Modification of the structural stability of human serum albumin in rheumatoid arthritis.

Differential scanning calorimetry (DSC) can indicate changes in structure and/or concentration of the most abundant proteins in a biological sample via heat denaturation curves (HDCs). In blood serum for example, HDC changes result from either concentration changes or altered thermal stabilities for 7-10 proteins and has previously been shown capable of differentiating between sick and healthy human subjects. Here, we compare HDCs and proteomic profiles of 50 patients experiencing joint-inflammatory symptoms, 27 of which were clinically diagnosed with rheumatoid arthritis (RA). The HDC of all 50 subjects appeared significantly different from expected healthy curves, but comparison of additional differences between the RA and the non-RA subjects allowed more specific understanding of RA samples. We used mass spectrometry (MS) to investigate the reasons behind the additional HDC changes observed in RA patients. The HDC differences do not appear to be directly related to differences in the concentrations of abundant serum proteins. Rather, the differences can be attributed to modified thermal stability of some fraction of the human serum albumin (HSA) proteins in the sample. By quantifying differences in the frequency of artificially induced post translational modifications (PTMs), we found that HSA in RA subjects had a much lower surface accessibility, indicating potential ligand or protein binding partners in certain regions that could explain the shift in HSA melting temperature in the RA HDCs. Several low abundance proteins were found to have significant changes in concentration in RA subjects and could be involved in or related to binding of HSA. Certain amino acid sites clusters were found to be less accessible in RA subjects, suggesting changes in HSA structure that may be related to changes in protein-protein interactions. These results all support a change in behavior of HSA which may give insight into mechanisms of RA pathology.

Development of a continuous direct compression platform for low-dose drug products.

In this work a continuous direct compression process was developed for a low-dosed drug product. Each unit operation of the GEA CDC-50 system was thoroughly investigated. This paper aimed to tackle the macroscopic and microscopic blend uniformity challenges inherently associated with continuous direct compression of cohesive and agglomerated APIs formulated at low dose. Density, compressibility and flow were identified as key material properties at the feeding stage. The screw speed coupled with powder flow regulated the gravimetric feeding performance. The impact of process and design variables was elucidated at the blending stage. The impeller configuration (number and pattern of radial mixing blades) and speed were key variables to steer the residence time distribution at the blending stage. An impeller configuration with distributed radial mixing blades could sufficiently filter the steady state feeding variability at low mixer speed, but exerted limited strain and shear on the blend. Hence micro-agglomerates persisted through the blending process and occasionally resulted in super potent tablets. Therefore, a new configuration was evaluated with more radial mixing blades centered on the impeller. This configuration resulted in a long mixing time at high tip speed which induced a maximized strain and shear. Consequently, excellent uniformity of the blend and tablets at macroscopic and microscopic level was achieved. Besides, this impeller improved robustness towards feeding disturbances, changes in process settings and variable blend properties. Next, it was demonstrated that the lubrication step requires critical attention during the design of the equipment, formulation and process. This study provided abundant evidence that an optimized continuous direct compression process allows direct compression of challenging low-dose drug products.

Sixteen-week analysis of unaltered elastomeric chain relating in-vitro force degradation with in-vivo extraction space tooth movement.

INTRODUCTION: The purposes of this study were to evaluate whether unaltered elastomeric chain can continue to move teeth for 16 weeks and to relate it to the amount of force remaining for the same batch of elastomeric chains. METHODS: The in-vivo portion of the study had a sample of 30 paired extraction space sites from 22 subjects who were measured for closure of the space every 28 days. The altered side elastomeric chain served as the control and was replaced at 28-day intervals whereas the experimental side remained unaltered. In the in-vitro portion of the study, 100 each of 2-unit and 3-unit segments of the same batch of elastomeric chains were placed in a water bath, and the force was measured for 20 of each segment length at the 28-day measurement points. RESULTS: Statistically significant amounts of space closure occurred at both the altered and unaltered sites at all measurement time points. The mean space closure at the altered sites was minimally greater than that observed at the paired unaltered sites. The mean differences of space closure between the altered and unaltered sites ranged from a minimum of -0.05 mm at 4 weeks to a maximum of -0.14 mm at 8 weeks. The elastomeric chain force degraded rapidly by 4 weeks but continued a gradual diminution of force to 86 g at 16 weeks. CONCLUSIONS: Unaltered elastomeric chain continued to move teeth into extraction spaces for 16 weeks in this sample from both statistically and clinically significant standpoints. There were minimal and statistically insignificant differences in the mean space closure measurements between the paired altered and unaltered sites. The elastomeric chain force at 16 weeks was less than 100 g, yet at the same time point, teeth continued to move clinically.

Continuous direct compression as manufacturing platform for sustained release tablets.

This study presents a framework for process and product development on a continuous direct compression manufacturing platform. A challenging sustained release formulation with high content of a poorly flowing low density drug was selected. Two HPMC grades were evaluated as matrix former: standard Methocel CR and directly compressible Methocel DC2. The feeding behavior of each formulation component was investigated by deriving feed factor profiles. The maximum feed factor was used to estimate the drive command and depended strongly upon the density of the material. Furthermore, the shape of the feed factor profile allowed definition of a customized refill regime for each material. Inline NIRs was used to estimate the residence time distribution (RTD) in the mixer and monitor blend uniformity. Tablet content and weight variability were determined as additional measures of mixing performance. For Methocel CR, the best axial mixing (i.e. feeder fluctuation dampening) was achieved when an impeller with high number of radial mixing blades operated at low speed. However, the variability in tablet weight and content uniformity deteriorated under this condition. One can therefore conclude that balancing axial mixing with tablet quality is critical for Methocel CR. However, reformulating with the direct compressible Methocel DC2 as matrix former improved tablet quality vastly. Furthermore, both process and product were significantly more robust to changes in process and design variables. This observation underpins the importance of flowability during continuous blending and die-filling. At the compaction stage, blends with Methocel CR showed better tabletability driven by a higher compressibility as the smaller CR particles have a higher bonding area. However, tablets of similar strength were achieved using Methocel DC2 by targeting equal porosity. Compaction pressure impacted tablet properties and dissolution. Hence controlling thickness during continuous manufacturing of sustained release tablets was crucial to ensure reproducible dissolution.

Draft Genome Sequence of Erwinia billingiae OSU19-1, Isolated from a Pear Tree Canker.

Plant-associated Erwinia include pathogenic and nonpathogenic species. We report the 5.6-Mb genome sequence of Erwinia billingiae OSU19-1, isolated from a canker on a pear tree inoculated with Erwinia amylovora. OSU19-1 and a closely related European isolate, E. billingiae Eb661(T), share many similarities including 40 kb of plasmid sequence.

Alefacept combined with tacrolimus, mycophenolate mofetil and steroids in de novo kidney transplantation: a randomized controlled trial.

Memory T cells play a central role in mediating allograft rejection and are a rational target for immunosuppressive therapy. Alefacept is a recombinant LFA3/IgG1 fusion protein that reduces the number of memory T cells in both psoriatic lesions and the peripheral circulation of psoriasis patients. This study evaluated the efficacy and safety of alefacept compared with placebo when combined with tacrolimus, mycophenolate mofetil and corticosteroids in de novo renal transplant recipients. Between December 2007 and March 2009 patients were randomized in a double-blind fashion to receive alefacept (n = 105) or placebo (n = 107) for 3 months and were then followed for a further 3 months. The primary efficacy endpoint was the incidence of biopsy-confirmed acute T cell mediated rejection (Banff grade ≥ 1) through Month 6. Memory T cell counts were significantly reduced in the alefacept group from Week 3 to study end compared with placebo. However, there was no significant difference between the alefacept and placebo groups for the primary efficacy endpoint (alefacept, 11.0% vs. placebo, 7.0%, p = 0.3). Patient and graft survival as well as renal function was similar between treatment groups. Safety and tolerability were generally similar between the treatment arms. Malignancy was higher in the alefacept treatment arm.

A phase 1, randomized ascending single-dose study of antagonist anti-human CD40 ASKP1240 in healthy subjects.

This first-in-human, phase I study evaluated the safety, tolerability, pharmacokinetic and pharmacodynamic profile of ASKP1240 in healthy subjects. Twelve sequential groups (each 6 active and 3 placebo) were randomly assigned to placebo or single ascending doses of intravenous ASKP1240 (0.00003-10 mg/kg). ASKP1240 exhibited nonlinear pharmacokinetics, with mean maximal serum concentrations and area under the serum concentration-time curves ranging from 0.7 to 251.6 µg/mL and 6.5 to 55409.6 h·µg/mL following doses 0.1 mg/kg-10 mg/kg, respectively. CD40 receptor occupancy by ASKP1240, which was dose-dependent, reached a maximum at doses above 0.01 mg/kg. ASKP1240 was well tolerated, with no evidence of cytokine release syndrome or thromboembolic events. Treatment emergent antibodies to ASKP1240 were detected in 5/70 (7.1%) ASKP1240 recipients. In conclusion, antagonism of the CD40/CD154 interaction with ASKP1240 was safe and well tolerated at the doses tested.

Effect of corticosteroid withdrawal on tacrolimus and mycophenolate mofetil exposure in a randomized multicenter study.

As corticosteroid-sparing protocols are increasingly utilized in kidney transplant recipients, it is crucial to understand potential drug interactions between tacrolimus (TAC) and the effect of corticosteroid withdrawal as well as to characterize dose adjustments of mycophenolate mofetil (MMF) in this setting. This prospective, multicenter, randomized, double-blind study included 397 patients who were randomized on posttransplant day 8 to receive either placebo (CSWD) or corticosteroid continuance (CCS). TAC trough levels at week two posttransplant were significantly greater in the CSWD group whereas TAC doses were comparable to the CCS group. This interaction was not observed in the African American subgroup. Higher serum creatinine and potassium levels were also observed in the CSWD group. MMF dose was significantly reduced in the CSWD group by the investigators because of decreased WBC counts, mostly outside of study protocol criteria, despite similar incidence of neutropenia and reported cytomegalovirus infection. Understanding TAC and MMF exposure in the context of corticosteroid-sparing protocols should allow for improved dosing of immunosuppressants and better management of posttransplant patients.

Heparan sulphate, its derivatives and analogues share structural characteristics that can be exploited, particularly in inhibiting microbial attachment

Heparan sulphate (HS) and the related polysaccharide, heparin, exhibit conformational and charge arrangement properties, which provide a degree of redundancy allowing several seemingly distinct sequences to exhibit the same activity. This can also be mimicked by other sulphated polysaccharides, both in overall effect and in the details of interactions and structural consequences of interactions with proteins. Together, these provide a source of active compounds suitable for further development as potential drugs. These polysaccharides also possess considerable size, which bestows upon them an additional useful property: the capability of disrupting processes comprising many individual interactions, such as those characterising the attachment of microbial pathogens to host cells. The range of involvement of HS in microbial attachment is reviewed and examples, which include viral, bacterial and parasitic infections and which, in many cases, are now being investigated as potential targets for intervention, are identified.

Heparan sulphate, its derivatives and analogues share structural characteristics that can be exploited, particularly in inhibiting microbial attachment.

Heparan sulphate (HS) and the related polysaccharide, heparin, exhibit conformational and charge arrangement properties, which provide a degree of redundancy allowing several seemingly distinct sequences to exhibit the same activity. This can also be mimicked by other sulphated polysaccharides, both in overall effect and in the details of interactions and structural consequences of interactions with proteins. Together, these provide a source of active compounds suitable for further development as potential drugs. These polysaccharides also possess considerable size, which bestows upon them an additional useful property: the capability of disrupting processes comprising many individual interactions, such as those characterising the attachment of microbial pathogens to host cells. The range of involvement of HS in microbial attachment is reviewed and examples, which include viral, bacterial and parasitic infections and which, in many cases, are now being investigated as potential targets for intervention, are identified.

Dense graphlet statistics of protein interaction and random networks.

Understanding evolutionary dynamics from a systemic point of view crucially depends on knowledge about how evolution affects size and structure of the organisms' functional building blocks (modules). It has been recently reported that statistics over sparse PPI graphlets can robustly monitor such evolutionary changes. However, there is abundant evidence that in PPI networks modules can be identified with highly interconnected (dense) and/or bipartite subgraphs. We count such dense graphlets in PPI networks by employing recently developed search strategies that render related inference problems tractable. We demonstrate that corresponding counting statistics differ significantly between prokaryotes and eukaryotes as well as between "real" PPI networks and scale free network emulators. We also prove that another class of emulators, the low-dimensional geometric random graphs (GRGs) cannot contain a specific type of motifs, complete bipartite graphs, which are abundant in PPI networks.

[A case-control study on green tea consumption and the risk of adult leukemia].

OBJECTIVE: To investigate whether green tea consumption can reduce the risk of adult leukemia. METHODS: A hospital-based matched case-control study was conducted in 2005 - 2006. We recruited 107 confirmed leukemia cases and 110 inpatient controls with orthopedic disease without leukemia or other malignancy matched on gender, age and hospitals that patients stayed. Related information were gathered on quantity, duration and frequency of tea consumption, demographic characteristics, exposure to radiation and occupational hazards, medications, using a validated questionnaire by face-to-face interview. Univariate and multivariate unconditional logistic regression analysis were used to estimate odds ratios (ORs) and associated 95% confidence intervals (CIs) with SPSS 11.5 software. RESULTS: Compared with non-tea-drinkers, the OR of those who consumed green tea was 0.58 (95% CI:0.34-1.00, P< 0.05) under univariate statistical analysis. The OR was 0.52 ( 95% CI: 0.28- 0.98, P = 0.04), using logistic regression to count for age, gender, residential area, smoking, level of education, exposure to radiation, benzene and organo-phosphorus. Compared with non-drinkers, the risk of adult leukemia declined with increasing quantity, duration, and frequency of green tea consumption. Tests for trend on dose-response was statistically significant (P < 0.01). CONCLUSION: A higher consumption of green tea seemed to be associated with a declined risk of adult leukemia. Tea consumption might be of help to human health planning projects.

One-year results with extended-release tacrolimus/MMF, tacrolimus/MMF and cyclosporine/MMF in de novo kidney transplant recipients.

Once-daily tacrolimus extended-release formulation (Prograf XL, formerly referred to as MR or MR4) was compared with the twice-a-day tacrolimus formulation (TAC) and cyclosporine microemulsion (CsA), all administered in combination with mycophenolate mofetil (MMF), corticosteroids and basiliximab induction, in a phase 3, randomized (1:1:1), open-label trial in 638 de novo kidney transplant recipients. In combination with MMF and corticosteroids, XL had an efficacy profile comparable to TAC and CsA. XL/MMF and TAC/MMF were statistically noninferior at 1-year posttransplantation to CsA/MMF for the primary efficacy endpoint, efficacy failure (death, graft loss, biopsy-confirmed acute rejection (BCAR) or lost to follow-up). One-year patient and graft survival were 98.6% and 96.7% in the XL/MMF group, 95.7% and 92.9% in TAC/MMF group and 97.6% and 95.7% in CsA/MMF group. The safety profile of XL in comparison with CsA was similar to that observed with TAC in this study and consistent with previously published reports of TAC in comparison with CsA. The results support the safety and efficacy of tacrolimus in combination with MMF, corticosteroids and basiliximab induction, as well as XL as a safe and effective once-daily dosing alternative.

Laparoscopic treatment of post renal transplant lymphoceles.

BACKGROUND: Traditionally, a post transplant lymphocele (PTL) is drained by widely opening the wall connecting the lymphocele cavity to the intraperitoneal space via laparotomy. We hypothesize that laparoscopic techniques can be effectively used for the treatment of PTL. METHODS: Patients requiring intervention for PTL between 1993 and 2002 were identified via a retrospective review. Results of drainage via laparotomy and laparoscopy were compared. RESULTS: During the study period 685 renal transplants (391 cadaveric, 294 living) were performed. The incidence of lymphocele was 5% [34/685 (36 cases)]. The indications for surgical drainage were local symptoms (69%), graft dysfunction (14%), or both (17%). The mean time to surgical therapy was 4.9 months. Laparoscopic drainage was performed in 25 patients (74%) and open drainage in 9 patients (26%). Open procedures were performed in cases for: previous abdominal surgery (5), undesirable lymphocele characteristics or location (2), or with concomitant open procedures (3). There were no conversions or operative complications in either group. There was no difference in operative time for the laparoscopic group vs the open group (108 +/- 6 vs 123 +/- 18 min, p = 0.8). Hospital stay was significantly shorter for the laparoscopic group (1.7 +/- 0.8 vs 3.8 +/- 1.0, p = 0.0007), with 88% of laparoscopic patients being either overnight admissions or same day surgery. Two patients (5%) developed symptomatic recurrences requiring reoperation [1 laparoscopic (4%), 1 open (10%)]. CONCLUSIONS: Laparoscopic fenestration of a peritransplant lymphocele is a safe and effective treatment. The large majority of patients treated with laparoscopic fenestration were discharged within one day of surgery. Unless contraindications exist, laparoscopy should be considered first-line therapy for the surgical treatment of posttransplant lymphocele.

The influence of attractiveness factors and distance to general practice surgeries by level of social disadvantage and global access in Perth, Western Australia.

The impact on attendance of the distance to general practice surgeries, and the attributes offered by those surgeries, was investigated. One thousand and forty four subjects, selected at random from the metropolitan area of Perth, Western Australia, responded to a household interview survey concerning which attributes of general practice (GP) surgeries they found attractive and the identity of the surgery they preferred to attend. The sample was stratified by different levels of social disadvantage and by good and poor global levels of spatial accessibility of GP surgeries. In separate fieldwork, interviewers collected detailed environmental data from practice staff at 466 GP surgeries available to the community survey respondents within metropolitan Perth. Respondents living in areas of poor global access were more likely to attend their nearest surgery (25% vs. 6%) and to bypass fewer surgeries to attend a preferred surgery (median 2 vs. 20). Those who were most socially disadvantaged were less likely than those who were better off to bypass surgeries where global access was poor, but more likely to bypass nearby surgeries and to seek out a surgery that bulk billed in areas where global access was good. A number of attractiveness factors had an important influence on choice of surgery, including: 'easy to make an appointment'; 'generally sees patients on time'; 'pharmacy nearby'; 'bulk bills' and 'open at all on Sundays'. Respondents attending their nearest surgery were more likely to have all of their nominated 'very important' attributes satisfied at that surgery than non-attenders (40% vs. 16%). A logistic regression model, adjusting for distance effects and size of surgery, showed within each level of global access and social disadvantage a consistent increase in the odds of attending a surgery that satisfied the attributes desired by respondents.

Evaluation of an automated preanalytical robotic workstation at two academic health centers.

BACKGROUND: Purchase of automated systems in today's clinical laboratory needs justification based on demonstrable improvements in efficiency and a sound payback model. Few studies provide information on laboratory automation that focuses on the preanalytical portion of specimen processing. METHODS: We recently evaluated an automated preanalytical processing unit (GENESIS FE500) at two academic health centers. This preanalytical unit processes blood specimens through automated specimen sorting, centrifugation, decapping, labeling, aliquoting, and placement of the processed specimen in the analytical rack. We quantified the output of the FE500 by processing >3000 barcode-labeled specimens according to a protocol designed to test all of the features of this automated specimen-processing unit. RESULTS: Depending on the batch size, aliquot number requested, and percentage of tubes that required centrifugation, the mean system output performance varied between 93 and 502 total tubes/h. Throughput increased when the batch size expanded from 40 or 100 samples (mean = 211 total tubes processed/h) to batch sizes of 200 and 300 tubes (mean = 474 total tube processed/h). The GENESIS FE500 processed specimen tubes differing in size from 13 x 65 mm (width x height) to 16 x 100. At one site, the FE500 was operated by one person, compared with the three individuals required to perform the same tasks manually. Finally, the specimen-processing error rate determined at one of the institutions was significantly reduced. CONCLUSIONS: We conclude that the GENESIS FE500 effectively reduces the labor associated with specimen processing; decreases the number of laboratory errors that occur with specimen sorting, labeling, and aliquoting; and improves the integrity of specimen handling throughout the steps of specimen processing.

Oral glucose solution for analgesia in infant circumcision.

OBJECTIVES: Our objectives were to determine if a 50% dextrose solution would reduce the percentage of circumcision procedure time a neonate spent crying by 50% compared with water and whether it would be similar to a dorsal penile nerve block (DPNB). STUDY DESIGN: This was a randomized placebo-controlled blinded clinical trial. POPULATION: We included 71 patients who were recruited from the inpatient nursery of a military community hospital over a 5-month period. OUTCOMES MEASURED: The primary outcome was the percentage of the procedure time neonates spent crying. Secondary outcomes were the percentage change in heart rate from baseline, the percentage of oxygen saturation, and the score from the modified behavioral pain scale. RESULTS: There were no significant differences between the oral glucose and water groups among any of the pain-related measurements. The DPNB group had significantly lower pain-related measurements (P <.05). CONCLUSIONS: Concentrated glucose administered orally does not provide significant analgesia in neonatal circumcision. The use of DPNB significantly reduced objective measurements of pain and physiologic stress in infants undergoing circumcision.

Technical competency in flexible sigmoidoscopy.

BACKGROUND: Family practice residencies offer training in flexible sigmoidoscopy; however, there are no objective recommendations for determining competency in learners. We describe a longitudinal study designed to determine the mean procedure time and depth of insertion for family practice residents. METHODS: During a 21-month period, data were collected for 421 patients undergoing flexible sigmoidoscopy. Second- or third-year family medicine residents supervised by family medicine faculty did all procedures. The data were analyzed with simple descriptive statistics, t test, and linear and logistic regression. RESULTS: The mean procedure time was 18+/-9.3 minutes (17.2 - 19.6, 95% confidence interval [CI]). The mean insertion depth was 51.4+/-12.4 cm (50.4 - 52.6 cm, 95% CI). Performing a biopsy was associated with an increase in procedure time (17.0 vs 27.3 minutes, P < .0001). Women with a history of pelvic surgery had less depth of insertion than did those who had no history of pelvic surgery (47 vs 53 cm, P = .002, t test). CONCLUSION: Procedural competency consists of knowledge, technical skills, and attitudes. Knowledge and attitudes can be assessed with other items, such as examinations and observation. Primary care faculty can now use these standards of insertion depth and procedure time when determining technical skill proficiency for their residents in flexible sigmoidoscopy.

Completed suicides and emergency psychiatric evaluations: the Louisville experience.

Suicide is one of the most serious outcomes of psychiatric illness, and the most extreme intervention (involuntary hospitalization) can be exercised if this event is likely. Despite this, the rate of suicide has remained fairly consistent at 1.1-1.4%. In an ongoing effort of identifying factors that can predict subsequent suicides, we retrospectively examined the records of individuals who completed suicides in Jefferson County, January 1997 through September 1998, and who were evaluated at the Emergency Psychiatric Service (EPS) at University of Louisville Hospital. Fifteen of the 132 (11.4%) subjects who completed suicide were evaluated at some point in time at EPS. Only 8 (6.1%) were seen within 60 days of the fatal event. This represents less than 0.1% of the total 9,469 patients seen at the EPS during this time period. No specific factors could be identified that predicted imminent suicide. Given the inaccuracy in being able to predict suicide, clinicians need to continue to be vigilant when assessing acutely distressed substance abusing or psychiatric patients.