Mechanical loading on the low back in three methods of refuse collecting.

The mechanical loading on the low back was studied in three different current methods of refuse collecting: in polythene bags, two-wheeled mini-containers and large four-wheeled containers. To this end the most prominent activities of each collecting method were performed in a laboratory. On the basis of movement analysis, force measurements and biomechanical modelling, spinal compressive and shear forces were estimated. From these forces and from the frequency of activities during the working day (assessed in a preliminary field study) the low-back stress in each collecting method was evaluated. In the bag-method, peak forces when throwing the bags ranged from 3341 to 5179 N (average compression) and from 284 to 673 N (shear) among the different conditions studied. The act of picking up bags also showed rather high forces (exceeding the NIOSH limit for disc compression in most cases). The frequency of exposure to these forces in the field is rather high (workers pick up and throw on average 807 times each day). The mini-container method compares favourably to the bags method. Peak compressive and shear force in tilting/pushing and pulling mini-containers ranged from 1657 to 2654 N and from 123 to 248 N respectively. Also, the frequency of stressful events in the field is lower in this method. In the large container method extremely high peak forces (e.g. compression ranged from 4991 to 5810 N) were observed in the task of putting the empty container back from street level to sidewalk level (surmounting the kerb). The frequency of activities like pushing, pulling and lifting the large container in the field is much lower compared with activities in the other methods. On the basis of the frequency and magnitude of spinal forces it was concluded that the mini-containers should be preferred to the bags. If kerbs are removed at container places and tasks are performed by two instead of a single person, the large container method would form another good alternative to the stressful task of collecting refuse in bags.

The suitability of saliva for detection of glucose-6-phosphate dehydrogenase deficiency.

Saliva was investigated for its suitability as a biopsy tissue for the determination of glucose-6-phosphate dehydrogenase deficiency. It appears that there is a significant difference between the activity of the enzyme in patients and controls. However, some controls have very low values making discrimination between patients and controls using a qualitative method impossible. Glucose-6-phosphate dehydrogenase deficiency is a relevant clinical problem in many rural areas in developing countries. Existing methods for determination of the deficiency in blood and hair follicles do not meet the criteria necessary for their large scale introduction in the areas of the world that are concerned by the problem. The present study shows that saliva is not a suitable alternative. Between the three biopsy tissues compared: blood, hair follicles and saliva, hair follicles remain most attractive since their isolation hardly involves the risk of infection. A simplified method for the detection of glucose-6-phosphate dehydrogenase activity in hair follicles that would allow health service workers in the field to determine the carrier status of pregnant women might form the basis for a future kernicterus prevention programme.

Psoriatic hair follicle cells. III. Characterization of aberrant morphology in differentiating cultures.

Psoriatic and control human hair follicle keratinocytes were cultured on bovine eye lens capsules in Epicult dishes for a period of 5-6 weeks and examined using light microscopy. The following morphological differences between cultures were observed: 1. The lower cell layers contained predominantly flattened cells in psoriatic cultures instead of roundish in control cultures. 2. The differentiation pattern was irregular in psoriatic cultures instead of regular in control cultures. 3. The differentiated zone of psoriatic cultures was more compact and thicker in comparison to normal cultures. These differences might allow discrimination between normal and psoriatic cultures.

Substrate specific sulfatase activity from hair follicles in recessive X-linked ichthyosis.

Recessive X-linked ichthyosis (RXLI) has its biochemical basis in a defect of the enzyme steroid sulfatase. Since several studies have reported a simultaneous deficiency of arylsulfatase C and steroid sulfatase it has been hypothesized that both enzymes are identical. In human hair follicles, however, hydrolytic activity for 4-methylumbelliferone sulfate, the substrate for arylsulfatase C, is found, while dehydroepiandrosterone sulfate is not hydrolyzed at all. These findings suggested the possible existence of two different enzymes. In the present paper structure-activity studies and molecular energy calculations are used for the demonstration that the remaining sulfatase activity in hair follicles of RXLI patients can be explained on the basis of the assumption that the enzyme has not lost its total function but has become less efficient.

Determination of glycogen and enzymes of glycogen metabolism in human hair follicles.

The skin epithelium and its organelles use glycogen as well as glucose as source of energy. Therefore the characterisation of glycogen metabolism and the enzymes involved is important in the study of mechanisms regulating the normal or abnormal differentiation of skin organelles such as sebaceous glands and hair follicles. The present paper describes fluorimetric methods for the determination of glycogen and for the measurements of phosphorylase and phosphorylase kinase activity in one and the same lysate of minute tissue samples. The methods were tested for their suitability on freshly isolated human hair follicles and cultured hair follicle cells. The possible use of these techniques for studies on the pathophysiology of acne and hirsutism is discussed.

Phosphoglycerate kinase deficiency: biochemical studies on hair follicles.

A fluorimetric procedure for the determination of phosphoglycerate kinase in single human hair follicles is described. Enzyme studies on different parts of hair follicles after dissection show that the distribution of glucose-6-phosphate dehydrogenase matches that of phosphoglycerate kinase. Glucose-6-phosphate dehydrogenase can therefore be used as a reference enzyme to compensate for differences in hair follicle sizes. It was shown that the variation in the values found in individual hair follicles is improved by relating phosphoglycerate kinase to glucose-6-phosphate dehydrogenase activity. In areas of the world where glucose-6-phosphate dehydrogenase deficiency occurs frequently, an autosomally inherited reference enzyme may be preferred. It is shown that 6-phosphogluconate dehydrogenase is useful in this respect. Upon storage a gradual drop in the activity of all three enzymes was observed, but the rate of decrease was about equal: the enzyme activity ratio was, therefore, almost unaffected for a period of one week. This allows the determination of phosphoglycerate kinase even in mailed hair follicles.

Studies on the local activity of antiandrogens at the molecular and histological level.

Androgen dependent skin disorders are important in clinical practice. Effective topical antiandrogens would lead to a breakthrough in their treatment. Although many attempts have been performed to develop such compounds, major successes have not been forthcoming. In the present study three existing antiandrogenic molecules have been compared with regard to their effect on androgen metabolism, receptor competition and on histological parameters in the hamster flank organ test. It appears that the effect on the hamster pigmented spot can be predicted on the basis of molecular mechanism. However, the effects on histological parameters are apparently dependent on additional factors such as metabolism of the active substance before reaching the sebaceous structure or limited penetration through the skin surface. The results indicate that in the development of new antiandrogens pre-screening can be performed with the aid of metabolic and receptor studies, while the histological parameters in the hamster flank organ test provide an animal model with a good predictive value.

Actin distribution patterns in patients with adenomatosis of colon and rectum.

The actin cytoskeleton was examined in cultured skin fibroblasts of control individuals and patients with adenomatosis of colon and rectum using a fluorescent dye specific for F-actin, NBD-phallacidin. We confirmed that the actin distribution pattern differs significantly between the group of controls and the group of patients. However, the method is not practicable for the diagnosis of individual patients due to the number of false positives and negatives to be expected.

Differentiation of keratinocytes in vitro: a new culture vessel mimicking the in vivo situation.

A culture vessel consisting of two independent chambers separated only by the growth substrate is described. Cells may be cultured on both sides of the growth substrate. Culture medium and gas exposure can independently be controlled in both compartments. Human hair follicles have been used as source of keratinocytes and the bovine eye lens capsule has been explored as growth substrate. The presence of 5% CO2 in air in the lower compartment appears to have a significant effect on the morphology of the cultures. When the cultures are being exposed to air with 5% CO2, the culture medium being applied in the lower compartment, formation of corneocytes characteristic for adult stratum corneum is induced, as evidenced by light and electron microscopy. To the knowledge of the authors, this stage of differentiation in vitro has not been obtained with previously described systems. Differentiation of the lower cell layers has been characterised with specific antibodies. The possible use of the system for applied and pure scientific research is discussed.

Psoriatic hair follicle cells. I. Biochemistry and behavior in culture.

It is generally accepted that in psoriasis there is an alteration of epidermal cell proliferation. It has been reported that an increased rate of thymidine incorporation into keratinocytes is found in the upper part of the hair follicle in involved skin, but this is not the case in the lower part. Here we show that cells from psoriatic hair follicles could be brought in culture under the same conditions as those of normal hair follicles. Cells, whether originating from the upper or lower part of the hair follicle sheath either from involved or uninvolved psoriatic skin, show a faster rate of outgrowth in the first days of culture. Moreover, a large number of psoriatic cells have an increased motility in the early stages of culture, as compared to control cells. These properties can no longer be observed after several days in culture. The activity of glucose-6-phosphate dehydrogenase known to be increased in psoriatic plaques is normal in hair follicles isolated from these plaques. Protein gel electrophoretic investigations showed that there is no difference in gel patterns between normal and psoriatic hair follicles. In conclusion, the isolation of human hair follicles represents a simple method that allows psoriatic keratinocytes to be brought in culture and permits the study of certain aspects of the disease.

The use of liposomes in the topical application of steroids.

The effect of topical application of the androgen 5 alpha-dihydrotestosterone (DHT), both encapsulated in liposomes and solved in acetone, has been evaluated using the female hamster flank organ as a model system. Systemic absorption of DHT was significant from the acetone solution, but negligible from the liposome system. The topical biological effect is, however, proportionally diminished when the liposome system is used. Under the experimental conditions used, the lited using the female hamster flank organ as a model system. Systemic absorption of DHT was significant from the acetone solution, but negligible from the liposome system. The topical biological effect is, however, proportionally diminished when the liposome system is used. Under the experimental conditions used, the liposome system had no advantages over application in acetone in this model.

Population screening for glucose-6-phosphate dehydrogenase deficiency on the Baleares.

Two thousand people on the Isles of the Baleares were screened for glucose-6-phosphate dehydrogenase deficiency using a commercially available kit. Among the thousand males tested, five were found deficient; of the thousand women, one had low enzyme activity according to this test. Diagnosis of glucose-6-phosphate dehydrogenase deficiency could be verified using hair follicle analysis on mailed hair samples. The same technique also allowed heterozygotes to be identified unequivocally.

Human hair follicles may be used for population screening of heterozygotes of glucose-6-phosphate dehydrogenase deficiency.

The carrier status for glucose-6-phosphate dehydrogenase deficiency can still be detected with the use of hair follicles after a period up to 10-14 days after plucking, even under conditions of temperature and humidity that are encountered in tropical countries, where the disease is most common. Furthermore, it is shown that the hair colour or age of the donor has no influence on the enzyme activity. As a consequence, the hair follicle is indeed suitable for screening in rural areas since mailing to existing medical centres is possible.

A simple fluorimetric microassay for DNA in hair follicles or fractions of hair follicles.

Human hair follicles may be useful for determining individual differences in the metabolism of polycyclic aromatic hydrocarbons in epithelial tissues. for quantitative measurements of carcinogen metabolism, determination of the amount of DNA in the hair follicles is necessary in order to correct for individual size variation. A simple method for DNa determination in hair follicles is described, based on the use of a hypotonic pronase solution to solubilise DNA which is then available for complex formation with mithramycin or 4,6-diamidino-2-phenylindole. 2HCI (DAPI). The mithramycin method permits the measurement of DNA in a small number of hair follicles, while the DAPI method is sensitive for as little as one single bulb.

Urinary oxalate estimation.

A method of estimating urinary oxalate is described. Impurities which might otherwise interfere with the estimation are removed by means of an ion-exchange resin. The oxalate is then reduced to glycolic acid, which forms a coloured compound with chromotropic acid. It can then be estimated colorimetrically. No special equipment is required, oxalate recovery is good, and the method is reproducible and precise.