Assessment of mould remediation in a healthcare setting following extensive flooding.

BACKGROUND: A new hospital building was close to completion when a large pipe carrying clean water broke, causing extensive flooding. AIM: To determine the flood-associated fungal risk to susceptible patients who would use that building. METHODS: Though standard flood remediation by the builders was relatively straightforward, there was no model for specialist assessment of patient risk due to the flood-associated mould growth. As levels of background airborne fungal spores can be expected to vary significantly over time, we could not use absolute levels to indicate either an excess of airborne fungal spores or successful remediation. Therefore it was decided to use weekly settle plates, exposed at the same time in flooded (test) and equivalent non-flooded (control) areas to compensate for variations in background levels. Flood-related risk was estimated by the ratio between fungal colonies on the test and control sets of settle plates, rather than absolute number. FINDINGS: Whereas the physical flood remediation, including the use of 'anti-fungal' treatments, was completed in three weeks post flooding, fungal contamination in flooded areas took 38 weeks to return to control levels and remained so for a further six weeks of observation. CONCLUSION: By the use of this method, we were able to assure the absence of flood-associated fungal risk to susceptible patients who would use that building. We recommend that infection prevention and control teams consider using this approach should they be faced with similar situations.

Comparative study on the deamidation of three recombinant human insulins using capillary electrophoresis.

The applicability of capillary zone electrophoresis (CZE) for the separation of different recombinant human insulins and their deamidated isoforms was studied. The high resolving power of CZE is demonstrated by its ability to separate insulin isoforms differing only by 0.984 Da (different-fold deamidated forms) and even components having the exacts same mass but slightly different shapes (same-fold deamidated forms). From among the several insulins available, humulin, glargine and glulisine were selected for our study because their sequences and chemical parameters are quite similar, however, the small differences present in their amino acid sequences influence the deamidation processes. Using a background electrolyte with basic pH was favourable not only for the separation of the different types of insulin but also for the separation of deamidated protein forms even in a bare fused silica capillary. The LOD values ranged between 0.6 - 0.93 mg/L and 2.17 - 4.37 mg/L for UV and ESI-MS detection, respectively. At -20 - -80 °C, the deamidation is minimal, but at temperatures above +5 °C deamidation is accelerated. At +5 °C only 1-fold deamidation forms could be observed for each insulin. Acidified samples incubated for 1-month at room temperature showed varying levels of deamidation: 1-fold, 1-2-fold and 1-2-3-fold forms for glargine, glulisine and humulin, respectively.

Analysis of intact proteins with capillary zone electrophoresis coupled to mass spectromery using uncoated and coated capillaries.

Although, in general, the application of coated capillaries is recommended for the separation of intact proteins, bare silica capillary is still the most often used capillary due to its simplicity and cheapness. In this work, the performance of bare fused silica capillary for intact protein analysis was compared to that of different (dynamically coated polybrene (PB) and permanently coated linear polyacrylamide (LPA)) coated capillaries using capillary zone electrophoresis - mass spectrometry (CZE-MS). In cases where low pH (pH=1.8) was used in bare silica capillaries, good precision (0.56-0.78 RSD% and 1.7-6.5 RSD% for migration times and peak areas, respectively), minimal adsorption and separation efficiency (N= 27 000/m - 322 000/m) similar to or even better than those obtained with the coated capillaries (created by an intricate multi-step process) was achieved. The PB and the LPA capillaries demonstrated their slightly better resolving power in terms of separating the different forms/variants of the same protein (e.g., hemoglobin subunits). Among the studied capillaries the one with LPA coating showed the most stable separations in the long term (n=25: 0.18-0.49 RSD% and 3.1-4.9 RSD% for migration times and peak areas, respectively). For the separation of a few proteins or even a larger number of proteins in biological samples (e.g., snake venom) the application of the simple and cheap bare fused silica capillary can be considered as an efficient choice.

Examining the role of mother-child interactions and DNA methylation of the oxytocin receptor gene in understanding child controlling attachment behaviors.

This study examined mother-child interactions and DNA methylation of the oxytocin receptor (OXTR) gene in the child, in relation with controlling-attachment behaviors at early preschool age. Maternal interactive behaviors were coded using the Emotional Availability Scales, and child attachment behaviors were assessed with the Separation-Reunion procedure and coded with the Preschool Attachment Rating Scales. DNA methylation data were captured from exon 3 of the OXTR. Results indicated that lower maternal sensitivity was associated with more controlling-caregiving behaviors, and that less maternal structuring was associated with more controlling-punitive behaviors. Hypomethylation of the OXTR gene was associated with greater maternal structuring behaviors, and with more child controlling-caregiving behaviors. The moderating role of the OXTR gene was examined in the association between interactive behaviors and child controlling behaviors, but no interaction effect was found. These results suggest that maternal interactive behaviors and OXTR methylation are independently associated with child controlling attachment.

Determination of deamidated isoforms of human insulin using capillary electrophoresis.

The applicability of capillary zone electrophoresis (CZE) for the separation of the deamidated forms of insulin has been studied. 50 mM NH4Ac (pH=9) with 20 % v/v isopropylalcohol was found optimal for efficient separation of insulin from its even 10 deamidated forms. The developed method was efficiently applied for monitoring the degradation rate of insulin and the formation of different deamidation isoforms. Two months after the acidification more than thirty peaks can be observed in the electropherogram, because degradation products other than deamidated components were formed as well. The recorded mass spectra enabled us to assign the exact mass of the components, and thus the identification of insulin isoforms could be accomplished. We think that this study provides useful information on how the determination of several deamidation forms can be carried out with CE-MS, but the identification of the exact position of deamidation sites in the insulin molecule remains a challenge.

Sleep quality is associated with vasopressin methylation in pregnant and postpartum women with a history of psychosocial stress.

BACKGROUND: The relationship between disturbed sleep and stress is well-documented. Sleep disorders and stress are highly prevalent during the perinatal period, and both are known to contribute to a number of adverse maternal and foetal outcomes. Arginine vasopressin (AVP) is a hormone and a neuropeptide that is involved in stress response, social bonding and circadian regulation of the sleep-wake cycle. Whether the AVP system is involved in regulation of stress response and sleep quality in the context of the perinatal mental health is currently unknown. The objective of the present study was to assess the relationship between levels of cumulative and ongoing psychosocial risk, levels of disordered sleep and AVP methylation in a community sample of pregnant and postpartum women. METHODS: A sample of 316 participants completed a battery of questionnaires during the second trimester of pregnancy (PN2, 12-14 weeks gestation), third trimester (PN3, 32-34 weeks gestation), and at 7-9 weeks postpartum (PP). Disordered sleep was measured using the Sleep Symptom Checklist at PN2, PN3 and PP; cumulative psychosocial risk was assessed with the Antenatal Risk Questionnaire (ANRQ) at PN2; salivary DNA was collected at the follow-up (FU, 2.9 years postpartum); and % methylation were calculated for AVP and for two of the three AVP receptor genes (AVPR1a and AVPR1b). Women were separated into high (HighPR) and low (LowPR) psychosocial risk groups, based on their scores on the ANRQ. RESULTS: Women in the HighPR group had significantly worse sleep disturbances during PN2 (p < .001) and PN3 (p < .001), but not at PP (p = .146) than women in the LowPR group. In HighPR participants only, methylation of AVP at intron 1 negatively correlated with sleep disturbances at PN2 (rs=-.390, p = .001), PN3 (rs=-.384, p = .002) and at PP (rs= -.269, p = .032). There was no association between sleep disturbances and AVPR1a or AVPR1b methylation, or between sleep disturbances and any of the AVP methylation for the LowPR group. Lastly, cumulative psychosocial stress was a moderator for the relationship between AVP intron 1 methylation and disordered sleep at PN2 (p < .001, adjusted R2 = .105), PN2 (p < .001, adjusted R2 = .088) and PP (p = .003, adjusted R2 = .064). CONCLUSIONS: Our results suggest that cumulative psychosocial stress exacerbates sleep disorders in pregnant women, and that salivary DNA methylation patterns of the AVP gene may be seen as a marker of biological predisposition to stress and sleep reactivity during the perinatal period. Further research is needed to establish causal links between AVP methylation, sleep and stress.

A role for activity-dependent epigenetics in the development and treatment of major depressive disorder.

Chronic stressors, during developmental sensitive periods and beyond, contribute to the risk of developing psychiatric conditions, including major depressive disorder (MDD). Epigenetic mechanisms including DNA methylation and histone modifications, at key stress response and neurotrophin genes, are increasingly implicated in mediating this risk. Although the exact mechanisms through which stressful environmental stimuli alter the epigenome are still unclear, research from the learning and memory fields indicates that epigenomic marks can be altered, at least in part, through calcium-dependent signaling cascades in direct response to neuronal activity. In this review, we highlight key findings from the stress, MDD, and learning and memory fields to propose a model where stress regulates downstream cellular functioning through activity-dependent epigenetic changes. Furthermore, we suggest that both typical and novel antidepressant treatments may exert positive influence through similar, activity-dependent pathways.

Microarchitectural study of the augmented bone following ridge preservation with a porcine xenograft and a collagen membrane: preliminary report of a prospective clinical, histological, and micro-computed tomography analysis.

Socket preservation using a combination of porcine xenograft and collagen membrane maintains the vertical and horizontal dimensions of the ridge. The aim of this study was to evaluate the microarchitecture of the grafted area by histological analysis and micro-computed tomography. Patients in the test group (group 1; nine patients) underwent socket preservation, while the sockets in the control group (group 2; eight patients) were allowed to heal without preservation. After a 6-month healing period, bone core biopsy samples were obtained and implants were placed in the augmented sites in the test group (12 biopsy samples) and the non-augmented sockets of the control group (12 biopsy samples). Analysis of the biopsy samples obtained from group 1 revealed that particles of the graft were surrounded by newly formed bone in eight cases and by granulation tissue in four cases. Micromorphometric data showed statistically significant differences in several parameters between the microarchitecture of the native bone and the newly formed bone within the augmented sites, which suggests that the xenograft particles interfere with the bony healing of the alveoli.

Animal-to-Human Dose Translation of Obiltoxaximab for Treatment of Inhalational Anthrax Under the US FDA Animal Rule.

Obiltoxaximab, a monoclonal antibody against protective antigen (PA), is approved for treatment of inhalational anthrax under the US Food and Drug Administration's (FDA) Animal Rule. The human dose was selected and justified by comparing observed obiltoxaximab exposures in healthy and infected New Zealand White rabbits and cynomolgus macaques to observed exposures in healthy humans, to simulated exposures in healthy and infected humans, and to serum PA levels in infected animals. In humans, at 16 mg/kg intravenous, obiltoxaximab AUC was >2 times that in animals, while maximum serum concentrations were comparable to those in animals and were maintained in excess of the concentration required for PA neutralization in infected animals for 2-3 weeks. Obiltoxaximab 16 mg/kg in humans provided exposure beyond that of 16 mg/kg in animals, ensuring a sufficient duration of PA neutralization to allow for adaptive immunity development. Our approach to dose translation may be applicable to other agents being developed under the Animal Rule.

Glial fibrillary acidic protein is differentially expressed across cortical and subcortical regions in healthy brains and downregulated in the thalamus and caudate nucleus of depressed suicides.

There is mounting evidence to suggest aberrant astrocytic function in depression and suicide. Independent studies have reported astrocytic abnormalities in certain brain regions, but it remains unclear whether this is a brain-wide phenomenon. The present study examined this question by measuring glial fibrillary acidic protein (GFAP) expression in postmortem brain samples from suicide completers and matched non-psychiatric controls. Suicide completers were selected based on their recent characterization as low GFAP expressors in the prefrontal cortex, (Brodmann areas 8/9 and 10). Real-time PCR and immunoblotting were used to measure GFAP gene expression and protein levels in BA4 (primary motor cortex), BA17 (primary visual cortex), cerebellar cortex, mediodorsal thalamus and caudate nucleus. We found downregulation of GFAP mRNA and protein in the mediodorsal thalamus and caudate nucleus of depressed suicides compared with controls, whereas GFAP expression in other brain regions was similar between groups. Furthermore, a regional comparison including all samples revealed that GFAP expression in both subcortical regions was, on average, between 11- and 15-fold greater than in cerebellum and neocortex. Examining astrocyte morphology by immunohistochemistry showed that astrocytes in both thalamus and caudate displayed larger cell bodies and extended more ramified processes across larger domains than the previously described cortical astrocytes. This study reveals that astrocytic abnormalities are not brain wide and suggests that they are restricted to cortical and subcortical networks known to be affected in mood disorders. Additionally, our results show a greater diversity in human astrocytic phenotypes than previously thought.

Astrocytic abnormalities and global DNA methylation patterns in depression and suicide.

Astrocytes are glial cells specific to the central nervous system and involved in numerous brain functions, including regulation of synaptic transmission and of immune reactions. There is mounting evidence suggesting astrocytic dysfunction in psychopathologies such as major depression, however, little is known about the underlying etiological mechanisms. Here we report a two-stage study investigating genome-wide DNA methylation associated with astrocytic markers in depressive psychopathology. We first characterized prefrontal cortex samples from 121 individuals (76 who died during a depressive episode and 45 healthy controls) for the astrocytic markers GFAP, ALDH1L1, SOX9, GLUL, SCL1A3, GJA1 and GJB6. A subset of 22 cases with consistently downregulated astrocytic markers was then compared with 17 matched controls using methylation binding domain-2 (MBD2) sequencing followed by validation with high-resolution melting and bisulfite Sanger sequencing. With these data, we generated a genome-wide methylation map unique to altered astrocyte-associated depressive psychopathology. The map revealed differentially methylated regions (DMRs) between cases and controls, the majority of which displayed reduced methylation levels in cases. Among intragenic DMRs, those found in GRIK2 (glutamate receptor, ionotropic kainate 2) and BEGAIN (brain-enriched guanylate kinase-associated protein) were most significant and also showed significant correlations with gene expression. Cell-sorted fractions were investigated and demonstrated an important non-neuronal contribution of methylation status in BEGAIN. Functional cell assays revealed promoter and enhancer-like properties in this region that were markedly decreased by methylation. Furthermore, a large number of our DMRs overlapped known Encyclopedia of DNA elements (ENCODE)-identified regulatory elements. Taken together, our data indicate significant differences in the methylation patterns specific to astrocytic dysfunction associated with depressive psychopathology, providing a potential framework for better understanding this disease phenotype.

Carbon multi-shell cages.

Complex chemical compounds found as minerals or synthesized in labs evidenced a multi-shell structure. Also, fullerenes aggregate, randomly or following a well-defined geometry, in multi-shells. The way of space filling differs as a function of the dimensions and shape of the composing small cages. In this paper an attempt to build and evaluate the stability of several fullerene aggregates is made. The results show multi-shell covalently bonded structures with stability comparable to that of C60, the reference fullerene in nanoscience. The calculations were made at the DFTB and DFT levels of theory.

Antiinflammatory effect of BPC 157 on experimental periodontitis in rats.

The pentadecapeptide BPC 157 has been shown to have anti-inflammatory and wound healing effects on multiple target tissues and organs. The purpose of the present study was to investigate the effect of BPC 157 on inflammation and bone resorption in experimental periodontitis in rats. First the acute effect of BPC was tested on gingival blood flow by laser doppler flowmetry. Then periodontitis was produced by a silk ligature placed around the lower left first molar. Rats were treated with BPC 157 (once daily for 12 days) or vehicle. At day 13, the gingivomucosal tissues encircling the molars were removed on both sides. Inflammation was assessed by Evans blue plasma extravasation technique and by histology. Alveolar bone loss was analyzed by microCT. BPC 157 had no effect on gingivomucosal blood flow. Twelve day ligature caused a significantly increased Evans blue extravasation in the gingivomucosal tissue, histological signs of inflammation, and alveolar bone destruction. BPC 157 treatment significantly reduced both plasma extravasation, histological alterations and alveolar bone resorption. In conclusion, systemic application of BPC 157 does not alter blood circulation in healthy gingiva. Chronic application of the peptide has potent antiinflammatory effects on periodontal tissues in ligature induced periodontitis in rats. Taken together, this proof of concept study suggests that BPC 157 may represent a new peptide candidate in the treatment of periodontal disease.

Acute normovolemic hemodilution, intraoperative cell salvage and PulseCO hemodynamic monitoring in a Jehovah's Witness with placenta percreta.

A Jehovah's Witness who had had four previous cesarean deliveries was referred to our institution for management of a complete placenta previa at 34 weeks of gestation. A subsequent ultrasound scan was suggestive of placenta percreta with bladder involvement. After erythropoietin and iron supplementation, cesarean hysterectomy was performed. Using PulseCO technology for continuous hemodynamic monitoring, preoperative acute normovolemic hemodilution and intraoperative cell salvage were used resulting in a successful cesarean hysterectomy with a 5500-mL estimated blood loss. The PulseCO system provides continuous, real-time hemodynamic data by applying pulse power analysis to the arterial pressure waveform. A bolus of oxytocin given after delivery produced profound hypotension, the hemodynamics of which were elucidated with the PulseCO system. To our knowledge, the combined use of acute normovolemic hemodilution, intraoperative cell salvage and PulseCO hemodynamic monitoring for cesarean hysterectomy has not been reported previously. These techniques may be particularly useful in managing patients who refuse blood products and/or in whom the baseline hemoglobin is suboptimal.

The influence of root canal shape on the sealing ability of two root canal sealers.

AIM: To evaluate the influence of root canal form on the sealing ability of two root canal sealers. METHODOLOGY: Twenty radiographically confirmed straight and 20 curved root canals were prepared with a stepback hand filing technique. Root canal aberrations created during preparation were determined by the use of double exposure radiographic technique. The prepared canals were filled with lateral condensation of gutta-percha and one or other of two root canal sealers (Pulp Canal Sealer and Sealapex). Leakage along the apical 10 mm of roots was measured with a fluid transport model at 1, 3, 6, 9 and 12-month intervals. RESULTS: There were no statistically significant differences between straight and curved root canals (P > 0.05) for prevalence of root canal transportation. The prevalence of apical transportation was 80% in the straight and 85% in the curved root canals. A complete seal was more frequently observed in straight canals compared with curved canals. Utilizing the pi* index, analysis showed the filling with Sealapex allowed more leakage than Pulp Canal Sealer at 1 year. CONCLUSION: Under the conditions of the study, root canal form influenced short-term sealing ability. In the long-term the seal was affected by the sealer rather than root canal form.

Endobronchial pseudo-tumour caused by herpes simplex.

Herpes simplex virus (HSV) causes tracheobronchitis and pneumonitis; however, to date, there has only been one report of an endobronchial mass caused by HSV type II. This case study describes a 68-yr-old female with severe kyphoscoliosis who was intubated for acute on chronic hypercapnic respiratory failure and developed blood-tinged endotracheal secretions. Fibreoptic bronchoscopy demonstrated an endobronchial mass in the right middle lobe. Cultures grew HSV type I and biopsy specimens demonstrated cytopathological changes consistent with HSV infection. This is the first reported case of HSV type I presenting as an endobronchial tumour.

Measurement of periapical pressure created by occlusal loading.

AIM: To develop an in vitro model in which the pressure in the periapical tissues can be measured during loading. METHODOLOGY: Extracted human maxillary central incisors were embedded in resin blocks that had physical characteristics similar to those of bone and periodontal ligament. Each tooth was loaded with 20, 40, 50, 60, 75, 85, 100, 200, 300 and 450 N vertical forces from the incisal edge of the crown on three consecutive occasions. A minute resistor embedded in the periapical space was used to detect apical pressure changes during occlusal loading. The ratio of apical pressure changes (DeltaP) to the loading force changes (DeltaF) was calculated. RESULTS: The periapical pressure detected was in direct proportion to the loading force. The mean value of DeltaP/DeltaF was 5.994 kPa N-1 (SD = 2.04). CONCLUSIONS: Direct proportionality was found between the coronal loading and the apical hydrostatic pressure. The DeltaP/DeltaF ratio determined in this study makes it easier to estimate the apical hydrostatic pressure values during occlusal loading of single-rooted teeth. In this study, the apical pressure generated under occlusal loading was the same magnitude as that estimated with the finite element method.

A comparison of the shaping characteristics of two nickel-titanium endodontic hand instruments.

AIM: The purpose of this study was to compare the shaping characteristics of Ni-Ti K-files and Ni-Ti S-files manipulated by hand. METHODOLOGY: A total of 60 extracted human roots were embedded in resin blocks. The embedded roots were divided into three groups: (i) roots with straight; (ii) apically curved; and (iii) continuously curved canals. Each of the three groups was randomly divided into two subgroups; one subgroup in each group was prepared with Ni-Ti K-files and the other with Ni-Ti S-files. The files were used with a step-back technique and enlarged so that the master apical file was size 30, and the canals were stepped back to size 40. The performance of the files was assessed by the superimposition of projected radiographs taken in bucco-lingual and mesio-distal directions before and after the preparation. The results were analysed statistically using analysis of covariance and Duncan's multiple range test. RESULTS: Although canal preparation using Ni-Ti K-files was quicker, there were no statistically significant differences between file types. The Ni-Ti S-file removed significantly more material at the most coronal level (P < 0.05). There were minor differences between instruments at the apical level. Only in the proximal view of apically curved canals prepared with Ni-Ti S-files was significantly more dentine removed from the outer aspect of the curvature (P < 0.05). At the middle level (wide danger zone) the Ni-Ti S-files removed more dentine from the inner aspect of the curvature in those roots with apically curved canals. CONCLUSIONS: Under the conditions of this study, preparation with Ni-Ti K-files produced more appropriate shapes in roots with apically curved canals than Ni-Ti S-files.

Crystal chemistry of the 4,4'-dimethyl-2,2'bipyridine/copper bromide system.

The reaction of 4,4'-dimethyl-2,2-bipyridine (henceforth dmbp) with copper(I) and/or copper(II) bromide under a wide variety of conditions has led to the isolation of 10 different crystalline materials. These include one Cu(I) salt, [Cu(dmbp)(2)]Br (a distorted tetrahedral Cu species and a lattice Br(-) ion); two mixed valence Cu(I,II) compounds, [Cu(dmbp)(2)Br][CuBr(2)] (discrete 5-coordinated Cu(II) and linear Cu(I) species) and Cu(dmbp)(2)BrCu(2)Br(3) (linked 5-coordinate Cu(II) and trigonal planar Cu(I) species); and seven Cu(II) compounds, (dmbp)CuBr(2) (stacked planar monomers), [(dmbp)CuBr(2)](2)(five coordinate bibridged dimers), (dmbp)Cu(2)Br(4) (stacked planar bibridged dimers), (dmbp)CuBr(2)(DMSO) (five coordinate monomers), [Cu(dmbp)(2)Br]OH.5(1)/(2)H(2)O and [Cu(dmbp)(2)Br](Br/OH).5(1)/(2)H(2)O (five coordinate monomers), and (dmbpH(2))CuBr(4).H(2)O (distorted tetrahedral monomers). The crystal structure determinations of these materials are reported. A common thread in their structural chemistry is the supramolecular architecture developed through interdigitation of the dmbp rings on neighboring molecular species. The interdigitation leads to layer structures in many of the materials. The distances between the interdigitated dmbp rings are in the range 3.4-3.7 A. The Cu(dmbp)(2)Br(+) species exhibits an exceptionally large distortion from tetrahedral geometry due to deviation of the dihedral angle between the mean planes of the Cu(dmbp) fragments from 90 degrees. The Cu(dmbp)(2)Br(+) cations have distorted trigonal bipyramidal geometry, the Br(-) ion occupying an equatorial position. The length of the Cu-Br bond in the Cu(dmbp)(2)Br(+) species is correlated with the change in dihedral angle between the planes of the two dmbp ligands. The mono-dmbp complexes show a greater variation in coordination geometry for the Cu(II) species, including distorted trigonal bipyramidal and augmented square planar 4 + 1 and 4 + 2 coordination.

Reduced developmental potential in oocytes from women with endometriosis.

PURPOSE: To study retrospectively the outcome of intracytoplasmatic sperm injection (ICSI) in women with endometriosis compared with women with no known female infertility factor. METHODS: All couples treated with ICSI because of male infertility plus verified endometriosis (n = 26) and all couples treated with ICSI because of male infertility only (n = 125) during the period January 1995 to June 1999 were included. Data were collected from patient files and ICSI protocols. RESULTS: The time to complete down regulation was significantly longer (p = 0.0108), the dose of FSH significantly higher (0.0247), the day for oocyte pickup significantly later (p = 0.0091), and the cleavage rate of oocytes significantly lower (p = 0.0011) in women with endometriosis compared with controls. There was no significant difference in implantation rate or pregnancy rate between the groups. CONCLUSIONS: Women with endometriosis presented significantly reduced follicular response and oocyte cleavage rate, two mechanisms that might be related to a disturbed oogenesis.