Vasopressin contributes to hypertension caused by nucleus tractus solitarius lesions.
Lesions of the nucleus tractus solitarius (NTS) were studied to determine whether they elevate plasma vasopressin levels and, if so, whether these elevated levels of vasopressin contribute to the hypertension caused by NTS lesions. Bilateral electrolytic lesions of the NTS caused acute, severe hypertension in rats anesthetized with chloralose and in conscious, freely moving rats. After placement of the NTS lesions there was a greater than tenfold elevation in plasma vasopressin levels. Administration of an antagonist of the vasoconstrictor action of vasopressin markedly diminished the hypertension in both conscious and anesthetized rats. Following ganglionic blockade with chlorisondamine, NTS lesions still elicited hypertension, and the magnitude of the hypertension was not different from that observed in rats not treated with chlorisondamine. The hypertension produced by lesions of the NTS in ganglionic-blocked rats was completely abolished by administration of a vasopressin antagonist. These results indicate that (1) NTS lesions elevate plasma vasopressin levels and (2) elevated plasma vasopressin contributes to the hypertension produced by such lesions.


Immunological evidence for multiple forms of alpha-melanotropin (alpha-MSH) in the pars intermedia of the Australian lungfish, Neoceratodus forsteri.
Acid extracts of individual pars intermedia from the Australian lungfish, Neoceratodus forsteri, were fractionated by gel filtration chromatography and analyzed for alpha-melanotropin (alpha-MSH) immunoreactivity. In these studies a C-terminal-specific alpha-MSH radioimmunoassay (RIA) was used. Following gel filtration chromatography on a Sephadex G-75 column, a major peak of immunoreactive alpha-MSH-sized material was detected. On the average there was 338 +/- 72 pmol (SD) of immunoreactive alpha-MSH per lungfish pars intermedia (n = 3). Following gel filtration the immunoreactive alpha-MSH was further analyzed by reverse-phase high-performance liquid chromatography (HPLC). Three peaks of immunoreactivity were detected. These peaks were designated Peaks 1, 2, and 3. The retention times of these peaks corresponded to, respectively, mammalian ACTH(1-13)amide, N-acetyl-ACTH(1-13)-amide, and N,O-diacetyl-ACTH(1-13)amide. Peaks 2 and 3 represented approximately 95% of the immunoreactive alpha-MSH recovered. Analysis of immunoreactive Peaks 2 and 3 by cation-ion-exchange indicated that both peaks had a net charge of +3 at pH 2.5. Since O-acetyl groups are sensitive to high pH, Peak 3 was incubated for 1 hr at 37 degrees in 0.01 N NaOH, pH 12. Under these conditions, Peak 3 eluted with the same retention time as untreated Peak 2. Collectively, these results indicate that Peaks 2 and 3 correspond to mono- and diacetylated lungfish alpha-MSH, respectively.


A mechanism of skin appendage macropores electroactivation during iontophoresis.
A physical mechanism of activation of skin appendage macropores under the influence of an electric field is considered theoretically. The macropore is considered as a long cylinder tube which is closed and flattened out before the electric field is applied. The charging of the capacitance of the macropore walls is the driving force of electroactivation. During this process the free energy of the system decreases, which is energetically favourable and results in water pulled into the tube and to a gradual opening of the tube. It is shown that consideration of the macropore wall conductance leads to a considerable slowdown of electroactivation. The opening time of a separate macropore is estimated. It is equal to 30 min for a macropore 4 mm in length. The dependence of the surface density of activated macropores on time is calculated theoretically. The obtained theoretical results are in a good agreement with the literature data.


Performance evaluation method for read mapping tool in clinical panel sequencing.
In addition to the rapid advancement in Next-Generation Sequencing (NGS) technology, clinical panel sequencing is being used increasingly in clinical studies and tests. However, tools that are used in NGS data analysis have not been comparatively evaluated in performance for panel sequencing. This study aimed to evaluate the tools used in the alignment process, the first procedure in bioinformatics analysis, by comparing tools that have been widely used with ones that have been introduced recently. With the accumulated panel sequencing data, detected variant lists were cataloged and inserted into simulated reads produced from the reference genome (h19). The amount of unmapped reads and misaligned reads, mapping quality distribution, and runtime were measured as standards for comparison. As the most widely used tools, Bowtie2 and BWA-MEM each showed explicit performance with AUC of 0.9984 and 0.9970 respectively. Kart, maintaining superior runtime and less number of misaligned read, also similarly possessed high level of AUC (0.9723). Such selection and optimization method of tools appropriate for panel sequencing can be utilized for fields requiring error minimization, such as clinical application and liquid biopsy studies.


Deiodinases and Cancer.
Hormones are key drivers of cancer development, and alteration of the intratumoral concentration of thyroid hormone (TH) is a common feature of many human neoplasias. Besides the systemic control of TH levels, the expression and activity of deiodinases constitute a major mechanism for the cell-autonomous, prereceptoral control of TH action. The action of deiodinases ensures tight control of TH availability at intracellular level in a time- and tissue-specific manner, and alterations in deiodinase expression are frequent in tumors. Research over the past decades has shown that in cancer cells, a complex and dynamic expression of deiodinases is orchestrated by a network of growth factors, oncogenic proteins, and miRNA. It has become increasingly evident that this fine regulation exposes cancer cells to a dynamic concentration of TH that is functional to stimulate or inhibit various cellular functions. This review summarizes recent advances in the identification of the complex interplay between deiodinases and cancer and how this family of enzymes is relevant in cancer progression. We also discuss whether deiodinase expression could represent a diagnostic tool with which to define tumor staging in cancer treatment or even a therapeutic tool against cancer.


North Carolina Latino Farmworkers' Use of Traditional Healers: A Pilot Study.
Farmworkers in the United States experience high rates of injury and illness but have limited access to conventional health care. Farmworkers are often from countries that have active traditional healers, so understanding the use of traditional healers among farmworkers is important. This pilot study (1) describes the use of traditional healers among farmworkers and (2) compares the use of traditional healers by farmworkers with other Latino immigrants. Interviews were conducted in 2015 with 100 Mexican farmworkers (80 men, 20 women) and 100 Mexican immigrant non-farmworkers (50 men, 50 women) in North Carolina. Most farmworkers (78%) had H-2A visas. More farmworkers (64%) than non-farmworkers (41%) had ever used traditional healers. Among farmworkers, 21% (vs. 11% of non-farmworkers) had used curanderos, 54% (vs. 32%) sobadores, 43% (vs. 21%) hueseros, 11% (vs. 13%) yerberos, and 4% (vs. 6%) espiritualistas. More farmworkers had used a traditional healer in the past year (16% vs. 8%), but fewer had used this healer in the United States (4% vs. 8%). Among all participants, males (58.5%) more than females (41.4%) (P = .0214), and returning to Mexico annually (64.1%) more than who do not (45.1%) (P = .0086) had ever used any traditional healer. This pilot study indicates the need for further research that documents the use of traditional healers by Latino farmworkers with diverse visa statuses, from countries in addition to Mexico, and in other regions in the United States. This research should also delineate the specific illnesses and injuries for which Latino farmworkers use traditional healers.


Brief Report: Estimated Incidence of Perinatally Acquired HIV Infection in the United States, 1978-2013.
An incidence of perinatally acquired HIV infection less than 1:100,000 live births is one of the Centers for Disease Control and Prevention (CDC) goals of the United States. Such an estimate has only been possible in recent years because regular nationwide data were lacking. Using previously published CDC estimates of the number of infants born with HIV infection in the United States (interpolating for years for which there was no published estimate), and census data on the annual number of live-born infants, estimated incidence was calculated for 1978-2013. Exact 95% confidence intervals (CIs) were calculated using the Poisson distribution. Estimated incidence of perinatally acquired HIV infection peaked at 43.1 (95% CI: 41.1 to 45.1) in 1992 and declined rapidly after the use of zidovudine prophylaxis was recommended in 1994. In 2013, estimated incidence of perinatally acquired HIV infection in the United States was 1.8 (95% CI: 1.4 to 2.2), a 96% decline since the peak. Estimated incidence of perinatally acquired HIV infection in the United States in 2013 was 1.8/100,000 live births.


Accidental orthodontic elastic band-induced periodontitis: orthodontic and laser treatment.
The periodontal effects of elastic bands in the subgingiva have been reported since 1870. Case reports illustrate the treatment of elasticband-induced periodontitis in two children with a combination of laser treatment, antibiotics, splinting and orthodontics. Follow-up evaluations show reattachment, maturation of the apical portion of the roots, improved radiographic bone levels and minimal sulcus depth, mobility and recession. Continued orthodontic and periodontic care is planned.


Accountability in the UK healthcare system: an overview.
Recent changes in the English National Health Service (NHS) have introduced new complexities into the accountability arrangements for healthcare services. This commentary describes how the new organizational structures have challenged the traditional centralized accountability structures by creating a more dispersed system of governance for local health-care commissioners. It sets the context of discussions about accountability in the UK NHS and then describes the key changes in England following the implementation of the NHS reforms in April 2013. The commentary concludes that while there is increased complexity of accountability within a more decentralized and fragmented healthcare system, the government's goal of achieving increased local autonomy and greater control by general practitioners (GPs) will probably not be realized. In particular, the system will continue to have strongly centralized aspects, with increased regulation and central political responsibility.


Small intestinal vascular malformation bleeding: a case report with imaging findings.
The small intestine is approximately 5-6 m long and occupies a large area in the abdominal cavity. These factors preclude the use of ordinary endoscopy and X-ray to thoroughly examine the small intestine for bleeding of vascular malformations. Thus, the diagnosis of intestinal bleeding is very difficult. A 47-year-old man presented at the hospital 5 mo ago with dark stool. Several angiomas were detected by oral approach enteroscopy, but no active bleeding was observed. Additionally, no lesions were detected by anal approach enteroscopy; however, gastrointestinal tract bleeding still occurred for an unknown reason. We performed an abdominal vascular enhanced computed tomography examination and detected ileal vascular malformations. Ileum angioma and vascular malformation were detected by a laparoscopic approach, and segmental resection was performed for both lesions, which were confirmed by pathological diagnosis. This report systemically emphasizes the imaging findings of small intestinal vascular malformation bleeding.


Exosomes: Biological Pharmaceutical Nanovectors for Theranostics.
Exosomes are natural cell-derived nanovesicles of endocytic origin that enable cellular crosstalk by transferring encapsulated molecular cargos across biological barriers, thereby holding significantly complex implications in the etiology and progression of diverse disease states. Consequently, the development of exosomes-based nano-theranostic strategies has received immense consideration for advancing therapeutic interventions and disease prognosis. Their favorable biopharmaceutical properties make exosomes a unique nanoparticulate carrier for pharmaceutical drug delivery. This review provides an update on the contemporary strategies utilizing exosomes for theranostic applications in nanomedicine. In addition, we provide a synopsis of exosomal features and insights into strategic modifications that control in vivo biodistribution. We further discuss their opportunities, merits and pitfalls for cell/tissue targeted drug delivery in personalized nanotherapy.


Evolution of mitochondrial DNA content after planned interruption of HAART in HIV-infected pediatric patients.
HAART-related long-term toxicities, many of them ascribed to mitochondrial (mt) toxicity of the nucleoside analogues, are being increasingly reported in HIV-infected children. HIV infection can also cause mt damage. Case series include 13 vertically HIV-infected pediatric patients (9 girls, median age 10.5 years) with optimal long-term response to a first-line HAART regimen who underwent planned treatment interruption (PTI). MtDNA content from peripheral blood mononuclear cells was assessed by means of a real-time PCR technique at PTI and 12 months later and expressed as an mtDNA/nuclear DNA ratio, together with lactate levels. At PTI, patients had remained a median time of 4.7 years on HAART and 4.3 years with complete suppression of viral replication. The main reason leading to PTI was treatment fatigue. One month after PTI, HIV plasmatic viral load had increased to 4.8 log copies/ml and stabilized thereafter. During the 12-month study period, all children remained free from any HIV-related clinical event. A progressive and significant decrease in median CD4 cell counts and percentages was observed 12 months after PTI. One year after PTI, the median mtDNA/nuclear DNA ratios had increased from 0.76 to 1.08 (p = 0.002) and lactate levels had decreased (from 1.12 to 0.73 mmol/liter; p = 0.019). Changes in mtDNA did not correlate with changes in lactate levels. No relationship was found between the evolution in mt toxicity markers and the rest of the clinical, immunological, and virological variables. In this series, PTI led to a partial restoration of mtDNA levels and a significant decrease in lactate values.


Single-port laparoscopic surgery by use of a surgical glove port: initial experience with 25 cases.
single-port laparoscopic surgery has gained popularity over the last decade. This technique is used for several surgical procedures. This paper documents a new and cheaper access method known as "surgical glove port" or "homemade single-port", and describes our initial experience with 25 cases. Eleven cholecystectomies, eight totally extraperitoneal inguinal hernia repairs, three splenectomies, two gastric wedge resections, and one anti-reflux procedure were performed. Twenty-four procedures out of twenty-five were completed without conversion to open or multiple port techniques. An individual patient who had both cholelithiasis and gastrointestinal stromal tumor underwent both cholecystectomy and gastric wedge resection in the same session with surgical glove port technique. In another case two additional 5-mm incisions were made for hemostasis. The additional operation was required and performed by using surgical glove port for a hiatal hernia repair case, because of postoperative hemorrhage. Single-port laparoscopic surgery is an applicable technique for plenty of surgical procedures. Also, surgical glove port is a newer technique and some initial experience shave already published all over the world. Surgical glove port has advantages such as cost-effectiveness, easy and safe port implantation and specimen extraction. SGP can also be used for treating post-operative complications.


Acute abdomen resulting from concurrent thrombosis of celiac trunk and superior mesenteric artery.
Mesenteric ischemia is one of the most mortal diseases of the gastrointestinal system causing acute abdomen. In most of the patients, the etiological factor is the obstructive embolism or thrombosis of superior mesenteric artery. In the literature, there have been reports regarding also celiac trunk occlusion in rare situations. The gold standard treatment relies on early diagnosis. The originality of our report relies on the concurrent obstruction of both vascular structures.


Evaluation of accuracy and precision of a smartphone based automated parasite egg counting system in comparison to the McMaster and Mini-FLOTAC methods.
Fecal egg counts are emphasized for guiding equine helminth parasite control regimens due to the rise of anthelmintic resistance. This, however, poses further challenges, since egg counting results are prone to issues such as operator dependency, method variability, equipment requirements, and time commitment. The use of image analysis software for performing fecal egg counts is promoted in recent studies to reduce the operator dependency associated with manual counts. In an attempt to remove operator dependency associated with current methods, we developed a diagnostic system that utilizes a smartphone and employs image analysis to generate automated egg counts. The aims of this study were (1) to determine precision of the first smartphone prototype, the modified McMaster and ImageJ; (2) to determine precision, accuracy, sensitivity, and specificity of the second smartphone prototype, the modified McMaster, and Mini-FLOTAC techniques. Repeated counts on fecal samples naturally infected with equine strongyle eggs were performed using each technique to evaluate precision. Triplicate counts on 36 egg count negative samples and 36 samples spiked with strongyle eggs at 5, 50, 500, and 1000 eggs per gram were performed using a second smartphone system prototype, Mini-FLOTAC, and McMaster to determine technique accuracy. Precision across the techniques was evaluated using the coefficient of variation. In regards to the first aim of the study, the McMaster technique performed with significantly less variance than the first smartphone prototype and ImageJ (p<0.0001). The smartphone and ImageJ performed with equal variance. In regards to the second aim of the study, the second smartphone system prototype had significantly better precision than the McMaster (p<0.0001) and Mini-FLOTAC (p<0.0001) methods, and the Mini-FLOTAC was significantly more precise than the McMaster (p=0.0228). Mean accuracies for the Mini-FLOTAC, McMaster, and smartphone system were 64.51%, 21.67%, and 32.53%, respectively. The Mini-FLOTAC was significantly more accurate than the McMaster (p<0.0001) and the smartphone system (p<0.0001), while the smartphone and McMaster counts did not have statistically different accuracies. Overall, the smartphone system compared favorably to manual methods with regards to precision, and reasonably with regards to accuracy. With further refinement, this system could become useful in veterinary practice.


Ischemia-reperfusion induced microvascular responses in LDL-receptor -/- mice.
The objective of this study was to determine whether the microvascular responses to ischemia and reperfusion (I/R) are altered in an animal model of atherosclerosis, the low-density lipoprotein-receptor knockout (LDLr -/-) mouse. Intravital video microscopy was used to monitor venular wall shear rate, leukocytes rolling velocity, the number of rolling, adherent and emigrated leukocytes, and albumin leakage in cremasteric postcapillary venules of wild-type (B6129) and LDLr -/- mice exposed to 60 min of ischemia and 60 min of reperfusion. The postcapillary venules of LDLr -/- mice exhibited two- to threefold larger increments in the number of adherent leukocytes and a more profound albumin leakage response to I/R than venules in wild-type mice. The exaggerated inflammatory responses noted in LDLr -/- mice placed on a normal diet were not exacerbated by a high-cholesterol diet. Treatment of LDLr -/- mice with either a platelet-activating factor (PAF) receptor antagonist (WEB-2086) or a monoclonal antibody (YN-1) against the endothelial cell adhesion molecule, intercellular adhesion molecule 1 (ICAM-1), markedly attenuated the I/R-induced leukocyte adherence and albumin leakage. These findings indicate that atherogenic mice are more vulnerable to the deleterious microvascular effects of I/R and that PAF-mediated, ICAM-1-dependent leukocyte adhesion contributes to this exaggerated response to I/R.


Serum HtrA1 is differentially regulated between early-onset and late-onset preeclampsia.
HtrA1 (high temperature requirement A1) is a serine protease of the HtrA family. We have previously shown that the placenta expresses the highest level of HtrA1 mRNA compared to other tissues in the human. Others have reported that placental HtrA1 is significantly up-regulated in preeclampsia (PE), a pregnancy-specific multi-systemic disorder associated with new onset hypertension and proteinuria. However, it is unclear how serum HtrA1 changes in a normal pregnancy and whether it is altered in PE pregnancies. A sandwich ELISA highly specific to human HtrA1 and suitable for serum detection was developed and thoroughly validated. This assay was then applied to serum samples from different stages of normal pregnancy, as well as early-onset (<34 weeks) and late-onset (>34 weeks) PE pregnancies. Serum HtrA1 increased progressively with increasing gestation in normal pregnancies. However, this trend was perturbed in women with PE. Compared to respective gestation-age-matched normal pregnancies, HtrA1 serum levels were significantly increased in early-onset PE, but significantly reduced in late-onset PE. This is the first report to show a clear increase of HtrA1 in the maternal circulation during normal pregnancy, consistent with HtrA1 being highly expressed in the placenta. Importantly, this study identified that serum HtrA1 was altered differently in early-onset and late-onset PE pregnancies, highlighting the complex regulation of HtrA1 in the different subtypes. The significant increase of serum HtrA1 in early-onset PE suggests that it may be a potential biomarker for the diagnosis of early-onset PE at disease presentation.


Expression analysis of immune genes in goldfish (Carassius auratus) infected with the monogenean parasite Gyrodactylus kobayashii.
Heavy infection with the ectoparasite Gyrodactylus kobayashii commonly leads to high mortality of ornamental goldfish Carassius auratus. To understand the underlying mechanisms of goldfish against infection of gyrodactylids, transcription responses of immune-related genes including IL-1β2, TNFα1, TNFα2, IFN-γ, TGFβ, MHC II β, TCRβ1 and complement factor C3 were studied by real-time quantitative PCR analysis. Significant increases of expression of inflammatory genes such as IL-1β2, TNFα1, TNFα2 and TGFβ were detected at days 7 and 14 post-infection (pi). No significant differences of G. kobayashii load were observed in response to primary infection and re-infection. In addition, the transcript levels of genes involved in adaptive immunity such as MHC II β and TCRβ1 remained unchanged. Curiously, fish moderately infected with gyrodactylid showed elevated expression IL-1β2, TNFα1 and TNFα2. In all the gyrodactylids-infected fish, expression of complement factor C3 was consistently inhibited. The results extend current knowledge to the understanding of gyrodactylid infection in fish and support the previous findings that innate immunity is indispensable for controlling parasite infection.


Recovery of avian schistosome cercariae from water using penetration stimulant matrix with an unsaturated fatty acid.
Avian schistosome cercariae that emerge from aquatic snails can penetrate human skin causing cercarial dermatitis resulting in serious skin disease in sensitized and immunocompromised people. A trap developed for Schistosoma mansoni cercariae was tested for recovery of avian schistosome cercariae. A matrix with an unsaturated fatty acid, linoleic acid stimulates attachment and penetration of Trichobilharzia spp. cercariae, and the immobilized larvae can be subsequently visualized. The number of trapped cercariae exceeded by 3 to 7 times the number of larvae expected on the surface of the trap, based on their random distribution in the water. Recognition, attachment, and penetration of Trichobilharzia spp. cercariae led to injection of more secretory products into the stimulant matrix than by Schistosoma mansoni cercariae. This method can assist in the identification of waters infected with avian schistosome cercariae so that human exposure to these parasitic larvae can be minimized.


UV-induced mutations of supF gene on a shuttle vector plasmid in p53-deficient mouse cells are qualitatively different from those in wild-type cells.
To verify the genetic instability of p53-deficient cells, UV-induced mutation of the supF gene on a shuttle vector was analyzed. UV-irradiated or non-irradiated shuttle vector plasmid carrying the supF gene as a target of mutation (pYZ289) was introduced into p53-deficient and p53-proficient mouse embryonic fibroblasts, and then the plasmid DNA replicated in mouse cells was recovered. Survival of UV-irradiated plasmid was almost equivalent in both p53-deficient and p53-proficient cells. The frequencies of UV-induced mutation of the supF gene were also the same in both types of cells. However, the distributions of base change mutations in the supF sequence were different between p53-deficient and p53-proficient cells; especially the locations of tandem CpC to TpT changes exhibited a marked difference. Since DNA repair activities of these two types of cell were almost the same, these qualitative differences in UV-induced mutations were probably caused by as yet unidentified differences in other than DNA repair activity.


Mutations in SOX9, the gene responsible for Campomelic dysplasia and autosomal sex reversal.
Campomelic dysplasia (CD) is a skeletal malformation syndrome frequently accompanied by 46,XY sex reversal. A mutation-screening strategy using SSCP was employed to identify mutations in SOX9, the chromosome 17q24 gene responsible for CD and autosomal sex reversal in man. We have screened seven CD patients with no cytologically detectable chromosomal aberrations and two CD patients with chromosome 17 rearrangements for mutations in the entire open reading frame of SOX9. Five different mutations have been identified in six CD patients: two missense mutations in the SOX9 putative DNA binding domain (high mobility group, or HMG, box); three frameshift mutations and a splice-acceptor mutation. An identical frameshift mutation is found in two unrelated 46,XY patients, one exhibiting a male phenotype and the other displaying a female phenotype (XY sex reversal). All mutations found affect a single allele, which is consistent with a dominant mode of inheritance. No mutations were found in the SOX9 open reading frame of two patients with chromosome 17q rearrangements, suggesting that the translocations affect SOX9 expression. These findings are consistent with the hypothesis that CD results from haploinsufficiency of SOX9.


Telerehabilitation for people with breast cancer through the COVID-19 pandemic in Chile.
Adapting face-to-face physical therapy consultations in cancer care to a model of telerehabilitation has been necessary, given the imminent spread of the COVID-19 pandemic. In this respect, the current model of telerehabilitation for people with breast cancer can be described as a method of continuing physical therapy treatment, in a public hospital with limited resources.


Female adult puncture-induced plant volatiles promote mating success of the pea leafminer via enhancing vibrational signals.
Herbivore-induced plant volatiles (HIPVs) synergize with the sex pheromones of herbivorous insects to facilitate mate location. However, the synergism of HIPVs and acoustic signals for sexual communication remains unknown. Here, we investigated the synergy between HIPVs and vibrational duets for sexual communication and mating in the pea leafminer ( Liriomyza huidobrensis). Our results indicated that adult leafminers do not produce species-specific pheromone, and female-puncture-induced plant volatiles facilitate the attraction of both sexes to host plant leaves and sexual encounters. Insect-derived cues do not participate in mate locations. Both sexes do not produce qualitatively different cuticular hydrocarbons (CHCs), and CHCs from females cannot elicit the antennal and behavioural responses of males. By contrast, induced green leaf volatiles, terpenoids and oximes elicit dramatic antennal responses in both sexes. Electrophysiological and behavioural tests consistently showed that the volatiles (Z)-3-hexenol and (Z)-3-hexenyl-acetate elicited the most intense gas chromatographic-electroantennographic responses, and attracted males and females. Remarkably, these volatiles significantly promoted the occurrence of vibrational duets between the sexes, thereby increasing the mating success of leafminers. Therefore, the synergism of HIPVs and vibrational signals largely promoted the mating success of leafminers, suggesting an alternative control strategy through precision trapping for non-pheromone-producing insects. This article is part of the theme issue 'Biotic signalling sheds light on smart pest management'.


Performance characteristics according to Commission Decision 2002/657/EC in the fluorimetric determination of tetracycline in the absence and in the presence of magnesium.
The fluorimetric determination of tetracycline is usually carried out in the presence of some metals that, through the formation of a complex with this antibiotic, enhance its fluorescence emission, giving more sensitive determination methods. It is well established that magnesium is one of these metals. However, it is possible that higher signals do not mean a real improvement in the quality of the analytical method. In this work, the univariate and multivariate fluorescence determination of tetracycline is performed in the presence and absence of Mg(2+), comparing the quality of the analyses through some performance characteristics that, according to Commission Decision 2002/657/EC define the functional qualities of analytical methods. The methods with the best performance characteristics were multivariate determinations carried out in the absence of Mg(2+), both when emission or excitation spectra were taken, the decision limits (CCalpha) being 13.1 and 20.1 microg/L and the detection capabilities (CCbeta) 25.3 and 38.5 microg/L, respectively. This study points out through a case study that higher analytical signals do not necessarily mean better performance characteristics of a method of analysis.


Suppressor of Ty 16 promotes lung cancer malignancy and is negatively regulated by miR-1227-5p.
Suppressor of Ty 16 (Spt16) is a component of the facilitates chromatin transcription (FACT) complex, which is a histone chaperone and involved in gene transcription, DNA replication, and DNA repair. Previous studies showed that FACT is highly expressed in cancer, and cancer cells are more reliant on FACT than normal cells. However, the relationship between Spt16 and lung cancer remains unclear. In this study, we explored the functions of Spt16 in lung cancer cells. The effects of Spt16 on lung cancer cell proliferation, cell cycle progression, apoptosis, migration, and invasion were examined. We found that knockdown of Spt16 led to obvious decreases of both Rb and MCM7, and further activated the DNA damage response (DDR) pathway. In addition, a novel micro-RNA, miR-1227-5p, directly targeted the 3'-UTR of Spt16 and regulated the mRNA levels of Spt16. Furthermore, we found that CBL0137, the functional inhibitor of FACT, showed similar effects as loss of Spt16. Together, our data indicated that Spt16 is likely to be an essential regulator for lung cancer malignancy and is negatively regulated by miR-1227-5p.


Evolution of infection control in Egypt: achievements and challenges.
The high prevalence of hepatitis C virus (HCV) infection in Egypt highlighted the urgent need for implementing infection control (IC) programs in Egypt. The Ministry of Health and Population (MOHP), in collaboration with the US Naval Medical Research Unit No. 3, and the World Health Organization (WHO), developed a national plan to initiate an IC program with the objectives of improving quality of care and reducing transmission of hospital-acquired infections. The strategic plan for this program included setting up an organizational structure, developing IC national guidelines, training health care workers, promoting occupational safety, and establishing a system for monitoring and evaluation. Implementation of the program started in late 2001. The achievements to date include developing a national organizational structure, IC guidelines, and a comprehensive IC training program. To date, a total of 72 hospitals in 13 governorates have been enrolled in the program, and 235 IC professionals have been trained. Many challenges were faced, including administrative, financial, and motivational difficulties. Future plans include expansion of the program to cover all 27 governorates of Egypt and establishment of a surveillance system for hospital-acquired infections. The process of developing the IC program in Egypt may serve as a model for other resource-limited countries that seek to initiate similar programs.


A new 2.5D representation for lymph node detection using random sets of deep convolutional neural network observations.
Automated Lymph Node (LN) detection is an important clinical diagnostic task but very challenging due to the low contrast of surrounding structures in Computed Tomography (CT) and to their varying sizes, poses, shapes and sparsely distributed locations. State-of-the-art studies show the performance range of 52.9% sensitivity at 3.1 false-positives per volume (FP/vol.), or 60.9% at 6.1 FP/vol. for mediastinal LN, by one-shot boosting on 3D HAAR features. In this paper, we first operate a preliminary candidate generation stage, towards -100% sensitivity at the cost of high FP levels (-40 per patient), to harvest volumes of interest (VOI). Our 2.5D approach consequently decomposes any 3D VOI by resampling 2D reformatted orthogonal views N times, via scale, random translations, and rotations with respect to the VOI centroid coordinates. These random views are then used to train a deep Convolutional Neural Network (CNN) classifier. In testing, the CNN is employed to assign LN probabilities for all N random views that can be simply averaged (as a set) to compute the final classification probability per VOI. We validate the approach on two datasets: 90 CT volumes with 388 mediastinal LNs and 86 patients with 595 abdominal LNs. We achieve sensitivities of 70%/83% at 3 FP/vol. and 84%/90% at 6 FP/vol. in mediastinum and abdomen respectively, which drastically improves over the previous state-of-the-art work.


[Effectiveness of pneumococcal polysaccharide vaccine in older patients with chronic respiratory diseases].
To evaluate effectiveness of pneumococcal polysaccharide vaccine, we recommended 1378 outpatients aged over 60 with chronic respiratory diseases to be vaccinated from August to October 2002, and 647 patients were vaccinated from August to November 2002. In the 1229 patients without respiratory failure, the incidence of antimicrobial treatment for bacterial respiratory infections in 547 vaccinated patients significantly decreased from 7.9% in the 2001/02 winter season to 5.7% in the 2002/03 winter season, although that in the 682 unvaccinated patients increased from 3.8% to 5.7%. The incidence of antimicrobial treatment for bacterial respiratory infections in 229 vaccinated patients with pneumococcal and influenza vaccines together significantly decreased from 10.5% in the 2001/02 winter season to 5.2% in 2002/03 winter season although that in 110 subjects vaccinated with influenza vaccine only increased from 2.7% to 7.2%. These findings suggest the effectiveness of the pneumococcal polysaccharide vaccine for the prevention of bacterial respiratory infections and the additive effectiveness of pneumococcal and influenza vaccines together.


The Complex Regulatory Role of Cytomegalovirus Nuclear Egress Protein pUL50 in the Production of Infectious Virus.
The regulation of the nucleocytoplasmic release of herpesviral capsids is defined by the process of nuclear egress. Due to their large size, nuclear capsids are unable to traverse via nuclear pores, so that herpesviruses evolved to develop a vesicular transport pathway mediating their transition through both leaflets of the nuclear membrane. This process involves regulatory proteins, which support the local distortion of the nuclear envelope. For human cytomegalovirus (HCMV), the nuclear egress complex (NEC) is determined by the pUL50-pUL53 core that initiates multicomponent assembly with NEC-associated proteins and capsids. Hereby, pUL50 serves as a multi-interacting determinant that recruits several viral and cellular factors by direct and indirect contacts. Recently, we generated an ORF-UL50-deleted recombinant HCMV in pUL50-complementing cells and obtained first indications of putative additional functions of pUL50. In this study, we produced purified ΔUL50 particles under both complementing (ΔUL50C) and non-complementing (ΔUL50N) conditions and performed a phenotypical characterization. Findings were as follows: (i) ΔUL50N particle preparations exhibited a clear replicative defect in qPCR-based infection kinetics compared to ΔUL50C particles; (ii) immuno-EM analysis of ΔUL50C did not reveal major changes in nuclear distribution of pUL53 and lamin A/C; (iii) mass spectrometry-based quantitative proteomics showed a large concordance of protein contents in the NIEP fractions of ΔUL50C and ΔUL50N particles, but virion fraction was close to the detection limit for ΔUL50N; (iv) confocal imaging of viral marker proteins of immediate early (IE) and later phases of ΔUL50N infection indicated a very low number of cells showing an onset of viral lytic protein expression; and, finally (v) quantitative measurements of encapsidated genomes provided evidence for a substantial reduction in the DNA contents in ΔUL50N compared to ΔUL50C particles. In summary, the results point to a complex and important regulatory role of the HCMV nuclear egress protein pUL50 in the maturation of infectious virus.


Different growth hormone (GH) response to GH-releasing peptide and GH-releasing hormone in hyperthyroidism.
Altered GH responses to several pharmacological stimuli, including GHRH, have been found in hyperthyroidism. The mechanisms underlying these disturbances have not been fully elucidated. GH-releasing peptide-6 (GHRP-6) is a synthetic hexapeptide that specifically stimulates GH release both in vitro and in vivo. The mechanism of action of GHRP-6 is unknown, but it probably acts by inhibiting the effects of somatostatin on GH release. The aim of this study was to evaluate the effects of GHRP-6 on GH secretion in patients with hyperthyroidism (n = 9) and in control subjects (n = 9). Each subject received GHRP-6 (1 microg/kg, iv), GHRH (100 microg, iv), and GHRP-6 plus GHRH on 3 separate days. GH peak values (mean +/- SE; micrograms per L) were significantly lower in hyperthyroid patients compared to those in control subjects after GHRH alone (9.0 +/- 1.3 vs. 27.0 +/- 5.2) and GHRP-6 plus GHRH (22.5 +/- 3.5 vs. 83.7 +/- 15.2); a lack of the normal synergistic effect of the association of both peptides was observed in thyrotoxicosis. However, a similar GH response was seen in both groups after isolated GHRP-6 injection (31.9 +/- 5.7 vs. 23.2 +/- 3.9). In summary, we have shown that hyperthyroid patients have a normal GH response to GHRP-6 together with a blunted GH responsiveness to GHRH. Our data suggest that thyroid hormones modulate GH release induced by these two peptides in a differential way.


An interferon-independent lncRNA promotes viral replication by modulating cellular metabolism.
Viruses regulate host metabolic networks to improve their survival. The molecules that are responsive to viral infection and regulate such metabolic changes are hardly known, but are essential for understanding viral infection. Here we identify a long noncoding RNA (lncRNA) that is induced by multiple viruses, but not by type I interferon (IFN-I), and facilitates viral replication in mouse and human cells. In vivo deficiency of lncRNA-ACOD1 (a lncRNA identified by its nearest coding gene Acod1, aconitate decarboxylase 1) significantly attenuates viral infection through IFN-I-IRF3 (interferon regulatory factor 3)-independent pathways. Cytoplasmic lncRNA-ACOD1 directly binds the metabolic enzyme glutamic-oxaloacetic transaminase (GOT2) near the substrate niche, enhancing its catalytic activity. Recombinant GOT2 protein and its metabolites could rescue viral replication upon lncRNA-ACOD1 deficiency and increase lethality. This work reveals a feedback mechanism of virus-induced lncRNA-mediated metabolic promotion of viral infection and a potential target for developing broad-acting antiviral therapeutics.


Robust self-assembled monolayers of Ru(II) and Os(II) polypyridines on gold surfaces: exploring new potentials.
Metal complexes [M(phtpy)(pztpy)](PF(6))(2) (phtpy = 4'-phenyl-2,2':6',2''-terpyridine, pztpy = 4'-(N-piperazinyl)-2,2':6',2''-terpyridine, M = Ru, Os) were prepared and examined spectroscopically and electrochemically. The piperazine attachment was found to significantly modify the photophysical and electrochemical properties compared to the parent bis-terpyridine complexes, causing a red-shift of the (1)MLCT (23 nm, 53.9 eV) and a substantial cathodic shift of the redox potential (0.3 V for Ru, 0.23 V for Os). Self-assembled monolayers (SAMs) of the complexes on polished gold electrodes were generated simply by the in situ formation of a dithiocarbamate (DTC) anchoring group at the terminal piperazinyl nitrogen on the pztpy ligand. Cyclic voltammetry revealed that the monolayers show excellent reversible behaviour and exceptional stability. The high stability of the SAMs is attributed to the strong bidentate attachment to the gold surface of the DTC tether and the favourable low oxidation potentials of the complexes which result from the electron-rich piperazine nitrogen on the pztpy ligand. Such DTC-based SAMs demonstrate a substantial improvement over commonly-employed thiol-based systems, and offer new scope for future development.


Antenna-Biased Odorant Receptor HvarOR25 in Hippodamia variegata Tuned to Allelochemicals from Hosts and Habitat Involved in Perceiving Preys.
Odorant receptors (ORs) of ladybird Hippodamia variegata play vital chemosensory roles in searching and locating preys. In the current study, 37 ORs were initially identified from the antennal transcriptome of H. variegata. The quantitative polymerase chain reaction demonstrated that several HvarORs including HvarOR25 were specific or enriched in ladybird antennae. In two-electrode voltage clamp recordings, recombinant HvarOR25 was narrowly tuned to six chemical ligands including aphid-induced, aphid-derived, and plant-derived volatiles. In electroantennogram assays, all six volatiles elicited electrophysiological responses. Among the six volatiles, cis-3-hexenyl acetate, hexyl butyrate, hexyl hexanoate, and 3-methyl-3-buten-1-ol were attractive for both sexes of H. variegata. Additionally, molecular docking indicated that HvarOR25 was bound to all ligands with high binding affinities. Taken together, HvarOR25 facilitates perception of preys by recognizing relevant allelochemicals from hosts and habitat. Our findings provide valuable insights into understanding biological functions of HvarORs and help to develop a novel biocontrol strategy based on olfactory-active compounds.


[Levels of alpha-2-macroglobulin in blood serum of women giving birth to hypotrophic and eutrophic newborns].
We tested the concentration of alpha-2-macroglobulin in sera of 33 women bearing eutrophic newborns, 36 women bearing hypotrophic newborns and 30 healthy not pregnant women in reproductive age. The concentration of this inhibitor was measured using radial immunodiffusion method according to Mancini et al. We found distinct decrease of alpha-2-macroglobulin concentration in sera of bearing women. In women bearing eutrophic newborns we found 154 mg/dl, in women bearing hypo-trophic newborns 171 mg/dl whereas in controls 250 mg/dl. We have noted statistically significant differences between tested groups to controls and between investigated groups. Taking under consideration the role of alpha-2-macroglobulin as the modulator of immune system as well as the activity of several cytokins, therefore one can suppose that alpha-2-M may affect on cellular growth developed foetus in intrauterine.


The Medicinal Chemistry and Therapeutic Potential of LRH-1 Modulators.
The ligand-activated transcription factor liver receptor homologue 1 (LRH-1, NR5A2) is involved in the regulation of metabolic homeostasis, including cholesterol and glucose balance. Preliminary evidence points to therapeutic potential of LRH-1 modulation in diabetes, hepatic diseases, inflammatory bowel diseases, atherosclerosis, and certain cancers, but because of a lack of suitable ligands, pharmacological control of LRH-1 has been insufficiently studied. Despite the availability of considerable structural knowledge on LRH-1, only a few ligand chemotypes have been developed, and potent, selective, and bioavailable tools to explore LRH-1 modulation in vivo are lacking. In view of the therapeutic potential of LRH-1 in prevalent diseases, improved chemical tools are needed to probe the beneficial and adverse effects of pharmacological LRH-1 modulation in sophisticated preclinical models and to further elucidate the receptor's molecular function.


Topiramate for the treatment of alcohol dependence: comparison with naltrexone.
A 6-month naturalistic, randomized and open-label, trial of topiramate versus naltrexone was conducted, with assessments at enrollment and after 3 and 6 months of treatment. 182 alcohol-dependent patients who had been drinking heavily during the past month were included. Outcome was measured using tools that assessed alcohol intake, cravings, disability, and quality of life; changes in biomarkers of alcohol intake were also used. At the 6-month evaluation, patients taking topiramate had significantly lower scores on the OCDS (all subscales), the EuropASI (medical, alcohol, family/social, and psychiatric) and the WHO/DAS (employment/social). More patients taking topiramate remained in the abstinence group and the moderate drinking without problems group. Topiramate at a mean dose of 200 mg/day was better than naltrexone at a mean dose of 50 mg/day at reducing alcohol intake and cravings throughout the study.


Assessing Value in Child Health.
Assessing value in child health is increasingly important as health care systems face difficult choices with regard to what services and programs for children to fund and deliver [...].


Bulk Organic Aerosol Analysis by Proton-Transfer-Reaction Mass Spectrometry: An Improved Methodology for the Determination of Total Organic Mass, O:C and H:C Elemental Ratios, and the Average Molecular Formula.
We have recently shown in this journal (Müller et al. Anal. Chem. 2017 , 89 , 10889 - 10897 ) how a proton-transfer-reaction mass spectrometry (PTR-MS) analyzer measured particulate organic matter in urban atmospheres using the "Chemical Analysis of Aerosol Online" (CHARON) inlet. Our initial CHARON studies did not take into account fragmentation of protonated analyte molecules, which introduced a small but significant negative bias in the determination of bulk organic aerosol parameters. Herein, we studied the ionic fragmentation of 26 oxidized organic compounds typically found in atmospheric particles. This allowed us to derive a correction algorithm for the determination of the bulk organic mass concentration, mOA, the bulk-average hydrogen-to-carbon ratio, (H:C)bulk, the bulk-average oxygen-to-carbon ratio, (O:C)bulk, and the bulk-average molecular formula, MFbulk. The correction algorithm was validated against AMS data using two sets of published data. Finally, we determined MFbulk of particles generated from the reaction of α-pinene and ozone and compared and discussed the results in relation to the literature.


Familial hypercholesterolaemia with severe cardiac involvement in a boy: successful management and mid-term follow-up.
A 13-year-old boy with double heterozygosity for familial hypercholesterolaemia with a 90% left coronary artery main stem stenosis is reported. The patient's cholesterol levels were effectively controlled with weekly sessions of selective low-density lipoprotein cholesterol removal through immunoadsorption by use of an extracorporeal system. Left main coronary artery stenosis was successfully treated with percutaneous transluminal balloon dilation. At 30 months after the intervention and still under treatment with weekly sessions of low-density lipoprotein apheresis the patient is free of cardiac symptoms. He shows normal exercise capacity and normal myocardial perfusion. It is concluded that aggressive management is justified in such patients and may result in a near-normal quality of life.


Treatment of peri-anal fistula in Crohn's disease.
Anal fistulas are a common manifestation of Crohn's disease (CD). The first manifestation of the disease is often in the peri-anal region, which can occur years before a diagnosis, particularly in CD affecting the colon and rectum. The treatment of peri-anal fistulas is difficult and always multidisciplinary. The European guidelines recommend combined surgical and medical treatment with biologic drugs to achieve best results. Several different surgical techniques are currently employed. However, at the moment, none of these techniques appear superior to the others in terms of healing rate. Surgery is always indicated to treat symptomatic, simple, low intersphincteric fistulas refractory to medical therapy and those causing disabling symptoms. Utmost attention should be paid to correcting the balance between eradication of the fistula and the preservation of fecal continence.


Molecular phylogenies of plants and Frankia support multiple origins of actinorhizal symbioses.
Molecular phylogenetic trees were reconstructed from nucleotide sequences of nifH and 16S rDNA for Frankia and of rbcL for actinorhizal plants. Comparison of Frankia phylogenetic trees reconstructed using nifH and 16S rDNA sequences indicated that subgroupings of both trees correspond with each other in terms of plant origins of Frankia strains. The results suggested that 16S rDNAs can be utilized for coevolution analysis of actinorhizal symbioses. Frankia and plant phylogenetic trees reconstructed using 16S rDNA and rbcL sequences were compared. The comparison by tree matching and likelihood ratio tests indicated that although branching orders of both trees do not strictly correspond with each other, subgroupings of Frankia and their host plants correspond with each other in terms of symbiotic partnership. Estimated divergence times among Frankia and plant clades indicated that Frankia clades diverged more recently than plant clades. Taken together, actinorhizal symbioses originated more than three times after the four plant clades diverged.


Identifying the lowest electronic states of the chlorophylls in the CP47 core antenna protein of photosystem II.
CP47 is a pigment-protein complex in the core of photosystem II that tranfers excitation energy to the reaction center. Here we report on a spectroscopic investigation of the isolated CP47 complex. By deconvoluting the 77 K absorption and linear dichroism, red-most states at 683 and 690 nm have been identified with oscillator strengths corresponding to approximately 3 and approximately 1 chlorophyll, respectively. Both states contribute to the 4 K emission, and the Stark spectrum shows that they have a large value for the difference polarizability between their ground and excited states. From site-selective polarized triplet-minus-singlet spectra, an excitonic origin for the 683 nm state was found. The red shift of the 690 nm state is most probably due to strong hydrogen bonding to a protein ligand, as follows from the position of the stretch frequency of the chlorophyll 13(1) keto group (1633 cm(-)(1)) in the fluorescence line narrowing spectrum at 4 K upon red-most excitation. We discuss how the 683 and 690 nm states may be linked to specific chlorophylls in the crystal structure [Zouni, A., Witt, H.-T., Kern, J., Fromme, P., Krauss, N., Saenger, W., and Orth, P. (2001) Nature 409, 739-743].


Molecular Mechanism and Potential Targets for Blocking HPV-Induced Lesion Development.
Persistent infection with high-risk HPV is the etiologic agent associated with the development of cervical cancer (CC) development. However, environmental, social, epidemiological, genetic, and host factors may have a joint influence on the risk of disease progression. Cervical lesions caused by HPV infection can be removed naturally by the host immune response and only a small percentage may progress to cancer; thus, the immune response is essential for the control of precursor lesions and CC. We present a review of recent research on the molecular mechanisms that allow HPV-infected cells to evade immune surveillance and potential targets of molecular therapy to inhibit tumor immune escape.


Phenotypic and genotypic characterization of Vibrio cholerae clinically isolated in Surabaya, Indonesia.
The phenotypic and genotypic characteristics of 6 clinical strains of Vibrio cholerae isolated in Surabaya, Indonesia in 2009 were examined. The DNA fingerprints obtained suggested that these isolates were not from a single clone. Furthermore, all isolates produced cholera toxin and possessed the classical type of toxin B subunit gene, thus meaning that this is the first report of the occurrence of El Tor variants of V. cholerae in Indonesia. Although all isolates were sensitive to almost all antibiotics tested, including ampicillin, chloramphenicol, ciprofloxacin, gentamicin, levofloxacin, kanamycin, nalidixic acid, norfloxacin, streptomycin, trimethoprim-sulfamethoxazole, and tetracycline, and had no mutation in the gyrA and parC genes, they nevertheless possessed the class 1 integron that is a molecular vehicle for the acquisition of antibiotic resistance genes, suggesting that they have the potential to acquire the genetic element for drug resistance.


Frameworks for quality improvement in pediatric intensive care: A concise review.
Quality improvement programs focused on improving care in intensive care units have become standard at pediatric hospitals around the world over the past several decades. However, the methodology or framework by which these programs assess quality is not standard. This review describes the varying quality improvement frameworks that have been promoted by prominent pediatric and critical care societies and the strengths and limitations of these frameworks, as well as several notable international collaboratives in this domain.


Discordant hepatic uptake of Tc-99m HIDA and Tc-99m colloid in a patient with segmental biliary obstruction.
Discordant hepatic uptake between Tc-99m HIDA and Tc-99m colloid occurred in a 63-year-old female with segmental biliary obstruction due to cholangioma. Radiographic CT and a percutaneous transhepatic cholangiogram revealed the obstructed right hepatic duct as well as the dilated intrahepatic duct in the right lobe. At surgery this was confirmed, and a 2 cm mass encasing the right hepatic duct was identified. It should be included in the gamut of discordant hepatic uptake of Tc-99m IDA and Tc-99m colloid.


Fatty acid profile, oxidative status, and content of volatile organic compounds in raw and cooked meat of different chicken strains.
Chicken meat is rich in unsaturated fatty acids. Therefore, it is more susceptible to lipid oxidation and production of volatile organic compounds (VOC). In this study, we evaluated the fatty acids, antioxidants, and VOC profiles of raw and cooked meat samples derived from 4 strains of chicken differing in their growth rates, which were as follows: slow-growing (SG, Leghorn), medium-growing (MG, Hubbard and Naked Neck), and fast-growing (FG, Ross). The VOC profile of meat was measured using proton-transfer reaction-mass spectrometry (PTR-MS). The VOC were identified using PTR-time of flight-MS (PTR-ToF-MS). The data were analyzed using both univariate and multivariate models. Twenty main VOC were identified, which were classified into the following chemical categories: aldehydes, alkadienes, alkenes, furans, amides, alcohols, and other compounds. Our results revealed that the chicken genotype and the method of cooking strongly influenced the VOC profile of the meat. Identifying the relationships between these traits allowed us to highlight the trade-off of the main substrates such as n-3 and n-6 polyunsaturated fatty acids (PUFA), protective substances (antioxidants), and degradation products (VOC) of the poultry meat produced during cooking. The extent of VOC production and n-3 loss was found to be higher for the SG genotype. Reduction of n-6 was higher in MG, whereas small losses in antioxidants and PUFA were observed in the FG genotype, consequently, resulting in the lowest production of VOC. The SG and MG are genotypes more active from a kinetic point of view respect to the FG ones. For this reason, in the FG genotypes, the antioxidants are less involved in the oxidative stress induced by the movement; thus, they were available to protect the lipid of the meat during the cooking process. These results suggested that the use of SG and MG genotypes requires a specific dietary protocol (i.e., increasing the antioxidants content) to counteract the lipid oxidations in all the phases: in vivo, postmortem, and during/after cooking.


Moisturisers for the treatment of foot xerosis: a systematic review.
Xerosis, literally dryness of the skin, of the foot is a common condition encountered clinically, which can lead to discomfort and predisposition to infection. Currently, there are no evidence-based recommendations on which moisturiser formulations best alleviate xerotic symptoms. The aim of this review was to guide clinical practice in the treatment of primary and diabetes related foot xerosis, by identifying from the existing literature the most effective ingredient or formulation of topical treatments for symptoms of primary foot xerosis in the general population. A systematic review of published experimental trials was undertaken. Only studies pertaining to primary xerosis, classified within levels II - IV of the NHRMC hierarchy were reviewed. EMBASE, AMED, Cochrane, MEDLINE, CINAHL, Ageline and SCOPUS were searched using relevant search terms and keywords and pearling of reference lists was undertaken. Studies were evaluated for methodological quality using a critical appraisal tool. Individual active ingredients were identified from all studies, along with observed reported outcomes. A narrative synthesis was then conducted. A total of 22 experimental studies were included, from which 12 different active ingredients were identified. Study literature consisted of mainly comparative studies against other active interventions or controls, or pre-post-tests and was of a poor-to-moderate methodological quality as assessed by the Epidemiological Appraisal Instrument. Urea was the most researched active ingredient (14 studies), with ammonium lactate being next (7 studies). No conclusive recommendations were possible due to wide variation in study quality, methodologies and outcome measures. A synthesis of available literature suggests that treatments containing urea as a primary active ingredient have been the most researched. The poor quality of literature generally, however, precludes recommendation of any active ingredient over another.


Transgenerational inheritance of fetal alcohol effects on proopiomelanocortin gene expression and methylation, cortisol response to stress, and anxiety-like behaviors in offspring for three generations in rats: Evidence for male germline transmission.
Previously it has been shown that fetal alcohol exposure increases the stress response partly due to lowering stress regulatory proopiomelanocortin (Pomc) gene expression in the hypothalamus via epigenetic mechanisms for multiple generations in mixed-breed rats. In this study we assess the induction of heritable epigenetic changes of Pomc-related variants by fetal alcohol exposure in isogenic Fischer 344 rats. Using transgenerational breeding models and fetal alcohol exposure procedures, we determined changes in hypothalamic Pomc gene expression and its methylation levels, plasma corticosterone hormone response to restraint stress, and anxiety-like behaviors using elevated plus maze tests in fetal alcohol-exposed offspring for multiple generations in isogenic Fischer rats. Fetal alcohol-exposed male and female rat offspring showed significant deficits in POMC neuronal functions with increased Pomc gene methylation and reduced expression. These changes in POMC neuronal functions were associated with increased plasma corticosterone response to restraint stress and increased anxiety-like behavior. These effects of fetal alcohol exposure persisted in the F1, F2, and F3 progeny of the male germline but not of the female germline. These data suggest that fetal alcohol exposure induces heritable changes in Pomc-related variants involving stress hyperresponsiveness and anxiety-like behaviors which perpetuate into subsequent generations through the male germline via epigenetic modifications.


Protective effect of Urtica dioica methanol extract against experimentally induced urinary calculi in rats.
Renal calculi formation is one of the most common urological disorders. Urinary stone disease is a common disease, which affects 10‑12% of the population in industrialized countries. In males, the highest prevalence of the disease occurs between the age of 20 and 40 years, while in females, the highest incidence of the disease occurs later. Previous studies have shown that long‑term exposure to oxalate is toxic to renal epithelial cells and results in oxidative stress. In the present study, a methanolic extract of aerial parts of Urtica dioica was screened for antiurolithiatic activity against ethylene glycol and ammonium chloride‑induced calcium oxalate renal stones in male rats. In the control rats, ethylene glycol and ammonium chloride administration was observed to cause an increase in urinary calcium, oxalate and creatinine levels, as well as an increase in renal calcium and oxalate deposition. Histopathological observations revealed calcium oxalate microcrystal deposits in the kidney sections of the rats treated with ethylene glycol and ammonium chloride, indicating the induction of lithiasis. In the test rats, treatment with the methanolic extract of Urtica dioica was found to decrease the elevated levels of urinary calcium, oxalate and creatinine, and significantly decrease the renal deposition of calcium and oxalate. Furthermore, renal histological observations revealed a significant reduction in calcium oxalate crystal deposition in the test rats. Phytochemical analysis of the Urtica dioica extract was also performed using liquid chromatography‑electrospray ionization tandem mass spectrometry and high-performance liquid chromatography with photodiode array detection, to determine the chemical composition of the extract. The eight chemical constituents identified in the extract were protocatechuic acid, salicylic acid, luteolin, gossypetin, rutin, kaempferol‑3‑O‑rutinoside, kaempferol‑3‑O‑glucoside and chlorogenic acid. In conclusion, the results of the present study suggest that Urtica dioica has strong antiurolithiatic activity and may have potential as a natural therapeutic agent for various urological disorders.


[Research progress of pharmaceutical and personal care products on ecological and human health].
EU (European Union) and Center of Water Pollution and Control in the United States have continued to measure persistent bioaccumulate and toxic (PB&T) chemicals and persistent organic pollutants (POPs) since the 1970s. In resent years, Pharmaceutical and personal care products (PPCPs) have raised more concern among chemists and toxicologists. The present paper introduced the main properties of PPCPs and their present situation of research. Additionally, the potential toxicites of PPCPs to ecological and human health were detailedly reviewed in this article.


Hepatopulmonary fusion: a rare and potentially lethal association with right congenital diaphragmatic hernia.
Hepatopulmonary fusion is an extremely rare accompaniment of right congenital diaphragmatic hernia. It is associated with abnormal systemic arterial supply and venous drainage of the right lung along with congenital heart disease. Children with this condition have a comparatively poor prognosis. We report a case of right congenital diaphragmatic hernia with hepatopulmonary fusion along with review of the literature with stress on diagnosis and management.


Detection of clinically-relevant EGFR variations in de novo small cell lung carcinoma by droplet digital PCR.
Targeted therapy that utilizes tyrosine kinase inhibitors (TKIs), specific to epidermal growth factor receptors (EGFR) has changed the landscape of treatment of non-small cell lung cancer (NSCLC). The success or failure of this approach depends on presence of certain variations in the tyrosine kinase domain of EGFR gene. Generally, patients diagnosed with Small cell lung cancer (SCLC) are considered ineligible for TKI therapy owing to the absence of EGFR variations. . However, there is evidence of these variations being detected in SCLCs, both in de-novo and in transformed SCLCs (TKI-treated adenocarcinomas). Despite the presence of clinically-relevant EGFR variations in SCLCs, the response to TKIs has been inconsistent. Liquid biopsy is a well-established approach in lung cancer management with proven diagnostic, prognostic and predictive applications. It relies on detection of circulating tumor-derived nucleic acids present in plasma of the patient. In this study, a liquid biopsy approach was utilized to screen 118 consecutive lung cancer patients for four clinically-relevant variations in EGFR gene, which included three activating/sensitizing variations (Ex18 G719S, Ex19del E746-A750 and Ex21 L858R) and one acquired/resistance (Ex20 T790M, de novo) variation by droplet digital PCR, the most advanced third generation PCR technique. As expected, clinically-relevant EGFR variations were found in majority of the non-small cell lung cancer cases. However, among the handful of small cell lung cancer samples screened, sensitizing variations (Ex18 G719S and Ex21 L858R) were seen in almost all of them. Interestingly, Ex20 T790M variation was not detected in any of the cases screened. The results of our study indicate that EGFR variations are present in SCLCs and highly sensitive liquid biopsy techniques like ddPCR can be effectively utilized for this purpose of screening EGFR variations in such samples.


Echinococcus multilocularis metacestode metabolites contain a cysteine protease that digests eotaxin, a CC pro-inflammatory chemokine.
In many helminthic infections, eotaxin, a CC-chemokine, triggers the mobilization of eosinophils, thus, contributing to an elevated blood and periparasitic eosinophil level. Following an experimental intraperitoneal infection of C57BL6 mice with Echinococcus multilocularis metacestodes, however, we observed the absence of eosinophils in the peritoneal cavity and a low number of such cells in the blood of infected animals. Therefore, we carried out an explorative study to address the question why eosinophilia did not occur especially in the peritoneal cavity of such secondarily AE-infected mice. In an in vitro assay, we showed that metacestode antigens (in vitro generated vesicle fluid and E/S products) were able to proteolytically digest eotaxin. This effect was confirmed with semiquantitative Western blotting, which demonstrated a decreasing intensity of remaining eotaxin signals. Proteolysis of eotaxin was, thus, dose-dependent and proportional to the time of incubation with the metacestode antigens. Using appropriate inhibitors, the respective protease was identified as a cysteine protease, which required the presence of Ca(++) as co-enzyme. A chromatographic fractionation procedure by successive separation of VF molecules using a superpose column and subsequently a MonoQ column mounted on an FPLC system allowed to yield a fraction, referred to us as fraction 6; containing the enriched cysteine protease, this fraction will be used for further molecular studies. Eotaxin inactivation by VF and E/S products may contribute to explain the absence of eosinophils within the peritoneal cavity of AE-secondary infected mice. Absent eosinophils, thus, may be a part of a series of events that maintain a low level of inflammation displayed within the peritoneal cavity of experimentally infected mice.


Audit of ranitidine administration in parenteral nutrient solutions.
An audit of the use of ranitidine administered in parenteral nutrient (PN) solutions is reported. Pharmacists at a 747-bed teaching institution developed criteria addressing justification of the use of ranitidine in PN solutions, process indicators for monitoring, and outcome measures. All patients who received ranitidine in their PN solutions between October 29 and November 22, 1989, were included in the study. Data were collected on a standardized form. A total of 23 evaluable patients received ranitidine in PN solutions during the study period. No inappropriate uses of ranitidine were identified. Patients with renal impairment tended to be underdosed. There were no duplicate administrations of ranitidine by the i.v. minibag and PN solution routes. However, on four occasions the PN solution was interrupted for more than six hours; additional ranitidine by i.v. minibag was ordered for only one of these patients. Testing of nasogastric aspirates for pH was not performed in 17 of the 22 patients who needed this test. Gastrointestinal bleeding was observed in two patients, both of whom were not monitored for gastric pH and were underdosed. A program is being developed to promote the safe and effective use of ranitidine administered in PN solutions. An audit of the administration of ranitidine in PN solutions showed that the drug is usually used appropriately at the institution and that most of the patients had positive clinical outcomes.


Genetic analysis of KillerRed in C. elegans identifies a shared role of calcium genes in ROS-mediated neurodegeneration.
In C. elegans, neurodegeneration induced by excitotoxicity or aggregation of misfolded proteins is dependent on genes involved in calcium release from the endoplasmic reticulum. Reactive oxygen species (ROS) can also induce neurodegeneration, but the relationship between ROS-mediated neurodegeneration and calcium has not been established. We activated KillerRed in the GABA neurons of C. elegans to produce ROS that leads to functional loss and structural degeneration of these neurons and demonstrated that the severity of neurodegeneration was dependent on extent of KillerRed activation. To genetically examine the role of calcium in ROS-mediated neurodegeneration, we measured functional neurodegeneration in itr-1 (inositol trisphosphate receptor), crt-1 (caltreticulin), and unc-68 (ryanodine receptor) mutants. Similar to other neurotoxic conditions, neurodegeneration triggered by KillerRed was reduced in itr-1 and crt-1 mutants. Somewhat unexpectedly, genetic or pharmacological disruption of unc-68 had a minimal effect on neurodegeneration. Our results indicate ROS-mediated neurodegeneration occurs through a conserved calcium regulated mechanism and suggest that components of the degeneration process have different sensitivities to ROS.


Fabrication of Homogeneous High-Density Antibody Microarrays for Cytokine Detection.
Cytokine proteins are known as biomarker molecules, characteristic of a disease or specific body condition. Monitoring of the cytokine pattern in body fluids can contribute to the diagnosis of diseases. Here we report on the development of an array comprised of different anti-cytokine antibodies on an activated solid support coupled with a fluorescence readout mechanism. Optimization of the array preparation was done in regard of spot homogeneity and spot size. The proinflammatory cytokines Tumor Necrosis Factor alpha (TNFα) and Interleukin 6 (IL-6) were chosen as the first targets of interest. First, the solid support for covalent antibody immobilization and an adequate fluorescent label were selected. Three differently functionalized glass substrates for spotting were compared: amine and epoxy, both having a two-dimensional structure, and the NHS functionalized hydrogel (NHS-3D). The NHS-hydrogel functionalization of the substrate was best suited to antibody immobilization. Then, the optimization of plotting parameters and geometry as well as buffer media were investigated, considering the ambient analyte theory of Roger Ekins. As a first step towards real sample studies, a proof of principle of cytokine detection has been established.


Man vs. man-made marijuana: A case of drug-induced posterior reversible encephalopathy syndrome (PRES) due to K2, a ynthetic cannabinoid (SCB).
Synthetic Cannabinoids (SCB) are engineered chemical compounds that share a similar chemical structure with the active ingredient of marijuana, delta-9-tetrahydrocanabinol. Although the FDA has not approved the use of SCB without a prescription from a licensed health-care provider, the cost effectiveness and availability of SCB has made it a popular choice among recreational drug users. Manufacture of SCB as a street drug is not regulated. These SCB are highly potent chemicals that cause various severe toxicities. In this case report, we describe an adult who suffered from PRES after consuming K2, a synthetic cannabinoid.


Chicken gonadotropin-releasing hormones enhance soluble and insoluble fibronectin production by granulosa cells of the domestic fowl in vitro.
Experiments were conducted in vitro to examine the effect of two chicken gonadotropin-releasing hormones, cGnRH-I ([Gln8]-GnRH) and cGnRH-II ([His5,Trp7,Tyr8]-GnRH), on fibronectin (soluble and insoluble) production by chicken granulosa cells isolated from the largest (F1; about 35 mm in diameter), and third largest (F3; 15 to 20 mm in diameter) preovulatory follicles as well as from a pool of immature small yellow follicles (SYF; 6 to 8 mm in diameter). The amounts of soluble fibronectin (fibronectin secreted into the incubation medium) and insoluble fibronectin (fibronectin associated with cells plus fibronectin attached to culture substratum) were quantified with a specific ELISA. Fibronectin secreted into the incubation medium (soluble fibronectin) by unstimulated cells increased with advanced stages of follicular maturation. Addition of both cGnRH-I and -II increased the amount of fibronectin secreted into the incubation medium by all follicular cell types. The amount of insoluble fibronectin in culture wells that contained unstimulated cells also increased with advanced stages of follicle development. Both cGnRH-I and -II increased the quantity of insoluble fibronectin by granulosa cells from all follicle types. Total (soluble plus insoluble) fibronectin production was elevated when cGnRH-I or -II was added to F1, F3, and SYF granulosa cells. The magnitude of cGnRH-I or -II stimulation (percentage increase) of soluble, insoluble, or total fibronectin production was calculated as a multiple of the unstimulated (control) value for each follicle type, and they were greatest in cells derived from developing and immature follicles. These results indicate that homologous cGnRH-I and -II are capable of directly modulating the physiology of the avian ovary.


Risperidone Ameliorates Prefrontal Cortex Neural Atrophy and Oxidative/Nitrosative Stress in Brain and Peripheral Blood of Rats with Neonatal Ventral Hippocampus Lesion.
Reduction of the dendritic arbor length and the lack of dendritic spines in the pyramidal cells of the prefrontal cortex (PFC) are prevalent pathological features in schizophrenia (SZ). Neonatal ventral hippocampus lesion (NVHL) in male rats reproduces these neuronal characteristics and here we describe how this is a consequence of BDNF/TrkB pathway disruption. Moreover, COX-2 proinflammatory state, as well as Nrf-2 antioxidant impairment, triggers oxidative/nitrosative stress, which also contributes to dendritic spine impairments in the PFC. Interestingly, oxidative/nitrosative stress was also detected in the periphery of NVHL animals. Furthermore, risperidone treatment had a neurotrophic effect on the PFC and antioxidant effects on the brain and periphery of NVHL animals; these cellular effects were related to behavioral improvement. Our data highlight the link between brain development and immune response, as well as several other factors to understand mechanisms related to the pathophysiology of SZ.SIGNIFICANCE STATEMENT Prefrontal cortex dysfunction in schizophrenia can be a consequence of morphological abnormalities and oxidative/nitrosative stress, among others. Here, we detailed how impaired plasticity-related pathways and oxidative/nitrosative stress are part of the dendritic spine pathology and their modulation by atypical antipsychotic risperidone treatment in rats with neonatal ventral hippocampus lesion. Moreover, we found that animals with neonatal ventral hippocampus lesion had oxidative/nitrosative stress in the brain as well as in the peripheral blood, an important issue for the translational approaches of this model. Then, risperidone restored plasticity and reduced oxidative/nitrosative stress of prefrontal cortex pyramidal cells, and ultimately improved the behavior of lesioned animals. Moreover, risperidone had differential effects than the brain on peripheral blood oxidative/nitrosative stress.


Patterns of sexual risk-taking among heterosexual men.
Heterosexual male STD (sexually transmitted disease) clinic patients (N=146) were classified by cluster analysis into four groups based on their self-reported frequency of participation in eleven sexual behaviors that varied in risk for contracting an STD or HIV. Significant differences among the four groups were found in their participation in some sexual practices that were not used in the clustering, and men in the four groups differed significantly by ethnicity, education, type of relationship, and motivations for engaging in sex. Results of this study have implications for the design of effective AIDS prevention programs to encourage safer-sex among heterosexual men. Given the differences in men's sexual practices, safer-sex messages need to be tailored to reach subgroups of STD patients with similar risk-taking patterns.


Measurement of formaldehyde as dimedone adduct and potential formaldehyde precursors in hard tissues of human teeth by overpressured layer chromatography.
Simple, automatic overpressured layer chromatographic methods for the parallel determination of endogenous formaldehyde in the form of dimedone adduct and potential formaldehyde generators (betaines) in hard tissues of human teeth are described together with an efficient densitometric evaluation. These simple procedures involve the special preparation of teeth and extraction of formaldehyde, of different binding force in teeth, with methanol containing dimedone, and the isolation of betaines with an aqueous solution of methanol.


RGD-expressed bacterial membrane-derived nanovesicles enhance cancer therapy via multiple tumorous targeting.
Background: A tumor microenvironment is a complicated multicellular system comprised of tumor cells, immune cells and blood vessels. Blood vessels are the barriers for drug tissue penetration. Effectively treating a cancer requires drug delivery systems to overcome biological barriers present in tumor microenvironments (TMEs). Methods: We designed a drug delivery system made of bacterial (Escherichia coli) double layer membrane-derived nanovesicles (DMVs) with the expression of RGD peptides and endogenous targeting ligands of bacteria. The physical and biological characteristics of DMVs were assessed by cryogenic transmission electron microscopy, western blotting, flow cytometry and confocal microscopy. Doxorubicin (DOX) was loaded in DMVs via a pH gradient driven drug loading method. Therapeutical effects of DOX-loaded DMVs were studied in a melanoma xenograft mouse model. Results:In vitro and in vivo experiments showed that DMVs can target neutrophils and monocytes that mediated the transport of DMVs across blood vessel barriers and they can also directly target tumor vasculature and tumor cells, resulting in enhanced delivery of therapeutics to TMEs. Furthermore, we developed a remote drug loading approach to efficiently encapsulate DOX inside DMVs, and the drug loading was 12% (w/w). In the B16-F10 melanoma mouse model, we showed that DOX-RGD-DMVs significantly inhibited the tumor growth compared to several controls. Conclusion: Our studies reveal that DMVs are a powerful tool to simultaneously target multiple cells in TMEs, thus increasing drug delivery for improved cancer therapies.


Knowledge, attitude and practice regarding diabetes and hypertension among school students of Nepal: A rural vs. urban study.
The burden of non-communicable diseases like diabetes and hypertension is increasing worldwide including low-and middle-income countries. Good knowledge of such diseases among young people will make them adopt a healthy lifestyle from an early age, which will, in turn, prevent them from developing such non-communicable diseases. This study aimed to assess the knowledge, attitude, and practice of rural and urban school students regarding diabetes and hypertension. We also aimed to see the differences in the knowledge, attitude, and practice of students from rural vs. urban communities. A school-based cross-sectional study was conducted from May 1 2021 to June 30, 2021, in four schools in Nepal (1 from a metropolitan city, 2 from an urban municipality, and 1 from a rural municipality). The study was conducted among the secondary-level students of classes 9 and 10 in each school. The data were collected from the participants via pre-tested questionnaires and analyzed in the Statistical Packages for Social Sciences (SPSS) version 20.0. Logistic regression analysis was carried out to determine the determinants of knowledge and attitude regarding diabetes and hypertension. Of 380 respondents, 35.5% were residents of metropolitan city, 37.4% were from the urban municipality and 27.1% were from the rural municipality. The mean age of respondents was 15.61±0.99 years and 51.1% were male. Respondents having a family history of diabetes and hypertension were 21.1% and 37.9% respectively. Respondents from the metropolitan city had significantly higher mean knowledge scores than the respondents from the urban and rural municipality (p<0.001) while there was no significant difference in mean attitude scores. There was significantly higher daily consumption of fruits and vegetables among the participants from rural municipality (p<0.01) while no significant difference was seen in salt consumption and time spent on physical activity. In univariate regression analysis, place of residence, family occupation, parental education, and family history of diabetes and hypertension were significantly associated with good knowledge level. In multivariate analysis, only a higher grade of study (grade 10 in comparison to grade 9) was an independent predictor of a student's good attitude level. In general, there was a good attitude towards diabetes and hypertension despite poor knowledge. The mean knowledge scores were lower in urban municipality and rural municipality compared to metropolitan city. Low knowledge scores on diabetes and hypertension among the students show an urgent need for school-based interventional programs focusing on non-communicable diseases and lifestyle modification with more emphasis on rural communities.


Evaluation of Th-1 and Th-2 immune responses in the skin lesions of patients with Blau syndrome.
Blau syndrome is an autosomal dominant syndrome characterized by arthritis, uveitis, skin rash, granuloma, and camptodactyly. It has overlapping symptoms with sarcoidosis and rheumatoid arthritis. Our study was directed toward determining the role of cytokines in granuloma formation in Blau syndrome. Antigenic stimulation usually follows two pathways: Th-1, which activates macrophages and cytotoxic T-lymphocytes and produces interleukin (IL)-2, IL-3, interferon gamma, and tumor necrosis factor alpha, and Th-2, which activates the humoral immune system and produces IL-4, IL-5, and IL-10. The development of cytokine profiles may shed some light on our understanding of this illness. Therefore, we studied the relative roles of two opposing lymphocytes, Th-1 and Th-2, by analyzing their relative expression in the skin lesions of patients with Blau syndrome, using the in situ reverse transcription-polymerase chain reaction technique. Our data revealed a significant upregulation of IL-2, an event that appears to play an important role in the formation of granuloma and in the pathogenesis of Blau syndrome. Expression of IL-10, however, was downregulated, and this may have an inhibitory role in the development of the disease. Further studies would be necessary to confirm the presence of other cytokines and to establish the regulatory roles of Th-1 and Th-2 lymphocytes in the pathogenesis of Blau syndrome.


Airway responsiveness in infants: comparison of inhaled and nasally instilled methacholine.
Airway responsiveness of infants is evaluated during sleep and the infants inhale the bronchial challenge agent via the nasal airway. Since stimulation of the nasal airway may produce bronchoconstriction, it is unclear whether the observed response in the infants results from deposition of the aerosol in the lower airways or from stimulation of nasal receptors. Therefore, in 6 healthy infants we compared the changes in partial expiratory flow-volume (PEFV) curves produced by aerosol inhalation of methacholine and the changes produced by instillation of equivalent doses of methacholine liquid into the nares. Following aerosol, the peak expiratory flow and the flow at functional residual capacity decreased, PEFV curves became concave in shape, and the oxygen saturation (SaO2) decreased. The highest methacholine concentration inhaled by any infant was 1.25 mg/mL. In contrast to aerosol delivery, a maximal methacholine concentration of 10.0 mg/mL was instilled into the nares of all 6 infants without any change in maximal flow at functional respiratory capacity (VmaxFRC) or SaO2. There was a significant decrease in peak flow and flattening of the PEFV curves at higher lung volumes; however, the PEFV curve remained convex in shape at the lower lung volumes. The changes in the PEFV curve following nasal instillation of methacholine are consistent with an increase in nasal resistance and no change in the lower airways. We conclude that the bronchoconstriction observed following inhaled methacholine does not result from stimulation of nasal receptors.


Expolygalacturonase from Suspension Cultures of Marchantia polymorpha: Its Presence and Involvement in Pectic Polysaccharide Degradation.
Polygalacturonase was isolated from cell suspension cultures of a thalloid liverwort, Marchantia polymorpha. The enzyme in the ;buffer-soluble' protein fraction was dialyzed at pH 5.2 and further purified 91-fold by a combination of chromatographic techniques including CM-Sephadex, Sephacryl S-200, DEAE-Sephadex, and Sephadex G-200. The purified enzyme had an optimum activity in the pH range at 3.6 to 3.8 and molecular weight of 76,000 daltons, and its activity was not stimulated by cations. The enzyme was identified as an exohydrolase from viscometric data and chromatographic analysis of the reaction products.The polysaccharides extracted from the Marchantia cell walls with 2% (w/v) Na hexametaphosphate solution were separated into two fractions, neutral polysaccharides (fraction P-N) and acidic polysaccharides (fraction P-A) by a DEAE-Sephadex column. The fraction P-N was not susceptible to the purified exopolygalacturonase, whereas fraction P-A was partially degraded. This resulted in hydrolysis of 19.5% of the glycosyl linkages of fraction P-A with the release of galacturonic acids. The specific activity of exopolygalacturonase increased during the growth cycle.


The influence of human neutrophils on N-nitrosodimethylamine (NDMA) synthesis.
N-nitrozodimethyloamine (NDMA) is a carcinogenic compound that can be formed in vivo. NDMA is synthesized from precursors-amines and nitrosating agents. Nitrosating agents are formed through the reaction of oxide, reactive oxygen species and nitric oxide (NO). Human neutrophils (PMN) are an important source of the most reactive oxygen species as well as of the nitric oxide. The increase in oxygen metabolism of PMN can lead to the increase nitrosating agent and nitroso-forms. Inflammatory process is associated with locally decreased pH that may favor nitrosation reaction. In the present study, we estimated the NDMA synthesis by LPS-stimulated PMN in the presence of the iNOS inhibitor--N-nitro-L-arginine methyl ester (L-NAME). In the nitrosation reaction dimethylamine (DMA) was used as substrat. The viability of the cells was measured by cytometric method. NDMA concentrations the culture media was measured by GCMS method. NO production was estimated by Griess's method. Expression of iNOS was determined by western blotting. Results obtained showed that DMA nitrosation is most effective in pH between 3-4.5. Nonstimulated PMN produced lower concentrations of NO than LPS-stimulated cells (1.27 microg/cm3 and 1.57 microg/cm3, respectively). In the culture of nonstimulated PMN supplemented with DMA, there was NDMA (mean--0.99 ng/cm3). In the culture of LPS-stimulated PMN in the presence of DMA, the concentration of NDMA was higher than in the culture of nonstimulated PMN (median--1.45 ng/cm3). In the supernatants of cells incubated without DMA and with DMA, LPS and L-NAME, no NDMA was detected. These results indicate that PMN can be one of sources of nitrosating agents and can play a role in endogenous NDMA synthesis. Stimulation of PMN can lead to the increase of NDMA concentration following the increase of NO production. Different pathological conditions associated with PMN activation as well as the decreased pH may favor endogenous NDMA synthesis.


Behavior of Free Aromatic Amino Acid Pools in Rosmarinic Acid-Producing Cell Cultures of Anchusa officinalis L.
The pool sizes of free l-phenylalanine and l-tyrosine, the precursors of rosmarinic acid in Anchusa officinalis L. cell suspension cultures, fluctuated during the culture cycle. The major increase in pool sizes was preceded by a peak of prephenate aminotransferase activity, while the subsequent decrease coincided with the presence of high activities of phenylalanine ammonia-lyase and tyrosine aminotransferase, the two entrypoint enzymes of the rosmarinic acid biosynthesis pathway. Timecourse feeding studies with linear growth stage cells revealed that the tyrosine pool turned over rapidly, consistent with direct participation in rosmarinic acid synthesis. Since externally applied l-tyrosine was rapidly incorporated into rosmarinic acid with little evidence of radioactively labeled intermediates, it is suggested that there exists a close coupling between the l-tyrosine pool and the rosmarinic acid biosynthetic pathway, which may involve the channelling of intermediates both into and within the pathway.


Order No. 0020 for the establishment of methods of reforestation, 19 March 1987.
In light of the "Green Movement" policy and the development of the Colombia Major National Reforestation Plan, this Order establishes the following methods of promoting reforestation: 1) advice and guidance to those concerned with tree planting; 2) forestry planning for farms; and 3) grants of trees whenever possible in the proportions indicated in the Order. The Order classifies beneficiaries into small-size tree farmers (up to 20 hectares, and 80% of whose income derives from that activity); and medium- and large-scale tree farmers (more than 20 hectares). Other provisions of the Order deal with applications, dates of completion of requirements, and the actual methods of promoting reforestation.


Coatable 2D Conjugated Polymers Containing Bulky Macromolecular Guests for Thermal Imaging.
Dynamic properties are derived from the structural flexibility of 2D polymers. Softening layered structures has the potential for tuning and enhancing the dynamic properties. In the present work, the flexibility of layered polydiacetylene (PDA) is tuned by the interlayer polymeric guests with different branching structures. PDA shows thermoresponsive color-change properties through shortening the effective conjugation length with molecular motion. Whereas the blue-to-red color transition is observed at certain threshold temperatures for the layered PDA without the interlayer guest, the intercalation of the bulky polymer guests lowers the starting temperature and widens the temperature range for the thermoresponsive color changes. The resultant layered composite of PDA and bulky polymer affords the homogeneous coating on substrates on the centimeter scale. The thermoresponsive color-change coating is applied to temperature-distribution imaging. The specific heat of liquids is colorimetrically estimated using the coating on the bottle. The coating on a silk cloth visualizes the temperature distribution on a simulated tissue during surgical operation using an ultrasonic coagulation cutting device. The coating can be applied to thermal imaging in a variety of fields. Moreover, the softening strategy contributes to explore dynamic properties of soft 2D materials.


Electrochemical Sensor for the Detection of 1-Hydroxypyrene Based on Composites of PAMAM-Regulated Chromium-Centered Metal-Organic Framework Nanoparticles and Graphene Oxide.
A nanocomposite was formed by combining graphene oxide (GO) with chromium-centered metal-organic framework (Cr-MOF) nanoparticles regulated by the dendrimer polyamidoamine (PAMAM). PAMAM can successfully regulate the synthesis of Cr-MOF; in doing so, the size of Cr-MOF is reduced, its original morphology is maintained, and it has good crystallinity. A simple ultrasonication method was used to make the Cr-MOF/GO hybrid nanocomposite. Various characterization methods confirmed the successful synthesis of PAMAM/Cr-MOF/GO nanocomposites. The PAMAM/Cr-MOF/ERGO modified electrode could be used with cyclic voltammetry (CV) and differential pulse voltammetry (DPV) to study the electrochemical behaviors of 1-hydroxypyrene (1-OHPyr). The results indicated that the constructed PAMAM/Cr-MOF/ERGO electrochemical sensor had a significantly enhanced electrocatalytic effect on the electrochemical reduction of 1-OHPyr compared with the sensors with no PAMAM and the ERGO sensor, which could be ascribed to the synergetic effect from the high porosity of Cr-MOF and the high conductivity of ERGO, as well as the further electron transport action of the nanocomposite. Under the optimal conditions, the reduction peak current and concentration of 1-OHPyr showed a good linear relationship in the range of 0.1-1.0 and 1.0-6.0 μM, and the detection limit of 1-OHPyr was calculated to be 0.075 μM. Moreover, the PAMAM/Cr-MOF/ERGO electrochemical sensor constructed in this paper can be expected to provide some instructions for the construction of electrochemical sensing platforms and wider potential applications.


Littoral cell angioma of the spleen: A case report.
Spleen tumors are an uncommon disease. Littoral cell angioma belongs to the group of vascular tumors. It is believed that this tumor originates from the tissue of the red pulp sinuses, specifically from the cells that are lining the sinuses. If this rare tumor is diagnosed, it is necessary to search for synchronous or metachronous visceral neoplasia. Littoral cell angioma can also mimic metastatic lesion of the spleen. This case report wants to draw attention on this rare tumor of the spleen which is very often associated with other visceral malignancy.


Cell-dependent mechanisms restrict the HIV type 1 coreceptor activity of US28, a chemokine receptor homolog encoded by human cytomegalovirus.
Several members of the chemokine receptor family are used together with CD4 for HIV-1 entry into target cells. The human cytomegalovirus US28 gene encodes a chemokine receptor homolog that has been reported to function as an HIV-1 coreceptor. However, studies of US28 have given conflicting results regarding its ability to mediate HIV-1 entry. We examined the ability of US28 to function as an HIV-1 coreceptor in various cell lines and found that its coreceptor activity is highly cell dependent. US28 could function as a coreceptor for HIV-1 entry in HeLa and U87 cells but not in COS-1 and Cf2Th cells. In COS-1 cells, US28 was expressed on the cell surface and could mediate cell-cell fusion with HIV-1 Env-expressing cells, suggesting that the block to infection may result from a defect in virus internalization or postentry steps. In Cf2Th cells, US28 was expressed at high levels intracellularly but was not transported to the cell surface. The block in US28 coreceptor function in COS-1 and Cf2Th cells was coreceptor dependent, since CCR5, CXCR4, and other coreceptors can mediate HIV-1 entry in these cell lines. HIV-1 viruses pseudotyped with the MuLV or VSV Env entered and replicated at similar efficiency in COS-1 and U87 cells in single-cycle infections, suggesting that postentry and other early events in the HIV-1 life cycle are not intrinsically inefficient in COS-1 cells. These results identify two distinct mechanisms that can restrict the HIV-1 coreceptor activity of US28 in a cell- and coreceptor-dependent manner, and help to explain the existing controversy regarding the ability of US28 to mediate HIV-1 entry.