Optimization of immunohistochemical detection of collagen type II in osteochondral sections by comparing decalcification and antigen retrieval agent combinations

Bone containing tissues such as osteochondral joint are resistant to routine tissue processing, therefore require decalcification. This technique causes removal of mineral salts, but in the process may macerate the organic tissue, hence the need for tissue fixation. Such severe processing demands careful antigen retrieval to necessitate optimal staining. The aim of our study was to compare five different antigen retrieval protocols (heat retrieval and protein digestion) following decalcification of rabbit knee joints using two different techniques (20% formic acid and 10% ethylenediamine-tetra acetic acid: EDTA). Osteochondral sections were compared based on time required for decalcification, ease of sectioning, morphological integrity using HE staining and antigen preservation (Collagen type II) using immunohistochemistry. The two decalcification solutions did not impair the tissue morphology and ease of sectioning. Joints processed with formic acid decalcified four times faster than EDTA. Among the five antigen retrieval approaches, maximal collagen II uptake with minimal nonspecific staining was found with protein digestion (pronase and hyaluronidase) in both formic acid and EDTA sections. For osteo-chondral sections, we recommend using 10% EDTA for decalcification and pronase plus hyaluronidase for antigen retrieval if maintaining tissue morphology is crucial, whereas if time is of the essence, 20% FA with pronase plus hyaluronidase is the faster option while still preserving structural integrity. Clin. Anat. 33:343–349, 2020. © 2019 Wiley Periodicals, Inc.     

Potassium permanganate resistant amyloid in fine-needle aspirate of the liver

A case of primary amyloidosis, initially detected by fine-needle aspiration of the liver, is reported here. Amorphous acellular metachromatic material was seen extracellularly in between the hepatocytic cords compressing them. This material showed typical apple-green birefringence under crossed bipolars after alkaline Congo-red staining proved its amyloid nature. It was resistant to potassium permanganate pretreatment, indicating it to be of the AL type. © 1994 Wiley-Liss, Inc.     

Production and characterization of monoclonal antibodies to rat plasma kininogen

Five hybridoma cell lines were established that secrete monoclonal antibodies (MAbs) directed against rat plasma low molecular weight plasma kininogen. Two of the secreted antibodies were of the IgG1k isotype. The remaining three were of the IgG1 lambda, IgG2ak and IgMk isotypes, respectively. The dissociation constants (Kd) of these MAbs ranged from 0.58 X 10(-9) M to 5.4 X 10(-9) M. At least two distinct epitopes on the rat plasma kininogen were recognized by these MAbs. Further characterization of the MAbs showed that four MAbs cross-reacted strongly with kininogen from the Murphy-Sturm lymphosarcoma (MSLS) tumor while one of the MAbs cross-reacted weakly with MSLS tumor kininogen.     

Placenta membranacea, previa and accreta. A case report.

A case of placenta membranacea, previa and accreta was managed conservatively and resulted in delivery of a mature fetus. The diagnosis of placenta membranacea and previa was obtained with ultrasound. The placenta previa and accreta necessitated a cesarean delivery and hysterectomy.     

Preventive oral health related behaviour of dentally anxious schoolchildren aged 13-14 years in Lothian, Scotland.

The aim of this study was to determine the self-reported preventive oral health related behaviours of dentally anxious schoolchildren. 1103 children participated in the study, mean age 14 years (SD 0.35 years), and the prevalence of high dental anxiety was 7.1 per cent (95 per cent confidence interval = 5.6 per cent, 8.6 per cent). Children with high self reported dental anxiety were more likely to defer, cancel and or not attend dental appointments. In addition, for this group the last dental visit was more likely to be as a result of pain and less likely to have been for a dental examination only. Overall dentally anxious children did not help themselves by keeping their teeth clean. Fluorides were infrequently used by all the children, and only 12 per cent of all who participated in the study used fluoride supplements regularly. In this context it is not surprising that no differences in present or past use of fluoride supplements could be determined between high and low/moderate dental anxiety groups. The high dental anxiety group spend significantly more (median = 50p) on sweets per day and drank more cans of fizzy drinks (median = 2) compared with the low/moderate anxiety groups. These effects were significant after taking into account social class and gender differences. It was clear from the study that even when social class and gender are taken into account the children with high dental anxiety were not helped by their relatively poor attitudes towards preventing disease in their own mouths.     

Impact of therapeutic management on use of seclusion and restraint with disruptive adolescent inpatients

Use of seclusion and restraint on three inpatient adolescent psychiatric units was examined during two five-month periods before and after implementation of a "therapeutic management" protocol. Under the protocol, staff classified disruptive behaviors into four stages and provided verbal and behavioral interventions to control behavior at each stage. Patients who required seclusion and restraint had significant latitude to determine the timing of their release from the interventions and met with staff one hour and 24 hours after their release to explore alternatives to aggression. The number of episodes of seclusion and restraint fell by 64 percent after the protocol was adopted, and the number of patients who required those interventions dropped by 39 percent. The mean duration of episodes of seclusion and restraint was reduced by 59 percent. Therapeutic management provides a corrective experience by addressing the developmental needs, deficient cognitive skills, and poor internalized controls of disruptive adolescents.