The influence of synovial fluid on adenovirus‐mediated gene transfer to the synovial tissue

Objective

To determine the effect of synovial fluid (SF) from rheumatoid arthritis (RA) patients on adenovirus type 5 (Ad5)–mediated gene transfer to synoviocytes, and to explore new strategies for vector development based on the neutralization data obtained.

Methods

SF was derived from 63 randomly selected RA patients. Ten samples were used to study the effect of SF on Ad5‐mediated gene transfer in synoviocytes. IgG and <100‐kd fractions were purified from these 10 SF, and their effect on gene transfer was determined. Neutralizing activity against wild‐type Ad5 (wt‐Ad5), wt‐Ad26, wt‐Ad34, wt‐Ad35, and wt‐Ad48 was tested in the SF from the remaining 53 patients.

Results

Seven of 10 SF samples inhibited Ad5‐mediated gene transfer. Purified antibodies exhibited inhibition patterns similar to those seen with unfractionated SF. In 5 of 10 SF samples, low molecular weight fractions inhibited gene transfer at low dilutions. Neutralization of wt‐Ad35 by SF from RA patients was less frequent than neutralization of other wt‐Ad tested (4% versus 42–72%; n = 53).

Conclusion

SF from 70% of the RA patients contained neutralizing antibodies that hamper Ad5‐mediated gene transfer to synoviocytes. The activity of neutralizing antibodies may be circumvented in the majority of RA patients when vectors based on an Ad35 backbone are used.

     

Activity progression for anterior cruciate ligament injured individuals

Background

Functional exercises are important in the rehabilitation of anterior cruciate ligament deficient and reconstructed individuals but movement compensations and incomplete recovery persist. This study aimed to identify how tasks pose different challenges; and evaluate if different activities challenge patient groups differently compared to controls.

Methods

Motion and force data were collected during distance hop, squatting and gait for 20 anterior cruciate ligament deficient, 21 reconstructed and 21 controls.

Findings

Knee range of motion was greatest during squatting, intermediate during hopping and smallest during gait (P < 0.01). Peak internal knee extensor moments were greatest during distance hop (P < 0.01). The mean value of peak knee moments was reduced in squatting and gait (P < 0.01) compared to hop. Peak internal extensor moments were significantly larger during squatting than gait and peak external adductor moments during gait compared to squatting (P < 0.01). Fluency was highest during squatting (P < 0.01). All patients demonstrated good recovery of gait but anterior cruciate ligament deficient adopted a strategy of increased fluency (P < 0.01). During squatting knee range of motion and peak internal knee extensor moment were reduced in all patients (P < 0.01). Both anterior cruciate ligament groups hopped a shorter distance (P < 0.01) and had reduced knee range of motion (P < 0.025). Anterior cruciate ligament reconstructed had reduced fluency (P < 0.01).

Interpretation

Distance hop was most challenging; squatting and gait were of similar difficulty but challenged patients in different ways. Despite squatting being an early, less challenging exercise, numerous compensation strategies were identified, indicating that this may be more challenging than gait.     

Townes-Brocks syndrome: twenty novel SALL1 mutations in sporadic and familial cases and refinement of the SALL1 hot spot region

Townes-Brocks syndrome (TBS) is an autosomal dominant malformation syndrome characterized by renal, anal, ear, and thumb anomalies caused by SALL1 mutations. To date, 36 SALL1 mutations have been described in TBS patients. All but three of those, namely p.R276X, p.S372X, and c.1404dupG, have been found only in single families thereby preventing phenotype-genotype correlations. Here we present 20 novel mutations (12 short deletions, five short duplications, three nonsense mutations) in 20 unrelated families. We delineate the phenotypes and report previously unknown ocular manifestations, i.e. congenital cataracts with unilateral microphthalmia. We show that 46 out of the now 56 SALL1 mutations are located between the coding regions for the glutamine-rich domain mediating SALL protein interactions and 65 bp 3' of the coding region for the first double zinc finger domain, narrowing the SALL1 mutational hotspot region to a stretch of 802 bp within exon 2. Of note, only two SALL1 mutations would result in truncated proteins without the glutamine-rich domain, one of which is reported here. The latter is associated with anal, ear, hand, and renal manifestations, indicating that the glutamine-rich domain is not required for typical TBS.     

Workstyle and overcommitment in relation to neck and upper limb symptoms

Few studies have examined the concepts of workstyle and overcommitment in relation to the occurrence of neck and upper limb symptoms. The aim of this study was to examine whether a high-risk workstyle is a mediator in the relation of work-related exposure (job demands and computer work) and overcommitment to neck and upper limb symptoms. The study participants comprised 3,855 office workers of a European institute. The Sobel test was applied to test the intermediate effects of 4 workstyle dimensions and of the total workstyle score. The results show that most mediated effects were statistically significant, meaning that the workstyle dimensions acted as a mediator in the relation between work-related exposure and symptoms as well as in the relation between overcommitment and symptoms. Given the results with the total workstyle score, 34% of the effect of prolonged computer work, 64% of the effect of job demands, and 84% of the effect of overcommitment was mediated by workstyle. However, due to possible bias in the assessment of workstyle factor and the crosssectional design of the study, the conclusions should be drawn with care.     

Do prescribed drugs always follow the patients to hospital?

The reliability was measured with which out-patient medications were identified at the time of a patient's admission to a Dutch hospital. The hospital-pharmacy records and the community-pharmacy records of 205 patients have been reviewed with the help of an expert panel. We established that 15 inadvertent drug discontinuations of a serious nature occurred among 12 patients. While the results are not necessarily representative of the overall Dutch situation, they indicate errors that can be reduced. Improvement is suggested by transmission of computerized data from the community pharmacy to the hospital whenever a patient is admitted and agrees with this transmission.     

Coevolution of gene families in prokaryotes

We study gene family coevolution on a tree of life based on a large-scale ancestral gene content reconstruction, which includes gene duplication and deletion events. The insights obtained from this study are threefold: (1) Global properties, such as the distribution of coevolution partners and the formation of disconnected clusters of coevolving families, can be an inevitable consequence of evolution along a tree. (2) Concerted family expansion (gene duplication) and contraction (gene deletion) reflect functional constraints and therefore lead to better function prediction. (3) "Long-range" coevolutionary relationships, caused mostly by large family expansions or contractions, reveal high-level evolutionary organization of cellular processes in prokaryotes.     

Leucocyte depletion during cardiac surgery: a comparison of different filtration strategies

The results of leucocyte filtration during cardiac surgery are conflicting. This may be due to timing and duration of the filtration procedure, and to flow and pressure conditions in the filter. Therefore, we prospectively compared three major leucocyte filtration strategies in cardiac surgical patients. Forty patients were randomly divided into four groups. Group I: leucofiltration of arterial blood throughout cardiopulmonary bypass (CPB) (associated with high-flow and pressure gradients), Group II: leucofiltration of a part of the venous return blood in the rewarming phase during CPB (associated with intermediate flow, but high pressure), Group III: leucofiltration of residual heart-lung machine blood during transfusion into the patient after CPB (associated with low flow and low pressure), Group IV: control group without leucofiltration. We measured circulating leucocyte counts, plasma elastase levels and arterial blood oxygenation. Filters were postoperatively examined using scanning electronmicroscopy (SEM). Leucocyte counts increased over time and oxygenation decreased in all groups, without significant differences between the groups. SEM demonstrated extensive protein deposits and damaged leucocytes in the deeper layers of the filters from Group I. This was not observed in the filters from Group III. The postoperative plasma elastase levels increased in Groups II and IV and decreased in Groups I and III. In conclusion, we could not demonstrate a clinical difference among the three leucocyte depletion strategies. However, our laboratory results suggest that leucocyte filtration at low flow and pressure conditions is associated with less leucocyte damage and less release of elastase.     

Design and conduct considerations for studies in patients with impaired renal function

An impaired renal function, including acute and chronic kidney disease and end‐stage renal disease, can be the result of aging, certain disease conditions, the use of some medications, or as a result of smoking. In patients with renal impairment (RI), the pharmacokinetics (PKs) of drugs or drug metabolites may change and result in increased safety risks or decreased efficacy. In order to make specific dose recommendations in the label of drugs for patients with RI, a clinical trial may have to be conducted or, when not feasible, modeling and simulations approaches, such as population PK modeling or physiologically‐based PK modelling may be applied. This tutorial aims to provide an overview of the global regulatory landscape and a practical guidance for successfully designing and conducting clinical RI trials or, alternatively, on applying modeling and simulation tools to come to a dose recommendation for patients with RI in the most efficient manner.