Specific 125I-iodination of cell surface lipids: plasma membrane alterations induced during humoral immune attack.

A method whereby lactoperoxidase-catalyzed 125I-iodination of plasma membrane lipids can be achieved is described. The reaction results in a uniform and stable labeling of neutral lipids, phospholipids, lysophosphatides, free fatty acids, and triacylglycerides. By the use of this method, the action of antibody plus complement (C) on the specific release of lipid from the plasma membrane of line-10 tumor cells was studied. Within 15 min after the addition of C to antibody-sensitized cells, the enhanced release of specific lipid classes from the cell surface was observed; these lipids included sphingomyelin, phosphatidylserine, and phosphatidylcholine. The release of phosphatidylethanolamine and, in some instances, triglycerides, was reduced after antibody-C attack. Neither the specificity of the antibody used to sensitize the cells nor the ability of the antibody plus C to be cytotoxic to the cells appeared to affect the identity or amounts of lipids released from the cells.


Deficient production of the lymphokine eosinophil stimulation promoter in chemically induced and mutation diabetes mellitus in mice.

The cellular defects possibly responsible for diminished in vivo granuloma formation in diabetic Schistosoma mansoni-infected mice were investigated. Diabetic and control animals develop a similar degree of eosinophilia. Eosinophils obtained from diabetic mice also respond normally to the lymphokine eosinophil stimulation promoter (ESP). Lymphoid cells of chemically induced (streptozotocin) and mutation diabetic (db/db) mice, however, have a decreased capacity to produce/secrete ESP in response to soluble egg antigens of S. mansoni. Administration of insulin to diabetic mice is associated with a partial reversal of the decreased ability of their lymphoid cells to generate ESP. These findings show that defective cellular immunity in diabetic animals may be partially explained by the failure of their lymphocytes to produce the soluble mediators involved in recruitment of target cells.


The beige mutation in the mouse. I. A stem cell predetermined impairment in natural killer cell function.

A point mutation, called beige, on linkage group 14 in C57BL/6 mice leads to a marked impairment in natural killing and antibody-dependent, cell-mediated cytolysis (ADCC) of tumor cells. The defect in NK cytolysis was predetermined at the level of progenitor cells in the bone marrow as revealed in radiation chimeras. This impairment in NK function could not be accounted for by an altered organ distrubution, target selectivity, or ontogenesis. Interferon did not fully restore the response, which suggests that the defect may not result solely from a lack of endogenous interferon stimulation in beige mice. The frequency of target-binding cells was normal in all lymphoid organs, which suggests that the defect is intrinsic to the NK cell and does not involve an altered population size or an inability to recognize and interact with the target. Rather, the defect may lie within the lytic pathway subsequent to target cell contact. These mice should provide a useful NK-deficient system for studies of T cell or macrophage immunity and in addition they provide a means for testing the in vivo significance of NK cells in resistance to tumors and virus-infected cells.


In vitro killing of S. mansoni schistosomula by eosinophils from infected rats: role of cytophilic antibodies.

The cytotoxic effect of peritoneal cells from Schistosoma mansoni-infected rats against antibody-opsonized or nonopsonized schistosomula in vitro has been studied during the course of infection. Eosinophil-enriched cell preparations were shown to have a high cytotoxic effect on schistosomula in the absence of antibody. The killer cells were identified as eosinophils. As in the ADCC mechanism previously described, mast cell-eosinophil interaction was required for eosinophil cytotoxicity. Rosette formation using S. mansoni antigen-coated erythrocytes was used to demonstrate the presence of anti-S. mansoni IgG2a antibody at the surface of infected eosinophils. Passive sensitization of normal eosinophils with ultracentrifugation pellets of immune rat serum resulted in a significant cytotoxicity of sensitized eosinophils. A close relationship was found between the cytotoxic activity of infected cells and the ability of the corresponding infected serum to arm normal eosinophils. At certain periods after infection, eosinophils from infected rats were less effective than normal eosinophils on antibody-coated schistosomula. EA- (rat) rosetting assay and blockade experiments with homologous immune complexes have revealed in a kinetic study that the blocking of cytotoxic activity of infected eosinophils was related to heat-stable circulating immune complexes. The possible role of immune complexes either in arming or inhibiting effector cells is suggested.


Isolation of the fourth component (C4) of rat complement.

The fourth component of rat complement was purified to homogeneity by sequential chromatography of rat plasma in benzamidine on QAE-A50, SP-C50, hydroxyapatite, and gel filtration on Bio-Gel A 1.5. The final material was homogeneous on SDS-PAGE analysis and had a calculated m.w. of 198,000. A monospecific antibody against rat C4 was obtained from immunized rabbits. The concentration of rat C4 in the plasma of normal 4-month-old Wistar rats was 190 +/- 34 microgram/ml (mean +/- 1 S.D.).


Macrophages/monocytes require cell-to-cell contact in order to regulate the growth of a murine lymphoma cell line.

The mechanism of the macrophage-regulated proliferation of the murine lymphoma cell line FIO 30 has been further investigated. It appears that the macrophage is alone in its ability to support FIO 30 growth; a macrophage-like cell line, however, is unable to provide the stimulus required by the FIO 30 cells for their proliferation. Investigations into the nature of this stimulus indicate that the serum factor pro-MaSF and a macrophage cell surface component act synergistically to support the growth of the FIO 30 cell, but only when the two cell types are in close promixity. MaSF has also been further characterized and is shown to be closely associated with serum albumin.


Preservation of C3, C5, and C5a functional activities by a new radiolabeling method; demonstration of C5 products in complement-activated serum.

A new method is described for the radiolabeling of C3, C5, and C5a. Using a methylation procedure we have labeled C3, C5, and C5a with 14CH20, achieving high specific activity, without loss of hemolytic activity in the case of C3 and C5, and in the case of C5a, full retention of chemotactic and enzyme releasing activities. The distribution of radiolabel in the alpha and beta chains of C5 was 75 and 25%, respectively. Using intact labeled C5 the fragmentation pattern in zymosan-activated human serum, assessed by gel filtration and electrophoresis, is complex, yielding multiple fragmentation products. The labeling method described in this paper yield materials that have many advantages over the currently used labeling procedures.


Characterization of the IgE receptor isolated from human basophils.

The IgE receptor of human basophils was purified by using simple and repetitive affinity chromatography on human IgE-Sepharose. Basophils were partially purified from peripheral blood of patients with chronic myelogenous or basophilic leukemia. Cells were labeled with 125I by using the lactoperoxidase method and were solubilized with nonionic detergent. Elution of IgE-Sepharose with 0.5 N acetic acid, 1% NP-40 allowed recovery of active IgE receptor. Analysis of human IgE receptor by SDS polyacrylamide gel electrophoresis with 10% gels demonstrated one major radioactive peak with an apparent m.w. of 58,000 to 68,000, somewhat larger than rat IgE receptor. The purified human IgE receptor was active since approximately 10 to 42% of labeled receptor could specifically rebind to insolubilized human IgE. Rebinding was blocked by nanomolar concentrations of soluble human IgE or rat IgE but not by human or rat IgG, heat-inactivated human IgE, or heat-aggregated human IgG; thus it appears that rat IgE receptor. The relative abilities of active rat IgE and active human IgE to inhibit human IgE receptor rebinding could not be precisely determined because of the limitations in assessing the proportion of human IgE that retains receptor-binding activity.


Translocation of dimeric IgA through neoplastic colon cells in vitro.

We studied the translocation of dimeric IgA across epithelium, using neoplastic human colon cells in culture as a source of epithelial cells, and immunoelectronmicroscopy with peroxidase-labeled antigens and antibodies. The cells had some of the ultrastructural characteristics of normal, mature epithelial cells, i.e., polarity, desmosomal junctions, and secretory component on their basal and lateral plasma membranes. Horseradish peroxidase-labeled dimeric IgA, exposed to the cells at 0 degrees C, bound selectively to secretory component on the cell surfaces. At 37 degrees C, the bound dimeric IgA was taken into the cells by endocytosis and transported apically through the cytoplasm in vesicles. After 30 min, IgA was discharged across the apical surface. Neither colchicine (10(-4) M) nor cytochalasin B (10(-5) M) interfered with binding or endocytosis of dimeric IgA, but colchicine inhibited intracellular transport of the IgA-containing vesicles. These experiments demonstrated that dimeric IgA can be transported through living intestinal epithelial cells in vitro. The transport includes 1) specific binding of IgA dimers to secretory component on plasma membranes, 2) endocytosis of IgA in vesicles, 3) transcytoplasmic transport of the IgA-containing vesicles by a process involving microtubules, and 4) discharge of IgA at the apical surfaces.


Separation of human lymphocytes and monocytes by velocity sedimentation at unit gravity.

An improved method of separating human peripheral blood lymphocytes and monocytes by velocity sedimentation at unit gravity is described. Cell separation capacity is considerably increased, while the time required for optimal separation is markedly reduced. Simple devices make it possible to layer and fractionate the gradients within 15 min. Overall recovery is 85%. On a small scale, monocytes were isolated greater than 80% pure from 4--20 X 10(7) mononuclear leukocytes. The average yield was 40% of all monocytes recovered after fractionation. In the same sedimentation runs 60% of all recovered lymphocytes were obtained strongly depleted of monocytes (monocyte contamination less than 0.2%). On a large scale 3--6 X 10(8) monocytes with an average purity of 85% were isolated from 3--6 X 10(9) mononuclear leukocytes in 2 subsequent runs. All cells recovered from the gradient were functionally active according to rosette formation, phagocytosis, stimulation by mitogens and in cytotoxicity tests.


A modified quantitative precipitin test which is rapid, sensitive and reproducible.

A modification of the standard quantitative precipitin test is described. The new procedure, which utilizes polyethylene glycol (PEG) at both the 37 degrees C and 4 degrees C incubation stages, provides a convenient quantitative precipitin test assay which is more reliable and much faster than the standard assay. Moreover, the modified or PEG assay gives a quantitative measure of antibody concentration even when antisera of low titre are tested. The sensitivity of the capillary-tube test is also enhanced when the supernatant solutions contain PEG.


Enzyme-linked immunosorbent microassay for quantification of specific antibodies to collagen type I, II, III.

An enzyme-liked immunosorbent assay (ELISA) is described for determination of antibodies to collagen type I, II, III in serum or other body fluids. Polystyrene microplates are coated with either collagen I, II or III, and samples of body fluids being tested are incubated in the plates. Antibodies which attach specifically to the collagen are detected with anti-IgG antibody conjugated with peroxidase. Bound peroxidase is quantitatively estimated by the colour reaction produced with the substrate 5-aminosalicylic acid.


Isoelectric focusing in agarose.

We describe here the use of a new agarose for isoelectric focusing of body fluids and tissue proteins. Macromolecular proteins even when significantly greater in size than 1 X 10(6) mol.wt. were easily and rapidly focused.


Measurement of immunoglobulin binding to synovial fibroblast monolayers: comparison of staphylococcal protein A binding to cytotoxic assay methods.

We have studied the binding of rabbit antibody to cultured synovial fibroblasts by 3 methods: complement mediated cell lysis, 125I-labeled staphylococcal protein A ([125I]PrA) and antibody dependent cell mediated cytotoxicity (ADCC). The relative sensitivities of these 3 methods is 1 : 33 : 3300 respectively. The [125I]PrA binding method is shown to quantitatively detect cell bound IgG with a molar stoichiometry of approximately 1 : 1, protein A (PrA) to IgG if PrA is present in excess.


Quantitation of immune complexes by nephelometry.

Laser nephelometry was used to quantitate immune complexes and IgG aggregates in native sera. Sensitivity limits and calibration curves were in good agreement with the C1q binding assay frequently used for measuring immune complexes. Distinction between sera containing lipids or immune complexes was achieved by polyethylene glycol precipitation. Although not superior to other methods for quantitating immune complexes, the nephelometric assay may prove useful as a screening test and has the potential of an antigen-specific immune complex assay.


A simple solid-phase radioimmunoassay for the measurement of IgG secreted in vitro by human lymphocytes.

A radioimmunoassay is described for measuring IgG, based on the ability of immunoglobulins of this class to inhibit the binding of radioiodinated staphylococcal protein A to IgG linked to a solid phase. The solid phase is represented by ox erythrocytes coated with anti-ox erythrocyte rabbit IgG, a reagent used for detecting cells equipped with receptors for the Fc fragment of IgG. By this assay the IgG secreted in vitro by human peripheral blood lymphocytes stimulated with PWM and those present in samples of very diluted human sera were measured. It was found that the assay is a very rapid, simple and reproducible procedure for the detection of IgG immunoglobulin at the nanogram level.


Technical improvements in the clotted plasma droplet MIF assay in vitro.

The migration inhibitory activity of culture supernatants of rat and human lymphocytes stimulated with PHA and Con A-Sepharose was tested on cell migration from clotted plasma droplets. This technique was improved by using homologous as well as heterologous plasma and fibrinogen solutions for suspending the migratory cells, automatic micropipettes for performing the technique and purified cell populations as target. The effects of calcium chloride and of the cell concentration in the plasma droplets on the migration indices obtained in the MIF assay were tested.


Comparison of the haplotypes of the major histocompatibility complex in the rat. IV. The six original Ag-B haplotypes.

The six original haplotypes described in the Ag-B system were compared with their counterparts in the H-1 system. Antisera to the Ag-B haplotypes raised in inbred rats and antisera to H-1 haplotypes raised in congenic lines were tested against various panels of cell from inbred and congenic lines by the dextran and Ficoll haemagglutination methods. The private and strong public specificities detected in both systems were the same, but there were some minor differences in the intermediate and weak reactivities detected. The cross-reactivity of the antisera raised in inbred rats was broader than that of antisera raised in congenic lines. The identity of the antigenic products detected in the two systems by the dextran and Ficoll tests was further confirmed by a variety of absorption analyses and by F1 tests. This study completes the systematic serological comparison of the haplotypes of the major histocompatibility complex of the rat described originally in the Ag-B and H-1 systems.


A search for association between IgD concentrations and immunoglobulin allotypes in 936 sera from a genetic isolate in Newfoundland.

Immunoglobulin allotype (Gm) data has been analysed agaonst immunoglobulin D (IgD) concentrations in a population study in Newfoundland. There was no significant difference between the distribution of IgD concentrations in people homozygous for the alleles G1m(f) and G3m(b) when compared with people homozygous for the alleles G1m(a) and G3m(g). These findings, involving 573 homozygous individuals as opposed to ninety-eight in an earlier study on a New York population, do not confirm the earlier findings. Thus a genetic influence on IgD concentration by Gm genes or genes closely linked to them is not universally demonstrable by typing for these four markers and by using the Mancini technique for mearusing IgD concentration.


Frequency distribution of histocompatibility-2 antigenic specificities in the Japanese wild mouse genetically remote from the European subspecies.

Fifty-eight Japanese wild mice, Mus musculus molossinus, collected from twenty-eight localities were surveyed for twelve H-2 antigens using the haemagglutination method. Significantly higher frequencies of H-2.3 and H-2.5 specifities and relatively lower frequencies of the other public specificities were observed. This was confirmed by examining four specificities, H-2.3, 5, 13 and 23, in 370 mice. Quantitative absorption of the alloantisera by erythrocytes and spleen lymphocytes of molossinus mouse revealed definite absorption of H-2.3, 5 and 8 antigens, though their antigenic strength was apparently weaker than the controls, B10 congenic mice. Comparative assay of the strength of H-2.5 antigen in either homozygous and heterozygous conditions distinctly eliminated the possibility of gene-dose effect for the reduced strength of the public antigens in molossinus mouse. To explain this, divergence time between molossinus and domesticus was computed based on the allelic frequency data already reported on ten loci in both subspecies. It is roughly 1.5 x 10(6) years, which could allow for the occurrence of considerable genetic changes in many public antigens, except those conserved through selection.


[A clinical study on the development of the acetabulum after various treatment in congenital dislocation of the hip (author's transl)].

In the treatment of congenital dislocation of the hip, prognosis varies greatly according to the status of the development of the acetabulum. In most of the patients with osteoarthritis of the hip primary causes are found. In this series, 50% had congenital dislocation of the hip and 35% had acetabular dysplasia. In order to identify the conditions of hip joints in which the acetabular development can be expected from conservative treatment and to estimate the limit of age for indication of this treatment, the status of the development of the acetabulum was investigated after various treatment. This is expected to provide valuable data for the establishment of the method of treatment for cases without hope for the development of the acetabulum. According to the results, patients with acetabular angle of more than 30% at the age of 3 years have less possibility of normalization in the future and therefore active means for formation of the acetabulum should be considered in these patients. As such active means included are formation of the acetabulum through the pelvic osteotomy and the derotation-varus osteotomy are available in order to improve concentric position of the femoral head. It was also indicated that more favourable results could be obtained in patients who had undergone the pelvic osteotomy. The most favourable results were obtained in patients in whom both operations were used in combination. It was concluded from the results of the study that the best method is to perform the pelvic osteotomy first and to add the derotation-varus osteotomy as indicated.


[Quantitative measurement of the tendon reflexes. A study on Achilles tendon reflex (author's transl)].

Quantitative analyses of tendon reflexes were performed for the purpose of clinical application. Tendon tap was marked with a WNS-5D semiconductor load cell inserted in the hammer to measure tapping force. Achilles tendon reflex (ATR) was measured with a LAS-0.1 displacement transducer in kneeling position on the chair. In ATR, six factors, i.e. duration of reaction time, the height of the wave, the time from the hammer tap to the top of the wave, the maximum speed of contraction, the time from the hammer tap to the maximum speed and half relaxation time, were analyzed. Correlation between these six factors were discussed. From the analyses of these factors, half relaxation time may serve as the best indicator of clinical status among the factors obtained by analysis of ATR.


[Clinical study on coxa magna during the treatment in congenital dislocation in the hip (author's transl)].

Enlargement of the femoral head is frequently observed in roentgenographs during the treatment of congenital dislocation of the hip. Clinical study was performed to clarify the definition, etiology and clinical entity of coxa magna. Three hundreds and three hip joints in 247 patients were examined roentgenographically and follow up for an average of 3.1 years. By measuring the longitudinal and cross-sectional lengths of both the femoral heads and necks, we felt that "coxa magna" should be defined as the condition with enlargement of all of these parameters. The incidence of coxa magna was 47% after open reduction, but only 5% after closed reduction. In most cases, coxa magna was roentgenographically evident approximately 10 months after open reduction. On the other hand, it was noticeable 18 months after closed reduction. In the hips without coxa magna, the acetabular index improved remarkably for 6 months following open reduction, but in the hips with coxa magna, it hardly improved. Coxa magna frequently developed femoral head deformity, unless persistent acetabular dysplasia had improved. The incidence of coxa magna appeared to be closely related to surgical intervention performed on acetabular dysplasia.


[Clinical evaluation on patients with lumbar intervertebral disc hernia treated in our clinic (author's transl)].

A few reports are available on the results of conservative therapy of lumbar intervertebral disc hernia. In our Department, conservative therapy of 3 month course is carried out under a determined guideline. The cure rate obtained from this conservative therapy is 83.1% so far as the results reviewed by us concern. In order to determine the indication of nucleous pulposus resection, the remote results have been traced on patients in whom the myelography, which had been performed for the purpose of operation, gave positive findings but operation was not carried out. It was found that in all cases ADL and employment were disturbed. In the group of patients, where resection of nuclear pulposus were carried out by the same surgeon by osteoplastic partial leminectomy, the results were found to be excellent in 72.1% and good in 21.2% of all cases. When excellent and good results are combined, the positive effects have been obtained in as many as 93.3%, indicating an extremely favourable results. In 6.7% of the cases, ADL and employment were restricted, with persistent complaints and clinical symptoms. In none of the cases aggravation through operation was noted. As described above, in patients with lumbar intervertebral disc hernia, better results of operation can be expected if resections of nucleus pulposus are carried out only on patients selected strictly after a careful conservative therapy, with a broad field through osteoplastic partial laminectomy by a skilled surgeon, minimizing the difference in technique.


Comparison of roentgenological and histological features of bone changes in primary hyperparathyroidism.

Fifteen cases of bone disease in primary hyperparathyroidism were investigated by comparing X-ray findings with metabolic data and histological features. Besides the usual features of subperiosteal resorption, diffuse demineralization and localized destruction, the less common features of osteosclerosis, and the infrequently described response of the growth plate were also demonstrated. The bone changes in roentgenographs were graded on a five point scale, 0-IV. All cases in Grade IV had parathyroid cancers and extremely high serum and urinary calcium values.


[Innominate osteotomy (Salter) related to the correction of the acetabular abnormality and the prediction of the acetabular angle after operation (author's transl)].

The purpose of this paper is to know how to correct the maldirection of the acetabulum. The author devised a new system for the measurement of the degree of the forward rotation of the acetabulum. The acetabular improvement after the osteotomy is classified into two types, namely the primary and secondary improvement. The primary improvement is seen immediately after the osteotomy and the secondary improvement follows gradually. The effective factors for each of them were analyzed. The primary improvement is influenced mostly by the lateral inclination of the distal fragment. The secondary improvement was intimately correlated to the forward rotation of the acetabulum at the operation. The development of the acetabulum after operation can therefore be predicted from the post-operative X-ray film. In the case of the residual subluxation of the hip, there is about 10 degrees of acetabular elevation (frontalization), so the acetabulum should be rotated forward by more than 10 degrees in this operation.


[An experimental study on the regional blood flow in the cervical spinal cord --the influence of respiration (author's transl)].

The responses of cervical spinal cord blood flow (C-SCBF) to alteration in respiration were examined in gallamine immobilized and respiration controlled cats. Alveolar PCO2 was continuously monitored with an infrared CO2 analyzer. Mean arterial blood pressure (BP) was measured through femoral canula, which also allowed collection of arterial samples for blood gas analysis. C-SCBF was measured using thermoelectrical element which placed on the dorsal surface of the C6 segment. Simultaneously, changes in blood flow of cerebral cortex, L4 segment of spinal cord, heart and skeletal muscles, skin, kidney and spleen were also measured and compared with that of C-SCBF. Cerebrospinal fluid pressure (CSFP) was also measured on C5 segment. Changes in the arterial or alveolar PCO2 increased about 20--30 mmHg following respiration arrest or 10% CO2 inhalation, and decreased about 20 mmHg following hyperventilation during 2 minutes, respectively. Respiratory arrest or 10% CO2 inhalation induced an increase in C-SCBF, and a rise in BP and CSFP. Hyperventilation induced a decrease in C-SCBF, a fall in BP and a rise in CSFP. Effect of the respiration on blood flow of cerebral cortex, L4 segment of spinal cord or heart muscle was similar to the effect on C-SCBF in direction, but that of skin, skeletal muscle, kidney or spleen was conversely. C-SCBF increase by respiratory arrest or 10% CO2 inhalation was incompletely suppressed by pretreatment of propranolol 7 mg/kg i.v., C-SCBF decrease by hyperventilation was also incompletely suppressed by pretreatment of phentolamine 5 mg/kg i.v.. Furthermore, these C-SCBF changes were almost completely suppressed by pretreatment of phentolamine 5 mg/kg, propranolol 7 mg/kg and acetazolamide 250 mg/kg i.v. These results suggest that response of C-SCBF to alteration in respiration is chiefly induced by changes in PCO2, that CO2 acts directly on vascular smooth muscle and also on alpha and beta adrenergic receptors in the cervical spinal cord, and that CO2 has a similar action on the C-SCBF as on the blood flow of cerebral cortex, lumbar spinal cord and heart muscle.


[Mechanical behavior of the subchondral bone in the experimentally induced osteoarthritis].

In order to evaluate the role of the subchondral bone (cancellous bone) in the development and progression of the joint degeneration, osteoarthritis of the knee joint was produced experimentally in the rabbits and viscoelasticity and strength of the subchondral bone from the femoral medial condyle have been investigated along with the pathological, histological study of the joint. The viscoelastic spectrometer and the Instron type testing machine were used. As the first change after operation, osteophyte formation around the joint margin has been observed before the initiation of the degeneration of articular cartilage and there is a possibility that mechanical properties of subchondral bone such as high deformability and low elasticity to the mechanism of osteophyte formation. Subchondral bone softening with marked increase of ultimate strain and phase lag, marked decrease of compressive elastic modulus and ultimate stress precedes or occurs concurrently with the degeneration of the articular cartilage. These facts indicate the relationship between the mechanical properties of the subchondral bone and joint degeneration. Once the joint degeneration starts, degeneration continues progressively while the subchondral bone tends to become brittle. These changes may be considered as a kind of functional adaptation to the damage or denudation of articular cartilage. It is postulated that some architectural changes of the subchondral bone may provide alterations of the mechanical properties. Biomechanical roles of the subchondral bone is suggested as one of the factors in the joint degeneration.


A case of bilateral primary posterior dislocation of the shoulder.

We present a case of bilateral primary dislocation of the shoulder, which was treated operatively on the left and conservatively on the right side. Primary posterior dislocation of the shoulder is rarely seen and bilateral case is extremely rare. For the diagnosis it is essential to have a knowledge of this dislocation. As to roentgenographic examination, the axillary projection is most helpful but can't be routinely taken because of severe pain and limited abduction of the shoulder. So special roentgenographic techniques are necessary.


Delayed hypersensitivity reaction in patients with bone and soft tissue sarcoma.

In patients with sarcoma of the extremity, the tuberculin and dinitrochlorobenzene (DNCB) skin test were performed to correlate the results with the extent and prognosis of disease. Of the tumor-bearing patients, 60.0 per cent gave a positive tuberculin test and 48.9 per cent a positive DNCB test, with a significant difference from the control group with regard to the incidence of positive results in the later test (P less than 0.03). There was no difference between the different stages in the incidence of positive reactions to tuberculin but a marked difference in that to DNCB (78.6%, 40.7% and 0%). Between survivors and those who died, there was no difference in the incidence of a positive tuberculin test but a significant difference was present in that of a positive DNCB test (P less than 0.04), a difference which may be considered to reflect that in stage of disease process at the time of first examination.


Pathology of the ossicular chain in various chronic middle ear diseases.

In a collection of 1,100 operated ears, 426 of which had cholesteatoma and 674 had not, the various defects of the ossicular chain are described and related to the nature of the disease and the site of perforation. The analysis showed marked differences between the various diseases and in the frequency of the individual ossicular defects or combinations of defects. Defects of the head of the malleus and of the body of the incus were found exclusively in chiolesteatomas, most often those affecting the attic. Isolated defects of the malleus handle were most common in cholesteatoma of the parts tensa and in total perforations. Defects of the long process of the incus occurred in 74--88 per cent of cholesteatomatous diseases, defects of the stapedial arch in 47 per cent of ears with sinus cholesteatoma. In granulating otitis without cholesteatoma and in sequelae to otitis there was less ossicular pathology, and 57 per cent of these ears had an intact ossicular chain. Total or posterior perforations were associated with pathology of the ossicles more often than inferior or anterior perforations. All cases with destruction of the body of the incus and the head of the malleus showed squamous epithelium in close relation to the ossicular defect, indicating a marked--presumably enzymatic--influence by the squamous epithelium upon the bone resorption.


Clearance by the mucociliary system in 'simple chronic otitis media'.

This is a study of the rate of middle ear clearance in chronic otitis media and atelectatic ears. The clearance rate was measured with the aid of non-soluble saccharin, introduced into the middle ear through an existing perforation in 'chronic ears', or through a ventilating tube in atelectatic ears. Cholesteatomatous ears were excluded. Altogether 122 ears were examined and about two-thirds of the subjects felt the sweet taste in their mouths thereafter. The average time it took for the saccharin to be transported from the middle ear to the taste buds was 33' 22" minutes in non-infected (i.e. dry) chronic ears (38 patients or 66 per cent). Wet or infected ears with chronic otitis media (30 patients, or 58 per cent) showed a slower transport rate, averaging 54' 22" minutes--the difference was significant at the 0.01 level. 8 (63 per cent) of the atelectatic ears transported the saccharin at an average rate of 50' 25". This study demonstrated that most ears with 'simple' chronic otitis media and atelectatic ears have a patent eustachian tube and that their mucociliary system can transport foreign particles through it. When the ear is infected, transport tends to be slowed down.


The ceruminous glands in chronic suppurative otitis media. A histological and histochemical study.

The ceruminous glands have been studied histologically and histochemically in twenty patients suffering from chronic suppurative otitis media with discharging ears, and in ten patients with dry central perforations, in comparison with normal ears. The present study revealed hyperactivity of the glands initially; later on, atrophy and fibrosis took place. Thus the diminution of wax may be due to: (1) The washing effect of the discharge. (2) Atrophy of the glandular structures. (3) Periglandular fibrosis. As the discharge stops, regeneration of the glands occurs.


Chloroquine ototoxicity--a reversible phenomenon?

The case of a six-year-old girl suffering from severe cochleo-vestibular dysfunction following a series of chloroquine phosphate injections is presented in this paper. Prompt institution of therapy with steroids and vaso-dilators has helped to restore the hearing to a serviceable (socially acceptable) level. A surmise is made about the pathogenesis of chloroquine ototoxicity and how prompt appropriate therapy may be able to reverse the process. A plea is made for early referral of these cases so that prompt attention may be paid to them.


Malignant tumours of the nasal septum.

(1) Primary malignant tumours of the nasal septum are rare. (2) Forty-three cases of primary malignant septal tumours are presented and analyzed. (3) This present series is compared with the 97 previously reported cases. (4) The commonest primary malignant tumour at this site is squamous cell carcinoma. (5) The aid of a computer has been utilized to assess survival trends. (6) The overall 5-year survival in this group is 58 per cent. (7) Prognosis is not significantly affected by different treatment protocols. (8) Prognosis is significantly worse in patients with late stage tumours and in patients who have radiological abnormalities in their paranasal sinus x-rays.


Chemical synthesis of three 14 alpha-hydroxymethyl cholestenols.

Reported herein are chemical syntheses of 14 alpha-hydroxymethyl-5 alpha-cholest-8-en-3 beta-ol, 14 alpha-hydroxymethyl-5 alph-cholest-7-en-3 beta-ol, and 14 alpha-hydroxymethyl-5 alpha-cholest-6-en 3 beta-ol. These compounds were obtained in pure form after repeated medium-pressure column chromatography of the mixture obtained by treatment of 3 beta-acetoxy-7 alpha,32-epoxy-14 alpha-methyl-5 alpha-cholestane with pyridine hydrochloride in refluxing acetic anhydride followed by reduction with lithium aluminum hydride. The compounds were characterized by their chromatographic properties and by the results of infrared, optical rotation, nuclear magnetic resonance, and low and high resolution mass spectral studies.


A convenient method for the separation of molecular species of sphingomyelin by reversed-phase high-performance liquid chromatography (HPLC) is described. Sphingomyelin species from bovine brain and sheep and pig erythrocytes were resolved into 10-12 separate peaks on a micro -BondaPak C(18) or Nucleosil-5-C(18) reversedphase column with methanol-5 mM potassium phosphate buffer, pH 7.4, 9:1 (v/v) as the solvent. Detection was at 203-205 nm. The sphingomyelin species were primarily resolved due to specific hydrophobic interaction of their fatty acid and sphingoid chains with the alkyl ligand of the stationary phase. The retention time of the sphingomyelin species increased progressively as the number of carbon atoms in the hydrophobic chains increased in the homologous series. The presence of one double bond in the molecule reduced the retention time significantly. Introduction of a second double bond in the fatty acid side chain did not reduce the retention time to the same extent as the first double bond. The presence of a trans double bond in the sphingoid moiety increased the retention time of sphingomyelin more than did a cis double bond in the fatty acid side chain. The differential hydrophobic interaction observed between the ligand of the stationary phase and different alkyl chains of the sphingomyelin species illustrates that reversed-phase HPLC technique can be conveniently used to study the extent of relative hydrophobicity of different types of alkyl chains.-Jungalwala, F. B., V. Hayssen, J. M. Pasquini, and R. H. McCluer. Separation of molecular species of sphingomyelin by reversed-phase high-performance liquid chromatography.


Enzyme--substrate interaction in lipid monolayers. III. A study of the variation of the surface concentration with lipolysis.

Monolayers of a diacylglycerol were submitted to the action of lipase, keeping the area constant. The variation of lipase, keeping the area constant. The variation of the surface concentration gamma of the substrate with time was derived from the recorded reduction of the surface pressure pi (the isotherm of the monolayer being previously established). The rate -d gamma/dt was determined both as a function of the surface concentration gamma of the substrate and as a function of the bulk concentration C of the enzyme in the underlying solution. The rate depends on the quantity of enzyme ze adsorbed on the monolayer and on the enzymatic specific activity alpha of these adsorbed enzyme molecules. Both ze and alpha vary with gamma. The two variations have been quantitatively dissociated. The curves of ze and of alpha as functions of gamma coincide with those previously established in the study of hydrolysis under constant surface pressure.


Mitochondria from the 7777 hepatoma incorporate substantial amounts of l-[U-(14)C]serine into phospholipid by a Ca(2+)-dependent base-exchange reaction. This reaction is virtually absent in normal liver mitochondria. The finding cannot be attributed to microsomal contamination of the sucrose gradient-purified 7777 hepatoma mitochondria. The reaction is also absent in the rapid-growth controls, fetal rat liver and regenerating rat liver. [(14)C]Serine incorporation into 7777 hepatoma mitochondrial phospholipid by base-exchange requires Ca(2+) and is inhibited by EDTA. Ca(2+) cannot be replaced by Mg(2+), Mn(2+), or Co(2+). The reaction is inhibited by a sulfhydryl reagent and by detergents and is abolished by heating to 70 degrees C for 10 min. Product analysis indicates that phosphatidylserine and its decarboxylation product, phosphatidylethanolamine, are formed by 7777 hepatoma mitochondria, while phosphatidylserine is the sole product with microsomes. The conversion of phosphatidylserine into phosphatidylethanolamine in 7777 hepatoma mitochondria is inhibited by KCN. This study provides further evidence of abnormal mitochondrial biogenesis in the 7777 hepatoma. Our earlier study indicated a greatly increased mitochondrial activity of CTP:phosphatidate cytidylyltransferase in the 7777 hepatoma (Hostetler, Zenner, and Morris. 1978. J. Lipid Res. 19: 553-560). The presence in mitochondria of these two enzymes, which are primarily microsomal in normal liver, does not appear to be due to rapid growth alone, since their intracellular distribution was not altered in fetal or regenerating rat liver.-Hostetler, K. Y., B. D. Zenner, and H. P. Morris. Phosphatidylserine biosynthesis in mitochondria from the Morris 7777 hepatoma.


Effects of high-glucose and high-fat diets on concanavalin A binding to rat liver plasma membranes and on the amount and pattern of their glycoprotein carbohydrates.

Purified liver plasma membranes were prepared from rats fed a high-fat, carbohydrate-free diet or a high-glucose, fat-free diet. Membranes from rats fed the high-fat diet bound significantly less 125I-labeled concanavalin A (Con A) than did those from rats fed the fat-free diets. The magnitude of the binding difference increased with increasing concentrations of Con A. Neither association nor dissociation rates of the lectin-receptor complex was affected by diet. The extent of degradation of Con A by liver plasma membrane preparations from rats fed either diet was the same. Chemical analysis of delipidated liver plasma membrane showed that membranes prepared from high-fat diet-adapted rats had significantly lower values for all carbohydrate components measured with the exception of galactose. The results indicate that, in liver cells, a change in plasma membrane glycoproteins is part of the complex adaptation to altered diet composition.


A high-performance liquid chromatographic method for the determination of tocopherol in plasma and cellular elements of the blood.

A rapid sensitive, and reproducible procedure is described for the analysis of alpha-tocopherol in blood cells and plasma using high-performance liquid chromatography and fluorometric detection. The cardinal feature for the increased sensitivity of this high-performance liquid chromatographic procedure is that the fluorometric analysis was carried out at a short excitation wavelength (205 nm) which increased the sensitivity of 20-fold over the usual excitation wavelength of 295 nm. Tocopherol levels can be measured in as little as 50 microliters of plasma and 200 microliters of erythrocytes. The tocopherol contentof plasma, red blood cells, platelets, polymorphonuclear leukocytes, and lymphocytes of normal subjects and subjects ingesting additional quantitites of vitamin E are reported. The values for the white cells are approximately 30 times higher than those of the red blood cells (polymorphonuclear leukocytes 4.47 +/- 0.62 micrograms/10(9), lymphocytes 3.89 +/- 0.85 micrograms/10(9), and erythrocytes 1.40 +/- 0.14 micrograms/10(10) cells). The tocopherol contents of the plasma and all the cellular elements of the blood were increased by oral feeding with vitamin E.


The intestinal absorption of cholesterol and beta-sitostanol (the saturated analogue of beta-sitosterol) were measured and their absorptions compared in the presence and absence of cholestyramine. After test meals containing [(3)H]cholesterol and [(14)C]beta-sitostanol without added cholestyramine, 4-day fecal collections yielded an average of 51% of the fed cholesterol and 83% of the fed beta-sitostanol. In separate lymph transport studies without cholestyramine, 36% of the fed cholesterol was recovered in lymph in 24 hours compared to only 2% of the fed beta-sitostanol. Thus, while total recoveries of the two labeled compounds in feces plus lymph were nearly identical (51% + 36% = 87% for cholesterol and 83% + 2% = 85% for beta-sitostanol) their distribution in the two compartments was markedly different, reflecting the relative nonabsorbability of beta-sitostanol. Adding cholestyramine to the test meal caused fecal excretion of cholesterol to increase to 73%, independent of the dose of cholestyramine used. Cholestyramine had no effect on the fecal excretion of beta-sitostanol (average excretion after cholestyramine, 85%). The relative non-absorbability of beta-sitostanol compared to cholesterol is clearly evident in this study and leads us to suggest its possible use as a lipid-soluble, nonabsorbable reference compound for measurement of the absorption of cholesterol and other lipids. Further data are presented to justify its use for this purpose.-Hassan, A. S., and A. J. Rampone. Intestinal absorption and lymphatic transport of cholesterol and beta-sitostanol in the rat.


Several aspects of lipid metabolism in the soleus and diaphragm muscles of streptozotocin-diabetic and control rats were investigated. The triglyceride content of both muscles was elevated in the diabetic state and the presence of increased intracellular lipid was confirmed by electron microscopy. In vitro glucose and palmitate oxidation studies showed that both types of muscle from the diabetic animals metabolized more fat than did the soleus and diaphragm from control rats. While isoproterenol alone produced a significant lipolytic response in both the soleus and diaphragm from control and diabetic animals, there was no difference in the percent increase in fatty acids released from muscles of diabetic rats compared to controls. However, the absolute difference was greater when the diaphragms were compared. Muscles from experimental and control animals showed a marked reduction in the amount of free fatty acids released in response to insulin. In addition, in the presence of the hormone, both the absolute and percent isoproterenol-stimulated increases in fatty acids were significantly greater for both diaphragm and soleus muscles from diabetic rats. The effects of insulin, isoproterenol, and the combination of these two hormones on the amount of glycerol released into the incubation medium were similar to those found on free fatty acid release. The results of these experiments show that there is an apparent increase in fat utilization in skeletal muscle of diabetic rats. Furthermore, measurements of triglyceride concentration and the enhanced response to isoproterenol stimulation in the muscles from these animals suggests that they may have an increased capacity for mobilization of intracellular lipids. Finally, in the diabetic state, both the soleus and diaphragm appear to demonstrate an increased response to the antilipolytic effect of insulin as measured by the decreased amount of fatty acid released into the incubation medium, the percent change also being significant for the soleus muscle.-Stearns, S. B., H. M. Tepperman, and J. Tepperman. Studies on the utilization and mobilization of lipid in skeletal muscles from streptozotocin-diabetic and control rats.


Isolation and characterization of a heptaglycosylceramide from bovine erythrocyte membranes.

A heptaglycosylceramide was isolated from bovine erythrocyte membranes. The structure was characterized to be Gal(alpha 1-3)Gal(beta 1-4)GlcNAc(beta1-3)Gal(beta 1-4)Glc-NAc(beta 1-4)al(beta 1-4)GlcCer. A hexaglycosylceramide that has the same sequence except for the terminal alpha-galactosyl unit has also been isolated. We have previously found that gangliosides isolated from bovine erythrocyte membranes contain a keratan sulfate type repeating unit --[3Gal(beta 1-4)-GlcNAc beta]--n. This study shows that the keratan sulfate type repeating unit is also present in the neutral glycosphingolipids of bovine erythrocyte membranes.


A convenient synthesis of phosphatidylcholines: acylation of sn-glycero-3-phosphocholine with fatty acid anhydride and 4-pyrrolidinopyridine.

A high-yield synthesis of saturated, unsaturated, and short chain phosphatidylcholines from sn-glycero-3-phosphocholine is described. The procedure offers advantages over other reported procedures for the synthesis of phosphatidylcholine in that the large-scale synthesis and purification can be achieved in a minimum time. The procedure utilizes 4-pyrrolidinopyridine as a catalyst and moderate amounts of fatty acid anhydride (2 mol eq. of fatty acid anhydride per mol of OH) in a 1:1 mixture of benzene-dimethylsulfoxide (DMSO) at 40 degrees--42 degrees C (oilbath) for 2--5 hr. At the end of the reaction, the phosphatidylcholine can be purified in the usual manner or by using a Waters Prep LC/500 with a radially compressed silica gel column eluted with chloroform-methanol-water 60:30:4. At a flow rate of 200 ml/min, the phospholipid elutes in 10--15 min, depending on the chain length and unsaturation.


Preparation of radiolabeled GM2 and GA2 gangliosides.

GM2 and GA2 gangliosides from the brain of a patient who died of Sandhoff's disease were purified by solvent partition, silicic acid and silica gel column chromatography, and silica gel preparative thin-layer chromatography. They were tritiated in the terminal N-acetylgalactosamine residue using galactose oxidase and sodium [3H]borohydride with the inclusion of catalase and peroxidase into the oxidation reaction. The specific activities were 4.62 X 10(8) dpm/mumol of GM2 ganglioside and 5.54 X 40(7) dpm/mumol of GA2 ganglioside. The addition of catalase and peroxidase to the tritiation procedure is recommended.


Sterol synthesis. High-pressure liquid chromatography of C27 sterol precursors of cholesterol.

Conditions have been described which permit the rapid (approximately 3.5 hr) column chromatographic separation of the acetate derivatives of a number of C27 sterol precursors of cholesterol differing only in the number and position of double bonds in the sterol nucleus. On columns containing muPorasil with hexane-benzene 9:1 as the eluting solvent, acetate derivatives of the delta5, delta8(14), delta8, delta7, delta8,7, delta 8,14, and delta7,14 sterols were separated.


Isotopic labeling of DNA in rat adipose tissue: evidence for proliferating cells associated with mature adipocytes.

The intraperitoneal administration of [3H]thymidine to adult rats resulted in the rapid appearance of label in the adipocyte fraction of collagenase digests of adipose tissue. Low-speed centrifugation followed by freezing and slicing showed the label to be uniformly distributed in the adipocyte fraction. The presence of label in DNA was confirmed by hydrolysis with deoxyribonuclease and by inhibition of incorporation with hydroxyurea. Organelle fractionation revealed that the label was predominantly in nuclei, and radioautography showed that only a few adipocyte nuclei were labeled. The label in the adipocyte fraction could not be reduced by increased collagenase digestion or by trypsin treatment. Mixing of labeled adipocytes with unlabeled stroma did not result in decrease of label and addition of labeled stroma to unlabeled adipocytes did not cause significant transfer of radioactivity. Addition of [3H]thymidine to the collagenase digestion medium of unlabeled adipose tissue resulted in more incorporation by adipocytes than by stroma, suggesting the presence of a very rapidly proliferating cell type associated more with adipocytes than with stroma. In vivo turnover studies of labeled DNA indicated that there are two components in both adipocytes and stroma, a rapidly labeled component with a half-life of only several days and another with a half-life of several months. These experiments suggest that there is a rapidly proliferating cell type in adipose tissue, closely associated with mature adipocytes, that may be an adipocyte progenitor or may have some other unknown function.


Increased cellular proliferation in adipose tissue of adult rats fed a high-fat diet.

The feeding of a high-fat diet to adult rats was shown to increase the incorporation of [3H]thymidine into DNA of the adipocyte and stromal fractions. After only 2 days on a high-fat diet there was a marked increase in the incorporation of label. When a 2-week period was interposed between [3H]thymidine administration and determination of DNA specific activity, the greatest increase in incorporation of label was found after 1 week on the diet, when incorporation increased 6-fold or more in both adipocytes and stroma and subsequently decreased to stabilize at a level two or three times that of chow-fed rats in the adipocyte fraction. Rats labeled when young and later placed on a high-fat diet showed a decrease in DNA specific activity in both adipocytes and stroma, confirming that cellular proliferation had occurred in both fractions. The specific activities of both stromal and adipocyte DNA were very similar at all time points studied. An attempt to increase the difference in specific activities by waiting many weeks after [3H]thymidine injection before isolating DNA was not successful. This may be because the total amount of DNA in the stromal and adipocyte fractions increases in parallel on the diet. The significance of these findings in terms of the normal turnover of adipose tissue DNA and the responsiveness to diet is discussed.


Absolute rates of cholesterol synthesis in extrahepatic tissues measured with 3H-labeled water and 14C-labeled substrates.

This study was undertaken to develop techniques for measuring absolute rates of sterol synthesis in extrahepatic tissues in vitro and to estimate the magnitude of the errors inherent in the use of various 14C-labeled substrates for such measurements. Initial studies showed that significant errors were introduced when rates of synthesis were estimated using [3H]water since about 20 nmol of water were bound to each mg of tissue cholesterol isolated as the digitonide. This source of error could be eliminated by subtracting apparent incorporation rates obtained at 0 degrees C from those obtained at 37 degrees C or by regenerating and drying the free sterol. In a second set of experiments, the H/C incorporation ratio in cholesterol was determined in the liver by measuring the absolute rates of hydrogen and acetyl CoA flux into sterols. The ratio of 0.69 +/- 0.03 was found to be independent of the rate of hepatic cholesterol synthesis, the rate of hepatic acetyl CoA generation, or the source of the acetyl CoA. In a third set of studies, rates of incorporation of [3H]water or 14C-labeled acetate, octanoate, and glucose into digitonin-precipitable sterols were simultaneously measured in nine different extrahepatic tissues. Assuming that the H/C ratio measured in the liver also applied to these tissues, the [3H]water incorporation rates were multipled by the reciprocal of the H/C ratio to give the absolute rates of sterol synthesis in each tissue. When these were compared to the incorporation rates determined with the 14C-labeled substrates the magnitude of the errors in the rates of sterol synthesis obtained with these substrates in each tissue could be assessed. Only [14C]octanoate gave synthesis rates approaching 100% of those obtained with [3H]water and this occurred only in the intestine and kidney; in the other extrahepatic tissues this substrate gave rates of 6--66+ of the absolute rates. Rates of [14C]acetate incorporation in sterols varied from 4 to 62% of the [3H]water incorporation rates while those obtained with [14C]glucose were only 2--88% of the true rates. These studies document the large and highly variable errors inherent in estimating rates of sterol synthesis in extrahepatic tissues using 14C-labeled substrates under in vitro conditions.


Crystal structure of cholesteryl decanoate.

Cholesteryl decanoate (C37H64O2) is monoclinic, space group P2I, with cell dimensions a = 12.931 (6), b = 9.066 (2), c = 30.22 (1) A, beta = 91.14 (4) degrees, and Z = 4. The atomic coordinates from cholesteryl laurate were used in an initial trial structure which was refined by block diagonal least-squares methods with 1846 observed X-ray reflections (R = 0.129). Molecules A and B have almost fully extended conformations, except at the ester bonds and towards the end of the decanoate B chain. The molecules are arranged in antiparallel array forming monolayers of thickness d001 = 30.22 A, with the molecular long axis making an angle of about 67 degrees with the layer interface. The crystal structure is very similar to that of cholesteryl nonanoate and cholesteryl laurate.


Unusual properties of retinyl palmitate hydrolase activity in rat liver.

These studies report the hydrolysis of retinyl palmitate with liver homogenates and homogenate fractions from retinol-depleted rats. The studies utilized an effective in vitro assay for retinyl palmitate hydrolase (RPH) activity, in which microgram amounts of retinyl palmitate were employed as substrate, followed by the chromatographic separation and fluorescence assay of free and esterified retinol. RPH activity was maximal near pH 8 in Tris-maleate buffers, and required a bile salt for stimulation. Both cholate and taurocholate stimulated the reaction, whereas a number of other detergents tested were ineffective. The enzymatic activity showed an unusual subcellular distribution, with about 40% of total RPH activity recovered in the washed "nuclear" fraction (1,500 g pellet) and about 30--35% in the 105,000 g supernatant. This unusual distribution was not observed for marker constituents for plasma membranes, nuclei, mitochondria, lysosomes, Golgi apparatus, or endoplasmic reticulum. Despite its enrichment in the "nuclear" fraction, RPH activity was not enriched in purified preparations of nuclei or plasma membranes. Thus, RPH activity was not localized in any single, characterized subcellular structure. Another striking feature of the hepatic RPH activity was its extreme variability from rat to rat as assayed in vitro. Both the unusual subcellular distribution and the marked variability in activity were not observed for a variety of other hepatic ester hydrolase activities examined. Of ten lipid and nonlipid esters tested as substrates, only the hydrolytic activities against cholesteryl oleate and phytyl oleate correlated with, and partly resembled, RPH activity in these respects. The results suggest that the observed RPH activity is relatively specific for the hydrolysis of retinyl palmitate, and may therefore be significantly involved in hepatic retinyl ester metabolism.


Lipid structure and the behavior of cholesteryl esters in monolayer and bulk phases.

The behavior of cholesteryl esters at the air-buffer interface was studied as a function of molecular area and the presence of noncholesterol-containing lipids (colipids). The data obtained indicate that cholesteryl esters with other than long, saturated acyl groups can be present in surface phases up to packing densities approximately those in natural membranes. Their apparent molecular areas in such phases, which are largely determined by colipid structure, suggest their orientation with the ester function toward the interface. The extent of miscibility in the surface phase is also a strong function of colipid structure. Reversibility of the monolayer to bulk phase transition is determined exclusively by the acyl structure of the cholesteryl ester. Of the esters examined, only those with cis unsaturation collapsed reversibly. Our data predict that cholesteryl esters should be present in small, but finite amounts on the surface of arterial lipid deposits and that a prerequisite for the removal of such deposits is that the bulk lipid phase be in a liquid or liquid crystalline state.


Separation of individual sulfated bile acid conjugates as calcium complexes using reversed-phase partition thin-layer chromatography.

A method for separating individual monosulfated primary bile acid conjugates by reversed-phase partition thin-layer chromatography on octadecyl-bonded silica gel is described. The solvent system is acetonitrile containing calcium, probably as calcium carbamate. Excellent resolution of the 3- and 7-monosulfated glycine conjugates, as well as 3- and 7-monosulfated taurine conjugates of cholic and chenodeoxycholic acids is reported. A convenient class separation of sulfated from nonsulfated primary bile acid conjugates by adsorption thin-layer chromatography on low-polarity silica gel is also described.


Effect of glucocorticoids on the oxidative desaturation of fatty acids by rat liver microsomes.

The effect of glucocorticoids on the oxidative desaturation of fatty acids by liver microsomal preparations of rats has been studied. Hydrocortisone produced a significant decrease in the conversion of [1-14C]linoleic acid to gamma-linolenic acid and [1-14C]eicosa-8, 11, 14-trienoic acid to arachidonic acid. Triamcinolone and dexamethasaone were more active than hydrocortisone in depressing delta 6 and delta 5 fatty acid desaturating activity in liver microsomes. The glucocorticoids evoked a maximal response approximately 24 hr after admission. Palmitic acid conversion to palmitoleic acid showed no statistically significant changes by any of the glucocorticoids. The mechanism of action of glucocorticoids is apparently different from other hyperglycemic hormones that produce similar effects.


The stereoconfiguration of bis(monoacylglycero)phosphate synthesized in vitro in lysosomes of rat liver: comparison with the natural lipid.

A procedure for stereoanalysis of radiochemically labeled glycerophospholipids is described. It is based on the study of the labeled alpha-glycerophosphate which retains its original configuration when liberated upon alkaline hydrolysis of the lipids. The labeled alpha-glycerophosphate is oxidized enzymatically with sn-3-glycerophosphate dehydrogenase and the product, dihydroxyacetone phosphate, is degraded with alkali to inorganic phosphate. The nonoxidizable alpha-glycerophophate (sn-1-glycerophosphate), the beta-glycerophosphate, and the inorganic phosphate derived from sn-3-glycerophosphate are quantitated after separation by thin-layer chromatography. The procedure gave the expected results when applied to [3H]glycerol-and 32P-labeled phosphatidylcholine, bis( monoacylglycero)phosphate, and phosphatidylglycerol from natural resources. Bis(monoacylglycero)phosphate, known also as lysobisphosphatidic acid, was synthesized from ]32P]diphosphatidylglycerol and from phosphatidyl[1',3'-3H]glycerol in lysosomal preparations of rat liver according to Poorthuis and Hostetler (1978. J. Lipid Res. 19: 309-315). Stereoanalysis proved that the product was in both cases a derivate of sn-1-glycerophospho-sn-1'-glycerol.


Distribution of 3-hydroxy-3-methylglutaryl coenzyme A reductase and alkaline phosphatase activities in isolated ileal epithelial cells of fed, fasted, cholestyramine-fed, and 4-aminopyrazolo[3,4-d]pyrimidine-treated rats.

3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase, E.C. 1.1.1.34), the major rate-limiting enzyme of the sterol biosynthetic pathway, was studied in ileal epithelial cells isolated in a villus-to-crypt gradient according to Weiser (Weiser, M. M. 1973. J. Biol. Chem, 248:2536-2541). Alkaline phosphatase (E.C. 3.1.3.1) served as a marker for the mature villus cells. Protease effects on activity determinations were negligible. The intracellular location of HMG-CoA reductase could not be precisely determined. The activity of ileal reductase was predominantly associated with the less differentiated lower villus and crypt cells, while the reverse gradient occurred with alkaline phosphatase. This distribution of enzymes persisted in both fed and fasted rats injected with control saline-phosphate, although fasting decreased total reductase units in the ileum by 86% in 72 hr. Treatment with cholestyramine and with 4-aminopyrazolo[3,4-d]pyrimidine (APP) enhanced reductase activity in ileal cells. The percent stimulation in both cases was higher in the upper villus cells than in the crypt cells, leading to abolition of the gradient in enzyme activity. However, APP treatment caused a 98% loss in total alkaline phosphatase units and a 55% loss in total epithelial cell protein in 72 hr. Thus, there was no increase in total reductase units. These data show that APP affects ileal cell metabolism directly. Furthermore, it appears that the regulation of sterol synthesis in the intestinal mucosa, via HMG-CoA reductase, involves a complex interplay of the effects exerted by the level of alimentation, the enterohepatic circulation of bile, and the levels of plasma lipoproteins.


Free ceramide, sphingomyelin, and glucosylceramide of isolated rat intestinal cells.

Free ceramide, glucosylceramide, and sphingomyelin were isolated from mature cells of adult rat small intestine. Free ceramide and ceramide cleaved from sphingomyelin by enzymatic hydrolysis were fractionated by thin-layer chromatography on borate-impregnated silica gel plates. Sphingoid bases were characterized by gas-liquid chromatography of aldehydes formed upon periodate oxidation. Fatty acids were quantified as methyl esters. Ceramide structures were confirmed by direct-inlet mass spectrometry. Free ceramide was found to contain two major long-chain bases in nearly equal quantity: sphingosine, mainly linked to palmitic acid, and 4D-hydroxysphinganine associated with C20 to C24 fatty acids, 22% being hydroxylated. Sphinganine occurred as a minor component linked to nonhydroxy fatty acids. Sphingomyelin contained the three long-chain bases and 63% of its ceramide was N-palmitoyl-sphingosine. Mass spectrometry of glucosylceramide confirmed 4D-hydroxyshingamine as the major sphingoid base associated preferentially with longer chain hydroxy fatty acids.


Measurement of arachidonic acid in the plasma by gas-liquid chromatography-flame ionization using dihomo-gamma-linolenic acid as an internal standard.

A gas-liquid chromatography-flame ionization method is described for measuring arachidonic acid in plasma using dihomo-gamma-linolenic acid as an internal standard. We found this technique to -e reproducible, and quicker and superior to previously described techniques because of the similar physico-chemical properties of the unsaturated fatty acid internal standard and arachidonic acid. In addition, we observed that the use of the saturated fatty acid, n-tricosanoic acid, was unsatisfactory as an internal standard because of its poor extractability from plasma as compared to arachidonic acid.


Measurement of cholic acid synthesis and secretion by isolated rat hepatocytes.

Liver cells isolated from normal and cholestyramine-treated rats were incubated as cell suspensions for up to 4 hr in a simple, defined medium. The bile acid concentration in cells plus cell medium was determined by gas-liquid chromatography. Normal hepatocytes synthesized cholic acid at an initial rate of 0.25 nmol/mg cell protein per hr, which is comparable to rates reported from in vivo methods. This rate was increased more than 4-fold when rats were fed a cholestyramine-containing diet for 7 days prior to liver cell isolation. Although cholic acid was secreted into the cell medium during the incubation, it could not be assayed reliably by the hydroxysteroid dehydrogenase assay method, contrary to the reports of Anwer et al. 1975. Biochem. Biophys. Res. Commun. 64: 603 and Gardner and Chenouda 1978. J. Lipid Res. 19: 985.


A threshold theorem for the general epidemic in discrete time.

A theorem, analogous to the continuous time Threshold Theorem of Kermack and McKendrick, is proved for a certain discrete time epidemic model. This model, in contrast to its continuous time analogue, leads to some solutions in which the total population of susceptibles may become infected in a finite time.


A stochastic Gompertz model with hereditary effect.

In this paper we have studied a stochastic version of the Gompertz model for population growth of a single species after incorporating the aspect of heredity. Various statistical characteristics--the mean-value function, covariance-kernel, etc.--are evaluated for a delta-correlated process and their asymptotic values obtained. The effect of the hereditary kernel on the various statistics is discussed and it is found that it is to shift the distribution towards the origin.


Effect of chicken hypothalamus on prolactin and growth hormone secretion in male chickens.

The effects of a chicken hypothalamic extract (HE) on the secretion of prolactin and growth hormone (GH) in vivo have been investigated by radioimmunoassay in the domestic fowl. Different i.v. doses of HE (0.25--25 HE equivalents/kg body weight) had no effect on GH secretion in conscious or anaesthetized cockerels. In both groups of birds the concentration of plasma prolactin was significantly increased within 10 min of administration of the extract. Extracts of other brain tissues (cerebral cortex, cerebellum and medulla oblongata) had no stimulatory effect on prolactin or GH secretion. Release of both prolactin and GH by dispersed pituitary cells and by hemipituitary glands in vitro was enhanced following incubation with HE (5 hypothalami equivalents/ml) or with single whole hypothalami respectively. Other brain tissues (cerebellum, optic lobes and medulla oblongata) had no effect on the concentration of prolactin or GH released by incubated hemipituitary glands.


Prolonged intravenous infusion of labelled iodocompounds in the rat: [125I]iodide metabolism and effects of moderate dietary iodine deficiency.

Radio-iodide was administered by prolonged continuous intravenous infusion to rats maintained under iodine-replete conditions and in moderate iodine deficiency. A close approximation to equilibrium labelling was thereby achieved. Labelled iodocompounds extracted from various tissues were analysed by thin-layer chromatography. Moderate iodine deficiency resulted in a slight increase in the ratio of mono-iodotyrosine to di-iodotyrosine in the thyroid. No change in the ratio of tri-iodothyronine (T3) to thyroxine (T4) was found in thyroid, plasma or skeletal muscle. Faecal excretion of T3 declined appreciably relative to that of T4. Under iodine-replete conditions the ratio of thyroidal secretion rates of T3 and T4 was estimated to be more than three times higher than the ratio of these iodocompounds within the thyroid. Heterogeneity of thyroglobulin structure and function may explain these observations.


Prolonged intravenous infusion of labelled iodocompounds in the rat: [125I]thyroxine and [125I]tri-iodothyronine metabolism and extrathyroidal conversion of thyroxine to tri-iodothyronine.

Extrathyroidal tissues of the rat were labelled to steady state by prolonged continuous intravenous infusion of 125I-labelled thyroxine (T4) or tri-iodothyronine (T3). Labelled iodocompounds extracted from various tissues were analysed by thin-layer chromatography. Signifficant amounts of labelled T3 were found in all tissues examined after infusion of [125I]T4, confirming that conversion of T4 to T3 occurs in extrathyroidal tissues of the rat. Faecal excretion of labelled T3 after [125I]T4 infusion provided an assessment of the extent of extrathyroidal conversion: about a third of the T4 was metabolized by this pathway. Extrathyroidal conversion was independently estimated to account for about a third of the total production of T3. The site of extrathyroidal conversion was established by comparing the distribution of labelled T3 after the two types of infusion: kidney and liver were both prominent sites of conversion of T4 to T3.


A sensitive and specific assay for 5-methoxytryptophol in plasma.

5-Methoxytryptophol (ML) is found in the pineal gland and is known to have biological activity especially as an antigonadotrophic agent, but methods have been lacking for its measurement in the circulation. A gas chromatography-mass spectrometry assay using a trimethylsilyl derivative has been developed for the routine measurement of ML in plasma. The assay is of great specificity and has a sensitivity of 20 pmol/l. Studies on the levels of pineal indoles in the circulation, however, have been hampered by the possibility that extraneous compounds are being cross-measured. Thus the specificity of the routine assay has been further validated by comparing it with an alternative assay system where all the major parameters were changed, i.e. derivatizing reagent, internal standard and mass number. Results that were obtained using both assay systems were closely comparable.


Synthesis of prolactin by human decidua in vitro.

To determine whether human decidua and/or chorion synthesizes and secretes prolactin, explants of decidua obtained at Caesarian section and explants of chorion from the membranes separating dizygotic twins were cultured for periods of up to 6 days. The decidual explants released 366 +/- 37 ng prolactin/100 mg tissue (mean +/- S.D.) during each day in culture and incorporated 3H-labelled amino acids into immunoprecipitable prolactin. In the radioimmunoassay for prolactin, serial dilutions of incubation medium displaced 125I-labelled prolactin parallel to the displacement by pituitary prolactin and the prolactin in the medium eluted from Sephadex G-150 in a position indentical to that of pituitary prolactin. Chorionic explants released prolactin into the incubation medium during day 1 of culture only and did not incorporate 3H-labelled amino acids into prolactin. These results demonstrate that prolactin is  synthesized by the decidua and not by the chorion and suggest that the decidua is the source of prolactin in amniotic fluid.


Changes in the concentration of 5-methoxytryptophol in the circulation at different phases of the human menstrual cycle.

The pineal indole 5-methoxytryptophol (ML) has been shown to have an antigonadal activity when administered to experimental animals, but data on its normal pattern of secretion have been lacking. Using a new gas chromatography-mass spectrometry assay, the concentration of ML at various phases of the human menstrual cycle has been studied. Daily samples were obtained throughout the month from five women with a normal cycle and two women taking an oral contraceptive. In women with a normal cycle levels of ML were found to be significantly lower in the last third of their cycle; this change was not seen in women taking an oral contraceptive who had low levels throughout the month. The changes in concentration of ML did not correlate with the changes in concentration of gonadotrophins.


Influence of the pineal gland, olfactory bulbs and photoperiod on surges of plasma prolactin in the female rat.

The role of the pineal gland, olfactory bulbs and photoperiod in the regulation of the two daily surges of plasma prolactin in the pseudopregnant rat has been investigated. Pinealectomy had no effect on the surges of prolactin in pseudopregnant rats maintained on either a long (14 h light : 10 h darkness; 14L : 10D) or a short (2L : 22D) photoperiod, but olfactory bulbectomy decreased the nocturnal surge in animals maintained on 14L : 10D. This effect of bulbectomy was eliminated if the rats maintained on 14L : 10D were also pinealectomized. After cervical stimulation, bulbectomized rats maintained on a 2L : 22D photoperiod had nocturnal-type prolactin surges similar to those of intact rats maintained on the same photoperiod. These results indicate that the pineal gland andlength of photoperiod are not involved in the regulation of the surges of plasma prolactin in pseudopregnant rats but that the olfactory bulbs may enhance the nocturnal surge.


Epitestosterone in the plasma of the goat during pregnancy and at parturition.

Epitestosterone, a product of the metabolism of androstenedione by the caprine placenta in vitro, is present in the plasma of the pregnant goat. The maternal concentrations of both epitestosterone and unconjugated oestrogens (mostly oestradiol-17 alpha) in the blood increased before parturition and dropped post partum. Measurement of arteriovenous differences at term indicated that epitestosterone was secreted by the uterus; its production was not dependent on the presence of corpora lutea. It is suggested that the concentration of epitestosterone (+ androstenedione + oestrogens) in maternal plasma may be used as an indicator of placental C-17,20 lyase activity; the slight rise in the concentration of these compounds prepartum suggests a relatively small increase in flow through this enzyme.


Influence of androgens on the weights of the male accessory reproductive organs and on the activities of mitochondrial enzymes in the epididymis of the rat.

The influence of androgens on the male accessory glands of the rat was assessed in terms of changes in weight and of the specific activity of the mitochondrial enzymes, succinate dehydrogenase, glycerolphosphate dehydrogenase and pyruvate carboxylase, in the epididymis. In some instances, the activity of the cytoplasmic enzymes, hexokinase and phosphofructokinase, was also measured and the influence of androgens on these enzymes was found to be similar to that on the mitochondrial enzymes. After the administration of androgen to castrated rats the specific activity of enzymes reached a new steady state sooner than did epididymal weight. The time taken for the specific activity of the enzymes to reach a new steady state after the removal of androgen was variable, depending on the enzyme and the region of the epididymis. This time was generally longer, however, than the time taken for induction, and in the case of glycerolphosphate dehydrogenase, the decline of activity was slower in the cauda than in the caput. In castrated animals, about 100 times as much androgen was required to attain maximum tissue weight as was required to attain maximum enzyme activity. The epididymis, prostate and seminal vesicles responded similarly to androgen in terms of the dose-response pattern and the time taken for tissue weight to attain a new steady-state value, although the gain in weight of the epididymis relative to its weight in unstimulated control animals was less than the relative gain of the other accessory glands. Enzymes in the cauda epididymidis required lower amounts of androgen to elicit maximum activity than were required by those in the caput. The rate of change in the accessory glands in attaining new steady-state levels of tissue weight and enzyme activity was independent of the dose of androgen except during the first few days of hormone administration. Androgens were the most effective steroids in stimulating an increase of tissue weight and enzyme activity, although some changes were induced by oestradiol-3-benzoate and progesterone.


Failure of melatonin to increase serum prolactin levels in ovariectomized rats subjected to superior cervical ganglionectomy or pinelaectomy.

The effects of melatonin on serum prolactin levels were examined in ovariectomized rats primed with oestradiol and progesterone, and subjected to bilateral superior cervical ganglionectomy or pinealectomy. Ganglionectomy resulted in a significant depression of the serum prolactin concentrations, as well as in impairment of the prolactin release evoked by administration of steroid. Treatment with melatonin increased serum prolactin in control but not in ganglionectomized rats. Injection of melatonin protentiated the steroid-induced release of prolactin in control rats; this effect of melatonin was not detected in ganglionectomized rats. Pinealectomy did not affect basal prolactin levels, nor impair the release of prolactin evoked by steroid treatment; however, it was effective in blocking the melatonin-induced release of prolactin in vehicle-treated rats, as well as the potentiation of steroid-induced prolactin release by melatonin. Intracranial surgery by itself increased prolactin release. These results suggest that systemically administered melatonin needs an intact pineal gland to augment serum prolactin levels.


Multiple sclerosis in southern Europe. I: Prevalence in Sicily in 1975.

Previous reports on large population groups have suggested that the prevalence of multiple sclerosis (MS) in Sicily and southern Italy is low, of the order of 4-8 per 100 000. In contrast, immigrants from Italy resident in Greater London, many of whom are from southern Italy and Sicily, had a hospitalised MS prevalence similar to that found among people born in the United Kingdom (1960-72). The present study shows that in Enna city (population 29 000) in central Sicily, the prevalence of probable MS was 53 per 100 000, which is of the same order of magnitude as has been reported from the United Kingdom and northern Europe. The high prevalence of MS found in Enna city may be due, at least in part, to the fact that the population studies was small. Because Enna is on high ground, similar studies are being undertaken in two small coastal towns of Sicily.


Multiple sclerosis in southern Europe. II: Prevalence in Malta in 1978.

After an intensive survey only 14 patients have been found with a diagnosis of probable multiple sclerosis (MS) in the islands of Malta. This is a low prevalence of 4.2 per 100 000. The low prevalence of MS is confirmed by the small number of deaths certified as due to MS--six in 11 years--and by the absence of Maltese MS patients resident in England among the MS patients admitted to hospital in Greater London and the West Midlands (1960-72). The low prevalence of MS found in Malta can be contrasted with the high prevalence found in Enna city in central Sicily. The genetic and environmental reasons for this difference in MS prevalence between the neighbouring islands of Sicily and Malta require further study.


Surprising decline of cardiovascular mortality in Switzerland: 1951-1976.

In Switzerland the age-standardised death rate for 'all diseases of the circulatory system' decreased by 22% in males and by 43% in females between 1951 and 1976, which represented a third and nearly a half respectively of the decrease in all deaths. Mortality was reduced by 13% in males and by 40% in females for non-rheumatic heart disease and hypertension, and by 36% and 47% respectively for cerebrovascular disease. These reductions were on the whole greater than those observed in the 13 other developed countries studied. From 1951 to 1976 consumption of animal fats per caput increased by 20% in Switzerland, in spite of a 46% reduction in milk intake. Throughout the same period, smoking increased among women while it remained steady or possibly declined in men. During the last decade a threefold rise in the sales of antihypertensive drugs was observed. Oral contraceptives seem to have been widely used by Swiss women since the 1960s. There were also increases in the proportion of women in professional occupations, and in urbanisation. The advent of the economic recession was associated with a sudden rise in mortality from non-rheumatic heart disease and hypertension in 1974-76.


Sickness absence after inguinal herniorrhaphy.

Eight hundred and ninety-nine men were studied, aged 16-65 inclusive, who underwent an elective inguinal herniorrhaphy during 1970 and 1971 in eight hospitals in Wessex, and under nine consultant surgeons. There was a significant variation in postoperative inpatient stay and total sickness absence between hospitals and between consultants. The physical activity involved in the patient's occupation, his age at operation, previous sickness absence, bilateral herniorrhaphy, attendance at follow-up outpatients' clinic, type of repair, and the influence exerted by three hospitals and two consultants accounted for only 21% of the variation in total sickness absence. The general practitioners who had referred patients to the hospitals for herniorrhaphy, and the consultant surgeons who carried out the operations, were sent a questionnaire to ascertain their attitudes towards follow-up outpatient appointments and the various factors identified in the first part of the study as significantly influencing total sickness absence. A higher proportion of GPs who felt that an outpatient appointment was necessary before return to work was found in relation to the patients who had the longest mean total sickness absences than among the GPs who looked after the patients with shorter total sickness absences.


Tuberculosis among immigrants in the United Kingdom: the role of occupational health services.

Immigrant workers in the United Kingdom account for 7.8% of the working population. Their health problems fall into three groups--imported diseases, occupational accidents, and acquired diseases. In the latter group, tuberculosis is still a major problem. A retrospective study tracing notifiable cases of tuberculosis to points of entry at Heathrow Airport indicates that the majority of immigrants acquire the disease after entry. The occupational health services have a great role to play in detecting the new cases very early and in providing appropriate screening and follow-up. Proper co-ordination between occupational and community health services will give the best results and lead to a rapid decline of the disease.


Alcohol and oesophageal cancer: an assessment of the evidence from routinely collected data.

Although various factors have been implicated in the aetiology of oesophageal cancer, one factor common to many countries is the consumption of alcoholic beverages. In England and Wales mrtality from oesophageal cancer declined rapidly during the early part of this century but both mortality and incidence have increased in recent years. The generation of males born in 1906 had lower mortality than any preceding or succeeding generation. It is suggested that ages 20 to 30 may be critical in the development of oesophageal cancer and that the 1906 cohort was less exposed to alcohol than other generations. The international analysis suggests that ethyl alcohol itself rather than any specific alcoholic beverage is associated with this cancer.


Self-reported morbidity data as an indicator of regional resource requirement.

In this paper five variables are taken from the General Household Survey and the statistical technique of multidimensional scaling is used to estimate the relationships between the standard economic regions of England and Wales in terms of their self-reported morbidity. An estimate is then made of the proportion of resources that must be allocated to each region in order to eliminate the differences found. The five variables measuring longstanding illness; limiting longstanding illness; restricted activity; certificated absence; and general practitioner consultation were standardised, and input to a multidimensional scaling programme. This showed that the English regions were very similar to each other, although a morbidity continuum could be identified which corresponded to the North-South variation in socioeconomic conditions found in England. Wales, however, shows considerable difference from the English regions, a difference which cannot be explained purely in terms of the socioeconomic conditions in the region. For this reason a far more detailed investigation is proposed.


The impact of training traditional birth attendants on the utilisation of maternal health services.

The impact of training traditional birth attendants (TBAs) on the utilisation of maternal health services at a rural health training centre in India has been studied. Records of the utilisation of the services for two periods of six months each (one before training and one after) have been analysed. A marked improvement in the registration of antenatal cases at an earlier date in pregnancy has been observed. The average number of visits to antenatal clinics, the immunisation against tetanus, and the presence of trained personnel at the time of delivery all showed considerable improvement after the local TBAs had been trained and motivated. The differences observed in the two series were statistically significant.


The role of dispensaries in community health care in the Kainji Lake area of Nigeria.

Dispensaries have been the most common providers of health services in the Kainji Lake rural area of Nigeria and the only form of health facility in this area. A survey of the working conditions of the dispensary assistants showed that 70% of the respondents are already frustrated because of lack of support, supervision, and prospects for further training. A proposal is made for further training of the dispensary assistants and for improving their conditions of service. The seasonal distribution of the common diseases (malaria, gastroenteritis, chest infections, skin diseases, gonorrhoea, and schistosomiasis) shows that infection follows the pattern of rainfall, and that there are some differences between the pattern of these common diseases and the common ailments reported by the villagers located away from the dispensaries. A suggestion is made for the introduction of primary health workers to meet the health needs of the villagers, 60% of whom have not received dispensary services.


The cost of treatment of dermatological disorders in hospital versus health centre.

Well-run outpatient services and reasonable charges to patients for drugs have led to reduced hospitalisation of dermatological patients and hence a reduction in the total cost of nursing and treating such patients in comparison with previous years, when there was greater emphasis on hospital treatment. There is at present no waiting list for dermatological beds at the National Hospital.


On the estimation of relative risk from vital statistical data.

A method is described for the determination of a measure of relative risk from vital statistical data. If the frequency of disease in a population is linearly related to the level of exposure to a given factor, then a measure of the relative risk can be estimated from the slope and intercept of the regression line. For example, when the exposure is measured in terms of the proportion of the population exposed to the factor, then the relative risk is equal to (Formula: see text). This offers an indirect but simple and inexpensive method for estimating relative risk. It should be used with caution, particularly where confounding factors may be responsible for the apparent association between disease and factor. Applications of the method to estimate the relative risk of (a) circulatory diseases in women using oral contraceptives and (b) ovarian cancer in women with different average family sizes, both yielded relative risk estimates comparable with those obtained from case-control and prospective studies.


Deviations from matching as a measure of preference for alternatives in pigeons.

Preferences for larger or smaller defined response classes were investigated in a concurrent schedule precedure. Twelve pigeons were run on a series of concurrent variable-interval reinforcement schedules, from which baseline matching functions were obtained. An experimental phase followed, in which a second response key was available in one concurrent schedule alternative. For half the birds, the second key was programmed identically with the first; for the other half, the added key was programmed for extinction, with position irrelevant. Comparison of baseline and experimental matching functions revealed no systematic changes in either slope or intercept for birds in the latter group.


Acquisition of stimulus control while introducing new stimuli in fading.

After establishing discrimination between a red positive stimulus and a green negative stimulus, the lowest intensity colors that restricted all responding to the positive stimulus were determined. Then, two new white lines differing in terms of line orientation were each superimposed on one of the colors and were increased in intensity. Thereafter, the intensity of the colors was decreased and eventually eliminated. Probe stimuli consisting of the lines presented against dark backgrounds were presented before each change of stimulus intensity, and probe responding was used to assess the control acquired by various dimension of the new stimuli during the course of fading. The lines acquired control of responding while they were being introduced, and control was strengthened as the colors were attenuated. Such a locus of acquisition was attributed to the starting intensity of the original controlling stimuli and was explained in terms of stimulus blocking. Finally, using probes while introducing the new stimuli enhanced the acquisition of control by the new stimuli.


Positive behavioral contrast in 3-month-old infants on multiple conjugate reinforcement schedules.

Positive behavioral contrast was assessed in two experiments with young infants using multiple conjugate reinforcement schedules. Reinforcement was produced by footkicks which activated the objects of an overhead crib mobile in a manner proportional to the vigor and rate of responding. Distinctive color/pattern cues on the sides of the objects served as discriminative stimuli for components of the multiple schedule. In Experiment 1, infants were trained with one cue (S+) only before insertion of S+ into a multiple schedule with an extinction component. A control group received S+ throughout all sessions. In Experiment 2, a multiple schedule was introduced at the outset, and responses in both components were reinforced before the introduction of extinction in the second component. In a final phase, reinforcement was reintroduced into the second component. Positive behavioral contrast occurred in both experiments. Response reduction in the extinction component was seen only in individual relative response curves. In both experiments, negative emotional behaviors accompanied the extinction component, and in Experiment 1, cooing accompanied presentations of S+.


Child abuse: an approach for early diagnosis.

The importance of early diagnosis in child abuse cases has long been recognized and its importance, as well as the potential sequelae of undiagnosed abuse, is addressed here. Because the family physician is in the unique position of treating a family as a unit, he/she has access to a variety of data that should aid in early identification of child abuse cases. By drawing on studies detailing profiles of the abusing parent, the abused child, their interactions, and the injuries associated with child abuse, a model for such early diagnosis is presented.


Maternal attitudes toward circumcision.

Two hundred questionnaires were completed anonymously by mothers to determine the nature of their attitudes and knowledge of neonatal circumcision. Neonatal circumcision had been performed on 95 percent of the sons. A wide variety of reasons were given for circumcising newborns; few were medically valid. Eighty-seven percent of mothers considered circumcision to be without risk, and 80 percent of mothers stated that the risks of circumcision had not been explained to them. This retrospective study revealed a lack of maternal understanding regarding neonatal circumcision.


Guidelines for family interviewing and brief therapy by the family physician.

Psychosomatic and behavioral problems are commonly seen in the practice of family medicine. If these problems are viewed as difficulties with family interaction, rather than as difficulties of an individual family member, intervention may be more successful. Treatment of families with problems involves interviewing the family unit, identifying and altering dysfunctional behavioral patterns within the family which serve to maintain the problem, and making selected referrals to experienced family therapists.


Predoctoral education in family medicine: a ten-year perspective.

The educational goals chosen by teachers of family medicine for predoctoral students a decade ago differ substantially from the present goals. The original goals were to increase the number of family physicians, provide them with the basic knowledge and skills to practice, integrate the concepts of family medicine into the total medical school curriculum, and develop the "attitudes and ideals" of the good family physician. A series of basic learning principles were evolved to attain these goals. These included: teach in a practice setting, focus on persons rather than disease processes, use a team approach to teaching, involve students in the planning and evaluation process, and recognize curriculum development as a dynamic process. The present curriculum has an increased emphasis on clinical skills in family practice and on integration of behavioral science; there is a new emphasis on the role of the physician in the community and a better understanding of health care systems. Future directions for family medicine include increasing the emphasis on interpersonal communications, clinical synthesis, and clinical assessment.


Graduate education in family practice: a ten-year review.

Assessment of the progress of graduate education in family practice after ten years shows that the original goals established for residency training in this specialty are being effectively met. There are now more than 360 approved family practice residencies in the United States with over 6,000 residents in training. Student interest in these programs has remained at a high level, and attrition has been low. Graduates of these programs have favored partnership and group family practice, and are well distributed in rural, suburban, and metropolitan areas. Heavy emphasis has been placed upon quality control mechanisms for both internal and external review of family practice residency programs. This paper outlines some concerns regarding the present status of family practice residencies, and suggests some directions for future development of these programs.