bluuebunny/crossref_metadata_2025 huggingface dataset:
Bacterial origin recognition complexes direct assembly of higher-order DnaA oligomeric structures
Authors: Miller Diana T. , Grimwade Julia E. , Betteridge Thu , Rozgaja Tania , Torgue Julien J.-C. , Leonard Alan C.
Published: October 2009

Eukaryotic initiator proteins form origin recognition complexes (ORCs) that bind to replication origins during most of the cell cycle and direct assembly of prereplication complexes (pre-RCs) before the onset of S phase. In the eubacterium
Escherichia coli
, there is a temporally similar nucleoprotein complex comprising the initiator protein DnaA bound to three high-affinity recognition sites in the unique origin of replication,
oriC
. At the time of initiation, this high-affinity DnaA–
oriC
complex (the bacterial ORC) accumulates additional DnaA that interacts with lower-affinity sites in
oriC
, forming a pre-RC. In this paper, we investigate the functional role of the bacterial ORC and examine whether it mediates low-affinity DnaA–
oriC
interactions during pre-RC assembly. We report that
E. coli
ORC is essential for DnaA occupation of low-affinity sites. The assistance given by ORC is directed primarily to proximal weak sites and requires oligomerization-proficient DnaA. We propose that in bacteria, DnaA oligomers of limited length and stability emerge from single high-affinity sites and extend toward weak sites to facilitate their loading as a key stage of prokaryotic pre-RC assembly.
Linear group II intron RNAs can retrohome in eukaryotes and may use nonhomologous end-joining for cDNA ligation
Authors: Zhuang Fanglei , Mastroianni Marta , White Travis B. , Lambowitz Alan M.
Published: October 2009

Mobile group II introns retrohome by an RNP-based mechanism in which the excised intron lariat RNA fully reverse splices into a DNA site via 2 sequential transesterification reactions and is reverse transcribed by the associated intron-encoded protein. However, linear group II intron RNAs, which can arise by either hydrolytic splicing or debranching of lariat RNA, cannot carry out both reverse-splicing steps and were thus expected to be immobile. Here, we used facile microinjection assays in 2 eukaryotic systems,
Xenopus laevis
oocyte nuclei and
Drosophila melanogaster
embryos, to show that group II intron RNPs containing linear intron RNA can retrohome by carrying out the first step of reverse splicing into a DNA site, thereby ligating the 3′ end of the intron RNA to the 5′ end of the downstream exon DNA. The attached linear intron RNA is then reverse transcribed, yielding an intron cDNA whose free end is linked to the upstream exon DNA. Some of these retrohoming events result in the precise insertion of full-length intron. Most, however, yield aberrant 5′ junctions with 5′ exon resections, 5′ intron truncations, and/or extra nucleotide residues, hallmarks of nonhomologous end-joining. Our findings reveal a mobility mechanism for linear group II intron RNAs, show how group II introns can co-opt different DNA repair pathways for retrohoming, and suggest that linear group II intron RNAs might be used for site-specific DNA integration in gene targeting.
Multidrug-Resistant Gram-Negative Bacteria at a Long-Term Care Facility: Assessment of Residents, Healthcare Workers, and Inanimate Surfaces
Authors: O'Fallon Erin , Schreiber Robert , Kandel Ruth , D'Agata Erika M. C.
Published: October 2009

Objective.

To characterize the clinical and molecular epidemiology of multidrug-resistant (MDR) organisms in residents, in healthcare workers (HCWs), and on inanimate surfaces at a long-term care facility (LTCF).

Design.

Point-prevalence study in 4 separate wards at a 600-bed urban LTCF that was conducted from October 31, 2006 through February 5, 2007.

Participants.

One hundred sixty-one LTCF residents and 13 HCWs.

Methods.

Nasal and rectal samples were obtained for culture from each resident, selected environmental surfaces in private and common rooms, and the hands and clothing of HCWs in each ward. All cultures were evaluated for the presence of MDR gram-negative bacteria, methicillin-resistantStaphylococcus aureus, and vancomycin-resistant enterococci. Clinical and demographic information were collected for each enrolled resident. Molecular typing was performed to identify epidemiologically related strains.

Results.

A total of 37 (22.8%), 1 (0.6%), and 18 (11.1%) residents were colonized with MDR gram-negative bacteria, vancomycin-resistant enterococci, and methicillin-resistantS. aureus, respectively. MDR gram-negative bacteria were recovered from 3 (1.8%) of the 175 environmental samples cultured, all of which were obtained from common areas in LTCF wards. One (7.7%) of the 13 HCWs harbored MDR gram-negative bacteria. Molecular typing identified clonally related MDR gram-negative strains in LTCF residents. After multivariable analysis, length of hospital stay of at least 4 years, fecal incontinence, and antibiotic exposure for at least 8 days were independent risk factors associated with harboring MDR gram-negative bacteria among LTCF residents.

Conclusions.

The prevalence of MDR gram-negative bacteria is high among LTCF residents and exceeds that of vancomycin-resistant enterococci and methicillin-resistantS. aureus. Common areas in LTCFs may provide a unique opportunity for person-to-person transmission of MDR gram-negative bacteria.
Indications for Percutaneous Endoscopic Gastrostomy and Procedure‐related Outcome
Authors: Srinivasan Ramesh , Irvine Tracey , Dalzell Mark
Published: October 2009

Objective:

To describe the indications and practice of percutaneous endoscopic gastrostomy (PEG) device insertion for nutritional support and to ascertain procedure‐related complications and outcome.

Methods:

Observational study with prospectively collected data on children who underwent PEG‐related procedures (PEG insertion, removal, or change to low profile button devices [LPBD]) for a 5‐year period (2002–2006) at our centre.

Results:

Six hundred one PEG‐related procedures were performed during the study period (384 insertions, 165 conversions to LPBD, 49 permanent PEG removals, and 3 PEG reinsertions). The main indications for PEG insertion were feeding difficulties associated with neurodisability (160/384; 41.6%) and congenital heart disease (CHD 115/384; 30%). Children with CHD required feeding nutritional support at an earlier age than children with neurodisability regardless of underlying cardiac condition. The age of PEG insertion was significantly different between the 2 main groups (neurodisability 3.56 years vs CHD 0.39 years; P < 0.001 [t test]). Fifty‐nine patients were discharged on the same day as PEG insertion without complications (day case gastrostomy). The median time between PEG insertion to LPBD conversion was 0.83 year (0.12–3.86). Twenty‐four of the 49 children having permanent PEG removal had CHD.

Conclusions:

Neurodisabilty and CHD were the main indications for PEG insertion, those with CHD requiring feeding support at an earlier age. Although CHD was an indication for PEG insertion in 30% of instances, a greater percentage of PEG removals (50%) were seen in this group indicating resolution of feeding difficulties.
Safety, Efficacy, and Pharmacokinetics of Balsalazide in Pediatric Patients With Mild‐to‐Moderate Active Ulcerative Colitis: Results of a Randomized, Double‐blind Study
Authors: Quiros J Antonio , Heyman Melvin B , Pohl John F , Attard Thomas M , Pieniaszek Henry J , Bortey Enoch , Walker Kelli , Forbes William P
Published: October 2009

Objectives:

A multicenter, double‐blind study was conducted to evaluate the safety, efficacy, and pharmacokinetics of balsalazide in pediatric patients with mild‐to‐moderate ulcerative colitis (UC).

Patients and Methods:

Sixty‐eight patients, 5 to 17 years of age, with mild‐to‐moderate active UC based on the modified Sutherland UC activity index (MUCAI) were randomized to receive oral balsalazide 2.25 or 6.75 g/day for 8 weeks. The primary endpoint was clinical improvement (reduction of the MUCAI score by ≥3 points from baseline). Clinical remission (MUCAI score of 0 or 1 for stool frequency) and histological improvement after 8 weeks were also assessed. Pharmacokinetic parameters for balsalazide, 5‐aminosalicylic acid, and N‐acetyl‐5‐aminosalicylic acid were determined at 2 weeks. Adverse events and laboratory changes were monitored throughout the study.

Results:

Clinical improvement was achieved by 45% and 37% of patients and clinical remission by 12% and 9% of patients receiving 6.75 and 2.25 g/day, respectively. Improvement in histologic grade was achieved by 8 of 16 (50%) and 3 of 10 (30%) patients receiving 6.75 and 2.25 g/day, respectively. No significant differences were seen in efficacy. Pharmacokinetics in 12 patients were characterized by large interpatient variability and low systemic exposure. Adverse events were similar between the treatment groups, the most common being headache and abdominal pain. No clinically significant changes were observed in laboratory values, including those indicative of hepatic or renal toxicity.

Conclusions:

Balsalazide is well tolerated and improves the signs and symptoms of mild‐to‐moderate active UC in pediatric patients 5 to 17 years of age.
Rotavirus Gastroenteritis: Precursor of Functional Gastrointestinal Disorders?
Authors: Saps M , Pensabene L , Turco R , Staiano A , Cupuro D , Di Lorenzo C
Published: October 2009

Background and Aims:

Abdominal pain‐related functional gastrointestinal disorders (AP‐FGIDs) following bacterial acute gastroenteritis (AGE) have been demonstrated in adults and children. An adult study demonstrated AP‐FGIDs resulting from an outbreak of viral AGE. Viral AGEs are common in children. Thus, the demonstration of AP‐FGIDs occurring after a viral infection in children could constitute a significant finding. The aim of the study was to investigate the development of FGIDs following an episode of acute rotavirus gastroenteritis in children. This is the first pediatric multicenter study designed to assess postviral AP‐FGIDs.

Patients and Methods:

It is a cohort study. Inclusion criteria of the study are children ages 4 to 18 years with history of AGE secondary to rotavirus. Sample size is 44 exposed and 44 controls (unidirectional α of 0.05, power of 0.80). Children consulting at 2 hospitals (Chicago, IL, and Naples, Italy) for AGE (2002–2004) who tested positive for rotavirus were randomly contacted by telephone >2 years after the episode. Each exposed child who visited the emergency department or outpatient site for acute trauma or well‐child visit within 4 weeks of the index case was matched with a control of the same age and sex. Gastrointestinal symptoms and disability were evaluated with a validated pediatric questionnaire.

Results:

Eighty‐eight patients (46 boys, mean age 5.3 years) were recruited. Contacted patients presented with AGE in 2002 (9), 2003 (11), and 2004 (24). Seven (16%) exposed patients and 3 (7%) controls reported AP‐FGIDs (P = 0.31).

Conclusions:

Our study suggests that rotavirus infection does not seem to place children at increased risk for AP‐FGIDs at long‐term follow‐up. Larger, prospective studies should be conducted to evaluate whether rotavirus gastroenteritis leads to AP‐FGIDs in children.
Protein Content and Fortification of Human Milk Influence Gastroesophageal Reflux in Preterm Infants
Authors: Aceti Arianna , Corvaglia Luigi , Paoletti Vittoria , Mariani Elisa , Ancora Gina , Galletti Silvia , Faldella Giacomo
Published: October 2009

Objectives:

Preterm human milk (HM) may provide insufficient energy and nutrients and thus may need to be fortified. Our aim was to determine whether fat content, protein content, and osmolality of HM before and after fortification may affect gastroesophageal reflux (GER) in symptomatic preterm infants.

Methods:

Gastroesophageal reflux was evaluated in 17 symptomatic preterm newborns fed naïve and fortified HM by combined pH/intraluminal‐impedance monitoring (pH‐MII). Human milk fat and protein content was analysed by a near‐infrared reflectance analysis. Human milk osmolality was tested before and after fortification. Gastroesophageal reflux indexes measured before and after fortification were compared and were also related to HM fat and protein content and osmolality before and after fortification.

Results:

An inverse correlation was found between naïve HM protein content and acid reflux index (RIpH: P = 0.041, ρ =−0.501). After fortification, osmolality often exceeded the values recommended for infant feeds; furthermore, a statistically significant (P < 0.05) increase in nonacid reflux indexes was observed.

Conclusions:

Protein content of naïve HM may influence acid GER in preterm infants. A standard fortification of HM may worsen nonacid GER indexes and, due to the extreme variability in HM composition, may overcome both recommended protein intake and HM osmolality. Thus, an individualised fortification, based on the analysis of the composition of naïve HM, could optimise both nutrient intake and feeding tolerance.
Prevalence of Metabolic Bone Disease in Children With Celiac Disease Is Independent of Symptoms at Diagnosis
Authors: Turner Justine , Pellerin Genevieve , Mager Diana
Published: October 2009

Objectives:

Given dietary gluten exposure, growing children with celiac disease may experience malabsorption of nutrients, negatively affecting bone health. The purpose of this study was to determine the prevalence of low bone mass in children with celiac disease, according to the presence of symptoms at diagnosis.

Materials and Methods:

A retrospective chart review of the Stollery Children's Hospital Celiac Clinic charts (April 1989–September 2007) was conducted. Bone mineral density (BMD) of the spine was measured using dual energy x‐ray absorptiometry. Demographics, symptoms at presentation, and anthropometric and biochemical data relevant to bone health were recorded.

Results:

Seventy‐four children (9.6 ± 3.7 years; range 3.3–16.0 years) were included. Lumbar BMDzscores more than or equal to −1 were observed in 58 cases (65%),zscores below −1 but above −2 were observed in 14 cases (19%) andzscores less than or equal to −2 were observed in 12 cases (16%). There was no significant difference in mean lumbar BMDzscores between symptomatic and asymptomatic children (P= 0.34). When adjusted for bone age and bone surface area, BMD lumbarzscore was inversely correlated with age at diagnosis (P< 0.05).

Conclusions:

An equivalent reduction in spine bone mass was observed in children with celiac disease at diagnosis regardless of the presence of symptoms. Delayed diagnosis of children with celiac disease may increase the risk of adult osteoporosis. Appropriate screening of children at risk of celiac disease for the purpose of early diagnosis, as well as routine evaluation of bone mineral density in such children, are important to prevent long‐term complications associated with poor bone health.
Teaching and Tomorrow: A Novel Recruitment Program for a Pediatric Subspecialty
Authors: Graham Rose C , Heubi James E , Cohen Mitchell B , Li BUK
Published: October 2009

Objectives:

In 2001, the North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition created Teaching and Tomorrow, a novel pediatric subspecialty recruitment program to provide prospective fellows the opportunity to participate in the society's annual meeting and to interact with current trainees and members. The aims of this report are to describe participant survey data from the first 4 years of the program and to explore potential influences of participation on outcomes and proposed career choices.

Materials and Methods:

Pre‐ and postprogram surveys were administered during opening and closing breakfast meetings, respectively, to ascertain demographics, certainty of career intent, factors influencing choice of specialty, and anticipated benefits from participation.

Results:

Among 189 applications, 107 (57%) participated and 107 (57%) matriculated into gastroenterology(GI) fellowship. The matriculation rate was similar between participants and nonparticipants. Among applicants who became GI fellows, there was a 42% lower odds of being female than male (OR 0.42; χ2 = 8.48, P < 0.01). Among participants who completed both pre‐ and postprogram surveys (n = 91) and who reported certainty about entering GI both before and after, 79% entered GI fellowship. Among participants who reported uncertainty about entering GI both before and after, 8% entered GI. More participants were interested in jobs as clinician‐investigators and funded clinical or translational investigators.

Conclusions:

In a selected population of pediatric residents with high interest in pediatric gastroenterology, there was no difference in the matriculation rate into pediatric GI fellowship training among those who participated compared with those who did not. However, those who eventually matriculated had a higher odds of being male than female. There was a high retention rate of participants who were initially certain and a modest conversion rate of those who were initially uncertain about pursuing a career in pediatric GI.
Parenteral Lipid Emulsions Based on Olive Oil Compared With Soybean Oil in Preterm (<28 Weeks' Gestation) Neonates: A Randomised Controlled Trial
Authors: Deshpande Girish C , Simmer Karen , Mori Trevor , Croft Kevin
Published: October 2009

Background:

New olive oil–based (OL) lipid emulsions (olive:soy oil = 4:1) have lower polyunsaturated fatty acid (PUFA) (20% vs 60%) and higher vitamin E content (an antioxidant) compared with traditional soybean oil (SO) emulsions.

Objective:

Compare efficacy and safety of OL with SO emulsions in preterm neonates (<28 weeks) at high risk for oxidative stress.

Patients and Methods:

Preterm neonates (gestation 23–<28 weeks) were randomised to receive OL or SO emulsion for 5 days using a standard protocol in a tertiary perinatal centre (King Edward Memorial Hospital for Women, Perth, Western Australia). Investigators and outcome assessors were masked to allocation. Plasma F2‐isoprostanes (lipid peroxidation marker), plasma, and red blood cell fatty acids were measured before and after the study. Safety was monitored by liver function tests.

Results:

Forty‐four of 50 participants (OL‐23, SO‐21) completed the study. Both emulsions were well tolerated with no significant adverse events. F2‐isoprostane levels were comparable at baseline and study end. Oleic and linoleic acid levels were significantly high on day 6 in OL and SO groups, respectively. Long‐chain PUFA levels were similar between groups despite the lower PUFA content of OL. The olive oil–based group had significantly higher levels of C18:4n‐3, suggesting Δ6‐desaturase enzyme inhibition in the SO group.

Conclusions:

Olive oil–based emulsion was safe and well tolerated by preterm neonates. Similar long‐chain PUFA levels were achieved in the OL group despite significantly lower amount of PUFA content; however, there was no difference in lipid peroxidation (F2‐isoprostane levels). Large trials are needed to confirm these benefits.
Pro‐ and Prebiotic Supplementation Induces a Transient Reduction in Hemoglobin Concentration in Infants
Authors: Kuitunen Mikael , Kukkonen Kaarina , Savilahti Erkki
Published: October 2009

Objectives:

Regarding safety, we investigated the effect of prenatal probiotic and 6 months of pro‐ and prebiotic supplementation of infants on their hematologic values at 6 months and 2 years and factors affecting these values.

Patients and Methods:

In a prospective randomized controlled probiotic intervention trial in infants at high risk for allergy, we obtained blood samples consecutively from 98 infants at 6 months and from 658 children at 2 years to measure hematologic values. We collected fecal samples at 3 and 6 months to measure immunologic development by calprotectin, α‐1‐antitrypsin, tumor necrosis factor‐α, and immunoglobulin A.

Results:

At 6 months, infants in the probiotic group had significantly lower hemoglobin (Hb) values than did the placebo group, mean (SD): 119.8 g/L (6.3) versus 123.3 g/L (8.4), P = 0.025. Adjustment for factors that might affect Hb values (breast‐feeding duration, solid‐food introduction, and sex), revealed no need for adjustment. A significant negative correlation emerged between Hb values at 6 months and fecal calprotectin at age 3 months r = −0.301, P = 0.009, which was affected neither by breast‐feeding, sex, nor study group. At 2 years, hematologic values in both groups became similar.

Conclusions:

Probiotics cause a gut mucosal inflammation with decreased Hb values during intervention, corrected after halting the supplementation.
Impact of Anxiety in the Puerperium on Breast‐feeding Outcomes: Role of Parity
Authors: Zanardo Vincenzo , Gasparetto Sara , Giustardi Arturo , Suppiej Agnese , Trevisanuto Daniele , Pascoli Irene , Freato Federica
Published: October 2009

Objective:

Studies in animals indicate that stress and anxiety can depress lactation, but there is much less information available concerning humans. We aimed to determine whether maternal anxiety in puerperium, potentially exacerbated by the primiparity inexperience, has a negative impact on breast‐feeding outcomes.

Subjects and Methods:

The state‐trait anxiety inventory Y form (Spielberger, 1983) was administered to a total of 204 mothers, 101 primiparae, and 103 pluriparae on the third to fourth day postpartum.

Results:

We found that primiparae had state anxiety (T) scores significantly higher than pluriparae (44.57 ± 5.85 vs 43.28 ± 7.10, P = 0.03). Increased state anxiety levels impaired breast‐feeding success. In addition, in the logistic regression analysis model used to assess factors predictive of breast‐feeding longer than 3 months, maternal state anxiety was the most significant risk factor (odds ratio 0.99; 0.88–0.98, P < 0.01).

Conclusions:

In the puerperium, anxiety, potentially exacerbated by primiparae inexperience, is associated with impaired lactation. Alleviating maternal anxiety could be beneficial for stimulating breast‐feeding in more vulnerable women.
Noninvasive Procedures to Evaluate Liver Involvement in HIV‐1 Vertically Infected Children
Authors: Rubio Amandine , Monpoux Fabrice , Huguon Emilie , Truchi Régine , Triolo Valérie , Rosenthal‐Allieri Maria‐Alessandra , Deville Anne , Rosenthal Eric , Boutté Patrick , Tran Albert
Published: October 2009

Objectives:

Progressive liver injury is a concern in HIV‐infected children exposed to long‐term antiretroviral drugs and to the cytopathic effect of HIV. Yet liver biopsy is usually considered too invasive to be repeated in these patients. The aims of this study are to evaluate the feasibility of noninvasive hepatic investigations in HIV‐1‐infected children, assess the prevalence of signs of liver affection, and analyse the influence of the HIV disease severity and the exposure to antiretroviral therapy.

Materials and Methods:

A cross‐sectional study conducted in 26 HIV‐1 vertically infected children ages 8 to 18 years old. Liver function was assessed with standard serum biochemical markers, FibroTest, ActiTest, SteatoTest, Forns index, aspartate aminotransferase to platelet ratio index, ultrasound, and Fibroscan.

Results:

Nineteen (>60%) children had signs of liver affection on at least 1 of the test results: 13 (50%) had elevated liver enzymes, 15 (63%), 8 (33%), 5 (21%), and 5 (21%) had abnormal FibroTest, ActiTest, Forns index, and aspartate aminotransferase to platelet ratio index results, respectively. Four children (17%) had mild liver steatosis on ultrasound. Fibroscan measures were significantly higher in patients than in age‐matched healthy children. Patients with elevated Fibroscan measures also had significantly higher FibroTest results. Age, HIV stage N in the Centers for Disease Control and Prevention classification and exposure duration to nucleoside reverse transcriptase inhibitor and non‐nucleoside reverse transcriptase inhibitor drugs were the main risk factors for hepatotoxicity.

Conclusions:

More than half of our population of HIV‐infected children had biological and/or radiological signs of liver affection. Regular follow‐up of liver function is necessary in these patients, which is now possible with noninvasive procedures.
Esophageal Impedance and Esophagitis in Children: Any Correlation?
Authors: Salvatore S , Hauser B , Devreker T , Arrigo S , Marino P , Citro C , Salvatoni A , Vandenplas Y
Published: October 2009

Aim:

The aim of this study was to correlate the data obtained with multiple intraluminal esophageal impedance and pH (MII‐pH) recordings in infants and children referred for suspected gastroesophageal reflux disease with esophageal histology.

Materials and Methods:

In a prospective study, results of esophageal biopsies and MII‐pH recording obtained in 45 children (mean age ± SD: 69 ± 55 months) were analyzed. Regarding the MII‐pH data, an automatic (Autoscan Bioview Analysis Software, version 5.3.4, Sandhill Scientific Inc, Highlands Ranch, CO) and a manual reading were performed; an automatic pH analysis (meal included) was also performed.

Results:

Acidic, weakly acidic, and alkaline reflux episodes accounted, respectively, for 48.7%, 49.5%, and 1.8% of the total number of reflux episodes detected by MII‐pH. Esophagitis was present in 25 (56%) children. Concordance between classic pH‐study analysis (alone) and esophageal histology was found in 19 of 45 (42%) children. According to the MII‐pH analysis, the mean and median value of the pH were significantly higher in the group with esophagitis than in the group with normal esophageal histology. A longer clearance time was found in the group with esophagitis than in subjects with normal histology. Gas reflux episodes represented 21% of the total reflux episodes and were comparable in both groups.

Conclusions:

Multiple intraluminal esophageal impedance and pH analysis does not provide a distinct parameter to predict esophageal mucosal injury in children. In our population, MII‐pH shows comparable acidic, weakly acidic, alkaline, and gas reflux in children with and without esophagitis. Further research is needed to analyze clearance parameters.
Analysis of Faecal Volatile Organic Compounds in Preterm Infants Who Develop Necrotising Enterocolitis: A Pilot Study
Authors: Garner Catherine E , Ewer Andrew K , Elasouad Kalifa , Power Francis , Greenwood Rosemary , Ratcliffe Norman M , Costello Ben de Lacy , Probert Chris S
Published: October 2009

Objective:

To determine differences in the profiles of volatile organic compounds (VOCs) in faecal samples from preterm infants who develop necrotising enterocolitis (NEC) compared with non‐NEC controls.

Materials and Methods:

Daily faecal samples from preterm infants were collected prospectively during an 8‐month period from a level 3 regional neonatal intensive care unit. Six infants subsequently developed NEC and were matched with 7 non‐NEC infants. Solid‐phase microextraction and gas chromatography and mass spectrometry were used to extract and identify the VOCs from the headspace above the faecal samples taken before the onset of NEC and after the disease was diagnosed. Faecal samples at similar ages were also studied from the control infants.

Results:

Two hundred twenty‐four different VOCs were extracted from 65 samples. Volatile organic compounds increased in number with age for non‐NEC infants. In the days before and after the diagnosis of NEC a reduction in the number of VOCs extracted was observed. In addition, 4 specific esters present in controls—2‐ethylhexyl acetic ester, decanoic acid ethyl ester, dodecanoic acid ethyl ester, and hexadecanoic acid ethyl ester—were consistently absent from all faecal samples in those infants who developed NEC in the 4 days before the onset of the disease.

Conclusion:

This pilot study shows that VOC extraction from faeces may be used to identify infants that are at risk of developing NEC.
Angiostatic activity of human plasminogen fragments is highly dependent on glycosylation
Authors: Santos Ivan Carlos , Silbiger Vivian Nogueira , Higuchi Débora Ayame , Gomes Maria Aparecida , Barcelos Lucíola Silva , Teixeira Mauro Martins , Lopes Mirian Teresa Paz , Cardoso Valbert Nascimento , Lima Mercia Paula , Araujo Ronaldo Carvalho , Pesquero João Bosco , Pesquero Jorge Luiz
Published: October 2009

To assess the importance of carbohydrate moieties to the anti‐angiogenic activity of plasminogen fragments, we cloned the fragment corresponding to amino acids Val79to Thr346(Kint3–4) that presents the three glycosylation sites. The activity of glycosylated and unglycosylated Kint3–4 was tested in murine sponge implant model. We observed a significant decrease in the neovascularization on the sponge after treatment with Kint3–4 by histological examination and determination of the hemoglobin levels. The effects were more intense with the glycosylated than the unglycosylated protein.99mTechnecium‐labeled red blood cells confirmed the inhibition of cell infiltration in the implanted sponge. Studies using melanoma B16F1 implanted in a mouse demonstrated that treatment with glycosylated Kint3–4 (0.15 nmol/48 h) during 14 days suppresses tumor growth by 80%. The vascular endothelial growth factor mRNA levels on the tumor were reduced after treatment. Kint3–4 is a potent plasminogen fragment that has been found to inhibit tumor growth. (Cancer Sci2009)
Activity of sulfotransferase 1A1 is dramatically upregulated in patients with hepatocellular carcinoma secondary to chronic hepatitis B virus infection
Authors: Wang Xiang‐Rui , Qu Zeng‐Qiang , Li Xiao‐Dong , Liu Hai‐Lin , He Ping , Fang Bei‐Xiong , Xiao Jie , Huang Wei , Wu Meng‐Chao
Published: October 2009

The phase I metabolizing enzyme and phase II metabolizing enzyme play vital roles in carcinogenesis, but little is known about the changes of their activities in patients with hepatocellular carcinoma (HCC) secondary to chronic hepatitis B virus (HBV) infection. In this study phenacetin, a probe drug (1 g for men and 0.85 g for women orally), was applied for the detection of sulfotransferase 1A1 (SULT1A1) and cytochrome P4501A2 (CYP1A2) activities in 82 healthy participants and 148 HCC, 106 cirrhosis, and 41 chronic hepatitis B patients. In addition, a prospective cohort study for susceptibility to HCC was performed in 205 patients with cirrhosis secondary to chronic HBV infection. Compared with the healthy participants, SLUT1A1 activity increased by 9.7‐fold in the HCC patients (P < 0.01). CYP1A2 activity did not significantly differ between the healthy participants and HCC patients. CYP1A2 activity decreased by 91.2% (P < 0.01) and 67.7% (P < 0.05) in the patients with cirrhosis and chronic hepatitis B, respectively; SULT1A1 activity did not increase significantly. During an approximate 2‐year follow up, three of the 46 cirrhosis patients with elevated SULT1A1 activity and normal CYP1A2 activity developed HCC, but none of the 159 cirrhosis patients used as parallel controls did (P = 0.012). These results indicate that SLUT1A1 activity is dramatically upregulated in patients with HCC secondary to chronic HBV infection. The upregulation of SULT1A1 activity is not caused by the tumor itself. The interaction between SULT1A1 and CYP1A2 can play an important role in hepatocarcinogenesis in the Chinese population. (Cancer Sci2009)
Prostaglandin EP2 receptor expression is increased in Barrett’s oesophagus and oesophageal adenocarcinoma
Authors: JIMÉNEZ P. , PIAZUELO E. , CEBRIAN C. , ORTEGO J. , STRUNK M. , GARCÍA‐GONZALEZ M. A. , SANTANDER S. , ALCEDO J. , LANAS A.
Published: October 2009

Aliment Pharmacol Ther 31, 440–451

Background  Accumulating evidence suggests that cyclooxygenase‐2 (COX‐2)‐derived prostaglandin E2 (PGE2) is involved in oesophageal adenocarcinogenesis. PGE2 exerts its biological action by binding to specific receptors (EP1, EP2, EP3 and EP4).

Aim  To investigate which PGE2 receptor subtypes regulate PGE2 signals in the oesophageal adenocarcinoma sequence.

Methods  Expression was determined in oesophageal biopsies from 85 patients with oesophagitis, Barrett’s metaplasia, intraepithelial neoplasia, oesophageal adenocarcinoma and normal oesophagus. Levels of mRNA and protein expression were determined by quantitative PCR, immunohistochemistry and western‐blot. Expression of EP receptors was also determined in response to acid and bile exposure in the Barrett’s adenocarcinoma cell line OE33.

Results  All four EP receptors subtypes were expressed in human oesophageal tissues. COX‐2 and, especially, EP2 were increased in the Barrett’s metaplasia‐intraepithelial neoplasia‐adenocarcinoma sequence. Expression of the EP4 receptor protein was increased in oesophageal adenocarcinoma. In contrast, expression levels of COX‐1 and EP3 receptor were decreased along the sequence. No differences in EP1 expression were found. Treatment with the bile acid deoxycholate increased COX‐2, EP1, EP2 and EP4 expression in OE33 cells.

Conclusions  Our data suggest that in addition to COX‐2, EP2 and EP4 receptors could be a selective target in the prevention and/or treatment of the Barrett’s‐associated adenocarcinoma.
Activation profile of Toll-like receptors of peripheral blood lymphocytes in patients with systemic lupus erythematosus
Authors: Wong C K , Wong P T Y , Tam L S , Li E K , Chen D P , Lam C W K
Published: October 2009

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease associated with aberrant activation of T and B lymphocytes for the production of inflammatory cytokines and autoreactive antibodies. Animal studies of SLE have indicated that Toll-like receptors (TLR) are important in the pathogenesis of murine lupus. In the present clinical study, differential protein expressions of TLR-1–9 of monocytes and different lymphocyte subsets from patients with SLE and normal control subjects were determined by flow cytometry. Results showed that the expression of intracellular TLRs (TLR-3, -8, -9) and extracellular TLRs (TLR-1, -2, -4, -5, -6) were elevated in monocytes, CD4+ T lymphocytes, CD8+ T lymphocytes and B lymphocytes of SLE patients compared to control subjects (all P < 0·001). Moreover, cell surface expression of TLR-4 on CD4+ T lymphocytes and CD8+ T lymphocytes, and TLR-6 on B lymphocytes, were correlated positively with SLE disease activity index (SLEDAI) (TLR-4 on CD4+ T lymphocytes and CD8+ T lymphocytes: r = 0·536, P = 0·04; r = 0·713, P = 0·003; TLR-6 in B lymphocytes: r = 0·572, P = 0·026). In concordance with the above results, there is an observable increased relative induction (%) of inflammatory cytokine interleukin (IL)-1β, IL-6, IL-10 and IL-12, chemokines CCL2, CXCL8, CCL5 and CXCL10 from peripheral blood mononuclear cells (PBMC) upon differential stimulation by PolyIC (TLR-3 ligand), lipopolysaccharide (TLR-4 ligand), peptidoglycan (TLR-2 ligand), flagellin (TLR-5 ligand), R837 (TLR-7 ligand) and CpG DNA (TLR-9 ligand) in SLE patients compared to controls. These results suggest that the innate immune response for extracellular pathogens and self-originated DNA plays immunopathological roles via TLR activation in SLE.
Aquaporins in the human testis and spermatozoa – identification, involvement in sperm volume regulation and clinical relevance
Authors: Yeung C. H. , Callies C. , Tüttelmann F. , Kliesch S. , Cooper T. G.
Published: October 2009

Despite the high water‐permeability of human spermatozoa, little is known about the identity and the role of aquaporins (AQP) in them or germ cells. Using ejaculates from donors, sperm AQPs were identified by western blotting followed by the analysis of mRNA with RT‐PCR. Protein expression in the testis and spermatozoa was localized by immunocytochemistry. Inhibitors were used to investigate the involvement of aquaporins in water transport when ejaculated spermatozoa were swollen in medium mimicking uterine hypo‐osmolality by quinine that blocks volume regulation. Sperm AQP7 and AQP8 in 39 infertile patients and 11 healthy donors were quantified by flow cytometry. AQP1 was absent from spermatozoa. Sperm and testicular AQP7‐9 had nucleotide sequences identical to those of somatic cells but AQP8 mRNA also showed shorter variants. AQP7 was expressed abundantly by round and elongated spermatids and ejaculated spermatozoa, AQP8 by all germ cells and spermatozoa, and AQP9 rarely by spermatocytes or Sertoli cells. Protein bands showed specificity by western blotting for AQP7 and AQP8 but not AQP9. The absence of sperm AQP9 was further suggested by the ineffectiveness of its inhibitor phloretin in blocking quinine‐induced swelling, but HgCl2, which inhibits AQP8, was effective. Sperm AQP7 expression was correlated with progressive motility and was lower in patients than in donors. Sperm AQP8 expression was inversely correlated with the extent of sperm coiling, which is a swelling phenomenon, but showed no difference between patients and donors. In conclusion, AQP7 and AQP8 were identified in human spermatozoa and could play a role in glycerol metabolism and water transport respectively.
Use of garlic, Allium sativum, to control Aeromonas hydrophila infection in rainbow trout, Oncorhynchus mykiss (Walbaum)
Authors: Nya E J , Austin B
Published: October 2009

Garlic, Allium sativum, which was fed at 0, 0.05, 0.1, 0.5 and 1.0 g per 100 g of feed for 14 days to rainbow trout, Oncorhynchus mykiss (Walbaum), led to control of experimental infection with Aeromonas hydrophila. At doses of 0.5 and 1.0 g garlic per 100 g of feed, there was a reduction in mortalities to 4% compared with the controls (88%). Moreover, there was a significant increase in growth, feed conversion and protein efficiency. There was stimulation of the number of erythrocytes and leucocytes, a significantly higher haematocrit, enhancement of phagocytic activity, respiratory burst, lysozyme, anti‐protease and bactericidal activities following feeding with garlic.
Use of dietary ginger, Zingiber officinale Roscoe, as an immunostimulant to control Aeromonas hydrophila infections in rainbow trout, Oncorhynchus mykiss (Walbaum)
Authors: Nya E J , Austin B
Published: October 2009

Ginger, Zingiber officinale, which was fed at 0, 0.05, 0.1, 0.5 and 1.0 g per 100 g of feed for 14 days to rainbow trout, Oncorhynchus mykiss (Walbaum), led to control of experimental infection with Aeromonas hydrophila. At 0.5 g ginger per 100 g of feed, there was a reduction in mortalities to 0% compared with the controls (64%). Moreover, there was a significant increase in growth, feed conversion and protein efficiency. There was proliferation in the number of neutrophils, macrophages and lymphocytes, and enhanced phagocytic, respiratory burst, lysozyme, bactericidal and anti‐protease activities compared with the controls.
Lagrange time delay estimation for scanning electron microscope image magnification
Authors: SIM K.‐S. , THONG L.W. , TING H.Y. , TSO C.P.
Published: October 2009

Interpolation techniques that are used for image magnification to obtain more useful details of the surface such as morphology and mechanical contrast usually rely on the signal information distributed around edges and areas of sharp changes and these signal information can also be used to predict missing details from the sample image. However, many of these interpolation methods tend to smooth or blur out image details around the edges. In the present study, a Lagrange time delay estimation interpolator method is proposed and this method only requires a small filter order and has no noticeable estimation bias. Comparing results with the original scanning electron microscope magnification and results of various other interpolation methods, the Lagrange time delay estimation interpolator is found to be more efficient, more robust and easier to execute.
The amplification of polymerized diaminobenzidine with physical developers: sensitizing effects of transition metal salts and sulphide
Authors: VON RUHLAND C.J. , JASANI B.
Published: October 2009

Amplification of metal‐complexed polymerized diaminobenzidine by two light‐insensitive physical developers was systematically examined in a dot blot model system following either polymerizing diaminobenzidine in the presence of transition metal salts or applying the metal salts post‐diaminobenzidine polymerization. The effect of sodium sulphide treatment on subsequent amplification was also investigated. Those metal–diaminobenzidine complexes that facilitated the most powerful amplification were subsequently tested in an immunohistochemical setting. The most dramatic amplification of polymerized diaminobenzidine was observed following its post‐polymerization treatment with salts of platinum alone, or gold or vanadium with subsequent sulphide treatment, and allowed previously invisible quantities of polymerized diaminobenzidine to be clearly seen. Three other transition metal salts also improved the amplification of polymerized diaminobenzidine but to a lesser degree, namely nickel alone, and silver or rhodium with subsequent sulphide treatment. Sensitivity was comparable with the colloidal gold–silver amplification system.
Automatic identification of Caenorhabditis elegans in population images by shape energy features
Authors: OCHOA D. , GAUTAMA S. , PHILIPS W.
Published: October 2009

Experiments on model organisms are used to extend the understanding of complex biological processes. In Caenorhabditis elegans studies, populations of specimens are sampled to measure certain morphological properties and a population is characterized based on statistics extracted from such samples. Automatic detection of C. elegans in such culture images is a difficult problem. The images are affected by clutter, overlap and image degradations. In this paper, we exploit shape and appearance differences between C. elegans and non‐C. elegans segmentations. Shape information is captured by optimizing a parametric open contour model on training data. Features derived from the contour energies are proposed as shape descriptors and integrated in a probabilistic framework. These descriptors are evaluated for C. elegans detection in culture images. Our experiments show that measurements extracted from these samples correlate well with ground truth data. These positive results indicate that the proposed approach can be used for quantitative analysis of complex nematode images.
Feeding‐induced changes in defence enzymes and PR proteins and their implications in host resistance to Nilaparvata lugens
Authors: Alagar M. , Suresh S. , Saravanakumar D. , Samiyappan R.
Published: October 2009

Six rice genotypes showing susceptible and resistant reactions to brown planthopper (BPH), Nilaparvata lugens were studied for feeding‐induced changes in defence enzymes and pathogenesis‐related (PR) proteins. The high resistant genotypes PTB 33, ADT 45 and ASD 7 and moderately resistant genotypes CO 43 and KAU 1661 recorded the greater expression of defence enzymes peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, total phenol and β‐1,3 glucanase in response to N. lugens feeding at 1 day after infestation (DAI) compared with susceptible genotype TN1. The greater activity of chitinase was observed in resistant cultivars at 3 DAI and the activity was sustained for more than 1 week compared with susceptible TN1. In conclusion, the current study revealed that these defence enzymes and PR proteins might attribute to the resistance mechanisms in rice plants against BPH infestation.
Orchidectomy enhances the effects of phenylephrine in rat isolated portal vein
Authors: De Souza Rossignoli Patrícia , Pereira Oduvaldo CM , Chies Agnaldo B
Published: October 2009

1. Orchidectomy results in long‐term testosterone deprivation similar to that observed in male clinical pathologies, such as hypogonadism and age‐related reductions in plasma testosterone concentrations. Although the vascular effects of these sorts of hormone deprivations are known in arteries, they have not been studied to the same extent in veins.

2. The aim of the present study was to determine the effect of orchidectomy, with or without subsequent testosterone replacement (started 23 days after orchidectomy; 10 mg/kg, i.m., testosterone propionate once every 5 days for 3 weeks), on responses of rat isolated portal veins and vena cavae to exogenous phenylephrine (PE). Isolated vessels were mounted in an organ bath and concentration–response curves constructed to PE (10−10–10−4 mol/L), endothelin (ET; 10−10–10−5 mol/L) and KCl (10−2–1.2 × 10−1 mol/L; as a control).

3. Orchidectomy had no effect on contractile responses of either the portal vein or vena cava to KCl. However, orchidectomy enhanced the maximum response (Rmax) of the portal vein, but not the vena cava, to PE. Testosterone replacement had no effect on these responses. The effects of orchidectomy on the Rmaxto PE in portal veins were not altered by the nitric oxide synthase inhibitorNG‐nitro‐l‐arginine methyl ester (10−4 mol/L) alone or combined with 10−5 mol/L indomethacin (a non‐selective cyclo‐oxygenase inhibitor), but they were abolished following treatment of isolated vessels with the ETAand ETBreceptor antagonists BQ‐123 and BQ‐788 (both at 10−6 mol/L). Orchidectomy did not alter portal vein responses to the application of exogenous ET.

4. The results of the present study indicate that orchidectomy‐induced decreases in plasma testosterone can increase the venoconstrictor effects of PE on the portal vein and that this effect involves activation of both ETAand ETBreceptors by locally produced ET.
Inhibitory effect of fucoidan on nitric oxide production in lipopolysaccharide‐activated primary microglia
Authors: Cui Yan‐Qiu , Zhang Li‐Juan , Zhang Ting , Luo Ding‐Zhen , Jia Yan‐Jun , Guo Zi‐Xuan , Zhang Quan‐Bin , Wang Xuan , Wang Xiao‐Min
Published: October 2009

1. Microglial activation plays an important role in the pathogenesis of neurodegenerative diseases by producing various pro‐inflammatory cytokines. Microglia‐derived nitric oxide (NO) is critical for the lipopolysaccharide (LPS)‐induced selective loss of dopaminergic neurons.

2. Fucoidan is a sulphated polysaccharide extracted from brown seaweeds. It has a variety of biological actions, including anticoagulant, antiviral and anti‐inflammatory effects. The aim of the present study was to investigate the effects of fucoidan on LPS‐induced cellular activation in microglia and to evaluate the inhibitory mechanisms involved.

3. To investigate the effects of fucoidan on LPS‐induced cellular activation in microglia, primary microglial cells were preincubated with fucoidan (31.25, 62.5 and 125 μg/mL) for 10 min, followed by stimulation with LPS (0.01 μg/mL). Then, cell shape and NO production were determined 24 h after LPS stimulation, whereas inducible nitric oxide synthase (iNOS) mRNA and protein expression were determined at 6 and 18 h after LPS stimulation, respectively. To evaluate the inhibitory mechanisms involved, mitogen‐activated protein kinase (MAPK) activation was also evaluated.

4. Lipopolysaccharide transformed cells into an amoeboid shape, whereas 62.5 μg/mL fucoidan inhibited this activation. Moreover, 125 μg/mL fucoidan significantly inhibited microglial NO production to 75% of that in LPS‐treated group and also significantly diminished the expression of iNOS mRNA and protein by nearly 50%. Fucoidan (125 μg/mL) also suppressed phosphorylation of p38 and extracellular signal‐regulated kinase (ERK) by approximately 50%, but not that of c‐Jun N‐terminal kinase.

5. The results provide the first evidence that fucoidan has a potent inhibitory effect against LPS‐induced NO production by microglia. The results also suggest that this inhibitory action of fucoidan involves suppression of p38 and ERK phosphorylation.