Ductal Carcinoma In Situ with Microinvasion on Core Biopsy: Evaluating Tumor Upstaging Rate,Lymph Node Metastasis Rate,and Associated Predictive Variables

Annals of Surgical Oncology - The role of sentinel lymph node biopsy (SLNB) when ductal carcinoma in situ with microinvasion (DCISM) is identified on core biopsy is unclear. Our aim was to assess...     

Physicians’ attitudes about interprofessional treatment of chronic pain: family physicians are considered the most important collaborators

Scand J Caring Sci; 2013; 27; 303–310 Physicians’ attitudes about interprofessional treatment of chronic pain: family physicians are considered the most important collaborators Aims and objectives: Interprofessional collaboration is the process in which different professional groups work together to positively impact health care. We aimed to explore physicians’ attitudes toward interprofessional collaboration in the context of chronic pain management with the implication that if attitudes are not positive, appropriate interventions could be developed. Design: A quantitative attitudes study. Ethical issues: The ethical committee approved the study. Methods: A web‐based survey about interprofessional treatment of chronic pain was administered to physicians. Outcome measures were as follows: physicians’ demographic and workplace information, previous experience of working within an interprofessional team, and attitudes towards interprofessional collaboration in chronic pain management. Results: There were 90 physicians who responded to the survey. Physicians had positive attitudes towards team work in the context of chronic pain, but they were undecided about sharing their role within an interprofessional team. The family physician was singled out as the most important as well as the most common collaborator in chronic pain treatment. Interprofessional educational seminars and workshops were suggested as methods for improving interprofessional collaboration. Conclusions: Interprofessional collaboration may be enhanced with continuing medical education that will bring together different healthcare professionals, enable them to exchange experiences and learn about their potential roles within a team.     

Success rates of nuclear short tandem repeat typing from different skeletal elements

Aim

To evaluate trends in DNA typing success rates of different skeletal elements from mass graves originating from conflicts that occurred in the former Yugoslavia (Bosnia and Herzegovina and Kosovo) during the 1990s, and to establish correlation between skeletal sample age and success of high throughput short tandem repeat (STR) typing in the large data set of the International Commission on Missing Persons.

Method

DNA extraction and short tandem repeat (STR) typing have been attempted on over 25 000 skeletal samples. The skeletal samples originated from different geographical locations where the conflicts occurred and from different time periods from 1992 to 1999. DNA preservation in these samples was highly variable, but was often significantly degraded and of limited quantity. For the purpose of this study, processed samples were categorized according to skeletal sample type, sample age since death, and success rates tabulated.

Results

Well-defined general trends in success rates of DNA analyses were observed with respect to the type of bone tested and sample age. The highest success rates were observed with samples from dense cortical bone of weight-bearing leg bones (femur 86.9%), whereas long bones of the arms showed significantly lower success (humerus 46.2%, radius 24.5%, ulna 22.8%). Intact teeth also exhibited high success rates (teeth 82.7%). DNA isolation from other skeletal elements differed considerably in success, making bone sample selection an important factor influencing success.

Conclusion

The success of DNA typing is related to the type of skeletal sample. By carefully evaluating skeletal material available for forensic DNA testing with regard to sample age and type of skeletal element available, it is possible to increase the success and efficiency of forensic DNA testing.The aftermath of the 1992-1995 conflict in the area of former Yugoslavia was marked with estimated 40 000 missing individuals. To address the issue of missing persons, the International Commission on Missing Persons (ICMP) was created in 1996 following the G-7 Summit in Lyon, France. ICMP’s mandate was expanded to also cover the DNA typing of missing persons resulting from 1999 conflict in Kosovo region.ICMP employs a “population based, DNA-led” identification system for the identification of missing persons in the region of former Yugoslavia. On a regional scale, DNA profiles from reference samples of living relatives of missing persons are continuously compared in a batch mode to the DNA profiles obtained from mortal remains of victims. To date, more than 84 000 blood samples representing over 28 000 missing individuals have been collected, analyzed, and entered into the database. Since 2001, short tandem repeat (STR) profiles from more than 21 000 skeletal samples, representing more than 15 000 different individuals, have also been entered into the ICMP database (1). DNA matching reports of greater than 99.95% probability of identity have been issued for over 11 600 individuals.ICMP DNA laboratories currently operate at a target rate of 105 bone or tooth extractions per day, using a silica-based extraction method (1-3). Bone and teeth samples tested are between 8 and 15 years post mortem. The quality of DNA preservation in these bones is highly variable and often substantially limited or/and degraded. This reflects the fact that the remains were buried or disposed in many different environmental contexts, with differential exposure to potentially harsh extrinsic factors such as temperature, UV radiation, humidity, and exposure to animals, insects, and microbes. Different disposal conditions are marked by burial in different soil types, complete or partial immersion of remains in water, contact with fire, or use of plastic sheeting. Microbial degradation is variably evidenced in these samples by both bone morphology and co-extraction of sometimes large amounts of microbial DNA (our unpublished observation). As always in this type of work, co-extraction of DNA inhibitors is a serious issue, and is also variable among samples.Bone and teeth samples clearly protect DNA through their physical and/or chemical robustness to environmental degradation and/or biological attack. An elementary manifestation of this is that bone and teeth are often the only surviving material that can be tested. However, the mechanisms by which DNA is preserved in bone are not very well characterized (4). Bone tissue is primarily composed of protein and mineral. The two most abundant proteins in bone tissue are collagen and osteocalcin. Approximately 70% of the mineral portion of the bone is composed of hydroxyapatite, which includes calcium phosphate, calcium carbonate, calcium fluoride, calcium hydroxide, and citrate. Structural arrangement of bone tissue is such that the mineral portion provides structural support to the protein portion in the bone and, by doing so, physically excludes exogenous/extracellular agents/enzymes that are potentially harmful to the protein portion of the bone (4). DNA has a very strong affinity for hydroxyapatite. DNA degradation is linked to the loss of crystallinity in the hydroxyapatite, but it may also be related to the loss of collagen (5).Overall, it seems reasonable to suppose that the characteristics of the bone that are correlated with its general long term survival, ie its resistance to morphological degradation at the macroscopic and microscopic level, would be those that contribute to the protection of DNA from degradation. Bone density, ie the extent of mineralization, is one of the most important intrinsic factors in survival of bone material. There is a significant difference in bone density between men and women, with the latter showing lower density values. The difference in bone density is also specific for different areas of the skeletal element morphology, with the highest density values noted for the mid-shaft region (6). Teeth are the hardest tissue in the human body because of the dental enamel (7).To know which bones best preserve DNA is of fundamental importance to DNA identification casework in mass fatality incidents and mass graves from armed conflict or genocide. The question equally applies to “ancient DNA” analyses in archaeological or human molecular evolutionary investigations. Despite the logical expectation that denser, more intact bones may be preferable, there is very little empirical data published on this issue (8). We also note that a successful recovery of DNA is linked not only to the degree of protection within the bone, but also the total amount of starting DNA. One reason for the lack of precise information on the best samples for DNA testing from degraded bone is the difficulty in performing controlled experiments, with regard to the effect of relevant environmental variables, inter-individual variation (related to for example sex or age), the long periods of time involved, and the need for large sample size.The aim of our study was to analyze DNA typing success rates from very large sample sizes of various skeletal elements from victims of conflict in the former Yugoslavia. Given the large number of variables affecting DNA preservation, a large sample size helps to average out the influences of a wide range of environmental contexts and permit general conclusions. Further, we divided our data into three time periods, with respect to time since death. This allows the analysis of the relative rate of degradation in different skeletal elements over time. These empirical data can serve as a useful guide to sampling strategies from degraded skeletal remains.     

Changes in cardiac innervation during maturation in long-term diabetes

Diabetic autonomic neuropathy being a common complication of diabetes mellitus (DM) is related to an increased risk of cardiovascular mortality. However, mechanisms underlying changes of innervation density in affected hearts remain insufficiently understood. Hence, the aim of this study was to describe quantitative changes of intra-myocardial nerve terminals in hearts of diabetic rats of various ages.     

Estimation of expiratory time constants via fuzzy clustering

Objective.In mechanically ventilated patients the expiratorytime constant provides information about respiratory mechanics. In thepresent study a new method, fuzzy clustering, is proposed to determineexpiratory time constants. Fuzzy clustering differs from other methodssince it neither interferes with expiration nor presumes any functionalrelationship between the variables analysed. Furthermore, time constantbehaviour during expiration can be assessed, instead of an average timeconstant. The time constants obtained with fuzzy clustering are comparedto time constants conventionally calculated from the same expirations.Methods.20 mechanically ventilated patients, including 10patients with COPD, were studied. The data of flow, volume and pressurewere sampled. From these data, four local linear models were detected byfuzzy clustering. The time constants () of the local linear models(clusters) were calculated by a least-squares technique. Time constantbehaviour was analysed. Time constants obtained with fuzzy clusteringwere compared to time constants calculated from flow-volume curves usinga conventional method. Results.Fuzzy clustering revealed twopatterns of expiratory time constant behaviour. In the patients withCOPD an initial low time constant was found (mean 1: 0.33 s, SD0.21) followed by higher time constants; mean 2: 2.00 s (SD0.91s), mean 3: 3.45 s (SD 1.44) and mean 4: 5.47 s (SD2.93). In the other patients only minor changes in time constants werefound; mean 1: 0.74 s (SD 0.30), mean 2: 0.90 s (SD 0.23),mean 3: 1.04 s (SD 0.42) and mean 4: 1.74 s (SD 0.78). Boththe pattern of expiratory time constants, as well as the time constantscalculated from the separate clusters, were significantly differentbetween the patients with and without COPD. Time constants obtained withfuzzy clustering for cluster 2, 3 and 4 correlated well with timeconstants obtained from the flow-volume curves. Conclusions.Inmechanically ventilated patients, expiratory time constant behaviour canbe accurately assessed by fuzzy clustering. A good correlation was foundbetween time constants obtained with fuzzy clustering and time constantsobtained by conventional analysis. On the basis of the time constantsobtained with fuzzy clustering, a clear distinction was made betweenpatients with and without COPD.     

Increased arginase activity underlies allergen-induced deficiency of cNOS-derived nitric oxide and airway hyperresponsiveness

1. A deficiency of constitutive nitric oxide synthase (cNOS)-derived nitric oxide (NO), due to reduced availability of L-arginine, importantly contributes to allergen-induced airway hyperresponsiveness (AHR) after the early asthmatic reaction (EAR). Since cNOS and arginase use L-arginine as a common substrate, we hypothesized that increased arginase activity is involved in the allergen-induced NO deficiency and AHR. 2. Using a guinea-pig model of allergic asthma, we addressed this hypothesis by examining the effects of the specific arginase inhibitor N(omega)-hydroxy-nor-L-arginine (nor-NOHA) on the responsiveness to methacholine of isolated perfused tracheae from unchallenged control animals and from animals 6 h after ovalbumin challenge. Arginase activity in these preparations was investigated by measuring the conversion of L-[14C]arginine to [14C]urea. 3. Airways from allergen-challenged animals showed a 2 fold (P<0.001) increase in responsiveness to intraluminal (IL) administration of methacholine compared to controls. A similar hyperresponsiveness (1.8 fold, P<0.01) was observed in control airways incubated with the NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 0.1 mM, IL), while L-NAME had no further effect on the airways from challenged animals. 4. Remarkably, 5 microM nor-NOHA (IL) normalized the hyperresponsiveness of challenged airways to basal control (P<0.001), and this effect was fully reversed again by 0.1 mM L-NAME (P<0.05). Moreover, arginase activity in homogenates of the hyperresponsive airways was 3.5 fold (P<0.001) enhanced compared to controls. 5. The results indicate that enhanced arginase activity contributes to allergen-induced deficiency of cNOS-derived NO and AHR after the EAR, presumably by competition with cNOS for the common substrate, L-arginine. This is the first demonstration that arginase is involved in the pathophysiology of asthma.     

Sorbitol Crystallization Can Lead to Protein Aggregation in Frozen Protein Formulations

Purpose This work examines the cause of aggregation of an Fc-fusion protein formulated in sorbitol upon frozen storage for extended periods of time at −30°C.Materials and Methods We designed sub-ambient differential scanning calorimetry (DSC) experiments to capture the effects of long-term frozen storage. The physical stability of formulation samples was monitored by size exclusion high performance liquid chromatography (SE-HPLC).Results DSC analysis of non-frozen samples shows the expected glass transitions (T_g′) at −45°C for samples in sorbitol and at −32°C in sucrose. In time course studies where sorbitol formulations were stored at −30°C and analyzed by DSC without thawing, two endothermic transitions were observed: a melting endotherm at −20°C dissipated over time, and a second endotherm at −8°C was seen after approximately 2 weeks and persisted in all later time points. Protein aggregation was only seen in the samples formulated in sorbitol and stored at −30°C, correlating aggregation with the aforementioned melts.Conclusions The observed melts are characteristic of crystalline substances and suggest that the sorbitol crystallizes over time. During freezing, the excipient must remain in the same phase as the protein to ensure protein stability. By crystallizing, the sorbitol is phase-separated from the protein, which leads to protein aggregation.     

Cystatin C as a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma

The concentration of cysteine protease inhibitor cystatin C was determined in sera from 59 patients with non-Hodgkin B-cell lymphoma using ELISA. The sera from 43 age and sex matched healthy blood donors served as controls. Cystatin C was significantly increased in sera of patients without therapy (mean 1136+/-SE 105.7ng/ml, p=0.00001) and with therapy (mean 1073+/-52ng/ml, p=0.001) compared to controls (mean 819+/-28ng/ml). The highest levels were determined in sera of patients with a relapse (mean 1680+/-196ng/ml). By using immunofluorescence staining and confocal microscopy we determined immature dendritic cells as a major population of cystatin C positive cells in affected lymph nodes. Our study reports for the first time that cystatin C is a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.     

Combined acute traumatic aortic injury and left main bronchus transection in a 5-year-old child

Blunt traumatic aortic and tracheobronchial injuries are extremely rare in children. We report a case of a 5 year old male who suffered both of these rare injuries (traumatic aortic rupture and left mainstem bronchus transaction). To our knowledge this combination of injuries has not been previously described in a child. CT with multiplanar images was critical for the detection of both injuries.