PubMed Health⌕ Search

Biomedical subjects

P Bolhuis

Publications and source records attributed to P Bolhuis.

10 recordsLinked to original sources
PubMed1997

Advances in Charcot-Marie-Tooth disease research: cellular function of CMT-related proteins, transgenic animal models, and pathomechanisms. The European CMT Consortium.

The First Workshop of the European Consortium on Charcot-Marie-Tooth (CMT) disease brought together neuroscientists, molecular and cell biologists, neuropathologists, neurologists, and geneticists with a common interest in the understanding of the fundamental mechanisms that underlie the pathogenesis of CMT. The interdisciplinary group of 25 expert scientists discussed recent advances in (i) molecular genetics and histopathology of CMT, (ii) development of suitable animal models, (iii) understanding of the cellular function of CMT-related proteins, and (iv) studies using nerve biopsies from CMT patients. In this minireview, we summarize the key findings presented and discuss their impact on CMT research.

Animals↗
PubMed1988

Penile studies and heterosexual partners. Peniscopy, cytology, histology, and immunocytochemistry.

The male partners of 20 women with human papilloma virus (HPV) infection (subtypes 6, 11, 16, and 18) were studied. These men were unsuspecting and allowed investigation "for scientific reasons" in the context of the abnormal findings in their female partners. Peniscopy (study of the penis with a colposcope) was abnormal in 17 cases. In the cytologic-brush preparations of the urethra, hyperkeratosis (visualized with the modified Papanicolaou technique) occurred in all cases, but koilocytosis in only one. Both urothelial cells and squamous cells in the urethral samples were positive for HPV, as shown in the immunocytochemistry. In the histologic sections taken from flat lesions there was hyperkeratosis, dyskeratosis, and abnormal nuclear maturation. The authors concluded that, using simple techniques, subtle disturbances in maturation of nuclei and cytoplasm can be established in penile epithelium of these sexual partners. Similar changes are established in other HPV lesions. It seems likely that a significant proportion of the male population is infected in countries were HPV an important factor in cervical carcinogenesis.

Condylomata Acuminata↗
PubMedDate not supplied

Brain enzyme histochemistry following stabilization by microwave irradiation.

The activities of various enzymes present in brain homogenates were assayed biochemically (a) with no pretreatment, (b) following a standard microwave treatment in saline and (c) after a standard microwave treatment in formalin. All enzyme activity was lost after the microwave - formalin in treatment. Following microwave - saline treatment, the activities of alkaline phosphatase, 5'-nucleotidase, isocitrate and succinate dehydrogenases were reduced. In contrast, the activities of lactate and malate dehydrogenases were unchanged, and that of acetylcholinesterase apparently increased. Analogous outcomes were seen following attempted histochemical demonstrations of these enzymes. Thus satisfactory histochemical demonstration of all enzymes was achieved (except with alkaline phosphatase, lactate and malate dehydrogenases) following the microwave-saline pretreatment. Since acid phosphatase, catalase and peroxidase were also successfully demonstrated, it seems that microwave-saline pretreatments permit both retention of sufficient enzyme activity for histochemical demonstration to occur and retention of sufficient structural integrity for critical morphological investigations. Since the failure to stain the sites of lactate and malate dehydrogenases is not due to microwave inactivation of these enzymes, their demonstration may be possible by varying the staining procedures.

Animals↗
PubMedDate not supplied

Microwave irradiation in label-detection for diagnostic DNA-in situ hybridization.

A DNA-in situ hybridization protocol was adapted for application to sections of routinely processed paraffin embedded material. This protocol was developed previously for detecting DNA-virus infected cells in whole cell preparations and employs biotinylated DNA as probe. Three different biotin detection methods were optimized and applied. The first uses streptavidin and a biotinylated complex of alkaline phosphatase, the second consists of an immunogold-silver staining, and the third of a peroxidase technique using a silver amplification. The alkaline phosphatase method was the most rapid, and as sensitive as the immunogold-silver staining. The peroxidase method was the most sensitive. Microwave irradiation was applied to the different incubation steps of these three detection methods. Short incubations with microwave irradiation gave results comparable to those obtained with conventional incubations, when streptavidin, antibiotin, complexed alkaline phosphatase, or gold labelled goat antirabbit were used. It was thus shown that microwave irradiation creates the possibility of a very rapid label-detection for nonradioactive DNA-in situ hybridization.

Alkaline Phosphatase↗
PubMedDate not supplied

A practical approach to routine immunostaining of paraffin sections in the microwave oven.

The Streptavidin-Biotin Complex (SABC) Immunostaining method can be carried out by performing the rinsing and blocking steps and in addition the incubation with the primary and the secondary antibody sera in the microwave oven. Irradiation of the Streptavidin-Biotin Complex reduces stain activity due to destruction of horseradish peroxidase (HRP) (Boon & Kok, 1988). Therefore, we decided not to perform this step in the microwave oven. The microwave incubations can be performed using antisera dilutions of 1:1000 (instead of 1:50) in tissue fixed with Kryofix, allowing staining in staining racks. This keeps hands-on time low and simplifies the microwave steps. It is very difficult to obtain reproducible results in the microwave oven using droplet incubations due to problems with hot spots and antenna effects. These problems are avoided when cuvettes are used and air is blown through the solutions during microwave incubations. With effective temperature control the method is highly reproducible, and takes at least 100 min less than the conventional SABC method. It is, in particular, attractive when large series of slides must be routinely immunostained and when reproducible results are desired.

Antibodies↗