantly decreased "all cause mortality during hospital stay" in the overall population of infants with sepsis [typical RR 0.40 (95%CI 0.20 to 0.77); typical RD -0.15 (95%CI -0.26 to -0.05); NNT 7 (95%CI 4 to 20)]. Subgroup analyses revealed significant reduction in mortality in preterm infants, infants with confirmed sepsis and gram-negative sepsis. Pentoxifylline treatment significantly decreased length of hospital stay [mean difference -11.20 [95%CI -22.09 to -0.31] but not development of NEC in neonates with sepsis [typical RR 0.29 (95%CI 0.07 to 1.24); typical RD -0.20 (95%CI -0.41 to 0.01)]. No adverse effects due to pentoxifylline were noted. No completed trial of treatment with pentoxifylline for treatment of NEC was identified. Current evidence from four small studies suggests that the use of pentoxifylline as an adjunct to antibiotics in neonatal sepsis decreases mortality without any adverse effects. Researchers are encouraged to undertake large well-designed multicenter trials to confirm or refute the effectiveness of pentoxifylline in reducing mortality and adverse outcomes in neonates with suspected or confirmed neonatal sepsis and NEC.


Nutritional Status in Patients with Esophageal Cancer Receiving Chemoradiation and Assessing the Efficacy of Usual Care for Nutritional Managements.
Malnutrition is prevalent in esophageal cancer patients which affects cancer prognosis. The purpose of this study was a comprehensive assessment of nutritional status during Chemoradiation (CRT). Newly diagnosed adults with esophageal cancer were recruited for this study. Patient-Generated- Subjective Global Assessment (PG-SGA), anthropometric indices, body composition, dietary intake, laboratory tests, and nutritional-related complications were assessed before, after, and 4 to 6 weeks after CRT. Seventy-one cases were enrolled. The mean age was 66.8±12 years. Patients' mean weight loss was 2.42±2.4 kilograms during treatment. A significant reduction observed in mean MUAC (26.68±4.9 vs. 25.42±5.1 cm), fat mass percentage (24.11±11.8 vs. 22.8±12.5), fat free mass index (16.87±2.4 vs. 16.47±2.6 kg/m2) and hand grip strength (43.2±19 vs. 36.1±20 kg) during CRT (all p-values <0.0001). We had also a non-significant change in mean energy intake (19.5±11 vs. 18.3±11 kcal/kgw. day) and protein intake (0.56±0.4 vs. 0.66±0.5 g/kgw.day) during CRT. In our assessment before, immediately after and 4-6 weeks following CRT, we recorded energy intake insufficiency in 55.7%, 58.7% and 27.3% and protein intake inadequacy in 89.8%, 89.1% and 72.7% of cases, respectively. The most common complications were dysphagia (56.7%), anorexia (25%), and constipation (47.9%) at admission. Dysphagia improved in some cases (42%), but anorexia (35%), early satiety (25%), Esophagitis (25%), dysosmia (21%) and dysgeusia (17%) were increased as CRT complication. yet, 25% of patients had dysphagia and 34.4% had constipation 4-6 weeks after CRT. The twelve-months mortality was significantly associated with lower BMI after CRT, primary PG-SGA score, weight loss, BMI<18.5, MUAC, physical performance, living in rural or urban areas, addiction. Our study demonstrated a high prevalence of malnutrition among esophageal cancer patients which worsened during Chemoradiotherapy. Our findings warrant early screening and monitoring of nutritional status and effective nutritional interventions with symptoms management during treatment in these patients.<br />.


Mechanical stretch induces osteogenesis through the alternative activation of macrophages.
For reconstructive surgeons, critically skeletal damage represents a major challenge. Growing evidence indicate that bone repair is dynamically regulated by the mesenchymal stem cell (MSC)-macrophage interaction. Mechanical strain plays a fundamental role in bone repair and regeneration by influencing MSCs differentiation. Recently, a few findings indicate that macrophages may be mechanically sensitive and their phenotype can be regulated, in part, by mechanical cues. However, how macrophages subjected mechanical stretch influence the osteogenic differentiation of MSCs remain unclear. Thus, the purpose of this study is to explore the effect of macrophages stimulated with mechanical stretch on MSCs osteogenesis. By using a coculture system, we discover that macrophages efficiently induce osteogenic differentiation of MSCs under specific stretch conditions. A synergy mechanism between M2 polarization and YAP/BMP2 axis are identified through molecular and genetic analyses. Macrophages are activated by cyclic stretch and polarized to M2 phenotype that produce anti-inflammatory cytokines such as IL-10 and TGF-β to regulate the local inflammatory microenvironment. Furthermore, mechanical stretch induces YAP activation and nuclear translocation, subsequently regulates downstream BMP2 expression to facilitate MSCs osteogenesis. These findings not only advance our understanding of the complex influence among the mechanical strain, macrophage inflammatory response as well as the osteogenic differentiation of MSCs, but also reveal a control system from mechanical signals to chemical response then to cell behaviors during bone repair and regeneration.


[Bilateral nodular pulmonary histoplasmosis: cytohistological correlation].
Nodular bilateral pulmonary histoplasmosis: cyto-histological correlation. A case of nodular pulmonary histoplasmosis is reported. A 29 year old man was admitted to hospital with temperature and general weakness following a short stay in a tropical country. Laboratory investigation showed an increased E.S.R. and a routine chest-roentgenogram revealed multiple bilateral nodular lesions confirmed by CT scan. Smears obtained from fine-needle-aspiration biopsy showed the presence of epithelioid cell's clusters with a few giant-cells in a background of inflammatory elements and necrotic debris. The cytological picture was consistent with an inflammatory process with necrotizing granulomatous features. The clinical evolution and the radiological picture progression caused, nevertheless, suspicion of a metastatic tumor. The histological examination of a resected peripheral nodule confirmed the inflammatory nature of the process, revealing the presence of multiple roundish encapsulated conidia 2-4 microns in diameter scattered within a granulomatous and necrotic tissue. The fungi are clearly pinpointed by using special stains like Grocott method. Serological and microbiological investigations are necessary in order to confirm the diagnosis.


Single-port access compared with three-port laparoscopic adnexal surgery in a randomized controlled trial.
Scar-related cosmetic outcomes were compared prospectively between conventional three-port and single-port access laparoscopic adnexal gynaecological surgery. Enrolled patients were randomly assigned to a single- or three-port surgery group. Scar-related outcomes were evaluated at 1 month, 6 months and 1 year. Scars were assessed by an independent observer using the modified Vancouver Scar Scale (mVSS). All patients were asked about pain related to the scar and scar satisfaction; results were recorded using a numerical rating scale. Seventy-three patients were enrolled between June 2010 and June 2011. Demographic and surgical outcomes did not differ between the groups. mVSS results were similar in the two groups at each follow-up point. The scar satisfaction profile measured at 1 month showed no significant difference between the groups, but the single-port access group had better results than the conventional group at all other follow-up timepoints. Cosmetic outcome was better for single-port than for three-port adnexal gynaecological surgery at 6-month and 1-year follow-up.


Pleckstrin Levels Are Increased in Patients with Chronic Periodontitis and Regulated via the MAP Kinase-p38α Signaling Pathway in Gingival Fibroblasts.
Chronic periodontitis (CP) is a bacteria-driven inflammatory disease characterized by the breakdown of gingival tissue, the periodontal ligament, and alveolar bone, leading ultimately to tooth loss. We previously reported the pleckstrin gene (PLEK) to be highly upregulated in gingival tissue of patients with CP and the only gene concurrently upregulated in other inflammatory diseases including rheumatoid arthritis and cardiovascular diseases. Using saliva from 169 individuals diagnosed with CP and healthy controls, we investigated whether pleckstrin could serve as a novel biomarker of periodontitis. Additionally, we explored signal pathways involved in the regulation of PLEK using human gingival fibroblasts (HGFs). Pleckstrin levels were significantly higher (p < 0.001) in the saliva samples of patients with CP compared to controls and closely associated with CP severity. Immunohistochemical analysis revealed the expression of pleckstrin in inflammatory cells and gingival fibroblasts of CP patients. To explore the signal pathways involved in pleckstrin regulation, we stimulated HGFs with either interleukin-1β (IL-1β) or lipopolysaccharides (LPS) alone, or in combination with inhibitors targeting c-Jun N-terminal kinase, tyrosine kinase, protein kinase C, or p38 MAP kinase. Results showed that IL-1β and LPS significantly increased PLEK mRNA and pleckstrin protein levels. VX-745, the p38 MAP kinase inhibitor significantly decreased IL-1β- and LPS-induced pleckstrin levels at both the mRNA and the protein level. Together, these findings show that pleckstrin could serve as a salivary biomarker for the chronic inflammatory disease periodontitis and a regulator of inflammation via the p38 MAP kinase pathway.


Cell Therapy Products in Menopausal Medicine.
The incidence of postmenopausal diseases increases with the age of women. In this review, we introduce cell therapy products, a new treatment for postmenopausal osteoporosis, which often occurs in postmenopausal women. We also figure out the trends of research on cell therapy products and emphasize the necessity and importance of this research for researchers and postmenopausal women. Finally, we suggest the direction for improvement of postmenopausal osteoporosis and research on cell therapy products. We investigated which medication have been used so far. We also examined the development and technical problems of technologies that are currently in use.


Granule-laden perivascular cells observed in rabbits with experimental cerebral atherosclerosis.
Granule-laden perivascular cells distributed around the fine vessels in brain of hypertensive rabbits fed a cholesterol diet were examined by electron microscopy. Perivenule granule-laden cells contained secondary lysosomes, residual bodies, Golgi vesicles, fusion vesicles, and vacuoles. In periarteriole granule-laden cells, secondary lysosomes and vacuoles were not prominent. Findings indicated that the granule-laden cell may be a histiocyte that appears during the vascular reaction to hypertension and hypercholesterolemia. This vascular reaction may occur more strongly in the veins than the arteries.


[Molecular cloning and identify expression of the novel human LNX gene in gliomas].
To clone and identify expression pattern of the novel LNX gene, and characterize its molecular mechanism in all grades of human gliomas. From a fetal brain cDNA library, we isolated the novel human LNX gene. The expression pattern of LNX gene in 16 normal tissues was examined by MTC panel (Clontech). Microarray were adopted and hybridized with LNX probes to examine the expression of novel gene in gliomas. Northern hybridization was used for verifying expression of LNX gene in gliomas. Two-hybrid screen in yeast was used to identify human LNX interacting proteins. Pull-down assays and Co-immunoprecipitation were transfected in HEK293 cells according to the lipofectAMINE protocol. We isolated a 3.7 kb cDNA containing an open reading frame (ORF) of 1,899 bp and a putative 632 amino acids protein, which was located on 4q12. cDNA microarray showed LNX was down-regulated in all 18 glioma samples and it was testified by Northern-blot. The MTC panel showed a ubiquitous expression pattern which highly expressed in adult brain, kidney and pancreas, while weak expression in heart, lung, etc. The two-hybrid screen in yeast revealed that LNX interacted with SKIP (Ski interacting protein) via PDZ domains. The co-immunoprecipation suggested that LNX interacted with SKIP in HEK293 cells and could affect the subcellular localization of Numb, which indicated that LNX might function as a molecular anchor that localized Numb to the subcellular site of its interaction with Notch. cDNA microarray technology is a powerful technique in screening and locking differentially expressed genes in gliomas, LNX was closely related to human gliomas and suggested playing an important role in gliomas by notch signal approach.


Use of a sestamibi-only approach to routine minimally invasive parathyroidectomy.
We performed this study to compare a sestamibi-only radio-guided approach (MIBI) versus using intraoperative parathyroid hormone monitoring (IOPTH) in the performance of minimally invasive parathyroidectomy (MIP) in patients with a clearly positive preoperative sestamibi scan from January 2000 to June 2010. Five of 81 patients in the MIBI group required additional surgery, three at the time of MIP when the intraoperative findings were in conflict with the preoperative sestamibi scan and two required a second operation as a result of an undiscovered second adenoma. In the IOPTH group, five patients had an unnecessary bilateral neck exploration as a result of an inadequate drop in PTH levels, whereas six had their disease cured because the PTH levels predicted additional pathology. One patient in the IOPTH group remains hypercalcemic and represents the only surgical failure in this study. The MIBI group had a shortened operating room time and less cost (P < 0.001). No deaths or complications, including recurrent laryngeal nerve injuries, occurred in this study. Although both strategies are effective in managing hyperparathyroidism, a MIBI-only approach is less expensive and has shorter operative times with an occasional need for reoperation, whereas the IOPTH group results in more extensive surgery that will occasionally be unnecessary.


Cardiac condition in young chagasic women.
Chagas disease is a complex disorder caused by Trypanosoma cruzi. Most patients remain asymptomatic for several years and 30% of them progress quietly to developing cardiomyopathy. The factors that lead to chronic myocardial lesions are not fully understood. To investigate the association between clinical symptoms and single nucleotide polymorphisms in chagasic and non-chagasic women younger and older than 55 years of age. we analyzed Ala-9Val and Ile58Thr polymorphisms of the SOD-Mn gene, 138ex1ins/del A of the endothelin-1 gene (ET-1) and H323H (T/C) of the endothelin receptor A gene (ETA), by PCR-RFLP using genomic DNA from leukocyte of 85 women. We also evaluated serum lipid profile, renal and liver function, chest X-rays, electrocardiograms (ECGs) and echocardiography (EchoCG). Biochemical profiling did not show differences between chagasic and non-chagasic patients. The polymorphisms analyses showed a significant association in the distribution of frequencies of the Mn-SOD Ile58Thr gene between both groups. Young chagasic patients had a significantly higher prevalence of abnormalities in X-rays, in ECGs and they showed grade II and III of NYHA functional classes. The chance of having an abnormal EchoCG was 5.87 higher in young chagasic patients (OR=5.87, 95% CI 1.47-23.4). We concluded that the parasite affects young females by accelerating the deterioration of cardiac function, independent of other cardiovascular risk factors and the cardioprotective action of estrogens present in the premenopausal stage.


Development of a three-dimensional human skin equivalent wound model for investigating novel wound healing therapies.
Numerous difficulties are associated with the conduct of preclinical studies related to skin and wound repair. Use of small animal models such as rodents is not optimal because of their physiological differences to human skin and mode of wound healing. Although pigs have previously been used because of their human-like mode of healing, the expense and logistics related to their use also renders them suboptimal. In view of this, alternatives are urgently required to advance the field. The experiments reported herein were aimed at developing and validating a simple, reproducible, three-dimensional ex vivo de-epidermised dermis human skin equivalent wound model for the preclinical evaluation of novel wound therapies. Having established that the human skin equivalent wound model does in fact "heal," we tested the effect of two novel wound healing therapies. We also examined the utility of the model for studies exploring the mechanisms underpinning these therapies. Taken together the data demonstrate that these new models will have wide-spread application for the generation of fundamental new information on wound healing processes and also hold potential in facilitating preclinical optimization of dosage, duration of therapies, and treatment strategies prior to clinical trials.


[Status epilepticus and its treatment].
Status epilepticus is an emergency situation that can often lead to death or neurocognitive deficits despite adequate therapy is conducted. Etiology and prognosis can be widely heterogeneous. Knowledge of the basic pathophysiologic mechanisms has altered the definition of status epilepticus and the emphasis of therapy has also changed according to that. Today, with neuroprotective approach and recognition of the importance of time-window the management of generalised tonic clonic status became an emergency or critical care task, because intratracheal narcosis is more often and earlier the therapy of choice and even the sufficient treatment can convey serious side effects. In the article, the authors describe the systemic and cerebral pathophysiologic factors during seizure. The authors offer flow-charts illustrating the treatment of patients with generalised tonic clonic status to make the daily work of physicians easier. Early on critical care treatment and multidisciplinary approach is the pledge of reduction in mortality and morbidity.


A distinct neurocognitive phenotype in female fragile-X premutation carriers assessed with visual attention tasks.
Premature ovarian failure (POF) and underlying hormonal changes are recognized as a distinct phenotype in female fragile-X premutation carriers. Neurocognitive deficits, in particular mental retardation, are associated with the full mutation in males and females. In female full mutation carriers this neurocognitive phenotype is expressed more mildly than in males. Research on whether the fragile-X premutation is associated with a particular neurocognitive phenotype or not has been equivocal. By means of the Sonneville Visual Attentions Tasks (SVAT) computer-based battery of neurocognitive tasks, we assessed reaction time on different tasks in three groups of subjects: female premutation carriers, female full mutation carriers, and female control subjects. The results show that a fraction of the female premutation carriers perform poorly on several selective attention tasks, but not on other tasks. Their neurocognitive profile is different from that of control subjects and of the majority of female premutation carriers. It may also be different from the phenotype of female full mutation carriers, though in that respect this study remains inconclusive. These findings support earlier findings that the fragile-X premutation may affect neurocognitive functioning, in particular aspects of attention.


1 alpha,25-Dihydroxyvitamin D3 increases transforming growth factor and transforming growth factor receptor type I and II synthesis in human bone cells.
To determine whether the inhibition of human osteoblast growth mediated by 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)D3) occurs as a result of changes in transforming growth factor (TGF) and TGF receptor synthesis, we examined the effects of 1 alpha,25(OH)2D3 on the synthesis of TGF beta and TGF-beta receptors. Treatment with 1 alpha,25(OH)2D3, but not vehicle, increased TGF-beta 2 concentrations in human osteoblast cell supernantants in a dose- and time-dependent manner. The increase in TGF-beta 2 concentrations was associated with an inhibition of osteoblast cell growth; antibodies directed against transforming growth factor beta partially blocked the inhibition of cellular growth mediated by 1 alpha,25-(OH)2D3 TGF-beta 2 gene transcription and TGF-beta 2 mRNA concentrations were increased in 1 alpha,25(OH)D3 but not in vehicle-treated cells. 1 alpha,25(OH)2D3 increased TGF-beta type I and type II receptor mRNA levels in osteoblasts. Increased expression of TGF-beta 2 and TGF-beta receptors by 1 alpha,25(OH)2D3 might account for the inhibition of human osteoblast growth seen following 1 alpha,25(OH)2D3 treatment.


Genetic dissection of the syndrome X in the rat.
In 1988, Reaven used the term syndrome X to describe a relation between several disorders including hypertension, dyslipidemia, impaired glucose tolerance, obesity, and coronary heart disease. Despite a number of studies dealing with syndrome X, its genetic basis remains poorly understood. Regarding the complexity of this syndrome, it is important to use animal models developing the traits of the disease. Here we show a genetic dissection of syndrome X in the WOKW rat, an animal model of genetically determined syndrome X. We found a major quantitative trait locus (QTL) for glucose metabolism on chromosome 3 and further QTLs influencing obesity and body weight on chromosomes 1 and 5. Genetic determinants of dyslipidemia were mapped to chromosomes 4 and 17. In addition, suggestive linkage for serum insulin was found on chromosome 1 to the region previously shown to be associated with type-1 diabetes mellitus. This is the first study demonstrating independent genetic factors influencing traits of the syndrome X in the rat as well as a possible genetic relationships between syndrome X and diabetes mellitus. Moreover, regarding the close similarities between WOKW rat and human syndrome X, the study could help in a search of genetic factors involved in this complex metabolic disorder in human.


Slug regulates proliferation and invasiveness of esophageal adenocarcinoma cells in vitro and in vivo.
Slug is a transcription factor and E-cadherin repressor, which has recently been demonstrated to be important for cancer cells to down-regulate epithelial markers and up-regulate mesenchymal markers in order to become motile and invasive. In the present study, we assessed the relevance of Slug for invasion and growth potential of esophageal adenocarcinoma (EA) cells in vitro and in vivo. Slug expression was detected in nine human esophageal cancer cell lines. OE33 cell line was infected with Slug siRNA to knockdown of Slug; TE7 cell line was infected with full Slug cDNA to increase Slug expression. Then, Bcl-2 and E-cadherin expression and Caspase-3 activity were analyzed. MTT assay was applied to detect growth curve. The flow cytometric and Hoechst33258 staining was performed to detect apoptosis. The cells invasion in vitro was detected with a Boyden chamber. A pseudometastatic model of OE33 and TE7 in immunodeficient mice was used to assess the effects of knockdown of Slug and Slug overexpression on metastasis development. A subcutaneously nude mice xenograft model of OE33 and TE7 was used to assess the effects of knockdown of Slug and Slug overexpression on tumor growth. Immunohistochemical staining was used to analyze the expression of Slug, bcl-2 and E-cadherin, and TUNEL was used to detected apoptosis in vivo. Western blotting and RT-PCR showed that Slug expression was detectable in 7 of 9 human esophageal cancer cell lines. Bcl-2 was down-regulated and E-cadherin was up-regulated significantly in Slug siRNA-infected OE33 cell line (P<0.01). Bcl-2 was upregulated and E-cadherin was downregulated significantly in Slug cDNA-infected TE7 cells (P<0.05). OE33 cells with Slug knockdown were shown to possess markedly decreased invasiveness (P<0.05) and markedly increased apoptosis (P<0.05). Slug cDNA-infected TE7 cells were shown only to possess markedly increased invasiveness (P<0.05). There was significant relationship between Slug knockdown or Slug overexpression and cells proliferation (respectively, P<0.05). Animals injected with Slug-silenced OE33 cells had fewer seeded tumor (P<0.01), more apoptosis cells (P<0.05) and significantly xenograft tumor growth regression (P<0.05). But in Slug cDNA-infected TE7 cells, more seeded tumor number and significantly xenograft tumor growth were found in xenograft tumor (respectively, P<0.05). It was showed in the subcutaneously nude mice xenograft model tumor tissue, bcl-2 expression was reduced followed by the decrease of Slug expression in Slug-silenced tumor, and bcl-2 expression was increased followed by the increase of Slug expression. In pseudometastatic model, E-cadherin overexpression was found in Slug siRNA tumor tissue, but less E-cadherin expression was found in Slug cDNA tissue. Slug is an important modulator of apoptosis, growth and invasion in EA in vitro and in vivo. Slug inhibition may represent a novel strategy for treatment of EA.


The role of alpha-adrenoceptors in the regulation of renal tubular sodium reabsorption and renin secretion in the rabbit.
A study was undertaken in the anaesthetized rabbit to classify the alpha-adrenoceptor subtypes responsible for increasing renal tubular sodium reabsorption and renin secretion. Intrarenal administration of noradrenaline, at doses which did not change renal blood flow or glomerular filtration rate, significantly decreased urine flow, absolute and fractional sodium excretion by between 26% and 29%. These renal responses to noradrenaline were abolished by the selective alpha 1-adrenoceptor antagonist, prazosin, but not by the selective alpha 2-adrenoceptor antagonist, idazoxan. Noradrenaline, given intrarenally, increased renin secretion between two and three fold and this response was not modified by either prazosin or idazoxan. Intrarenal administration of the selective alpha-adrenoceptor agonists, phenylephrine and methoxamine, at dose rates which did not change renal haemodynamics, significantly reduced urine flow, absolute and fractional sodium excretion by 15% to 33%, but at doses which reduced blood flow and filtration rate, by between 11% and 26%, urine flow, absolute and fractional sodium excretion decreased between 42% and 49%. Infusion of guanabenz (selective alpha 2-adrenoceptor agonist), at doses with no renal haemodynamic action, increased urine flow, absolute and fractional sodium excretion by 11% to 15%, while at doses which decreased blood flow by 7%, these other variables did not change. Administration of UK 14304 (selective alpha 2-adrenoceptor agonist) reduced blood flow and filtration rate by 3% and 9% respectively but had no other measurable action. At higher doses, which decreased blood flow by 14% and filtration rate by 24%, urine flow, absolute and fractional sodium excretion fell by between 27% and 50%. Renin secretion was significantly increased by the high doses of phenylephrine and UK 14304 but not by the low doses of these drugs. These studies show that adrenergic stimulation of renal tubular sodium reabsorption involves the activation of alpha 1- but not alpha 2-adrenoceptors. Further, adrenergic activation of the juxtaglomerular cells to release renin does not appear to involve either alpha 1- or alpha 2-adrenoceptors.


Study on negative incident photon-to-electron conversion efficiency of quantum dot-sensitized solar cells.
Recently, negative signals are frequently observed during the measuring process of monochromatic incident photon-to-electron conversion efficiency (IPCE) for sensitized solar cells by DC method. This phenomenon is confusing and hindering the reasonable evaluation of solar cells. Here, cause of negative IPCE values is studied by taking quantum dot-sensitized solar cell (QDSC) as an example, and the accurate measurement method to avoid the negative value is suggested. The negative background signals of QDSC without illumination are found the direct cause of the negative IPCE values by DC method. Ambient noise, significant capacitance characteristics, and uncontrolled electrochemical reaction all can lead to the negative background signals. When the photocurrent response of device under monochromatic light illumination is relatively weak, the actual photocurrent signals will be covered by the negative background signals and the resulting IPCE values will appear negative. To improve the signal-to-noise ratio, quasi-AC method is proposed for IPCE measurement of solar cells with weak photocurrent response based on the idea of replacing the absolute values by the relative values.


Identification and photocatalytic destruction of malodorous compounds in sewage.
Malodorous compounds were identified in sewage samples using two different types of pre-concentration procedures: (a) pre-concentration onto adsorbent column (Tenax), (b) liquid-liquid extraction, in conjunction with sensory analysis. Sulphur and nitrogen containing compounds, hydrocarbons, substituted benzenes and components of essential oils and aldehydes were identified. These compounds were considered to be responsible for the septic and grassy/earthy odour described by sensory panellists. Studies on photodestruction of malodorous compounds present in the sewage samples showed that the main compounds were destroyed. This destruction was confirmed by both sensory analysis and gas chromatography through abatement in the intensity of odour as well as chromatogram peak areas. Comparing odour destruction using two processes, photocatalysis and loss due to mass transfer, the conversion rate was 71% at the beginning, later reaching a plateau of about 65%. Thus photocatalysis seems to be a promising technology in the degradation of malodorous compounds stripped to the atmosphere from sewage, especially when they are present at low concentrations.


The United States perspective towards poultry slaughter.
Handling and immobilizing animals prior to slaughter can be viewed from three interrelated and sometimes competing perspectives: processing efficiency, worker safety, and humaneness. Efforts to pass laws requiring humane slaughter of livestock and poultry spanned over 60 yr. The first, The Humane Slaughter Act of 1958, Public Law 85-765, applicable only to livestock plants supplying meat to the Federal Government, passed because of the right mix of available technology, experience with stunning methods, and moral suasion. This proposed law was not supported by the Department of Agriculture because of concerns over anticipated problems with its administration. The 1958 law became effective in 1960 and did not include humane slaughter requirements for ritual handling. By 1977, 26 states had enacted their own humane slaughter laws to cover livestock packers not included under the federal statute. The Humane Methods of Slaughter Act of 1978 extended the 1958 requirements to all livestock slaughter facilities, both federal and state, including facilities in foreign countries exporting to the U.S. The Department of Agriculture supported the 1978 legislation. The poultry industry was not included in either law primarily because electrical stunning was being used for the majority of birds slaughtered. The technology to humanely slaughter poultry is widely used but not all poultry are stunned before slaughter. Experience with stunning technology has shown well-defined production efficiency. Expanded use of stunning methods and research to confirm effectiveness from a humane slaughter perspective is needed to assure consumers that the industry will voluntarily continue humane practices.


Adrenohepatic fusion in domestic ferrets (Mustela putorius furo).
We describe two cases of adrenohepatic fusion (AHF) in domestic ferrets (Mustela putorius furo). This condition is defined as the union of hepatic tissue with the adrenal gland with close fusion of the respective parenchymal cells and lack of a fibrous capsule between the two cell populations. AHF is believed to be a congenital anomaly caused by failure of retroperitoneal mesenchyme to stimulate capsule formation, promoting the fusion of the structures. Two male domestic ferrets had a mass adherent to the liver, comprising adrenal gland with areas of fusion between the liver parenchyma and adrenal cortex. There was no evidence of a capsule separating the hepatic and adrenal cell populations. Clinical signs related to either the liver or adrenal gland were not observed, so this was considered to be an incidental finding.


Comparison of outcomes and survivorship between patients of different age groups following TKA.
A total of 6275 consecutive TKA patients were divided into three groups based on their age: <55, 55-70 and >70 years. There was a statistically significant difference in the change in WOMAC score (32 vs 31 vs 26, P<0.001) and Knee Society Score (78.9 vs 76.0 vs 69.0, P<0.001) favoring the younger populations. However, the revision rate was higher in the younger patients with a Kaplan-Meier survivorship at 5 years of 95.5%, 97.2% and 98.1% and at 10 years of 92.2%, 95.9% and 97.6%. Infection was the most frequent cause for failure in all groups although aseptic loosening and instability accounted for the increased revision rate in the younger patients. Younger patients may experience slightly better clinical outcomes following TKA but lower survivorship, when compared to older patient populations.


Towards sound based testing of COVID-19-Summary of the first Diagnostics of COVID-19 using Acoustics (DiCOVA) Challenge.
The technology development for point-of-care tests (POCTs) targeting respiratory diseases has witnessed a growing demand in the recent past. Investigating the presence of acoustic biomarkers in modalities such as cough, breathing and speech sounds, and using them for building POCTs can offer fast, contactless and inexpensive testing. In view of this, over the past year, we launched the "Coswara" project to collect cough, breathing and speech sound recordings via worldwide crowdsourcing. With this data, a call for development of diagnostic tools was announced in the Interspeech 2021 as a special session titled "Diagnostics of COVID-19 using Acoustics (DiCOVA) Challenge". The goal was to bring together researchers and practitioners interested in developing acoustics-based COVID-19 POCTs by enabling them to work on the same set of development and test datasets. As part of the challenge, datasets with breathing, cough, and speech sound samples from COVID-19 and non-COVID-19 individuals were released to the participants. The challenge consisted of two tracks. The Track-1 focused only on cough sounds, and participants competed in a leaderboard setting. In Track-2, breathing and speech samples were provided for the participants, without a competitive leaderboard. The challenge attracted 85 plus registrations with 29 final submissions for Track-1. This paper describes the challenge (datasets, tasks, baseline system), and presents a focused summary of the various systems submitted by the participating teams. An analysis of the results from the top four teams showed that a fusion of the scores from these teams yields an area-under-the-receiver operating curve (AUC-ROC) of 95.1% on the blind test data. By summarizing the lessons learned, we foresee the challenge overview in this paper to help accelerate technological development of acoustic-based POCTs.


A helically twisted imine macrocycle that allows for determining the absolute configuration of α-amino carboxylates.
Binding of α-amino carboxylates to a helically twisted imine macrocycle based on the indolocarbazole scaffold gives rise to characteristic circular dichroism spectra, and the patterns of the Cotton effects are consistent with the absolute configuration of α-amino carboxylates.


[Changes in plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) by the interferon treatment for chronic hepatitis C].
To determine the effects of interferon (IFN) treatment for chronic hepatitis C on vascular endothelium, we measured the concentrations of tissue plasminogen activator (t-PA) and its inhibitor (PAI-1) in the plasma from patients before and after IFN treatment for 14 consecutive days. The plasma t-PA and PAI-1 levels were measured before and after treatment. The plasma t-PA level was significantly increased after IFN treatment (p < 0.01) but no significant difference in plasma PAI-1 level was observed before and after treatment. The ratio of t-PA/PAI-1 was significantly increased after IFN treatment (p < 0.05). These changes may be caused by the effect of IFN on endothelium, leading to an activation of the endothelium derived fibrinolysis factors. Increase in plasma t-PA concentration may induce hyperfibrinolysis which may be one of the causes of suborbital hemorrhage. Further study on the fibrinolysis pathway in the blood is necessary to elucidate the mechanisms of the many side effects observed during IFN treatment.


Current practice in laboratory diagnostics of autoimmune diseases in Croatia. Survey of the Working group for laboratory diagnostics of autoimmune diseases of the Croatian Society of Medical Biochemistry and Laboratory Medicine.
With the trend of increasing incidence of autoimmune diseases, laboratories are faced with exponential growth of the requests for tests relating the diagnosis of these diseases. Unfortunately, the lack of laboratory personnel experienced in this specific discipline of laboratory diagnostic, as well as an unawareness of a method limitation often results in confusion for clinicians. The aim was to gain insight into number and type of Croatian laboratories that perform humoral diagnostics with the final goal to improve and harmonize laboratory diagnostics of autoimmune diseases in Croatia. In order to get insight into current laboratory practice two questionnaires, consisting of 42 questions in total, were created. Surveys were conducted using SurveyMonkey application and were sent to 88 medical biochemistry laboratories in Croatia for the first survey. Out of 33 laboratories that declared to perform diagnostic from the scope, 19 were selected for the second survey based on the tests they pleaded to perform. The survey comprised questions regarding autoantibody hallmarks of systemic autoimmune diseases while regarding organ-specific autoimmune diseases was limited to diseases of liver, gastrointestinal and nervous system. Response rate was high with 80 / 88 (91%) laboratories which answered the first questionnaire, and 19 / 19 (1.0) for the second questionnaire. Obtained results of surveys indicate high heterogeneity in the performance of autoantibody testing among laboratories in Croatia. Results indicate the need of creating recommendations and algorithms in order to harmonize the approach to laboratory diagnostics of autoimmune diseases in Croatia.


Smartphone-based dispatch of community first responders to out-of-hospital cardiac arrest - statements from an international consensus conference.
Over the past decade Smartphone-based activation (SBA) of Community First Responders (CFR) to out-of-hospital cardiac arrests (OHCA) has gained much attention and popularity throughout Europe. Various programmes have been established, and interestingly there are considerable differences in technology, responder spectrum and the degree of integration into the prehospital emergency services. It is unclear whether these dissimilarities affect outcome. This paper reviews the current state in five European countries, reveals similarities and controversies, and presents consensus statements generated in an international conference with the intention to support public decision making on future strategies for SBA of CFR. In a consensus conference a three-step approach was used: (i) presentation of current research from five European countries; (ii) workshops discussing evidence amongst the audience to generate consensus statements; (iii) anonymous real-time voting applying the modified RAND-UCLA Appropriateness method to adopt or reject the statements. The consensus panel aimed to represent all stakeholders involved in this topic. While 21 of 25 generated statements gained approval, consensus was only found for 5 of them. One statement was rejected but without consensus. Members of the consensus conference confirmed that CFR save lives. They further acknowledged the crucial role of emergency medical control centres and called for nationwide strategies. Members of the consensus conference acknowledged that smartphone-based activation of CFR to OHCA saves lives. The statements generated by the consensus conference may assist the public, healthcare services and governments to utilise these systems to their full potential, and direct the research community towards fields that still need to be addressed.


An unexpected tenant: contamination in a Physeter catodon (Physeteridae, Artiodactyla) genome indicates undescribed species of Sarcocystis Lankester, 1882 (Sarcocystidae, Eucoccidiorida) in the marine environment.
Data mining animal of genomes has been used before to identify endoparasites, and may be a particularly useful tool to surpass some difficulties faced by studies in the marine environment. We detected a species of Sarcocystis Lankester, 1882, contamination in the sperm whale (Physeter catodon Linnaeus) reference genome available in the GenBank database. We identified and extracted multiple gene fragments and placed the sequences in a phylogenetic framework. Our results indicate that the sequences of Sarcocystis sp. found in the genome do not correspond to any currently described species, despite a few other similar sequences having been identified in fur seals (Pinnipedia) and another sperm whale. Including data from previous studies, we suggest there is enough evidence to support the occurrence of at least four species of Sarcocystis in marine mammals. We also demonstrate that the term "S. canis-like" has been used for samples not closely related to Sarcocystis canis Dubey et Speer, 1991.


Diagnosis of bubonic plague by PCR in Madagascar under field conditions.
The diagnostic value of a PCR assay that amplifies a 501-bp fragment of the Yersinia pestis caf1 gene has been determined in a reference laboratory with 218 bubo aspirates collected from patients with clinically suspected plague managed in a regional hospital in Madagascar. The culture of Y. pestis and the detection of the F1 antigen (Ag) by enzyme-linked immunosorbent assay (ELISA) were used as reference diagnostic methods. The sensitivity of PCR was 89% (57 of 64) for the Y. pestis-positive patients, and 80.7% (63 of 78) for the F1 Ag-positive patients. The specificity of PCR for the culture-, F1 Ag-, and antibody-negative patients (n = 105) was 100%. Because in Madagascar most patients with plague are managed and their clinical samples are collected in remote villages, the usefulness of PCR was evaluated for routine diagnostic use in the operational conditions of the control program. The sensitivity of PCR was 50% (25 of 50) relative to the results of culture and 35.2% (19 of 54) relative to the results of the F1 Ag immunocapture ELISA. The specificity of PCR under these conditions was 96%. In conclusion, the PCR method was found to be very specific but not as sensitive as culture or the F1 Ag detection method. The limitation in sensitivity may have been due to suboptimal field conditions and the small volumes of samples used for DNA extraction. This technique is not recommended as a routine diagnostic test for plague in Madagascar.


Chemoarchitecture of layer 4 isocortex in the American water shrew (Sorex palustris).
We examined the chemoarchitecture of layer 4 isocortex and the number of myelinated nerve fibers of selected cranial nerves in the American water shrew (Sorex palustris). This study took advantage of the opportunity to examine juvenile brain tissue, which often reveals the most distinctive cortical modules related to different sensory representations. Flattened cortical sections were processed for the metabolic enzyme cytochrome oxidase, revealing a number of modules and septa. Subdivisions related to sensory representations were tentatively identified by performing microelectrode recordings in a single adult shrew in this study, combined with microelectrode recordings and anatomical findings from a previous investigation. Taken together, these results suggest that characteristic chemoarchitectonic borders in the shrew neocortex can be used to delineate and quantify cortical areas. The most obvious subdivisions in the water shrew include a relatively small primary visual cortex which responded to visual stimuli, a larger representation of vibrissae in the primary somatosensory cortex, and a prominent representation of oral structures apparent in the more rostral-lateral cortex. A presumptive auditory area was located in the far caudal cortex. These findings for the cortex are consistent with counts from optic, auditory and trigeminal nerves, suggesting that somatosensory inputs dominate the shrew's senses whereas visual and auditory inputs play a small role in navigation and in finding prey. More generally, we find that shrews share unusual features of cortical organization with moles, supporting their close taxonomic relationship.


Expression of CCR2 on monocytes and macrophages in chronically inflamed skin in atopic dermatitis and psoriasis.
Monocytes form a significant component of the inflammatory reaction taking place in the skin of atopic dermatitis and psoriasis. Chemokines are pivotal in mediating the attraction of leucocytes to sites of inflammation. The CC-chemokine, monocyte chemotactic protein 1 (MCP-1/CCL2), is expressed by keratinocytes in both atopic dermatitis and psoriasis. MCP-1 binds to the chemokine receptor CCR2 which is known to be expressed on monocytes and macrophages. We examined the expression of CCR2 on peripheral blood monocytes from patients with psoriasis (n=8) and atopic dermatitis (n=7) and found it to be expressed on approximately 90% of the cells, whereas monocytes from healthy donors had a significantly lower CCR2 expression (p<0.05). Skin biopsies from patients suffering from atopic dermatitis and psoriasis revealed that CCR2-positive cells expressed CD163, a marker for monocytes/macrophages. However, not all CD163-positive cells expressed CCR2, which could be interpreted as a mechanism for retaining the macrophages in the skin. Furthermore, we found that keratinocytes are able to express MCP-1, when stimulated with tumour necrosis factor-alpha and/or interferon-gamma in a dose-dependent manner. Thus MCP-1 and CCR2 interaction is likely of importance for the monocyte/macrophage trafficking of inflammatory skin disorders.


Estimated tris(1,3-dichloro-2-propyl) phosphate exposure levels for US infants suggest potential health risks.
Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) has been widely used as a flame retardant and is commonly detected in environmental samples. Biomonitoring studies relying on urinary metabolite levels (i.e. bis(1,3-dichloro-2-propyl) phosphate (BDCIPP)) demonstrate widespread exposure, but TDCIPP intake is unknown. Intake data area critical component of meaningful risk assessments and are needed to elucidate the potential health impacts of TDCIPP exposure. Using biomonitoring data, we estimated TDCIPP intake for infants. Infants aged 2-18 months were recruited from central, North Carolina (n=43, recruited 2014-2015), and spot urine samples were analyzed for BDCIPP. TDCIPP intake rates were estimated using daily urine excretion and the fraction of TDCIPP excreted as BDCIPP in urine. Daily TDCIPP intake estimates ranged from 0.01-15.03 μg/kg-day for children included in our assessment, with some variation depending on model assumptions. The U.S. Consumer Products Safety Commission (CPSC) previously established an acceptable daily intake of 5μg/kg-day for non-cancer health risks. Depending on modeling assumptions, we found that 2-9% percent of infants had TDCIPP intake estimates above this threshold. Our results indicate that current TDCIPP exposure levels could pose health risks for highly exposed infants.


[Tuberculosis in the San José University Hospital in Popayán, Colombia, 1998-2000].
TB is a public health problem in the world. In Colombia the Health 100 Law caused changes in prevention programs. The TB control program was compromised too. The actions in relation with active looking and examination of respiratory symptomatic people has been lowering. This retrospective study describes the clinical, epidemiological and medical care characteristics of tuberculosis patients attended in the Hospital Universitario San José de-Popayán. The clinical expedients and laboratory registers were investigated. 120 cases of 187 registered patients were included; finally only 89 were tuberculosis cases: 39 pulmonary (43.8%) and 50 extrapulmonary (56.2%). The extrapulmonary forms were: miliar, 39 patients; pleural, 4: of lymph nodes, 3; osteoarticular, 2, and meningitis, 2 cases. 49% of patients were 15-59 years old, 28% more than 59; males 64% and 65% coming from country areas. Ziehl Neelsen (ZN) smear was positive in 22% of 59 sputums of pulmonary TB and 20.2% of 94 extrapulmonary TB samples: mycobacteria culture positive in 14.3% of 14 pulmonary TB sputum, and 29.6% of 54 extrapulmonary TB samples. The media days into hospital/yard was 4-26 and the media days to begin tuberculosis treatment/yard was 4-8. In summary there are important request for TB medical care to universitary hospital, however the diagnosis is difficult and takes a long time while mycobacteria can be transmitted to hospital contacts. It is necessary to get a better efficiency of diagnostic tests in the hospital and appropriated survey of the cases that begin treatment.


Resistance to rabies virus infection conferred by the PMLIV isoform.
Various reports implicate PML and PML nuclear bodies (NBs) in an intrinsic antiviral response targeting diverse cytoplasmic replicating RNA viruses. PML conjugation to the small ubiquitin-like modifier (SUMO) is required for its localization within NBs. PML displays antiviral effects in vivo, as PML deficiency renders mice more susceptible to infection with the rhabdovirus vesicular stomatitis virus (VSV). Cells derived from these mice are also more sensitive to infection with rabies virus, another member of the rhabdovirus family. Alternative splicing from a single gene results in the synthesis of several PML isoforms, and these are classified into seven groups, designated PMLI to -VII. We report here that expression of PMLIV or PMLIVa, which is missing exon 5, inhibited viral mRNA and protein synthesis, leading to a reduction in viral replication. However, the expression of other nuclear isoforms (PMLI to -VI) and cytoplasmic PMLVIIb failed to impair viral production. This antiviral effect required PMLIV SUMOylation, as it was not observed with PMLIV 3KR, in which the lysines involved in SUMO conjugation were mutated. Thus, PMLIV and PMLIVa may exert this isoform-specific function through interaction with specific NB protein partners via their common C-terminal region.


Assessing the potential regeneration ability of corpus spongiosum in rabbit models.
Most congenital or acquired urethral diseases are usually accompanied by corpus spongiosum (CS) defects. However, Substitution urethroplasty can only reconstruct urethral lumen, not the CS. Many long-term complications occur due to the lack of protection from CS. Is CS a kind of tissue that cannot be repaired by regeneration and self-healing? In this study, the CS defect with urethral mucosa intact model was established in rabbits by removing the ventral CS tissue. Based on this model, three groups of different CS defect sizes, with lengths of 0.5 cm (Group A), 1.0 cm (Group B) and 1.5 cm (Group C), were then constructed, respectively, to assess the potential regeneration ability of CS. Three months later, the entire urethra, including the CS defect, was assessed by histological staining. Results showed that the vascular sinusoids were completely removed from urethral mucosa. The rabbit model of CS defect was established successfully. Three months post-operatively, the CS defects in all the 3 groups were replaced by disordered collagen instead of regenerating typical sinusoid-like vascular structure, which is significantly different from the normal CS rich in vascular sinusoids. The CS defects could not be repaired through self-healing. The potential regeneration ability of CS is extremely poor.


Age-Related Uptake of Heavy Metals in Human Spinal Interneurons.
Toxic heavy metals have been implicated in the loss of spinal motoneurons in amyotrophic lateral sclerosis/motor neuron disease (ALS/MND). Motoneuron loss in the spinal anterior horn is severe in ALS/MND at the time of death, making this tissue unsuitable for examination. We therefore examined spinal cords of people without muscle weakness to look for any presence of heavy metals that could make these neurons susceptible to damage. Spinal cord samples from 50 individuals aged 1-95 y who had no clinical or histopathological evidence of spinal motoneuron loss were studied. Seven μm formalin-fixed paraffin-embedded sections were stained for heavy metals with silver nitrate autometallography (AMGHM) which detects intracellular mercury, silver or bismuth. Neurons in the spinal cord were classified as interneurons or α-motoneurons based on their site and cell body diameter. Spinal interneurons containing heavy metals were present in 8 of 24 people (33%) aged 61-95 y, but not at younger ages. These AMGHM interneurons were most numerous in the lumbar spinal cord, with moderate numbers in the caudal cervical cord, few in the rostral cervical cord, and almost none in the thoracic cord. All people with AMGHM interneurons had occasional AMGHM staining in α-motoneurons as well. In one man AMGHM staining was present in addition in dorsomedial nucleus and sensory neurons. In conclusion, heavy metals are present in many spinal interneurons, and in a few α-motoneurons, in a large proportion of older people. Damage to inhibitory interneurons from toxic metals in later life could result in excitotoxic injury to motoneurons and may underlie motoneuron injury or loss in conditions such as ALS/MND, multiple sclerosis, sarcopenia and calf fasciculations.


Reverse micelles as nanocarriers of nisin against foodborne pathogens.
Reverse micelles (RMs) as nanocarriers of nisin were optimized for the highest water and bacteriocin content. RMs formulated with either refined olive oil or sunflower oil, distilled monoglycerides, ethanol, and water were effectively designed. Structural characterization of the RMs was assessed using Electron Paramagnetic Resonance Spectroscopy and Small Angle X-ray Scattering in the presence and absence of nisin. No conformational changes occurred in the presence of nisin for the nanocarriers. To assess efficacy of the loaded systems, their antimicrobial activity against Staphylococcus aureus and Listeria monocytogenes was tested in lettuce leaves and minced meat, respectively. Antimicrobial activity was evident in both cases. Interestingly, a synergistic antimicrobial effect was observed in lettuce leaves and to a lesser extent in minced meat between nisin and some of the nanocarriers' constituents (probably ethanol). Our findings suggest complex interactions that take place when RMs are applied in different food matrices.


Rac1 is required for the formation of three germ layers during gastrulation.
The Rac1, a member of the Rho family proteins, regulates actin organization of cytoskeleton and cell adhesion. We used genetic analysis to elucidate the role of Rac1 in mouse embryonic development. The rac1 deficient embryos showed numerous cell deaths in the space between the embryonic ectoderm and endoderm at the primitive streak stage. Investigation of the primary epiblast culture isolated from rac1 deficient embryos indicated that Rac1 is involved in lamellipodia formation, cell adhesion and cell migration in vivo. These results suggest that Rac1-mediated cell adhesion is essential for the formation of three germ layers during gastrulation.


Homeopathic Symphytum officinale increases removal torque and radiographic bone density around titanium implants in rats.
This study evaluated the effect of Symphytum officinale in homeopathic potency (6cH), on the removal torque and radiographic bone density around titanium implants, inserted in rats tibiae. Implants were placed in male rat tibiae, and the animals randomized to two groups (Control and S. officinale 6cH treated), which were evaluated at 7, 14, 28 and 56 days post-implantation. Radiographic bone density was measured at 6 points around the implant, using digital radiographic images, when implants were inserted and at sacrifice. Removal torque of the implants was also evaluated. Both removal torque and radiographic bone density evaluation showed that S. officinale 6cH treatment enhanced bone formation around the micro-implants, mainly at 14 days. At 56 days, the radiographic bone density was higher in the treated group. We conclude that S. officinale 6cH enhances, principally at the early stages of osseointegration, bone formation around titanium implants in rats' tibiae, based on radiographic and mechanical analysis.


A method for preparing genomic DNA that restrains branch migration of Holliday junctions.
The Holliday junction is a central intermediate in genetic recombination. This four-stranded DNA structure is capable of spontaneous branch migration, and is lost during standard DNA extraction protocols. In order to isolate and characterize recombination intermediates that contain Holliday junctions, we have developed a rapid protocol that restrains branch migration of four-way DNA junctions. The cationic detergent hex-adecyltrimethylammonium bromide is used to lyse cells and precipitate DNA. Manipulations are performed in the presence of the cations hexamine cobalt(III) or magnesium, which stabilize Holliday junctions in a stacked-X configuration that branch migrates very slowly. This protocol was evaluated using a sensitive assay for spontaneous branch migration, and was shown to preserve both artificial Holliday junctions and meiotic recombination intermediates containing four-way junctions.


On the money: making forecasts what they should be.
A forecast is an extrapolation from data describing what has happened in the past. Most healthcare organizations can perform all of the forecasting they require on a spread-sheet without use of special software. To ensure objectivity, managers should create the forecasts, and executives should evaluate them.