Iron handling by the human kidney: glomerular filtration and tubular reabsorption both contribute to urinary iron excretion.
In physiological conditions, circulating iron can be filtered by the glomerulus and is almost completely reabsorbed by the tubular epithelium to prevent urinary iron wasting. Increased urinary iron concentrations have been associated with renal injury. However, it is not clear whether increased urinary iron concentrations in patients are the result of increased glomerular iron filtration and/or insufficient tubular iron reabsorption and if these processes contribute to renal injury. We measured plasma and urine iron parameters and urinary tubular injury markers in healthy human subjects ( n = 20), patients with systemic iron overload ( n = 20), and patients with renal tubular dysfunction ( n = 18). Urinary iron excretion parameters were increased in both patients with systemic iron overload and tubular dysfunction, whereas plasma iron parameters were only increased in patients with systemic iron overload. In patients with systemic iron overload, increased urinary iron levels were associated with elevated circulating iron, as indicated by transferrin saturation (TSAT), and increased body iron, as suggested by plasma ferritin concentrations. In patients with tubular dysfunction, enhanced urinary iron and transferrin excretion were associated with distal tubular injury as indicated by increased urinary glutathione S-transferase pi 1-1 (GSTP1-1) excretion. In systemic iron overload, elevated urinary iron and transferrin levels were associated with increased injury to proximal tubules, indicated by increased urinary kidney injury marker 1 (KIM-1) excretion. Our explorative study demonstrates that both glomerular filtration of elevated plasma iron levels and insufficient tubular iron reabsorption could increase urinary iron excretion and cause renal injury.


Clinical features of multiple myeloma patients with isolated extramedullary relapse.
This study sought to analyze the clinical features and prognosis of multiple myeloma with isolated extramedullary relapse and with the absence of systemic progression. The clinical features and outcome were retrospectively analyzed in six multiple myeloma patients. These patients had secretory multiple myeloma at diagnosis. When relapsed, the dissociation between medullary and extramedullary response was detected. The serum or urine monoclonal component was extremely low or absent. The plasma cells in bone marrow were <5%. All patients received new targeted therapies (thalidomide or bortezomib) before extramedullary relapse. It is difficult to achieve second remission for them. Even in those showing response, the duration of response was extremely short. The median of overall survival from diagnosis and from extramedullary relapse was 19 months and 6 months, respectively. The overall survival was significantly shorter compared to the patients without extramedullary involvement (84 months, P=0.001). These patients exhibited a special and rare relapse pattern. Patients with this relapse pattern were resistant to current therapies, including novel targeted agents and associated with poor prognosis.


Structural features of the first component of human complement, C1, as revealed by surface iodination.
Lactoperoxidase-catalysed surface iodination and sucrose-gradient ultracentrifugation were used to investigate the structure of human complement component C1. 1. Proenzymic subcomponents C1r and C1s associated to form a trimeric C1r2-C1s complex (7.6 S) in the presence of EDTA, and a tetrameric Clr2-C1s2 complex (9.1 S) in the presence of Ca2+. Iodination of the 9.1 S complex led to a predominant labelling of C1r (70%) over C1s (30%), essentially located in the b-chain moiety of C1r and in the a-chain moiety of C1s. 2. Reconstruction of proenzymic soluble C1 (15.2 S) from C1q, C1r and C1s was partially inhibited when C1s labelled in its monomeric form was used and almost abolished when iodinated C1r was used. Reconstruction of fully activated C1 was not possible, whereas hybrid C1q-C1r2-C1s2 complex was obtained. 3. Iodination of proenzymic or activated C1 bound to IgG-ovalbumin aggregates led to an equal distribution of the radioactivity between C1q and C1r2-C1s2. With regard to C1q, the label distribution between the three chains was similar whether C1 was in its proenzymic or activated form. Label distribution in the C1r2-C1s2 moiety of C1 was the same as that obtained for isolated C1r2-C1s2, and this was also true for the corresponding activated components. However, two different labelling patterns were found, corresponding to the proenzyme and the activated states.


Functional characterization of the rice SPX-MFS family reveals a key role of OsSPX-MFS1 in controlling phosphate homeostasis in leaves.
• Proteins possessing the SPX domain are found in several proteins involved in inorganic phosphate (Pi) transport and signalling in yeast and plants. Although the functions of several SPX-domain protein subfamilies have recently been uncovered, the role of the SPX-MFS subfamily is still unclear. • Using quantitative RT-PCR analysis, we studied the regulation of SPX-MFS gene expression by the central regulator, OsPHR2 and Pi starvation. The function of OsSPX-MFS1 in Pi homeostasis was analysed using an OsSPX-MFS1 mutant (mfs1) and osa-miR827 overexpression line (miR827-Oe). Finally, heterologous complementation of a yeast mutant impaired in Pi transporter was used to assess the capacity of OsSPX-MFS1 to transport Pi. • Transcript analyses revealed that members of the SPX-MFS family were mainly expressed in the shoots, with OsSPX-MFS1 and OsSPX-MFS3 being suppressed by Pi deficiency, while OsSPX-MFS2 was induced. Mutation in OsSPX-MFS1 (mfs1) and overexpression of the upstream miR827 (miR827-Oe) plants impaired Pi homeostasis in the leaves. In addition, studies in yeast revealed that OsSPX-MFS1 may be involved in Pi transport. • The results suggest that OsSPX-MFS1 is a key player in maintaining Pi homeostasis in the leaves, potentially acting as a Pi transporter.


Characterization of the proteolytic activity of commercial proteases and strained ruminal fluid.
The objective of this research was to formulate a mixture of commercial proteases that would mimic the rate and extent of protein degradation obtained using strained ruminal fluid. The proteolytic activity of strained ruminal fluid and several commercial proteases was characterized using 13 L-amino acid p-nitroanilides as artificial substrates. A mixture of Streptomyces griseus protease, chymotrypsin, and proteinase K at .042, 2.5, and .5 enzyme units/mL, respectively, was similar to the activity of strained ruminal fluid against the same artificial substrates. However, degradative activities were different in incubations with feed proteins as substrates. The rates of degradation of expeller soybean meal, solvent soybean meal, and casein were .08, .05, and .08/h, respectively, using the enzyme mixture and .03, .15, and .24/h using strained ruminal fluid. A second experiment compared degradative activity of S. griseus protease at .066 enzyme units/mL, ficin at .5 enzyme units/mL, and a mixture of trypsin, carboxypeptidase B, chymotrypsin, and carboxypeptidase A at 116.6, .5, 2.5, and .5 enzyme units/mL, respectively. Protein degradation rates obtained with strained ruminal fluid were two to six times faster than those obtained with the enzyme mixtures. A third experiment compared the degradability of 15 feed proteins with the mixture of trypsin, carboxypeptidase B, chymotrypsin and carboxypeptidase A to that with strained ruminal fluid. Degradation rates obtained using strained ruminal fluid ranged from .007 to .217/h; degradation rates using the enzyme mixture ranged from .010 to .079/h and were lower (P = .004) than with strained ruminal fluid. Overall, the experiments indicated that the commercial enzymes tested did not mimic the protein degradative activity of strained ruminal fluid.


Effect of Repeated Active Recovery During a High-Intensity Interval-Training Shock Microcycle on Markers of Fatigue.
To investigate the effect of repeated use of active recovery during a 4-d shock microcycle with 7 high-intensity interval-training (HIT) sessions on markers of fatigue. Eight elite male junior tennis players (age 15.1 ± 1.4 y) with an international ranking between 59 and 907 (International Tennis Federation) participated in this study. After each training session, they completed 15 min of either moderate jogging (active recovery [ACT]) or passive recovery (PAS) with a crossover design, which was interrupted by a 4-mo washout period. Countermovement-jump (CMJ) height, serum concentration of creatine kinase (CK), delayed-onset muscle soreness (DOMS), and perceived recovery and stress (Short Recovery and Stress Scale) were measured 24 h before and 24 h after the training program. The HIT shock microcycle induced a large decrease in CMJ performance (ACT: effect size [ES] = -1.39, P < .05; PAS: ES = -1.42, P < .05) and perceived recovery (ACT: ES = -1.79, P < .05; PAS: ES = -2.39, P < .05), as well as a moderate to large increase in CK levels (ACT: ES = 0.76, P > .05; PAS: ES = 0.81, P >.05), DOMS (ACT: ES = 2.02, P < .05; PAS: ES = 2.17, P < .05), and perceived stress (ACT: ES = 1.98, P < .05; PAS: ES = 3.06, P < .05), compared with the values before the intervention. However, no significant recovery intervention × time interactions or meaningful differences in changes were noted in any of the markers between ACT and PAS. Repeated use of individualized ACT, consisting of 15 min of moderate jogging, after finishing each training session during an HIT shock microcycle did not affect exercise-induced fatigue.


Physical fitness and performances of an amputee cycling world champion: a case study.
To describe the physical fitness of a top-level lower limb amputee (LLA) cyclist and paracycling time-trial (TT) race demands. The 40-y-old male unilateral transfemoral amputee TT World Champion was tested in a laboratory for peak oxygen uptake (VO2peak), ventilatory threshold (VT2), power output (PO), and hemoglobin mass (Hb-mass). Moreover, several measures (eg, PO, heart rate [HR], cadence) were collected during 4 international TT competitions in the same season. The races' intensity was evaluated as time spent below, at, or above VT2. The cyclist (1.73 m, 55.0 kg) had a VO2peak of 3.372 L/min (61.3 mL·kg(-1)·min(-1)). The laboratory peak PO was 315 W (5.7 W/kg). The maximal HR was 208 beats/min, and his Hb-mass was 744 g (13.5 g/kg). The TTs were meanly 18±4.5 km in length, and the mean PO was 248±8 W with a cadence of 92±1 rpm. During the TTs, the cyclist spent 23%±9% of total time at VT2, 59%±10% below, and 18%±5% above this intensity. The subject's relative VO2peak is higher than previously published data on LLA, and surprisingly it is even higher than "good" ACSM normative data for nondisabled people. The intensity of the races was found to be similar to cycling TTs of the same duration in elite female cyclists. These results might be useful to develop specific training schedules and enhance performance of LLA cyclists.


Redispersible Spray-Dried Powder Containing Nanoencapsulated Curcumin: the Drying Process Does Not Affect Neuroprotection In vitro.
A redispersible spray-dried formulation containing curcumin-loaded, lipid-core nanocapsules (LNC-C) was developed for oral administration. The neuroprotective activity of curcumin after the spray-drying process was evaluated in vitro. The spray-dried powder (SD-LNC-C) was produced using a drying adjuvant composed of a blend of maltodextrin and L-leucine (90:10 w/w). Acceptable process yield (~ 70%) and drug content (6.5 ± 0.2 mg g-1) were obtained. SD-LNC-C was formed by smooth, spherical-shaped particles, and confocal Raman analysis indicated the distribution of the LNC-C on the surface of the leucine/maltodextrin agglomerates. The surface of the agglomerates was formed by a combination of LNC-C and adjuvants, and laser diffraction showed that SD-LNC-C had adequate aqueous redispersion, with no loss of controlled drug release behaviour of LNC-C. The in vitro curcumin activity against the lipopolysaccharide (LPS)-induced proinflammatory response in organotypic hippocampal slice cultures was evaluated. Both formulations (LNC-C and SD-LNC-C) reduced TNF-α to similar levels. Therefore, neuroprotection of curcumin in vitro may be improved by nanoencapsulation followed by spray-drying, with no loss of this superior performance. Hence, the redispersible spray-dried powder proposed here represents a suitable approach for the development of innovative nanomedicines containing curcumin for the prevention/treatment of neurodegenerative diseases.


Insomnia in patients with depression: a STAR*D report.
Insomnia symptoms, which are common in depression, have a significant impact on function and quality of life. However, little is known about the prevalence and associated features of insomnia symptoms in representative treatment-seeking patients with depression. Data from the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) trial were analyzed. STAR*D recruited 3,743 adult outpatients diagnosed with nonpsychotic major depressive disorder (MDD) from primary (n=18) and psychiatric care (n=23) clinics across the United States. Baseline sociodemographic and clinical features were compared between those with insomnia symptoms (84.7%) and those without (15.3%). The most common presentation was the simultaneous presence of sleep onset, mid-nocturnal, and early morning insomnia symptoms (27.1%). Of these three types of insomnia symptoms, mid-nocturnal insomnia symptoms were the most commonly found alone (13.5%) and in combination with one or more other types (82.3%). Insomnia symptoms were associated with several indicators of a more severe depressive illness. Only a small proportion of participants with insomnia symptoms were receiving treatment for sleep disturbances at study initiation, and the vast majority of those receiving treatment still reported having insomnia symptoms. In outpatients who seek treatment for nonpsychotic MDD in typical clinical settings, insomnia symptoms are very common, undertreated, and indicative of a more severe depression.


Histone Variant and Cell Context Determine H3K27M Reprogramming of the Enhancer Landscape and Oncogenic State.
Development of effective targeted cancer therapies is fundamentally limited by our molecular understanding of disease pathogenesis. Diffuse intrinsic pontine glioma (DIPG) is a fatal malignancy of the childhood pons characterized by a unique substitution to methionine in histone H3 at lysine 27 (H3K27M) that results in globally altered epigenetic marks and oncogenic transcription. Through primary DIPG tumor characterization and isogenic oncohistone expression, we show that the same H3K27M mutation displays distinct modes of oncogenic reprogramming and establishes distinct enhancer architecture depending upon both the variant of histone H3 and the cell context in which the mutation occurs. Compared with non-malignant pediatric pontine tissue, we identify and functionally validate both shared and variant-specific pathophysiology. Altogether, we provide a powerful resource of epigenomic data in 25 primary DIPG samples and 5 rare normal pediatric pontine tissue samples, revealing clinically relevant functional distinctions previously unidentified in DIPG.


β-Cyclodextrin based Pickering emulsions for α-tocopherol delivery: Antioxidation stability and bioaccessibility.
β-Cyclodextrin (β-CD) Pickering emulsion and cinnamaldehyde/β-cyclodextrin (CIN/β-CD) Pickering emulsion were prepared and the influences of oxidation and digestion were investigated. CIN/β-CD composite was better dispersed at the oil-water interface than β-CD. Hydrophobic group of CIN anchored in the oil phase and Hydrophilic hydroxyl group of β-CD extended into the aqueous phase, which allowed CIN/β-CD composite to be oriented at the oil-water interface and formed a more stable oil-water interface layer. β-CD Pickering emulsion was more susceptible to oxidative deterioration than CIN/β-CD Pickering emulsion, its malondialdehyde (MDA) value was as high as 509.41 ± 9.37 nmol/L. Digestion experiment indicated that CIN/β-CD Pickering emulsion was released inner oil phase in the small intestine and free fatty acid (FFA) release rate was 44.32 ± 1.08%. Pharmacokinetic parameters manifested that α-tocopherol peak concentration (Cmax) was 64.32 ± 6.45 mg/L and the peak time (Tmax) appeared at 5 h after administration of CIN/β-CD Pickering emulsion.


Pulmonary aspergillosis as opportunistic mycoses in a cohort of human immunodeficiency virus-infected patients: Report from a tertiary care hospital in North India.
The incidence of pulmonary aspergillosis in human immunodeficiency virus (HIV)-infected persons is rising. This study was designed to determine the prevalence of pulmonary aspergillosis in a cohort of HIV-positive patients (n = 71) presenting with lower respiratory tract infection at a tertiary care medical center in India. Sputum samples were collected, and potassium hydroxide mount, cultural characteristics, and lactophenol cotton blue preparations were employed to aid in the identification of Aspergillus species. In addition, serum galactomannan antigen testing was also performed. Pulmonary aspergillosis was diagnosed in 7 patients, five of whom showed a positive antigenemia indicating invasive form of disease. The prevalence of pulmonary aspergillosis was highest in individuals 21-40 years of age (13.3%). The gender-wise prevalence of pulmonary aspergillosis was 18.7% and 7.7% in females and males, respectively. The common chest radiographic findings noted in patients with pulmonary aspergillosis included a normal chest radiograph in 3 (42.8%), infiltrates in 2 (28.6%), and pleural effusion in 2 (28.6%). The common Aspergillus species recovered from sputa of these patients were Aspergillus flavus (4; 57.1%); Aspergillus fumigatus (2; 28.6%), and Aspergillus niger (1; 14.3%). A predisposing lung condition in the form of pulmonary tuberculosis was identified in 2; Pneumocystis carinii pneumonia in 2 and a dual tubercular and P. carinii infection in one. The mean CD4 count of these patients was 155.86 ± 119.33 cells/µl (median = 117 cells/µl; range = 18-329 cells/µl). Our findings suggest that Aspergillus species be considered possible etiological agents in HIV-positive patients with pulmonary infection.


The composition of animal cells: solutes contributing to osmotic pressure and charge balance.
The cytoplasmic solutes of vertebrates and invertebrates, other than Na, K and Cl, are surveyed in relation to their influence on ionic regulation through osmolality and charge balance. The most abundant include MgATP, phosphagens, amino acids, various other nitrogen and phosphorus compounds and sometimes anaerobic end products and antifreeze agents. Differences in muscle osmolality, e.g. between marine and non-marine animals, affect mainly nitrogenous solutes of no net charge, such as certain amino acids, taurine, betaine, trimethylamine oxide and urea. The high osmolality of axoplasm in marine invertebrates is due more to anions such as aspartate, glutamate and isethionate.


Bcl-3 expression promotes cell survival following interleukin-4 deprivation and is controlled by AP1 and AP1-like transcription factors.
We have analyzed the interleukin-4 (IL-4)-triggered mechanisms implicated in cell survival and show here that IL-4 deprivation induces apoptotic cell death but does not modulate Bcl-2 or Bcl-x expression. Since Bcl-x expression is insufficient to ensure cell survival in the absence of IL-4, we speculate that additional molecules replace the antiapoptotic role of Bcl-2 and Bcl-x in an alternative IL-4-triggered pathway. Cell death is associated with Bcl-3 downregulation and Bcl-3 expression blocks IL-4 deprivation-induced apoptosis, suggesting that Bcl-3 acts as a survival factor in the absence of growth factor. To characterize the IL-4-induced regulation of murine Bcl-3 expression, we cloned the promoter of this gene. Sequencing of the promoter showed no TATA box element but did reveal binding sites for AP1, AP1-like, and SP1 transcription factors. Retardation gels showed that IL-4 specifically induces AP1 and AP1-like binding activity and that mutation of these binding sites abolishes the IL-4-induced Bcl-3 promoter activity, suggesting that these transcription factors are important in Bcl-3 promoter transactivation. IL-4 deprivation induces downregulation of Jun expression and upregulation of Fos expression, both of which are proteins involved in the formation of AP1 and AP1-like transcription factors. Overexpression of Jun family proteins transactivates the promoter and restores Bcl-3 expression in the absence of IL-4 stimulation. Taken together, these data describe a new biological role for Bcl-3 and define the regulatory pathway implicated in Bcl-3 expression.


RPB2 gene phylogeny in flowering plants, with particular emphasis on asterids.
Two, apparently functional, paralogues of the RPB2 gene, which encodes the second largest subunit of RNA polymerase II, are shown to be present in two major groups of asterid plants. Although all other land plants surveyed so far have been found to have only one of these two copies, the RPB2 gene phylogeny inferred from the 3' half of the gene for 35 angiosperm taxa and six other land plants indicates that the duplication of the RPB2 gene occurred earlier than the time for origin of the asterid group, probably near the origin of "core eudicots." The d copy is present in all plants which are unambiguously assigned to the core eudicots, whereas the I copy is retained only in the lamiid clade, Ericales, and Escallonia, all belonging to the asterid group of plants. Both parsimony and likelihood analyses of sequences from the 3' half of the gene give strong bootstrap support for these conclusions. There is no support for monophyly of the taxa having both copies. Thus, numerous losses of one of the copies must be inferred. Structurally, both paralogues appear functional, and transcription is demonstrated for both copies. In the lamiid group, the d copy has lost introns 18-23. The well supported phylogenetic relationships implied by the RPB2 gene phylogeny are largely congruent with well supported phylogenetic hypotheses based on other sequence data. However, Ilex, usually assigned to the campanuliid clade, is instead supported as being a member of the lamiid clade, both from sequence data and the presence of an I copy as well as the loss of introns 18-23 in the d copy. Escallonia, supported as a member of the campanuliid clade both by RPB2-d-sequences and previously published DNA sequence data, has all the introns 18-23 in its d copy, as do all other members studied from the campanuliid group. We used the Markov Chain Monte Carlo (MCMC) approach of the MrBayes program to implement Maximum Likelihood bootstrapping. Under the same model of evolution, bootstrapping frequencies are significantly lower than the Bayesian posterior probabilities inferred from the MCMC chain.


Health care providers' knowledge of, attitudes toward and provision of emergency contraceptives in Lagos, Nigeria.
Emergency contraception can play an important role in reducing the rate of unintended pregnancies in Nigeria. Although it is included in the national family planning guidelines, there is limited awareness of this method among clients. In 2003-2004, a sample of 256 health care providers within Lagos State were surveyed about their knowledge of, attitudes toward and provision of emergency contraceptives, using a 25-item, self-administered questionnaire. Frequencies were calculated for the various measures, and chi-square tests were used to determine significant differences. Nine in 10 providers had heard of emergency contraception, but many lacked specific knowledge about the method. Only half of them knew the correct time frame for effective use of emergency contraceptive pills, and three-fourths knew that the pills prevent pregnancy; more than a third incorrectly believed that they may act as an abortifacient. Fewer than a third of respondents who had heard of the pills knew that they are legal in Nigeria. Of those who had heard about emergency contraception, 58% had provided clients with emergency contraceptive pills, yet only 10% of these providers could correctly identify the drug, dose and timing of the first pill in the regimen. Furthermore, fewer than one in 10 of those who knew of emergency contraception said they always provided information to clients, whereas a fourth said they never did so. Nigerian health care providers urgently need education about emergency contraception; training programs should target the types of providers who are less knowledgeable about the method.


ANOVA model for network meta-analysis of diagnostic test accuracy data.
Procedures combining and summarising direct and indirect evidence from independent studies assessing the diagnostic accuracy of different tests for the same disease are referred to network meta-analysis. Network meta-analysis provides a unified inference framework and uses the data more efficiently. Nonetheless, handling the inherent correlation between sensitivity and specificity continues to be a statistical challenge. We developed an arm-based hierarchical model which expresses the logit transformed sensitivity and specificity as the sum of fixed effects for test, correlated study-effects to model the inherent correlation between sensitivity and specificity and a random error associated with various tests evaluated in a given study. We present the accuracy of 11 tests used to triage women with minor cervical lesions to detect cervical precancer. Finally, we compare the results with those from a contrast-based model which expresses the linear predictor as a contrast to a comparator test. The proposed arm-based model is more appealing than the contrast-based model since the former permits more straightforward interpretation of the parameters, makes use of all available data yielding shorter credible intervals, and models more natural variance-covariance matrix structures.


Rabies postexposure management of travelers presenting to travel health clinics in Auckland and Hamilton, New Zealand.
Rabies is a fatal disease, and travelers going to endemic areas need to take precautions. Little is known about the rabies postexposure management of travelers from New Zealand. A total of 459 post-travel records from October 1998 until February 2006 at two travel medicine clinics, in Auckland and Hamilton, were examined for those who reported postexposure management to animals while abroad. Fifty-four patients were included, 48 (88.9%) were New Zealand residents and 52.0% were male. The mean age of exposed travelers was 30.4 years (SD = 15.5). There was an adult to child ratio of 5:1. The highest exposure risk group was those aged 16 to 30 years. South and Southeast Asia were the most prominent geographical regions where exposure occurred, with 45 (83.3%) of subjects being potentially exposed to rabies. Dogs were the commonest animals involved, accounting for two thirds of incidents (36; 66.7%). The commonest sites of animal exposure on the body were the thigh and lower leg (26; 48.1%) and the hand (10; 18.5%). Forty-six (85.2%) of the animal exposures were graded as World Health Organization (WHO) category III. Forty-nine (90.7%) of the travelers had not had preexposure prophylaxis. Once in New Zealand, the correct WHO postexposure prophylaxis regime was applied on 44 of 52 (84.6%) occasions. However, overall, only 25% of the sample received postexposure treatment consistent with WHO guidelines, reflecting inappropriate management abroad. Post-travel consultations at two New Zealand travel clinics were analyzed for prophylactic rabies postexposure management. The majority were travelers aged 16 to 30 years, who sustained WHO category III exposures to the lower limb in Asia, predominantly from dogs. Few of these travelers had been immunized prior to travel, and only 25% of them received postexposure prophylaxis consistent with WHO guidelines. Thus, 75% of the study sample remains at theoretical risk of contracting rabies due to inappropriate management overseas.


[NMR screening in fragment-based drug discovery].
Nuclear magnetic resonance (NMR) is a versatile technique for the pharmaceutical industry. From organic chemistry to MRI, there are a number of applications of NMR. Among them, biomolecular NMR has been used for structure determination of biomolecules and analyzing the interaction between a target protein and its inhibitors. In the context of fragment-based drug discovery (FBDD), NMR has been known as a fragment screening technique, because NMR is good at detecting a weak binding compound in an accurate manner. Generally, the NMR technique for fragment screening is classified into two families: the ligand-based technique and the protein-based technique. The latter technique requires stable isotope labeled protein and also can be applied to a relatively small MW protein target. In the ligand-based technique such as saturation transfer difference (STD) and WaterLOGSY, only the NMR signals of the ligands are observed. The disadvantage of STD and WaterLOGSY is that the non-specific binding is also observed and a competition experiment is required in order to select the specific binding compound. Due to the difference in the consumption of the protein sample, the ligand-based technique has generally been used recently as a primary screening.


Mechanisms of in vitro resistance to dihydroartemisinin in Plasmodium falciparum.
The recent reports of artemisinin (ART) resistance in the Thai-Cambodian border area raise a serious concern on the long-term efficacy of ARTs. To elucidate the resistance mechanisms, we performed in vitro selection with dihydroartemisinin (DHA) and obtained two parasite clones from Dd2 with more than 25-fold decrease in susceptibility to DHA. The DHA-resistant clones were more tolerant of stressful growth conditions and more resistant to several commonly used antimalarial drugs than Dd2. The result is worrisome as many of the drugs are currently used as ART partners in malaria control. This study showed that the DHA resistance is not limited to ring stage, but also occurred in trophozoites and schizonts. Microarray and biochemical analyses revealed pfmdr1 amplification, elevation of the antioxidant defence network, and increased expression of many chaperones in the DHA-resistant parasites. Without drug pressure, the DHA-resistant parasites reverted to sensitivity in approximately 8 weeks, accompanied by de-amplification of pfmdr1 and reduced antioxidant activities. The parallel decrease and increase in pfmdr1 copy number and antioxidant activity and the up and down of DHA sensitivity strongly suggest that pfmdr1 and antioxidant defence play a role in in vitro resistance to DHA, providing potential molecular markers for ART resistance.


The relationship between asthma and smoking during pregnancy.
Asthma is the most common and potentially serious medical problem encountered in pregnancy. The prevalence of asthma during pregnancy is generally quoted as being between 1% and 4% which is much lower than general population estimates. The aim of this analysis was to estimate the prevalence of asthma and smoking during pregnancy; and to describe the relationship between the two. A cross-sectional random sample survey of recently delivered Western Australian mothers was carried out in 1995 and 82% of mothers responded. In total 21.3% of women reported ever having asthma at some stage in their life and 12.4% currently had asthma. These results are consistent with general population estimates. Overall 22.7% of women smoked at some stage in pregnancy, which is consistent with the limited comparison data available. Current asthmatics were more likely to smoke during pregnancy than non-asthmatics, although this difference was not statistically significant. Continued vigorous efforts are required to discourage all women and girls from taking up smoking and to encourage those who smoke to quit. This is particularly pertinent for asthmatic women who are pregnant or planning a pregnancy.


Priming protective CD8 T cell immunity by DNA vaccines encoding chimeric, stress protein-capturing tumor-associated antigen.
DNA vaccines encoding heat shock protein (hsp)-capturing, chimeric peptides containing antigenic determinants of the tumor-associated Ag (TAA) gp70 (an envelope protein of endogenous retrovirus) primed stable, specific, and tumor-protective CD8 T cell immunity. Expression of gp70 transcripts was detectable in most normal tissues but was particularly striking in some (but not all) tumor cell lines tested (including the adenocarcinoma cell line CT26). An approximately 200 residue gp70 fragment or its L(d)-binding antigenic AH1 peptide cloned in-frame behind an hsp-capturing (cT(272)) or noncapturing (T(60)) N-terminal large SV40 tumor Ag sequence was expressed as either hsp-binding or -nonbinding chimeric Ags. Only hsp-capturing, chimeric fusion proteins were expressed efficiently in transfected cell lines and primed TAA-specific CD8 T cell immunity. This immunity mediated protection in the CT26 and mKSA models. A vaccination strategy based on delivering antigenic, hsp-associated TAA fragments can thus prime protective CD8 T cell immunity even if these TAA are of low intrinsic immunogenicity.


Movement Disorders and Liver Disease.
The association of movement disorders with structural or functional hepatic disease occurs in three principal scenarios: (1) combined involvement of both organ systems from a single disease entity, (2) nervous system dysfunction resulting from exposure to toxic compounds in the setting of defective hepatic clearance, or (3) hepatic and/or neurological injury secondary to exposure to exogenous drugs or toxins. An important early step in the workup of any patient with combined movement disorders and liver disease is the exclusion of Wilson's disease. Diagnostic delay remains common for this treatable disorder, and this has major implications for patient outcomes. Thereafter, a structured approach integrating variables such as age of onset, tempo of progression, nature and severity of liver involvement, movement disorder phenomenology, exposure to drugs/toxins and laboratory/neuroimaging findings is key to ensuring timely diagnosis and disease-specific therapy. Herein, we provide an overview of disorders which may manifest with a combination of movement disorders and liver disease, structured under the three headings as detailed above. In each section, the most common disorders are discussed, along with important clinical pearls, suggested diagnostic workup, differential diagnoses and where appropriate, treatment considerations.


The significance of isolated elevation of serum aminotransferases in infants and young children.
The aim of this study was to assess the clinical significance and prognosis of a prolonged isolated elevation of serum aminotransferases without cholestasis (>3 months) in infants and young children, investigated for a variety of conditions, and to determine a protocol for their follow-up and investigation. A combined prospective-retrospective analysis of apparently healthy babies and young children with isolated elevation of serum aminotransferases of at least 1.5 times above the norm for age which persisted for at least 3 months and whose creatine phosphokinase (CK), gamma glutamyltransferase (GGT), alkaline phosphatase and bilirubin levels remained normal throughout the study duration. The children underwent the following investigations: abdominal ultrasound and infectious, metabolic and/or immunological investigation depending on the duration of the abnormality. Six children were eliminated following the finding of positive cytomegalovirus (CMV) antigen in the urine. 72 children were investigated (47 males and 25 females). The duration of serum aminotransferases elevation was 3-36 months (average 12.4, median 11.5 months). The initial, maximal and final alanine aminotransferase (ALT) values were 85.5, 140.5 and 39.8 IU/l, respectively. Of seven children who had liver biopsies performed, three (42.8%) were suspected of having a glycogen storage disease which was not confirmed enzymatically. Four biopsies revealed non-specific histological changes. Isolated elevation of serum aminotransferases in healthy looking young children is mostly a benign condition that usually resolves within a year. If no pathology is found during routine investigation, these children can be followed conservatively. Liver biopsy does not contribute much to the diagnosis and is probably unnecessary.


Impact of the CFTR-potentiator ivacaftor on airway microbiota in cystic fibrosis patients carrying a G551D mutation.
Airway microbiota composition has been clearly correlated with many pulmonary diseases, and notably with cystic fibrosis (CF), an autosomal genetic disorder caused by mutation in the CF transmembrane conductance regulator (CFTR). Recently, a new molecule, ivacaftor, has been shown to re-establish the functionality of the G551D-mutated CFTR, allowing significant improvement in lung function. The purpose of this study was to follow the evolution of the airway microbiota in CF patients treated with ivacaftor, using quantitative PCR and pyrosequencing of 16S rRNA amplicons, in order to identify quantitative and qualitative changes in bacterial communities. Three G551D children were followed up longitudinally over a mean period of more than one year covering several months before and after initiation of ivacaftor treatment. 129 operational taxonomy units (OTUs), representing 64 genera, were identified. There was no significant difference in total bacterial load before and after treatment. Comparison of global community composition found no significant changes in microbiota. Two OTUs, however, showed contrasting dynamics: after initiation of ivacaftor, the relative abundance of the anaerobe Porphyromonas 1 increased (p<0.01) and that of Streptococcus 1 (S. mitis group) decreased (p<0.05), possibly in relation to the anti-Gram-positive properties of ivacaftor. The anaerobe Prevotella 2 correlated positively with the pulmonary function test FEV-1 (r=0.73, p<0.05). The study confirmed the presumed positive role of anaerobes in lung function. Several airway microbiota components, notably anaerobes (obligate or facultative anaerobes), could be valuable biomarkers of lung function improvement under ivacaftor, and could shed light on the pathophysiology of lung disease in CF patients.


Hepatic prolyl hydroxylase activity in human liver disease.
The activity of prolyl hydroxylase was measured in liver tissue obtained from a small series of patients with a variety of liver disease. Enzyme levels were marginally elevated in patients with fatty liver and viral hepatitis, conditions not normally associated with progressive fibrosis. In some patients with alcoholic hepatitis and in all patients with cirrhosis and chronic active hepatitis, there was a marked increase in enzyme activity. Patients with conditions characterised by high liver prolyl hydroxylase levels showed histological evidence of extensive hepatic fibrosis and also significant increases in the serum values of glutamate dehydrogenase and gamma-glutamyl-transpeptidase. Prolyl hydroxylase activity was not detected in serum.


Intravenous secretin for autism spectrum disorder.
Secretin is a gastro-intestinal hormone which has been presented as an effective treatment for autism based on anecdotal evidence. To determine if intravenous secretin:1. improves the core features of autism (social interaction, communication and behaviour problems); 2. improves the non-core aspects of behaviour or function such as self injurious behaviour;3. improves the quality of life of affected individuals and their carers; 4. has short term and long term effects on outcome; 5. causes harm. Results of electronic searches of CENTRAL, MEDLINE, EMBASE, PsycINFO, CINAHL, ERIC, HealthStar and Sociofile (1998 - March 2005) were independently examined by two authors. Reference lists of trials and reviews were searched; experts and trialists were contacted to find unpublished studies. Randomised controlled trials of intravenous secretin comparing secretin with a placebo treatment in children or adults diagnosed with autism spectrum disorders, where at least one standardised outcome measure was reported. Fourteen studies met inclusion criteria. All outcome data were continuous. Where trials used cross-over designs, analysis was conducted on results from first treatment phase, allowing combined analysis with parallel design trials. Where standardised assessment tools generated scores as outcome measures, comparisons were made between means of these scores. Where baseline means were reported, differences between treatment and control were determined to assess possible bias. Where mean change from baseline was reported, this was used in preference to post-treatment scores for meta-analyses or forest plots. As meta-analysis was possible for only one outcome (Childhood Autism Rating Scale), it was impossible to use sensitivity or subgroup analyses to assess impact of study quality, clinical differences in the intervention, or clinically relevant differences between groups, such as age or presence of gastrointestinal symptoms. Twenty-five established standardised outcome measures were reported to assess core features of autism, communication, behaviour, visio-spatial skills, affect and adverse events within fourteen included studies. No more than four studies used any one outcome measure similarly. Outcomes were reported between three and six weeks. RCTs of efficacy of secretin in autism have not shown improvements for core features of autism. There is no evidence that single or multiple dose intravenous secretin is effective and as such it should not currently be recommended or administered as a treatment for autism. Further experimental assessment of secretin's effectiveness for autism can only be justified if methodological problems of existing research can be overcome.


Pathologic paediatric conditions associated with a compromised airway.
The purpose was to describe pathologic paediatric conditions associated with airway compromise adversely affecting dental treatment with sedation and general anaesthesia. A review of available literature was completed, identifying pathologic paediatric conditions predisposing to airway compromise. Airway-related deaths are uncommon, but respiratory complication represents the greatest cause of morbidity and mortality during the administration of general anaesthesia. Differences in anatomy and physiology of the paediatric and adult airway contribute to the child's predisposition to rapid development of airway compromise and respiratory failure; juvenile rheumatoid arthritis, cervical spine injury, morbid obesity, and prematurity represent only a few conditions contributing to potential airway compromise of which the paediatric clinician needs to be aware. In all cases, thorough physical examination prior to treatment is mandated to affect a positive treatment outcome. Successful management of children and adolescents with a compromised airway begins with identification of the problem through a detailed medical history and physical examination. Due to the likely fragile nature of many of these patients, and possibility of concomitant medical conditions affecting airway management, dental treatment needs necessitating pharmacological management are best treated in a controlled setting such as the operating room, where a patent airway can be maintained.


SlbHLH068 interacts with FER to regulate the iron-deficiency response in tomato.
Iron is an essential micronutrient for all organisms and its uptake, translocation, distribution and utilization are regulated in a complex manner in plants. FER, isolated from tomato (Solanum lycopersicum), was the first transcription factor involved in the iron homeostasis of higher plants to be identified. A FER defect in the T3238fer mutant drastically downregulates the expression of iron uptake genes, such as ferric-chelate reductase 1 (LeFRO1) and iron-regulated transporter 1 (LeIRT1); however, the molecular mechanism by which FER regulates genes downstream remains unknown. The aim of this work was therefore to identify the gene that interacts with FER to regulate the iron-deficiency response in tomato. The homologue of the Arabidopsis Ib subgroup of the basic helix-loop-helix (bHLH) proteins, SlbHLH068, was identified by using the program BLASTP against the AtbHLH39 amino acid sequence in the tomato genome. The interaction between SlbHLH068 and FER was detected using yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. In addition, virus-induced gene silencing (VIGS) was used to generate tomato plants in which SlbHLH068 expression was downregulated. The expression of genes was analysed using northern blot hybridization and multiple RT-PCR analysis. Seedlings of wild-type and mutant plants were grown under conditions of different nutrient deficiency. SlbHLH068 is highly upregulated in roots, leaves and stems in response to iron deficiency. An interaction between SlbHLH068 and FER was demonstrated using yeast two-hybrid and BiFC assays. The heterodimer formed by FER with SlbHLH068 directly bound to the promoter of LeFRO1 and activated the expression of its reporter gene in the yeast assay. The downregulation of SlbHLH068 expression by VIGS resulted in a reduction of LeFRO1 and LeIRT1 expression and iron accumulation in leaves and roots. The results indicate that SlbHLH068, as a putative transcription factor, is involved in iron homeostasis in tomato via an interaction with FER.


Trichophytobezoars in miniature swine.
Miniature swine confined indoors on concrete developed gastric trichophytobezoars that resulted in gradual weight loss and debilitation. The ability to maintain the swine on a chronic research project was threatened by premature deaths associated with the trichophytobezoar formation. The predisposing husbandry practices and history of the development of the trichophytobezoars included confinement for several years and consumption of pelleted feed directly from the concrete floor. Gastrotomy performed on 3 sows and 5 boars avoided the progression of complications leading toward starvation and resulted in clinical improvement.


[Assessment of the economic impact of the antiretroviral vaccine in Venezuela].
To assess the cost of medical care for rotavirus gastroenteritis and the cost-effectiveness of the antiretroviral vaccine in Venezuelan children under five. We used an economic model that comprises epidemiologic information, vaccine efficacy, and the cost of medical care in connection with rotavirus gastroenteritis, viewed from a social perspective. In order to determine the effectiveness of the vaccine, we estimated the number of hospitalized cases, of medical visits, and of deaths averted after vaccination. The cost-effectiveness of the vaccine was determined on the basis of the number of disability-adjusted life years (DALYs) and cases averted. In Venezuela, health services spend approximately 4.2 million US$ yearly on covering the costs of medical care for rotavirus-related disease. In a vaccinated cohort, an antiretroviral vaccination program would prevent around 52% (186) of the deaths, 54% (7,232) of the hospitalizations, and 50% (55,168) of the ambulatory visits that take place during the first five years of life. For an estimated cost of approximately 24 US$ per individual vaccination schedule, the cost-effectiveness ratio obtained is 1,352 US$ per DALY. The results of this study suggest that antiretroviral vaccination is a cost-effective strategy for preventing rotavirus gastroenteritis in Venezuela, since it can prevent deaths and DALYs in the population under five years of age.


The effect of iatrogenic vas deferens injury on fertility in an experimental rat model.
The long-term result of accidental crushing of the vas deferens during inguinal surgery is unpredictable. It is stated that even a slight disturbance in the muscular layer or mucosa may endanger fertility. This study was designed to investigate the early and late histopathologic changes of iatrogenic injury on the vas and its effect on fertility in a rat model. Both vasa deferentia of 54 male rats, divided into three groups (n = 18 each), were subjected to an operative manipulation. These manipulations consisted of digital compression for 45 seconds in group A, grasping with a mosquito clamp for 2 seconds in group B, and grasping with a mosquito clamp for 2 minutes in group C. In five rats from each group, the vasa were removed on the third and 21st postoperative day for histological evaluation. The remaining 13 rats in each group were allowed to mate for a period of 4 months. Bilateral vasa were analyzed for patency and histology. A spermatic granuloma was observed in 79% of the fertile and 80% of the infertile rats. Statistical analysis showed no significant fertility difference among the three groups. There was no statistical difference between groups B and C (P > .05) in terms of high and low flow patency rates in vitro, whereas a statistical significance was present between these two groups and group A (P< .001). It can be said that type rather than time of injury is important in this experimental model, because the results of the patency test alone are sufficient to prove the possibility of functional damage in vas deferens. The authors conclude that inguinal contents should be handled carefully and contralateral exploration in asymptomatic cases older than 1 year with inguinal pathology should be avoided.


Recombinant latent transforming growth factor beta 1 has a longer plasma half-life in rats than active transforming growth factor beta 1, and a different tissue distribution.
Transforming growth factor beta 1 (TGF-beta 1) is a key regulator of cell growth and differentiation. Under normal physiological conditions, it is made as a biologically latent complex whose significance is unknown. Previous work has indicated that active TGF-beta 1 has a very short plasma half-life in rats (Coffey, R. J., L. J. Kost, R. M. Lyons, H. L. Moses, and N. F. La-Russo. 1987. J. Clin. Invest. 80:750-757). We have investigated the possibility that latent complex formation may extend the plasma half-life of TGF-beta 1 and alter its organ distribution. Radiolabeled latent TGF-beta 1 was formed by noncovalent association of 125I-TGF-beta 1 with the TGF-beta 1 precursor "pro" region from recombinant sources. TGF-beta 1 in this latent complex had a greatly extended plasma half-life (greater than 100 min) in rats compared with active TGF-beta 1 (2-3 min). Whereas active TGF-beta 1 was rapidly taken up by the liver, kidneys, lungs, and spleen and degraded, TGF-beta 1 in the latent complex was largely confined to the circulation, and was less than 5% degraded after 90 min. The pharmacokinetics of TGF-beta 1 in the latent complex were shown to be critically dependent on the degree of sialylation of the complex. The results suggest that formation of latent complexes may switch endogenous TGF-beta 1 from an autocrine/paracrine mode of action to a more endocrine mode involving target organs distant from the site of synthesis.


[Use of the hormonal preparation prephyson in the treatment of epilepsy in children and adolescents].
Children and adolescents suffering from epilepsy were treated with anticonvulsants and prephyson. The paroxysmal and psychopathological symptomatology showed positive changes, more remarkable as compared to those seen as a result of administering anticonvulsants alone, accompanied by the improvement of endocrine system function. The characteristic features of the clinical symptomatology and course of epilepsy requiring the use of hormonal drugs were defined.


The Effect of Carbohydrate-Restricted Diets on the Skin Aging of Mice.
Low-carbohydrate, high-protein diets, known as carbohydrate-restricted diets, are in contrast to the carbohydrate-centric meals typical of the Japanese diet. Carbohydrate-restricted diets were reported to reduce visceral fat, owing to which they have attracted attention and been widely implemented. If, as proposed, carbohydrate-restricted diets are effective in delaying senescence, then Japanese diets have a hidden potential to evolve further. However, long-term carbohydrate restriction in mice was reported to have a negative effect on the cardiovascular system, with shortening of lifespan due to activation of mechanistic target of rapamycin (mTOR). As a result, the safety of long-term adherence to carbohydrate-restricted diets remains doubtful. Recently, we conducted a study using senescence-accelerated mouse-prone 8 (SAMP8) mice to examine the effects of a carbohydrate-restricted diet on aging and skin senescence, and to determine the effect of long-term carbohydrate restriction on the aging process in mice. Here, we discuss the safety of long-term carbohydrate restriction based on the findings obtained from animal studies.


[Ethical aspects of human lactation and breastfeeding].
Breastfeeding has experienced a surge in Puerto Rico thanks to the efforts of many women, as well as governmental, non-governmental and private organizations. Among the aspects which can affect the initiation, establishment and maintenance of successful breastfeeding one finds potential ethical conflicts among health professionals as well as the financial interests of the companies which produce and market artificial milks. These companies dedicate great marketing efforts and enormous sums of money in the sale of their products, in detriment of breastfeeding. The profit motive of these companies is in open contradiction to public health and to breastfeeding promotion. The international community has carried out efforts for more than two decades in the creation of structures which protect the rights of women and their children to ideal nutrition practices, free from commercial pressures, through breastfeeding. Possible ethical conflicts related to the new industry of breastfeeding paraphernalia are discussed, especially when these are marketed exclusively for profit. Recommendations are presented for health professionals in order for them to be able to face and respond to the unethical strategies practiced by the manufacturers of artificial milks in utilizing these professionals for legitimating and marketing their products.


Carotenoid transformations in the chloroplast envelope.
The envelope of the spinach chloroplast is a yellow membrane system with a unique carotenoid composition. Envelopes prepared from dark-treated leaves had a violaxanthin content up to 3.5 times the lutein plus zeaxanthin content, whereas in chloroplast envelopes from illuminated leaves this ratio was only 0.75. Light-catalyzed changes in violaxanthin content also occurred in the thylakoid fraction.The role of this reversible light-catalyzed de-epoxidation of violaxanthin in the function of the envelope of the chloroplast is discussed.


Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) in four calves.
Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA), or Bland-White-Garland syndrome, is a rare congenital malformation described in children and adults. In this condition, the left coronary artery, which normally originates from the left coronary sinus in the aorta, instead originates from the pulmonary trunk, which results in retrograde flow of blood away from the myocardium into the lower-pressure pulmonary artery. Myocardial hypoxic-ischemic injury results in cardiac dysfunction, failure, and eventually in patient death if not surgically repaired. This report describes gross and microscopic findings in 4 beef calves with ALCAPA. All the calves had a history of being found dead with few or no premonitory signs, 2 shortly after sudden strenuous exercise. Gross necropsy lesions suggestive of heart failure included cardiomegaly with atrial and ventricular dilation and/or ventricular hypertrophy, and hepatomegaly. Dissection of each heart revealed the origin of the left coronary artery arising in the pulmonary trunk above the anterior cusp of the pulmonic valve. No degeneration; mineralization; and fiber loss, with replacement by fibrous connective tissue, predominantly in the left ventricular papillary muscle and the interventricular septum. Changes observed in the liver and lungs, including hepatomegaly, sinusoidal congestion, centrilobular fibrosis, and pulmonary congestion, edema, and intra-alveolar pigment-laden macrophages were consistent with heart failure.


Phenolic acids and methylxanthines composition and antioxidant properties of mate (Ilex paraguariensis) residue.
Ilex paraguariensis is known to contain compounds with antioxidant properties, such as phenolic acids, and its stimulant properties are attributed to methylxanthines, such as caffeine. The aims of this study were to evaluate the phenolic, methylxanthinic, and tannin composition of a mate residue (mate powder), to compare the quali-quantitative phenolic composition and the antioxidant potential of extracts obtained from distinct solvent systems. Among the extracts prepared with different solvents, the 80% methanol extract showed the highest total polyphenol content (11.51 g/100 g) and antioxidant activity. HPLC analysis showed that 4,5 dicaffeoylquinic acid is the major component of the phenolic fraction of mate powder. The caffeine, theobromine, and tannin contents in mate powder were 1.01, 0.10, and 0.29 g/100 g, respectively. Consumption of mate powder would significantly contribute to antioxidant and stimulant intake, providing high amounts of phenolic acids, tannins, and methylxanthines with biological effects potentially beneficial for human health. This article contributes to the minimization of residues in yerba-mate processing.


Role of early hormonal and nutritional experiences in shaping feeding behavior and hypothalamic development.
Obesity in adults and children is increasingly becoming a major health problem worldwide. However, the precise biological mechanisms governing this disease have not been fully elucidated. Obesity involves the complex interaction of a wide range of environmental and genetic factors. Additionally, there is now a growing body of evidence suggesting that alterations in metabolic environment during important periods of organ development can predispose individuals to later development of obesity and diabetes. Maternal obesity or malnutrition during pregnancy increases the risk for metabolic disorders (including obesity) in the offspring. Similarly, early postnatal overnutrition also predisposes offspring to adult obesity. The hypothalamus appears to play an essential role in controlling appetite. It undergoes a tremendous growth beginning early in gestation and continuing during the postnatal period. These developmental windows represent periods of sensitivity for hypothalamic development during which alterations in the nutritional and/or hormonal environment may perturb hypothalamic development and subsequent function.


Applications of positron emission tomography/computed tomography image fusion in clinical positron emission tomography-clinical use, interpretation methods, diagnostic improvements.
Positron emission tomography (PET)/computed tomography (CT) scanners with combined dedicated high performance PET and CT scanners have been introduced recently in PET imaging. Oncological imaging with fluorodeoxyglucose (FDG) is currently the dominant application of PET. The addition of CT to PET offers many advantages, including obtaining a fast and relatively low-noise transmission scan, shortening the duration of the examination, adding precise anatomical information to FDG imaging, and providing additional diagnostic information. However, the use of CT for attenuation correction can lead to some artifacts that need to be considered when interpreting a PET/CT study: quantitative measurements may be altered, high density IV and oral and metallic objects may produce artifacts, and the registration of PET and CT may occasionally be suboptimal. Areas where using PET/CT offers particular potential advantages include the head and neck region, abdomen, and pelvis. Even in the thorax, PET/CT offers some advantages. Although clinical data evaluating the added value of PET/CT over PET are presently limited, preliminary results are very encouraging. More studies are warranted to clearly define the clinical impact of PET/CT over PET; however, it is clear this dedicated fusion technology will be very important for patient imaging in the coming years.


Frequency and syndrome specificity of antibodies to aquaporin-4 in neurological patients with rheumatic disorders.
A new autoantibody (termed NMO-IgG, or AQP4-Ab) has recently been described in patients with neuromyelitis optica (NMO) and its formes frustes, longitudinally extensive transverse myelitis (LETM) and recurrent optic neuritis (rON). However, AQP4-Ab has been found also in patients with co-existing rheumatic diseases such as systemic lupus erythematosus (SLE) or Sjögren's syndrome (SS), conditions which are characterized by broad, polyspecific B cell activation. In this study, we aimed at evaluating the syndrome specificity and frequency of AQP4-Ab in patients with rheumatic diseases and neurological symptoms. For this purpose, serum samples from 109 neurological patients with established connective tissue disorders (CTD) (n = 54), possible CTD (n = 42), or vasculitis (n = 13) were analysed for the presence of AQP4-Ab by a cell-based assay employing recombinant human AQP4. AQP4-Ab was detectable in 31/40 (78%) patients with CTD and NMO spectrum disorders (median titre, 1:1000) but in none of the samples obtained from patients with CTD or vasculitis and neurological disorders other than NMO, LETM, or rON (n = 69). The high syndrome specificity of the antibody for neuromyelitis optica spectrum disorders (NMOSDs) in patients with CTD supports the concept of AQP4-Ab being involved in the pathogenesis of these neurological conditions, and argues against AQP4-Ab simply being part of the polyclonal B cell activation generally associated with rheumatic diseases. Moreover, the finding that AQP4-Ab is present in patients with CTD and co-existing NMOSD with approximately the same frequency as in patients without CTD strengthens the case of CTD and AQP4-Ab positive NMOSD representing two co-existing yet distinct entities in the majority of patients.


Improved template representation in cpn60 polymerase chain reaction (PCR) product libraries generated from complex templates by application of a specific mixture of PCR primers.
Some classes of high G+C content organisms such as the Actinobacteria, which are known through culture-based studies to be present in large numbers in particular microbial communities, are under-represented or even absent from 16S rRNA or cpn60 polymerase chain reaction (PCR) product libraries derived from these templates. Using reference cpn60 sequence data from organisms with high G+C content genomes, a pair of PCR primers were designed which, when used in combination with the previously developed degenerate, universal cpn60 primers, improve the representation of templates with high G+C content. The primers were validated using a combination of traditional and quantitative real-time PCR on both manufactured template mixtures and biological samples. The development and optimization of this specific primer mixture represents an improvement of established methods and a significant advance in the ability to generate cpn60 PCR product libraries that more closely represent the sequence diversity in complex templates.


Fluoroquinolone-resistant Campylobacter isolates from conventional and antibiotic-free chicken products.
The use of fluoroquinolones (FQs) in poultry production is an important issue in public health today. In February 2002, two prominent U.S. poultry companies pledged to stop using FQs for flock-wide treatment. One year later, we began a survey of Campylobacter isolates on chicken products from these two companies and from two producers claiming total abstention from antibiotic use. Using both standard isolation methods and new methods modified to enhance detection of FQ-resistant Campylobacter, we compared rates of FQ-resistant Campylobacter among these products. Four major findings were drawn from this study: a) antibiotic-free brands were not more likely to be contaminated with Campylobacter; b) a high percentage of products from the two conventional brands were contaminated with FQ-resistant Campylobacter (43 and 96%); c) these conventional brands had significantly higher odds of carrying resistant strains compared with antibiotic-free products; and d) supplementing media with FQs increased the sensitivity of detecting FQ-resistant strains among mixed populations of Campylobacter, thus reducing a bias toward underestimating the prevalence of FQ-resistant Campylobacter on samples. These results suggest that FQ resistance may persist in the commercial poultry environment in the absence of FQ-selective pressure and that these strains contaminate a larger proportion of foods than reported previously.


EI24, as a Component of Autophagy, Is Involved in Pancreatic Cell Proliferation.
Autophagy is a highly conserved cellular process in which cytoplasmic materials are degraded and recycled as energy sources when nutrient supplies are lacking. Established tumor cells require autophagy for cell growth and tumor promotion. In particular, the survival of pancreatic tumor cells appears to be strongly dependent on autophagy, referred to as autophagy addiction. This dependency of pancreatic tumor cells on autophagy may be a candidate target for pancreatic tumor therapy. EI24 (etoposide-induced gene 2.4 kb; PIG8, p53-induced gene 8) acts as a tumor suppressor, inhibiting cell growth and inducing apoptosis in breast, cervical, and prostate cancer cells. However, recent papers have reported that EI24 is an essential component of the autophagy pathway. This newly discovered role of EI24 as a component of autophagy may act as a tumor promoter, which is contradictory to its known role as a tumor suppressor. We investigated the role of EI24 as a component of autophagy in pancreatic tumor cell proliferation. Here, we demonstrated that knockdown of EI24 using siRNA in pancreatic tumor cells led to impaired autophagy at a late step (increase in LC3-II and accumulation of p62 and autolysosomes). EI24 deficiency in pancreatic tumor cell lines inhibited cell proliferation. We confirmed that loss of EI24 inhibited pancreatic cell proliferation using the CRISPR-Cas9 system. However, loss of EI24 in other cell lines did not affect cell proliferation. Taken together, our results suggest that EI24 acts as a tumor promoter in pancreatic tumor cells, and studying the role of EI24 in reference to its cellular context may lead to a useful therapeutic target.


Effects of a synthetic progestogen, altrenogest, on oestrus synchronisation and fertility in miniature pigs.
Groups of six, six and eight miniature gilts, respectively, received 5, 10 or 15 mg/day of altrenogest for 18 days, and the numbers of corpora lutea and residual follicles were counted approximately 14 days after the treatment by an exploratory laparotomy. They were compared with the numbers in a control group of eight gilts which were examined six to eight days after oestrus. The interval between the last dose of altrenogest and the onset of oestrus increased with the dose of altrenogest, and was significantly longer after the treatments with 10 or 15 mg/day than after 5 mg/day (P < 0.01). Significantly more corpora lutea were observed in the gilts receiving 5 or 10 mg/day of altrenogest than in the control gilts (P < 0.1). Groups of six, seven and six miniature gilts that had respectively received 5, 10 or 15 mg/day of altrenogest were artificially inseminated; four, six and five of the gilts in these groups farrowed, and their mean (sd) litter sizes were 5.5 (2.4), 6.8 (1.2) and 5.0 (2.3), respectively. All six of a group of control gilts farrowed and their mean litter size was 5.8 (1.2).


The C-terminus of the P22 tailspike protein acts as an independent oligomerization domain for monomeric proteins.
TSP (P22 tailspike protein) is a well-established model system for studying the folding and assembly of oligomeric proteins, and previous studies have documented both in vivo and in vitro folding intermediates using this protein. Especially important is the C-terminus of TSP, which plays a critical role in the assembly and maturation of the protrimer intermediate to its final trimeric form. In the present study, we show that by grafting the C-terminus of TSP on to the monomeric MBP (maltose-binding protein), the resulting chimaera (MBP-537) is a trimeric protein. Moreover, Western blot studies (using an anti-TSP antibody) indicate that the TSP C-terminus in the MBP-537 chimaera has the same conformation as the native TSP. The oligomerization of the MBP-537 chimaera appears to involve hydrophobic interactions and a refolding sequence, both of which are analogous to the native TSP. These results underscore the importance of the TSP C-terminus in the assembly of the mature trimer and demonstrate its potential utility as a model to study the folding and assembly of the TSP C-terminus in isolation.


Kar9p is a novel cortical protein required for cytoplasmic microtubule orientation in yeast.
kar9 was originally identified as a bilateral karyogamy mutant, in which the two zygotic nuclei remained widely separated and the cytoplasmic microtubules were misoriented (Kurihara, L.J., C.T. Beh, M. Latterich, R. Schekman, and M.D. Rose. 1994. J. Cell Biol. 126:911-923.). We now report a general defect in nuclear migration and microtubule orientation in kar9 mutants. KAR9 encodes a novel 74-kD protein that is not essential for life. The kar9 mitotic defect was similar to mutations in dhc1/dyn1 (dynein heavy chain gene), jnm1, and act5. kar9Delta dhc1Delta, kar9Delta jnm1Delta, and kar9Delta act5Delta double mutants were synthetically lethal, suggesting that these genes function in partially redundant pathways to carry out nuclear migration. A functional GFP-Kar9p fusion protein localized to a single dot at the tip of the shmoo projection. In mitotic cells, GFP-Kar9p localized to a cortical dot with both mother-daughter asymmetry and cell cycle dependence. In small-budded cells through anaphase, GFP-Kar9p was found at the tip of the growing bud. In telophase and G1 unbudded cells, no localization was observed. By indirect immunofluorescence, cytoplasmic microtubules intersected the GFP-Kar9p dot. Nocodazole experiments demonstrated that Kar9p's cortical localization was microtubule independent. We propose that Kar9p is a component of a cortical adaptor complex that orients cytoplasmic microtubules.


Genetic determinism for within-litter birth weight variation and its relationship with litter weight and litter size in the Ripollesa ewe breed.
Birth weight plays a central role in lamb survival and growth, and the knowledge of its genetic determinism has become essential in worldwide selection programmes. Within this context, within-litter birth weight variation (BWV) has been suggested as an attractive trait to homogenise litters in prolific species, although it has not been analysed in sheep. The objective of this study was to ascertain whether maternal additive genetic variance exists for BWV in Ripollesa ewes, and to study its genetic, permanent environmental and residual relationships with litter weight (LW) and litter size (LS) at birth. Data were recorded in the Ripollesa experimental flock of the Universitat Autònoma of Barcelona, between 1986 and 2005, and included 1 662 litters from 380 ewes, with 712 records of BWV and 1 530 records of LW. Traits were analysed with a multivariate animal model solved through Bayesian methodologies, and with a threshold characterisation of LS. Additionally, the effect of BWV on lamb survival was studied. Additive genetic variance was observed for BWV (h2 = 0.061), as well as for LW (h2 = 0.200) and LS (h2 = 0.141). Nevertheless, genetic correlations among those traits were not substantial (BWV and LW = 0.151; BWV and LS = - 0.219; LW and LS = - 0.320) and suffered from a high degree of uncertainly, with the null correlation included within the highest posterior interval at 95%. Within-litter birth weight variation and LS showed a negative and large permanent environmental correlation ( - 0.872), and LW and LS were negatively correlated due to residual ( - 0.762) and permanent environmental ( - 0.449) random sources of variation. Within-litter birth weight variation influenced lamb mortality during the first 7 days of life (P < 0.05), increasing and decreasing survivability in heavier and lighter littermates, respectively. Nevertheless, stillbirths and lambs died after the 1st week of life were not affected by BWV (P>0.05). The low heritability found indicates that slow genetic progress may be expected from selecting for BWV. Close to zero genetic correlations suggest that this selection will probably not affect LS and LW, although some significant permanent and residual correlations must be taken into account. Further studies are needed to understand better the genetic architecture among these three reproductive traits.


T-2 mycotoxin intensifies iodine deficiency in mice fed low iodine diet.
Mice were depleted of iodine and fed a 125I-labeled low iodine diet. After they developed isotopic equilibrium they were given T-2 mycotoxin, in doses 34% to 14% of LD50/day for 4 to 12 days. In all cases T-2 toxin caused loss of thyroid iodine from which the animals recovered when the T-2 toxin was stopped. When iodine intake was adequate, the T-2 toxin had no statistically significant effect on the thyroid iodine content. Since T-2 toxin is known to block the initiation of some protein synthesis, it may block thyroglobulin synthesis, and, in stimulated thyroids, limited hydrolysis may continue. Therefore, when the thyroids were stimulated and thyroglobulin was depleted, a blockade of thyroglobulin synthesis may have caused further depletion of thyroidal iodine.


Mesenchymal stem cells preserve neonatal right ventricular function in a porcine model of pressure overload.
Limited therapies exist for patients with congenital heart disease (CHD) who develop right ventricular (RV) dysfunction. Bone marrow-derived mesenchymal stem cells (MSCs) have not been evaluated in a preclinical model of pressure overload, which simulates the pathophysiology relevant to many forms of CHD. A neonatal swine model of RV pressure overload was utilized to test the hypothesis that MSCs preserve RV function and attenuate ventricular remodeling. Immunosuppressed Yorkshire swine underwent pulmonary artery banding to induce RV dysfunction. After 30 min, human MSCs (1 million cells, n = 5) or placebo (n = 5) were injected intramyocardially into the RV free wall. Serial transthoracic echocardiography monitored RV functional indices including 2D myocardial strain analysis. Four weeks postinjection, the MSC-treated myocardium had a smaller increase in RV end-diastolic area, end-systolic area, and tricuspid vena contracta width (P < 0.01), increased RV fractional area of change, and improved myocardial strain mechanics relative to placebo (P < 0.01). The MSC-treated myocardium demonstrated enhanced neovessel formation (P < 0.0001), superior recruitment of endogenous c-kit+ cardiac stem cells to the RV (P < 0.0001) and increased proliferation of cardiomyocytes (P = 0.0009) and endothelial cells (P < 0.0001). Hypertrophic changes in the RV were more pronounced in the placebo group, as evidenced by greater wall thickness by echocardiography (P = 0.008), increased cardiomyocyte cross-sectional area (P = 0.001), and increased expression of hypertrophy-related genes, including brain natriuretic peptide, β-myosin heavy chain and myosin light chain. Additionally, MSC-treated myocardium demonstrated increased expression of the antihypertrophy secreted factor, growth differentiation factor 15 (GDF15), and its downstream effector, SMAD 2/3, in cultured neonatal rat cardiomyocytes and in the porcine RV myocardium. This is the first report of the use of MSCs as a therapeutic strategy to preserve RV function and attenuate remodeling in the setting of pressure overload. Mechanistically, transplanted MSCs possibly stimulated GDF15 and its downstream SMAD proteins to antagonize the hypertrophy response of pressure overload. These encouraging results have implications in congenital cardiac pressure overload lesions.


Associated and mediating variables related to quality of life among service users with mental disorders.
This study aimed to identify variables associated with quality of life (QoL) and mediating variables among 338 service users with mental disorders in Quebec (Canada). Data were collected using nine standardized questionnaires and participant medical records. Quality of life was assessed with the Satisfaction with Life Domains Scale. Independent variables were organized into a six-block conceptual framework. Using structural equation modeling, associated and mediating variables related to QoL were identified. Lower seriousness of needs was the strongest variable associated with QoL, followed by recovery, greater service continuity, gender (male), adequacy of help received, not living alone, absence of substance use or mood disorders, and higher functional status, in that order. Recovery was the single mediating variable linking lower seriousness of needs, higher service continuity, and reduced alcohol use with QoL. Findings suggest that greater service continuity creates favorable conditions for recovery, reducing seriousness of needs and increasing QoL among service users. Lack of recovery-oriented services may affect QoL among alcohol users, as substance use disorders were associated directly and negatively with QoL. Decision makers and mental health professionals should promote service continuity, and closer collaboration between primary care and specialized services, while supporting recovery-oriented services that encourage service user involvement in their treatment and follow-up. Community-based organizations should aim to reduce the seriousness of needs particularly for female service users and those living alone.


[The perinatal mortality in the Omskaya Oblast].
The study of the perinatal mortality was carried out in the Omskaya Oblast from 1996 to 2005. The ample decrease of the perinatal mortality from 14.64 per 1000 newborns in 1996 to 10.21 in 2005 is noted. At the same time, the decrease of the still-born rate was less marked in comparison with the earlier neonatal mortality. The assessment of the analyzed indicators in the urban and rural settlements testify to the usefulness of the activities applied by the urban medical care institutions in comparison with the rural ones. The analysis of the fetus loss rates before and during the delivery and the comparison of the levels of the newborns mortality during the first week of their life in the formation of the urban and rural perinatal mortality in the aggregative (i.e. Five-year) periods give grounds to consider that that further decrease of the perinatal mortality will be restrained by the antenatal losses. In the present-day decade a tendency in increasing the numbers of small-for-date fetuses both in the urban and rural areas is taken shape. More pronounced deterioration of the physical development of new generations to come is noticed among rural population. The full accounting of the factual losses of fetuses and children is needed to make optimal management decisions promoting the development of the perinatal service and the decrease of the morbidity and mortality of children.


Wiskott-Aldrich syndrome in a girl caused by heterozygous WASP mutation and extremely skewed X-chromosome inactivation: a novel association with maternal uniparental isodisomy 6.
Wiskott-Aldrich syndrome (WAS) is an X-linked disease characterized by microthrombocytopenia, eczema and immune deficiency, caused primarily by mutations in the WASP (Wiskott-Aldrich syndrome protein) gene. Female carriers are usually asymptomatic because of the preferential activation of the normal, nonmutated X-chromosome in their hematopoietic cells. We report our observations of a female child with WAS, who displayed symptoms of congenital thrombocytopenia. DNA sequencing analysis of the WASP gene revealed a heterozygous nonsense mutation in exon 10. The expressions of WASP and normal WASP mRNA were defective. We found preferential inactivation of the X-chromosome on which wild-type WASP was located. Single-nucleotide polymorphism microarray testing and the analysis of the polymorphic variable number of tandem repeat regions revealed maternal uniparental isodisomy of chromosome 6 (UPD6). Our results underscore the importance of WASP evaluation in females with congenital thrombocytopenia and suggest that UPD6 might be related to the pathophysiology of nonrandom X-chromosome inactivation.


Stepwise co-fermented traditional Chinese medicine byproducts improve antioxidant and anti-inflammatory effects in a piglet model.
Lianhua Qingwen capsule is a traditional Chinese medicine (TCM) formula having antiviral and anti-inflammatory activities. During capsule production, a large amount of byproducts will be yielded and disposed of as waste by burying. Resourceful utilization of these kinds of TCM byproducts as feed additives through stage-based co-fermentation using enzyme and probiotics could reduce environmental stress and resource shortage. The in vitro characterization and the supplementary effects of fermented TCM byproducts (FTCM) for weaned piglets (initial body weight: 7.23 ± 0.33 kg; dose: basal diet + 300 mg kg-1 FTCM) were investigated. Higher reducing sugar content, total flavonoid content, flavonoid compounds (e.g. tectoridin, tricetin, flavone, apigenin, naringenin) and total antioxidant activity were determined in the FTCM compared to spontaneously fermented and unfermented materials. Supplementation of the FTCM to piglets did not significantly affect the feed intake, body weight gain and feed/gain ratio, but significantly decreased a proinflammatory cytokine, IL-8, and increased intestinal total antioxidant activity (TAC) and superoxide dismutase (SOD) activity. Moreover, FTCM supplementation increased α-diversity of the colonic microbiota accompanied with increased abundance of Prevotella genus and Treponema berlinense species. Correlation analysis indicates that T. berlinense is responsible for the decreased IL-8 level and enhanced intestinal TAC and SOD activities which might be mediated by a homoserine lactone molecule (3-oxo-C14). Overall, the stepwise co-fermentation enriched bioactive compounds within the TCM byproducts and their dietary supplementation did not generate any side effect on growth performance but displayed beneficial effects on enrichment of potential probiotic T. berlinense and relevant functions. © 2023 Society of Chemical Industry.


Brca2 is required for embryonic cellular proliferation in the mouse.
Mutations of the tumor suppressor gene BRCA2 are associated with predisposition to breast and other cancers. Homozygous mutant mice in which exons 10 and 11 of the Brca2 gene were deleted by gene targeting (Brca2(10-11)) die before day 9.5 of embryogenesis. Mutant phenotypes range from severely developmentally retarded embryos that do not gastrulate to embryos with reduced size that make mesoderm and survive until 8.5 days of development. Although apoptosis is normal, cellular proliferation is impaired in Brca2(10-11) mutants, both in vivo and in vitro. In addition, the expression of the cyclin-dependent kinase inhibitor p21 is increased. Thus, Brca2(10-11) mutants are similar in phenotype to Brca1(5-6) mutants but less severely affected. Expression of either of these two genes was unaffected in mutant embryos of the other. This study shows that Brca2, like Brca1, is required for cellular proliferation during embryogenesis. The similarity in phenotype between Brca1 and Brca2 mutants suggests that these genes may have cooperative roles or convergent functions during embryogenesis.


Politics of a policy for aging in Canada.
Canada's health and social care system is paralyzed by our decentralized federalist governing structure. Public policy change, such as that suggested by Chappell and Hollander, will require a new political paradigm that recognizes the need for a multi-sectoral, co-operative approach to integrated systems of care delivery. The federal government must provide the necessary leadership, and the provinces and territories must show the political will to co-operate if Canada is to embrace the challenges of an aging population.


New insights regarding chronic antibody-mediated rejection and its progression to transplant glomerulopathy.
To discuss new insights regarding chronic antibody-mediated rejection (CAMR) and its progression to transplant glomerulopathy. We will describe the progression to transplant glomerulopathy from a histologic perspective and provide updates on what is known about its pathophysiology, prognosis, and potential therapy. Transplant glomerulopathy is a major contributor to long-term renal allograft loss and is most often associated with CAMR. On the basis of protocol biopsies, we have found that 3.5% of conventional transplants and 27.5% of positive crossmatch kidney transplants have transplant glomerulopathy at 1 year. The pathophysiology of the process is largely unknown, but complement activation was previously thought to be essential. However, CAMR appears to develop despite terminal complement blockade and many C4d negative cases of CAMR have been identified. Thus, complement independent mechanisms, such as direct endothelial cell activation and the infiltration of natural killer cells and monocytes, are likely key to the development of transplant glomerulopathy. Transplant glomerulopathy is often the result of CAMR and leads to allograft loss. It is characterized by distinctive histologic changes, and its pathophysiology is a multifaceted process involving both innate and adaptive immunity. Despite advances in the understanding of this condition, no effective therapy exists.


Unsatisfactory outcome of arthrodesis performed after septic failure of revision total knee arthroplasty.
Periprosthetic infection is one of the most dreaded orthopaedic complications. Current treatment procedures include one-stage or two-stage revision total knee arthroplasty. If the periprosthetic infection is no longer controllable after several revision total knee arthroplasties, many surgeons regard knee arthrodesis as a promising option. The aim of our study was to ascertain whether intramedullary nailing results in the suppression or eradication of an infection and to identify risk factors for persistent infection. All patients who had undergone intramedullary nailing following septic failure of revision total knee arthroplasty between 1997 and 2013 were included in the study. Pathogens, risk factors predisposing to persistent infection, and the rate of persistent infections were recorded. In addition, a visual analog scale (VAS) and Knee injury Osteoarthritis Outcome Score (KOOS), Knee Society Score (KSS), Lysholm, Short Form-36 (SF-36), and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaires were completed to assess clinical outcomes and quality of life. Twenty-six patients were included in the study. Thirteen (50%) had a persistent infection requiring additional revision surgery. Nineteen patients (73%) reported persistent pain (VAS score of >3). All scores showed marked impairment of quality of life. Intramedullary nailing following septic failure of revision total knee arthroplasty must be regarded with skepticism, and we cannot recommend it. Repeat revision total knee arthroplasty or amputation should be considered as an alternative in such difficult cases.


SymbioGBR: a web-based database of Symbiodinium associated with cnidarian hosts on the Great Barrier Reef.
The algal endosymbionts (genus Symbiodinium) associated with scleractinian corals (and other reef invertebrates) have received a lot of research attention in the past decade, particularly as certain host-symbiont associations appear more affected by increasing seawater temperatures than others. With the rapid accumulation of information on the diversity of Symbiodinium, it is becoming increasingly difficult to compare newly acquired Symbiodinium data with existing data to detect patterns of host-symbiont specificity on broader spatial scales. The lack of a general consensus on the classification of Symbiodinium species coupled with the variety of different markers used to identify the genus Symbiodinium (ITS1, ITS2, LSU D1/D2, chloroplast 23S rDNA and psbA minicircle) further complicate direct comparison. The SymbioGBR database compiles all currently available Symbiodinium sequences and associated host information of data collected from the Great Barrier Reef into a single relational database that is accessible via a user-friendly, searchable web-based application (http://www.SymbioGBR.org). SymbioGBR allows users to query Symbiodinium types or sequences sourced from various genetic markers (e.g. ITS1, ITS2, LSU D1/D2 and chloroplast 23S) and invertebrate host species to explore their reported associations. In addition, as the database includes sequence information of multiple genetic markers, it allows cross-referencing between conventional (e.g. ITS2 region) and novel markers that exhibit low intragenomic variability (e.g. psbA region). Finally, the database is based on the collection details of individual specimens. Such host-symbiont associations can be assessed quantitatively and viewed in relation to their environmental and geographic context. The SymbioGBR database provides a comprehensive overview of Symbiodinium diversity and host-associations on the Great Barrier Reef. It provides a quick, user-friendly means to compare newly acquired data on Symbiodinium (e.g. raw sequences or characterized Symbiodinium types) with previous data on the diversity of invertebrate host-symbiont associations on the GBR. The inclusion of psbAncr sequence information allows for validation of widely used ITS1/ITS2 markers and their ability to accurately identify relevant sequences. Most importantly, centralization of sequence information from multiple genetic markers will aid the classification of Symbiodinium species diversity and allow researchers to easily compare patterns of host-Symbiodinium associations.


Generation and characterization of monoclonal antibodies specific for the human IFN-gamma receptor.
Purified preparations of the human IFN-gamma R derived from placental membranes were used to produce receptor-specific murine mAb. Supernatants from growth-positive wells were screened for their ability to block binding of 125I-IFN-gamma to human placental membranes. Ten inhibitory cultures were identified. Two of these (GIR-208 and GIR-301) abrogated all binding of radioligand to either intact placental membranes or soluble, purified IFN-gamma R. Three others (GIR-72, 76 and 94) showed moderate blocking activity (65, 59, and 49%, respectively) whereas the remaining five (GIR-57, 67, 83, 109, and 153) blocked binding to a low but significant extent (20 to 40%). Specificity experiments demonstrated that the antibodies reacted with the receptor and not the ligand (IFN-gamma). None of the antibodies reacted with IFN-gamma by ELISA. Moreover, GIR-208 and GIR-301, but not isotype-matched controls, identified the receptor by Western blot analysis. GIR-208 and GIR-301 also completely abrogated binding of 125I-IFN-gamma to either mononuclear phagocytes (U937) or human fibroblasts (WISH). Competition experiments revealed that GIR-208 and GIR-301 recognized similar epitopes on the IFN-gamma R and that these (or this) epitopes were identical to or linked to the ligand binding site of the receptor. In addition, both antibodies inhibited development of IFN-gamma-dependent anti-viral activity in WISH cells in a dose-dependent fashion. These data thus indicate that the IFN-gamma R expressed on human placental cells, mononuclear phagocytes, and fibroblasts are similar.


Adenocarcinoma of the cecum with Crohn's-like features in baboons.
During the past 9.5 years, 51 baboons dying as a result of chronic diarrhea and poor health and having a chronic inflammatory process of the cecum (i.e. typhlitis) were recorded. The cecum was turgid, irregularly thickened the lumen reduced and the mesenteric lymph nodes enlarged. Of the 45 baboons with well preserved histologic material, 24 had shallow ulcerations and/or focal mucosal inflammation, 3 had crypt abcesses, 3 had mucosal fissures,l aphtoid ulcerations, and 1 pseudopyloric metaplasia (ulcer associated cell lineage-UACL-). The submucosa in all 45 animals was thick due to fibrosis, 35 had focal chronic inflammation with lymphoid aggregates, 12 submucosal abcesses, 6 epithelioid cell granulomas, 5 fissures reaching the submucosa, 3 neural hyperplasia of the myenteric plexus, and 2 lymphangiectasia. Focal chronic transmural inflammation with lymphoid aggregates was recorded in 44 animals. In addition, 2 animals showed deep fissure tracts. Ten of the 45 animals had in addition a cecal adenocarcinoma; one was overlayed by a villous adenoma with high grade dysplasia. Liver metastasis was found in another animal. The present study indicated that a) baboons may develop transmural chronic focal inflammation in the proximal colon (cecum), b) the inflammation in baboons closely mimic Crohn's disease of the cecum in humans c) adenocarcinomas in baboons with Crohn's-like inflammatory changes may originate in an adenomatous growth, and d) cecal adenocarcinomas in baboons with Crohn's-like inflammatory changes of the cecum, are able to metastasize.


Surgical treatment for suicidal jumper's fracture (unstable sacral fracture) with thoracolumbar burst fracture: a report of three cases.
Suicidal jumper's fracture (unstable sacral fracture) is characterized not only by multiple fractures including thoracolumbar fractures, but also major chest and abdominal injuries. Early stabilization of these fractures and early ambulation are required for the treatment and management of chest and abdominal injuries. We present 3 cases of suicidal jumper's fracture with thoracolumbar burst fracture, treated with minimally invasive posterior fixation surgery, which is a combination of percutaneous pedicle screws (PPS) and the mini-open Galveston technique. Case 1. A 50-year-old woman was injured by a fall from the 5th floor of a building as the result of a suicide attempt. Computed tomography revealed an H-shaped unstable sacral fracture and thoracolumbar fractures with major chest and abdominal injuries. For early stabilization of spinopelvic instability and early ambulation, we treated the patient with PPS and the mini-open Galveston technique. Her early postoperative emergence from bedrest contributed to the improvement of her general condition. One year after surgery at the final follow-up, she was able to walk with a T-cane without any motor, bladder, or bowel dysfunction (BBD) and achieved almost complete healing of the fractures. Cases 2 and 3. A 25-year-old woman (Case 2) and a 43-year-old woman were injured in falls. They had multiple injuries including unstable sacral fractures, and thoracolumbar fractures with major chest and abdominal injuries. We treated these patients with PPS and the mini-open Galveston technique. One year after surgery, they were able to walk with a T-cane and achieved almost complete healing of thoracolumbar fractures, but delayed healing of an unstable sacral fracture in Case 2, and remaining BBD in Case 3. PPS and the mini-open Galveston technique is a good approach to fixation because they are minimally invasive and provide moderately rigid fixation, especially in patients with multiple trauma whose general condition is poor.


The pharmacology of nedocromil sodium.
The pathophysiology of obstructive airways disease involves, in varying degree, components of reversible bronchoconstriction, inflammation and bronchial hyperreactivity. This multifactorial aetiology is the probable reason why no single animal model exists which is predictive of therapeutic efficacy in airway diseases, including asthma. Nedocromil sodium has therefore been profiled in a number of systems ranging from simple in vitro tests and in vivo models of passive anaphylaxis to complex models of anaphylactic bronchoconstriction in actively sensitized primates. Nedocromil sodium possesses efficacy and potency similar to those of sodium cromoglycate in classical passive models of immediate hypersensitivity in the rat. Preliminary studies have also shown that nedocromil sodium, like sodium cromoglycate, attenuates non-specific bronchial hyperreactivity in dogs exposed to sulphur dioxide. Extensive studies have been carried out in a model of airway disease in the primate Macaca arctoides infected with the nematode Ascaris suum. Bronchoalveolar cells obtained from these animals by lung lavage release substantial quantities of the inflammatory mediators histamine, leukotriene C4 (LTC4) and prostaglandin D2 (PGD2) on immunological activation. Nedocromil sodium significantly inhibits the release of histamine, LTC4 and PGD2 from these cells in response to stimulation with antigen or antibody to human IgE. Under identical conditions, sodium cromoglycate has less than 1/200th the potency of nedocromil sodium, producing only insignificant inhibition of mediator release even at concentrations greater than 10(-4) M. A similar difference between the two drugs has been observed in vivo, where nedocromil sodium produced significant inhibition of antigen-induced bronchoconstriction in the Ascaris-sensitized macaque but sodium cromoglycate did not.(ABSTRACT TRUNCATED AT 250 WORDS)