A Recurrent De Novo PACS2 Heterozygous Missense Variant Causes Neonatal-Onset Developmental Epileptic Encephalopathy, Facial Dysmorphism, and Cerebellar Dysgenesis.

Developmental and epileptic encephalopathies (DEEs) represent a large clinical and genetic heterogeneous group of neurodevelopmental diseases. The identification of pathogenic genetic variants in DEEs remains crucial for deciphering this complex group and for accurately caring for affected individuals (clinical diagnosis, genetic counseling, impacting medical, precision therapy, clinical trials, etc.). Whole-exome sequencing and intensive data sharing identified a recurrent de novo PACS2 heterozygous missense variant in 14 unrelated individuals. Their phenotype was characterized by epilepsy, global developmental delay with or without autism, common cerebellar dysgenesis, and facial dysmorphism. Mixed focal and generalized epilepsy occurred in the neonatal period, controlled with difficulty in the first year, but many improved in early childhood. PACS2 is an important PACS1 paralog and encodes a multifunctional sorting protein involved in nuclear gene expression and pathway traffic regulation. Both proteins harbor cargo(furin)-binding regions (FBRs) that bind cargo proteins, sorting adaptors, and cellular kinase. Compared to the defined PACS1 recurrent variant series, individuals with PACS2 variant have more consistently neonatal/early-infantile-onset epilepsy that can be challenging to control. Cerebellar abnormalities may be similar but PACS2 individuals exhibit a pattern of clear dysgenesis ranging from mild to severe. Functional studies demonstrated that the PACS2 recurrent variant reduces the ability of the predicted autoregulatory domain to modulate the interaction between the PACS2 FBR and client proteins, which may disturb cellular function. These findings support the causality of this recurrent de novo PACS2 heterozygous missense in DEEs with facial dysmorphim and cerebellar dysgenesis.

Genetic network modeling.

The inference of genetic interactions from measured expression data is one of the most challenging tasks of modern functional genomics. When successful, the learned network of regulatory interactions yields a wealth of useful information. An inferred genetic network contains information about the pathway to which a gene belongs and which genes it interacts with. Furthermore, it explains the function of the gene in terms of how it influences other genes and indicates which genes are pathway initiators and therefore potential drug targets. Obviously, such wealth comes at a price and that of genetic network modeling is that it is an extremely complex task. Therefore, it is necessary to develop sophisticated computational tools that are able to extract relevant information from a limited set of microarray measurements and integrate this with different information sources, to come up with reliable hypotheses of a genetic regulatory network. Thus far, a multitude of modeling approaches have been proposed for discovering genetic networks. However, it is unclear what the advantages and disadvantages of each of the different approaches are and how their results can be compared. In this review, genetic network models are put in a historical perspective that explains why certain models were introduced. Various modeling assumptions and their consequences are also highlighted. In addition, an overview of the principal differences and similarities between the approaches is given by considering the qualitative properties of the chosen models and their learning strategies.

Increased serum lactate dehydrogenase isoenzyme 1 and macro creatine kinase type 2 in a patient with lung cancer.

A 72-yr-old man, having a lung tumor with metastatic spread to liver and bone, was hospitalized with a lactate dehydrogenase (LD; EC 1.1.1.27) activity and a creatine kinase (CK; EC 2.7.3.2) activity in serum of 30 and 2 times the upper reference limit (URL), respectively. The LD isoenzyme-1/LD activity ratio was 62% (2 times URL). This ratio was maintained throughout hospitalization, during which LD activity increased up to 90 times URL. Macro CK type 2 activity represented almost all of the CK activity, which increased up to 4 times URL during hospitalization. The patient died the 29th day after admission. The enzyme abnormalities were thought to stem from tumor tissue.

Radial immunodiffusion and immunoelectrophoresis compared for identifying autoantibodies to lactate dehydrogenase in human serum.

Variant electrophoretic patterns of lactate dehydrogenase isoenzymes were studied. By radial immunodiffusion and immunoelectrophoresis, immunoglobulin and light chain class of autoantibodies to lactate dehydrogenase were identified in nine sera: seven of these sera demonstrated IgG (5 lambda, 2 kappa) autoantibodies to lactate dehydrogenase, the other two demonstrated IgA (both kappa) autoantibodies to lactate dehydrogenase, the other two demonstrated IgA (both kappa) autoantibodies to lactate dehydrogenase. We conclude that radial immunodiffusion and immunoelectrophoresis are equally effective for identifying auto-antibodies to lactate dehydrogenase in serum. Radial immunodiffusion, however, is easier to perform than immunoelectrophoresis.

Determination of urinary cystine for monitoring anticystinuric therapy.

Cystine is assayed in urine by a method adapted from the procedure of Haux and Natelson (Clin Chem 1970;16:366-369) and modifications of the method, which improved its specificity, are presented. Assay imprecision (CV) at four different cystine concentrations (range: 0.042-1.658 mmol/l) varied from 23.1% to 3.7%. Analytical recovery was 94.6 +/- 6.0%. D-Penicillamine, alpha-mercaptopropionylglycine, L-glutamine and metabolites, added to urine, had negligible effect on the results of the cystine assay. Samples were stable for 3 wk when stored at 4 degrees C or frozen. Cystine excretion (mmol/24 h) in apparently healthy persons ranged from 0.041 to 0.170 (mean +/- SD: 0.099 +/- 0.039). Cystine excretions are presented in patients treated with anticystinuric drugs.

Sorsby fundus dystrophy without a mutation in the TIMP-3 gene.

AIMS: To examine a large family with an autosomal dominant fundus dystrophy and to investigate whether or not mutations in TIMP-3 gene were involved. METHODS: A large family of 58 individuals with an autosomal dominant fundus dystrophy was examined ophthalmologically. A DNA linkage analysis in the 22q12.1-q13.2 region was performed. The TIMP-3 gene was screened for mutations in all five exons. RESULTS: In this large family 15 individuals were affected. All other individuals were found to be clinically unaffected. Pisciform flecks in the midperiphery and drusen-like deposits were the most typical ophthalmological finding in this family and were encountered from the fifth decade on. Chorioretinal atrophy and neovascularisation with disciform lesions characterised the disease from the sixth decade on. Linkage analysis using an affected only analysis, showed a maximum positive lod score of 3.94 at theta = 0.0 with marker D22S283. No mutations possibly causing Sorsby fundus dystrophy were found in either the exonic sequences, the promotor region, or the 3'UTR. CONCLUSION: The family in this pedigree has an autosomal dominant fundus dystrophy, which is most probably Sorsby fundus dystrophy. Although, in the linkage analysis, significant positive lod scores were found with the region 22q12.1-q13.2, no causative mutations could be identified in the TIMP-3 gene.

Detection of Bence-Jones protein in serum by immunoblotting.

To detect Bence-Jones protein (BJP) in serum we precipitated intact immunoglobulins (Ig) using polyethylene glycol (PEG) and subjected the BJP in solution to electrophoresis in agarose gel, followed by transfer to a polyvinylidene difluoride membrane, and immunoenzymatic staining (successively using rabbit anti-human light/heavy chain of Ig, biotinylated swine anti-rabbit Ig, and alkaline phosphatase-conjugated streptavidin). Treatment with PEG effectively reduced background staining of polyclonal Ig in the immunoblots, although intact monoclonal Ig was not always completely removed. To compare the present method with immunoelectrophoresis (IEP), we selected samples from patients demonstrating BJP by IEP in both serum and urine (n = 40), serum only (n = 18), and urine only (n = 32); 21 of these patients had BJP alone and 69 had BJP in addition to intact monoclonal Ig. Efficiency of detection of BJP in serum was increased by the present method: serum BJP was detected in 70 patients by the present method versus 58 by IEP. The present method demonstrated single BJP bands in the samples from 16 patients (kappa, n = 7; lambda, n = 9) and multiple BJP bands (range: 2-9) in the samples from 54 patients (kappa, n = 31; lambda, n = 23). This method could be useful for detecting BJP in serum from patients suspected of having light chain gammopathy (without the need for urine testing) and may complement urine testing in patients excreting polyclonal free light chains of Ig.

A case of recurrent adrenocortical carcinoma, with observations on long-term o,p'-DDD therapy and complications.

This report describes a patient with a recurring, one stemline-aneuploid, adrenocortical carcinoma. The condition showed a number of unusual characteristics over a period of 22 yr. It changed from a biochemically functioning, low-grade malignant tumour into a non-functioning malignancy with pronounced mitotic activity, accompanied by an ovarian carcinosarcoma 1 yr before death. Quality of life was reasonable for many years despite chemotherapy, consisting of a total of almost 10 kg of o,p'-DDD administered over a period of 8 yr, and the subsequent side effects (e.g. low T4; increased bleeding time). A reduced mineralocorticoid activity, induced by o,p'-DDD, was reversed after discontinuation of o,p'-DDD treatment. During o,p'-DDD administration the substitution requirements for both hydrocortisone and fludrocortisone acetate increased, leading to periods of hypoadrenocorticism with prerenal uraemia.

A clustering performance measure based on fuzzy set decomposition.

Clustering is primarily used to uncover the true underlying structure of a given data set and, for this purpose, it is desirable to subject the same data to several different clustering algorithms. This paper attempts to put an order on the various partitions of a data set obtained from different clustering algorithms. The goodness of each partition is expressed by means of a performance measure based on a fuzzy set decomposition of the data set under consideration. Several experiments reported in here show that the proposed performance measure puts an order on different partitions of the same data which is consistent with the error rate of a classifier designed on the basis of the obtained cluster labelings.

Segmentation of muscle cell pictures: a preliminary study.

This paper describes a procedure for segmenting muscle cell pictures. The segmentation procedure is broken into two logical parts. The first part segments the picture into regions composed of cells or clumps of cells using a number of low-level operations. The second part of the procedure involves the segmentation of the cell clumps into individual cells. This is done by using a hierarchical clustering algorithm to group together those boundary points of a cell clump that belong to the same globally convex sections of the boundary. The dissimilarity measure used by the clustering algorithm is based only on information about the shape of the boundary, where this information is derived from line segments interior to the boundary. This procedure has given us satisfactory results on a number of test pictures.

On the stability of shared near neighbor clustering.

This correspondence concentrates on the detection of stable clustering results when using the nonparmetric clustering technique of Jarvis and Patrick. This technique incorporates the concept of similarity based on sharing of near neighbors. Essentially, in this clustering scheme, two parameters are involved: neighborhood depth and similarity threshold. Combining a penalty for chaining with the detection of so-called hierarchical stable solutions in the field of all possible solutions, due to the setting of the two parameters, is experimentally shown to provide a powerful method for discriminating between reliable and unreliable results.

[Aponeurotic suspension of the bladder cervix: how much tension should be put on the sling and how should this be measured (author's transl)]. / Suspension aponévrotique du col vésical: quelle tension donner à la fronde et comment la mesurer

In the sling operation for stress incontinence, it is difficult to have an appropriate tension on the sling. The authors describe the use of a modified Spring-gauge to measure this tension: this should be of 170 grammes.

[Vesico-intestinal fistulas]. / Vesico-intestinale fistels.

A review is given on frequency, etiology, anatomopathology and physiopathology of vesico-intestinal fistulas. Symptomatology and diagnosis with stress on adjuvant technical investigations are studied. In conclusion the principles for a surgical treatment are discussed.

Congenital intranasal cyst in dacryocystocele.

The authors present a case of a newborn with bilateral dacryocystocele in whom repeated probing was unsuccessful. An associated intranasal cyst was diagnosed. Using fiberoptic nasal endoscopy, treatment consisted of probing, irrigation and marsupialization of the cyst. This rarely reported anomaly is a more common finding in dacryocystocele than previously recognized. Careful nasal examination in all infants with congenital dacryocystocele is recommended.

[Misleadingly high amylase and pancreatic amylase activity in the plasma of patients with macroamylasemia]. / Misleidende hoge amylase- en pancreasamylase-activiteit in het serum van een patiënt met macro-amylasemie.

A man aged 68 years with choledocholithiasis and cholangitis, with no clinical signs suggestive of acute pancreatitis and with a low excretion of amylase in the urine, showed persistent hyperamylasaemia which appeared to be caused by macroamylasaemia. The macroamylase (an IgA-lambda-amylase complex) accounted for nearly all (90%) of the amylase activity in the serum. The activity of pancreatic amylase in serum, determined by an immunoinhibition test which selectively blocks salivary amylase activity, constituted 99% of the amylase activity in serum (normal reference range 19-71). We showed, however, that complexed salivary amylase is not inhibited in the test, resulting in a falsely-increased activity of pancreatic amylase in serum. We conclude that macroamylasaemia can lead to a clinically misleading increase in the activity of pancreatic amylase in serum.

Use of office-based smoking cessation activities in family practices.

BACKGROUND: Smoking is the leading cause of morbidity and mortality in the United States. Recommendations for increasing physician effectiveness in smoking cessation through the use of office-based activities have been disseminated, but the extent of implementation is unknown. We describe the degree to which selected family practices in Nebraska have implemented 15 specific office-based activities. METHODS: We employed a cross-sectional integrated multimethod design. A research nurse observed a target physician and his or her staff during a 1-day visit in a random sample of 89 family practices. Data collection consisted of focused observation of the practice environment, key informant interviews, medical record reviews, and in-depth interviews with the physicians. RESULTS: The majority of the practices sampled had an office environment that restricted smoking, but few used visual cessation messages or information in the waiting room offering help and encouraging patients to quit. Most had educational materials that were supplied by pharmaceutical companies for promoting nicotine replacement systems. These materials were easily accessible in more than half of the practices. Smoking cessation activities were initiated and carried out by physicians with minimal use of their staff. Smoking status was documented in 51% of the medical records reviewed but seldom in a place readily accessible to the physician. All physicians were very aware of the importance of smoking cessation counseling, and most felt confident in their skills. CONCLUSIONS: Despite identification of patient smoking as a problem, most practices were not using office-based activities to enhance and support physician counseling. New perspectives for helping practices with this task need to be explored.

The value of pharmaceutical representative visits and medication samples in community-based family practices.

BACKGROUND: Interactions between the pharmaceutical industry and physicians have been discussed in numerous publications; however, most articles are limited to surveys and self-report data and often focus on academic or training contexts. We describe the role of pharmaceutical representatives and the use of samples in community-based family practices, using data obtained by directly observing clinical encounters. METHODS: We collected detailed descriptive field notes of the direct observations of 53 primary care clinicians and 1588 patient encounters in 18 purposefully selected Nebraska family practices. We used a comparative case study design, that used depth interviews of clinicians and office staff, and included details of the interactions with pharmaceutical representatives and the use of samples in clinical encounters. RESULTS: Individual providers and practices displayed noticeable variation in their approaches to drug representatives and samples. We found formal strategies and policies in a minority of practices. Generally there was little structure in the organization and distribution of sample medications at the office level, and detailed patient education regarding these drugs was rarely observed in patient encounters. Nevertheless, samples were used in almost 20% of observed encounters, at times as starter dosages, but often as complete courses of treatment. The benefits derived from contact with the pharmaceutical industry varied substantially, but most often included free medication samples, meals, and patient education materials. CONCLUSIONS: Clinicians have a complex symbiosis with the pharmaceutical industry and need to critically evaluate their handling of samples and their contact with pharmaceutical representatives to optimize this relationship and ensure quality patient care. Clinics with specific policies for interactions with drug companies appear to derive more satisfaction from their encounters.

Current trends in tobacco prevention and cessation in Nebraska physicians' offices.

BACKGROUND: Despite years of intervention, few studies describe the extent to which recommended tobacco use prevention and cessation activities occur in community-based family practices. This study was designed to discover current practice patterns in these areas and to describe physician outcome and efficacy expectations. METHODS: An exploratory comparative case study of 11 family practices used direct observation of practices and clinical encounters, chart reviews, and in-depth interviews. Qualitative and quantitative information was gathered on (1) intensity of tobacco use prevention and cessation; (2) physicians' attitudes and beliefs regarding outcome expectations; and (3) physicians' perceptions of their ability to counsel. Qualitative content analysis and descriptive statistics were used to construct case studies for comparisons. RESULTS: Themes common to most practices included the "provision of little prevention" and "a lack of perceived need to address smokeless tobacco." Responsibility for tobacco activities fell almost solely to physicians. Although physicians felt confident in their counseling skills, the skills they identified were fairly basic. Most physicians were pessimistic about the positive effects of these activities. None of the practices was using any specifically developed "package," and pharmaceutical companies provided almost all patient education material. There was considerable variation in intensity of activities because of differences in attitudes, expectation, and background. CONCLUSIONS: To increase tobacco control activities, practice systems need to be individually evaluated to identify what is needed, how it will fit within the practice culture, and how it can best be implemented in this specific practice. One-size-fits-all interventions probably will not be widely implemented.

In vitro activity of secnidazole against Atopobium vaginae, an anaerobic pathogen involved in bacterial vaginosis.

Bacterial vaginosis is a polymicrobial syndrome. The most important marker for bacterial vaginosis is the presence of Gardnerella vaginalis and Atopobium vaginae. In this study, the in vitro susceptibilities to metronidazole and secnidazole of 16 strains of A. vaginae were tested with the agar dilution method. We observed an MIC range for metronidazole of 4-64 mg/L (MIC(50), 8 mg/L; MIC(90), 32 mg/L) and an MIC range for secnidazole of 4-128 mg/L (MIC(50), 16 mg/L; MIC(90), 64 mg/L). According to these findings, we can conclude that the activity of secnidazole is similar to that of metronidazole.