Setup of a Contamination Control Strategy Using the Hazard Analysis Critical Control Point (HACCP) Methodology.

A Contamination Control Strategy (CCS) is a document that focuses on how to prevent contaminations with microorganisms, particles, and pyrogens within sterile and/or aseptic and preferably also in nonsterile manufacturing facilities. This document determines to what extent measures and controls in place are efficient in preventing contamination. In order to efficiently evaluate and control all potential hazards associated with sources of contamination within a CCS, the Hazard Analysis Critical Control Point (HACCP) methodology could be a useful tool to monitor all Critical Control Points (CCPs) related to various sources of contamination. This article describes a way to set up the CCS within a pharmaceutical sterile and aseptic manufacturing facility (GE HealthCare Pharmaceutical Diagnostics) by applying the HACCP methodology. In 2021, a global CCS procedure and a general HACCP template became effective for the GE HealthCare Pharmaceutical Diagnostics sites having sterile and/or aseptic manufacturing processes. This procedure guides the sites through the setup of the CCS by applying the HACCP methodology and helps each site to evaluate whether the CCS is still effective taking all (proactive and retrospective) data following the CCS into account. A summary of setting up a CCS using the HACCP methodology, specifically for the pharmaceutical company GE HealthCare Pharmaceutical Diagnostics Eindhoven site, is provided in this article. Use of the HACCP methodology enables a company to include proactive data within the CCS, making use of all identified sources of contamination, associated hazards, and/or control measures and CCPs. The constructed CCS allows the manufacturer to identify whether all included sources of contamination are under control and, if not, which mitigatory actions need to be performed. All current states are reflected by a traffic light color to reflect the level of residual risk, thereby providing a simple and clear visual representation of the current contamination control and microbial state of the manufacturing site.

Device and method for nonlinear ultrasonic measurements on highly irradiated 304 stainless steel specimens in a hot cell environment.

This paper reports on the first successful nonlinear ultrasonic measurement on highly irradiated specimens in a hot cell environment. The specimens are ANSI 304 stainless steel specimens for which the microstructure characterization and ultrasonic velocity measurement have been previously conducted. The critical part of this research is the development of an automatic fixture device that can facilitate repeatable loading and unloading to place the contact ultrasonic transducers on and off of the specimen. The key step to achieve high measurement repeatability is a careful adjustment of the support-spring constants such that the contact force at the interface between the transducer face and specimen surface is as uniform and constant as possible. The longitudinal ultrasonic velocities, which are obtained as a by-product of the nonlinear ultrasonic measurements, show a level of random variation in terms of (max-min)/average (%) below 0.2%, and the velocity distributions and magnitudes are in good agreement with those from the previous work. The ultrasonic nonlinearity parameters show the level of random variation below 4.7%, which is extremely low, considering that the measurements are conducted in a hot cell environment. The nonlinearity parameters also show a strong dependence on the measurement location in a particular specimen with respect to the radiation source, demonstrating a possible inhomogeneous microstructure evolution in these 12.7 mm thick specimens. This research demonstrates the feasibility of making nonlinear ultrasonic measurement on highly radioactive materials and/or in a highly radioactive environment using the device and procedure developed.

The transmission of OXPHOS disease and methods to prevent this.

Diseases owing to defects of oxidative phosphorylation (OXPHOS) affect approximately 1 in 8,000 individuals. Clinical manifestations can be extremely variable and range from single-affected tissues to multisystemic syndromes. In general, tissues with a high energy demand, like brain, heart and muscle, are affected. The OXPHOS system is under dual genetic control, and mutations in both nuclear and mitochondrial genes can cause OXPHOS diseases. The expression and segregation of mitochondrial DNA (mtDNA) mutations is different from nuclear gene defects. The mtDNA mutations can be either homoplasmic or heteroplasmic and in the latter case disease becomes manifest when the mutation exceeds a tissue-specific threshold. This mutation load can vary between tissues and often an exact correlation between mutation load and phenotypic expression is lacking. The transmission of mtDNA mutations is exclusively maternal, but the mutation load between embryos can vary tremendously because of a segregational bottleneck. Diseases by nuclear gene mutations show a normal Mendelian inheritance pattern and often have a more constant clinical manifestation. Given the prevalence and severity of OXPHOS disorders and the lack of adequate therapy, existing and new methods for the prevention of transmission of OXPHOS disorders, like prenatal diagnosis (PND), preimplantation genetic diagnosis (PGD), cytoplasmic transfer (CT) and nuclear transfer (NT), are technically and ethically evaluated.

Transmission and prenatal diagnosis of the T9176C mitochondrial DNA mutation.

A family presented with three affected children with Leigh syndrome, a progressive neurodegenerative disorder. Analysis of the OXPHOS complexes in muscle of two affected patients showed an increase in activity of pyruvate dehydrogenase and a decrease of complex V activity. Mutation analysis revealed the T9176C mutation in the mtATPase 6 gene (OMIM 516060) and the mutation load was above 90% in the patients. Unaffected maternal relatives were tested for carrier-ship and one of them, with a mutation load of 55% in blood, was pregnant with her first child. The possibility of prenatal diagnosis was evaluated. The main problem was the lack of data on genotype-phenotype associations for the T9176C mutation and on variation of the mutation percentage in tissues and in time. Therefore, multiple tissues of affected and unaffected carriers were analysed. Eventually, prenatal diagnosis was offered with understanding by the couple that there could be considerable uncertainty in the interpretation of the results. Prenatal diagnosis was carried out twice on cultured and uncultured chorion villi and amniotic fluid cells. The result was a mutation percentage just below the assumed threshold of expression (90%). The couple decided to continue the pregnancy and an apparently healthy child was born with an as yet unclear prognosis. This is the first prenatal diagnosis for a carrier of the T9176C mutation. Prenatal diagnosis for this mutation is technically reliable, but the prognostic predictions are not straightforward.

Pearson syndrome and the role of deletion dimers and duplications in the mtDNA.

Pearson syndrome is an often fatal multisystem disease associated with mitochondrial DNA rearrangements. Here we report a patient with a novel mtDNA deletion of 3.4 kb ranging from nucleotides 6097 to 9541 in combination with deletion dimers. The mutation percentage in different tissues (blood, muscle and liver) varied between 64% and 95%. After a remission period of about a year, the patient suddenly died at the age of 3 years owing to a severe lactic acidosis. A second patient with a previously reported deletion of 8 kb and a milder phenotype was found to have mitochondrial duplications and died at the age of 10 years. From these data and data from previous reports, we hypothesize that duplications might be beneficial in the clinical course of the disease and in life expectancy.

Crack characterization using guided circumferential waves.

This paper examines the propagation of guided circumferential waves in a hollow isotropic cylinder that contains a crack, with the goal of using these guided waves to both locate and size the crack. The crack is sized using a modified Auld's formula, which relates the crack's length to a reflected energy coefficient. The crack is then located by operating on the backscattered signal with a time-frequency digital signal processing (DSP) technique, and then comparing these results to those obtained if the cylinder is perfect. The guided circumferential waves are generated with a commercial finite element method (FEM) code. One objective of this work is to demonstrate the effectiveness of using sophisticated DSP techniques to describe the effect of scattering on dispersive waves, showing it is possible to characterize cracks systematically and accurately by quantifying this scattering effect. The results show that the need for high frequency signals to detect small cracks is significantly decreased by using these techniques.

Time-frequency representations of Lamb waves.

The objective of this study is to establish the effectiveness of four different time-frequency representations (TFRs)--the reassigned spectrogram, the reassigned scalogram, the smoothed Wigner-Ville distribution, and the Hilbert spectrum--by comparing their ability to resolve the dispersion relationships for Lamb waves generated and detected with optical techniques. This paper illustrates the utility of using TFRs to quantitatively resolve changes in the frequency content of these nonstationary signals, as a function of time. While each technique has certain strengths and weaknesses, the reassigned spectrogram appears to be the best choice to characterize multimode Lamb waves.

Multiplex PCR of polymorphic markers flanking the CFTR gene; a general approach for preimplantation genetic diagnosis of cystic fibrosis.

Cystic fibrosis (CF) is the first monogenic disorder for which single cell preimplantation genetic diagnosis (PGD) has been successfully applied. The spectrum of mutations in CF is extremely heterogeneous, and hence, the development of mutation-specific PGD protocols is impracticable. The current study reports the development and evaluation of a general multiplex marker polymerase chain reaction (PCR) protocol for PGD of CF. Four closely linked highly polymorphic (CA)(n) repeat markers D7S523, D7S486, D7S480 and D7S490, flanking the cystic fibrosis transmembrane regulator (CFTR) gene, were used. In 99% of the single cells tested (100 leukocytes and 50 blastomeres), multiplex PCR results were obtained and the overall allelic drop out (ADO) rate varied from 2 to 5%. After validation for the presence of ADO and additional alleles, 95% of the multiplex PCR results were accepted to construct the marker genotypes. Depending on the genotype of the couple, and taking into account the embryos lost for transfer due to validation criteria (5%), ADO (0-2%) and single recombination (1.1-3%), in general >90% of the embryos could be reliably genotyped by PGD using a single blastomere. The risk of misdiagnosis equals the chance of a double recombination between informative flanking markers and is <0.05%. Therefore, this polymorphic and multi-allelic marker system is a reliable and generally applicable alternative for mutation-directed PGD protocols. Furthermore, it provides a test for the origin of the detected genotype and also gives an indication of the chromosomal ploidy status of the blastomere tested.

Reversible dilated cardiomyopathy induced by methamphetamine.

This brief report details the case of a woman who was thought, after extensive evaluation, to have idiopathic congestive cardiomyopathy. It was subsequently found that systolic ventricular dysfunction was due to amphetamine abuse, and that ventricular function normalized after discontinuation of this drug.

Ventricular arrhythmias during Swan-Ganz catheterization of the critically ill.

The incidence of ventricular arrhythmias in critically-ill patients during bedside right-sided heart catheterization with a flow-directed balloon-tipped catheter was determined. Twenty-nine of 60 catheterizations (48 percent) were associated with premature ventricular contractions and 20 (33 percent) were associated with ventricular tachycardia. Two patients required antiarrhythmic therapy or a precordial thump to convert ventricular tachycardia. One patient developed ventricular tachycardia and fibrillation and died. Serious catheter-induced arrhythmias, including sustained ventricular tachycardia, may occur during Swan-Ganz catheterization of the critically ill.

Imitative behavior by Down's syndrome persons.

Three scales of imitative behavior (nonverbal, vocal, and verbal) were administered to 28 Down's syndrome subjects and 56 subjects with other diagnoses. All of the subjects were severely retarded and had at least partial vision and hearing, and the two groups were closely comparable with respect to sex, CA, and IQ. The results provided no support for the hypothesis--backed by a century of clinical observation--that individuals with Down's syndrome "are outstanding in their mimicry" (Belmont, 1971, p. 38).