Lymphocyte associated beta 2-microglobulin studied by crossed radioimmunoelectrophoresis. A sensitive radioimmunoelectrophoretic assay was employed in the study of beta 2-microglobulin (beta2m) from human blood lymphocytes, allowing the detection of 2.5 pmol of beta2m corresponding to 3 x 10(6) lymphocytes. The intrinsic (amphiphilic) membrane proteins were solubilized and examined in crossed immunoelectrophoresis against rabbit antisera in the presence of non-ionic detergent. The beta2m associated precipitate was traced by post-electrophoretic incubation with 125I-labelled antibodies to beta2m and autoradiography. A polyspecific rabbit antiserum to human lymphoid cells retained its capacity to precipitate lymphocyte beta2m after absorption with isolated beta2m (on an immunosorbent column), showing that lymphocyte beta2m is complexed to other molecules. No free beta2m was found on lymphocytes when examined against the absorbed antiserum to human lymphoid cells in an intermediate gel and anti-beta2m in a reference gel.


[Disseminated lupus erythematosus: an analysis of organ involvement]. The spectrum of involvement of organs in 70 patients with systemic lupus erythematosus has been assessed in a prospective study. All patients were admitted to hospital electively for 2 days and a complete clinical and laboratory assessment protocol completed. The overall mean observation period was 29 months. Widespread multisystem involvement was found in every patient. Subclinical abnormalities of respiratory and cerebral function were common. A more conservative approach than has been generally recommended was used for the management of systemic lupus erythematosus and is justified by 5-year survival of 98%.


[alpha 1-fetoprotein in neonatal jaundice]. The relationshop between the concentration of alpha1-fetoprotein (AFP) in cord blood and neonatal jaundice has been examined in 259 neonates. Cord blood AFP varies inversely with gestational age, as does the incidence of severe jaundice. Among babies born at term a higher mean AFP concentration in cord blood was found in the group which subsequently developed marked hyperbilirubinaemia. On average, the full-term babies with a cord blood AFP level above 130 mg/l developed more pronounced jaundice than the rest. We conclude that cord blood AFP concentrations reflect the maturity of the liver in both premature and full-term infants. It is a better criterion than the estimation of gestational period by physical examination of the baby.


Antenatal diagnosis in practice.

Antenatal diagnosis in practice. Over a period of 5 years, 434 women at risk of having abnormal babies have had antenatal daignostic tests carried out during the first half of their pregnancy by the laboratories of the Department of Human Genetics, University of Cape Town. From these investigations, it was predicted that 13 fetuses had chromosomal abnormalities, 6 had severe central nervous system defects and 4 had autosomal recessive metabolic disorders. In addition, 4 cases with X-linked recessive traits were monitored and 3 male fetuses were recognized. Affected pregnancies were terminated except for 1 with a fetal sex-linked disorder where the parents revoked their original decision. The diagnosis was confirmed by fetal autopsies in all cases except 4 (2 spontaneous abortions and 2 out-of-town terminations). There was only 1 case where culture failed and the pregnancy went to term with the birth of a baby with Down syndrome. Antenatal diagnosis is now an important part of normal clinical practice. The fact that the fetal abnormalities were recognized in 6% of pregnancies is justification for the use of this procedure.


Effects of IRA in vitro on T- and B-lymphocytes from peripheral blood of patients with connective tissue diseases. The effect of immunoregulatory alpha-globulin (IRA) in vitro on T-lymphocytes (rosette forming cells) and B-lymphocytes (surface IgG and IgM) in peripheral blood of healthy subjects and the patients with connective tissue diseases were investigated. Marked inhibitory effects of IRA were observed on T-lymphocytes and surface IgG-bearing lymphocytes from healthy subjects. The effect of IRA was almost in parallel with the amount of IRA and with the length of incubation period at 37 degrees C. The inhibitory effects of IRA were more remarkable on surface IgM-bearing lymphocytes from the patients with rheumatoid arthritis (RA) and those with systemic lupus erythematosus (SLE) than on those from healthy subjects. On the other hand, the effects of IRA were more remarkable on surface IgG-bearing lymphocytes from the cases of SLE than on those from healthy subjects.


Prostaglandin inhibition of testosterone production induced by luteinizing hormone, dibutyryl cyclic AMP or 3-isobutyl-1-methyl xanthine in dispersed rat testicular interstitial cells. Testicular interstitial cells were utilized to study the effects of prostaglandins (PG) on in vitro testosterone production and to examine the role of cyclic adenosine-3',5'-monophosphate (cAMP) in this process. Testosterone production was assessed after 3 hour incubations while cAMP accumulation was examined after a 0.5 hour incubation period. Testosterone and cAMP were measured by radioimmunoassay. None of the PGs tested (PGA, PGA2, PGB1, PGE1, PGE2, PGF1alpha PGF2alpha) altered basal testosterone production when present in incubates at concentrations of 1.3 X 10(-8) M to 1.3 X 10(-4). However, at concentrations of 1.3 X 10(-4) M all of these PGs were capable of decreasing Luteinizing Hormone (LH; 100ng)-induced testosterone production. The inhibition of LH-induced testosterone production by the B, E and F series PGs was less pronounced than that for the A series. PGA1 and PGA2 exhibited 80% and 95% inhibition, respectively, at 1.3 X 10(4) M. The inhibitory action of 4 X 10(5) M PGA1 or PGA2 was evident within 30 minutes. Preincubation of interstitial cells with indomethacin (10(-5) or 10(-6) M) for 30 minutes did not alter subsequent basal or LH (100ng)-induced testosterone production. Accumulation of cAMP was stimulated by LH (10 microgram) or by PGs (1.3 X 10(-4) M PGA1, PGA2, PGB1, PGE1 or PGF2alpha). The PG-induced cAMP accumulation thus occurred at concentrations where LH-stimulated testosterone production was inhibited. Furthermore, PGA1 and PGA2 (1.3 X 10(-4) M) inhibited testosterone production induced by either 3-isobutyl-1-methyl xanthine (MIX; 10(-4) M or 10(-3) M) or dibutyryl cAMP (dbcAMP; 10(-4) M or 10(-3) M). These results indicate that PGs can block testosterone production by a direct effect on testicular interstitial cells and suggest that PGs exert their inhibitory action distal to stimulation of cAMP formation. PGs do not appear to play a role in the mechanism of LH action.


Nonassociation of hyperamylasemia and prostatic disease. The possible correlation of hyperamylasemia and prostatic disease was investigated in patients with benign prostatic hyperplasia, adenocarcinoma of the prostate, and in patients without clinical evidence of prostatic lesion. Our results indicate that prostatic lesions whether benign or malignant are not associated with either elevation of serum amylase or abnormal isozyme zymograms.


[Surgical treatment of cancer of the extrahepatic bile ducts]. The authors present the 12 years' experience with the treatment of hepatocholedochal cancer (cancer of the supraduodenal end of the common bile duct and of the common hepatic duct included). 22 patients were operated upon. The authors managed to perform radical operations upon 5 out of 22 patients, when there were small tumors of the hepato-choledochus. The technic of operation and results of the treatment are described.


[Complications of a biliodigestive anastomosis in tumors of the organs of the pancreatoduodenal area]. To prevent from an ascendant infection of biliary tract the authors apply a maximally large (8--9 cm) cholecystojejunoanastomosis through the excision of a part of the gallbladder wall. Such an anastomosis eliminates the ballbladder cavity, and its function approximates that of an excretory canal. The patients operated upon by means of this surgical method developed cholangitis 4 to 5 times less than those who underwent cholecystojejunostomy through the apparatus PKS-25.


[Treatment of dolichosigmoid and dolichocolon]. The authors observed 591 patients with stretching of various portions of the colon. The patients were treated conservatively and surgically. 191 patients were operated upon: 21 palliative operations and 170 resections of the colon were carried out. The resections were performed by intraabdominal and abdomino-perineal methods. In authors' opinion the intraabdominal resection of the colon is the best method of the treatment of the disease.


[Inhibition of opsonization of E. coli by enzyme-treated gammaglobulin (author's transl)]. The influence of Cohn fraction II, pepsin- and plasmin-treated gammaglobulin on the opsonization of E. coli 0111 was investigated. Normal serum at different dilutions, as well as serum of patients with different forms of antibody deficiency syndromes were tested. No effect was observed by pre-incubation of the bacteria in these gammaglobulins if concentrated or 1:10 diluted serum was used for opsonization. In diluted serum Cohn fraction II potentiated and pepsin-treated gammaglobulin competitively inhibited opsonization, whilst plasmin-treated gammaglobulin potentiated at the low and competitively inhibited at higher dilutions. In serum from patients with antibody-deficiency syndromes, potentiation of opsonization by Cohn fraction II and competitive inhibition by pepsin- and plasmin-treated gammaglobulins was already seen at low serum dilutions.


[Ferrokinetic studies in haemoglobin Wien haemolytic anaemia (author's transl)]. Ferrokinetic studies in two patients with haemolytic anaemia due to haemoglobin Wien revealed increased haemoglobin synthesis and an increase in effective erythropoiesis. Haemolysis is due to an instability of the haemoglobin itself. The spleen does not appear to play any appreciable role in the aetiology of this condition. There was no extramedullary erythropoiesis in our patients.


[Dermatologic indications for anti-androgenic treatment]. In spite of remarkable therapeutic results obtained by gestagens with antiandrogenic activity, usually combined with estrogen, in oily seborrhea, acne, Fox-Fordyce disease, androgenetic alopecia and hirsutism many dermatologist still hesitate to treat the named disorders by hormones. The reason for their hesitation appears to be the erroneous belief, that the named disturbances represent hormonal disorders the treatment of which does not belong to dermatology. After a survey on the mechanism of action of antiandrogens the basic difference between androgen dependent skin disorders and endocrinopathies with manifestation on the skin and its appendages is explained. Androgen dependent skin disorders, like oily seborrhea and most cases of acne are not the result of endocrine disturbances in the sense of an pathologically increased or decreased production of sexual hormons. Administering sexual hormons the physician takes advantage of the sebosuppressive effect of female sexual hormons as he does of the antiallergic activity of the hormon cortisol (and related compounds) in the treatment of eczemas. The antiandrogenic treatment of androgenetic alopecia, hirsutism and androgenetic acne--with their underlying hormonal disturbance, consisting in an increased production of androgens, represents an quasi etiological therapy. As in these cases the hormonal disturbances finds its expression mainly or exclusively in disorders of the skin or hair growth, the dermatologist, preferentially in cooperation with endocrinogists and/or gynacologists remains entitled to take over the treatment. The available drugs are discussed and suggestions are made for their appropriate use.


[Cardiac arrhythmias and other complications in patients after myocardial infarction during a long term training program]. In 85 patients with myocardial infarction undergoing a physical training on a bicycle ergometer 3 times a week over a period of 18 months beginning 5 weeks after the acute infarction, telemetric and haemodynamic investigations were made. The telemetry was done during ergometer training, dynamic and isometric gymnastics and during work at home. The diastolic pulmonary arterial pressure under exercise was registered in 43 patients before the beginning of the training program; in 16 of these patients a control heart catheterization was made after a training period of 4 months. During the ergometer training program 32% of all patients had ventricular premature beats more than 1:10, 6% had paroxysmal ventricular tachycardias, 1 patient had supraventricular tachycardias and 1 patient died at home after having a tachycardia of unknown origin. During dynamic or isometric gymnastics or under work at home the patients had not more arrhythmias than in the telemetric controls during the ergometer training. Arrhythmias and other complications are very often found in patients with high diastolic pulmonary arterial pressure in the exercise test.


[Value and problems of alpha-fetoprotein determinations in testicular neoplasms]. In order to improve our diagnostic possibilities and the decisive therapeutic consequences resulting from this we began also to include the determination of the alpha-foetoprotein into our diagnostic spectre in tumours of the testicles. Our recent results and the problems appearing are described.


[Therapy of testicular neoplasms].

[Therapy of testicular neoplasms]. It can be established that it is nowadays possible by means of an interdisciplinary therapy to achieve a full remission of more than 2 years in 80% of the malignant teratomas of stages I and II and in up to 40% of the teratomas of stage III, which possibly means a cure of the patient.


[Rapid staining of cytologic smears with methylene blue]. It is described a handy staining of cytological smears suitable for out-patients department use. On a dry not fixed smear we put a drop of 0,25% of aqueous solution blue de methylene and we cover it with a cover slide. The smear is stained in a few seconds and the stain remains for half an hour.


[Effectiveness of surgical measures in progressive ovarian cancer]. A report is given about the value of the active large removing of tumour-tissue in cases of advanced ovarian cancers (89 patients suffering an ovarian in the stage T3/T4M1 in the time from 1970 to 1975). The survival chances of the 49 patients in which it was possible to remove greatest parts of the tumours, was nearly 20 per cent better than in 38 patients without such therapy.


Basic isotope thyroid parameters in Zambia. The paper analyses basic isotope thyroid parameters in 20 normal Zambians. The results of tests (excluding uptake studies) do not differ from those established elsewhere but the range of thyroid radio-iodine uptakes is wide. It may be explained by the mixed nature of the sample of patients with regard to iodine intake.


The constituents of keratohyalin granules of newborn rat epidermis under an electron microscopy. Three-day-old rat skins were treated either by the freezing--thawing method or by the tetraphenylboron technique. After the former procedure, keratohyalin granules were solubilized, leaving empty spaces with an electron-dense or less dense rim. Particles roughly 200 A in diameter, which were either electron-dense or less dense, were observed in the empty spaces and also seen detaching from the rim. After the freeze--thaw procedure, two types of keratohyalin granule were observed. The one is the same electron-dense granule as keratohyalin granules in vivo and the other is the granule, the electron density of which was identical with tonofilaments. Both granules were granular rather than fibrous in structure.


Chronic bullous dermatosis of childhood. Immunological and ultrastructural studies on the melanocyte--dermal junction. Ultrastructural study of a case of benign chronic bullous dermatosis of childhood revealed the presence of melanocytes in the base of the blister. The melanocyte anchorage mechanism is described. The absence of labelling by bullous pemphigoid antibodies on the base of the blister, where melanocytes persist, indicates that the antigens recognized by these autoantibodies do not exist in the melanocyte basal membrane-contact zones. This suggests that the chemical nature of the melanocyte dermal junction anchorage system is different from that of the hemidesmosomes.


Cellular aspects of phototoxic reactions induced by kynurenic acid. II. Fine structural analysis and cytochemistry on cultivated cells. A recent report described the establishment of an experimental model for studying photo-oxidative damage inflicted by kynurenic acid on cultivated cells. Preliminary findings by scanning electron microscopy (SEM) were reported. For the present paper supplementary SEM studies were performed, and it was found that maximum cell damage appears 2 to 4 hours post irradiation. With low concentrations of kynurenic acid and brief UVA exposure, repair mechanisms seem to occur. Damage in the form of cellular swelling and plasma membrane alterations is most easily recognized by SEM, which is far more sensitive than ordinary light microscopy. Transmission electron microscopy (TEM) reveals morphologic alterations, consisting of an enhanced formation of endocytotic vacuoles from the plasma membranes, swelling with disruptions of the cell sap continuity, mitochondrial swelling, dilatation of the endoplasmic net, and vesiculation of some of the microvilli. Lysosomal damage, which was not seen until very late post irradiation and only for high concentrations of kynurenic acid and long-term UVA exposure, seems to be a secondary phenomenon. This hypothesis is further supported by the cytochemical analysis. The primary event thus seems to affect the plasma membrane level and originate in the binding of kynurenic acid to the plasma membrane on which subsequent UVA irradiation inflicts secondary intracellular alterations due to increased permeability and swelling.


Inhibition of ultraviolet and phototoxic dermatitis in the mouse. The effect of inhibitors on the inflammatory oedema elicited by medium-wave ultraviolet radiation (UVB) and long-wave ultraviolet radiation (UVA) in combination with chlorpromazine has been studied in the mouse, by means of a quantitative technique. Inhibition of the UVB reaction was observed with indomethacin, acetylsalicylic acid and betamethasone valerate, whereas the latter compound only was effective in the phototoxic state. No inhibition was obtained with hydrocortisone, phenylbutazone, epsilon-aminocaproic acid, polyphloretin phosphate, clemastin, alpha-tocopherol or ascorbic acid. With indomethacin and betamethasone valerate there was no inhibition at high doses when the compound was administered before UVB irradiation. These results are in accordance with a proposed central role for the prostaglandins in UVB inflammation. It is suggested that the phototoxic reaction to chlorpromazine may not be due to mediator action but rather to the effect of toxic photoproducts.


Increased 133xenon washout from cutaneous tissue in generalized scleroderma indicates increased blood flow. Blood flow in cutaneous tissue of the dorsum of the hand was measured by the local 133Xenon washout technique in 18 normal persons and in 26 patients suffering from generalized scleroderma of the acrosclerosis types. Flow values calculated from the accepted blood--tissue solubility coefficient (lambda) were 8.6 +/- 0.4 S.E.M. ml/100 g X min in normals and 14.5 +/- 1.1 ml/100 g X min in sclerodermic skin (i.e. increased by 69%). The difference was significant (p less than 0.001). Blood flow values in sclerodermic skin were directly proportional to the degree of dermal sclerosis. As lambda in sclerodermic skin is probably decreased due to a decreased fat content, it may not be correct to interpret these results as evidence of increased blood flow. A "worst case" was constructed in calculation of lambda using zero fat content for sclerodermic skin. Flow value was now 12.3 +/- 1.1 S.E.M. ml/100 g X min for sclerodermic skin, an increase of 43% compared with normal skin. It is probably only justified to use this "worst case lambda" in the group of severely involved cases of scleroderma. In these patients it yielded a flow value of 17.8 +/- 3.1 (N = 6) which, when compared with the flow value for normal skin, indicates an increase by 100% of the blood flow in severely involved sclerodermic skin.


An exogenous variety of pseudoxanthoma elasticum in old farmers. Cutaneous lesions clinically similar to and histopathologically indistinguishable from pseudoxanthoma elasticum have been observed in the cubital folds of nine elderly farmers. In all cases the skin lesions were ascribed to exposure to salpeter during fertilization on a single occasion decades previously. Signs of systemic pseudoxanthoma elasticum were absent.


Sensitization capacity of epoxy resin hardeners in the guinea pig. Cold-curing resin hardeners were examined regarding their sensitization capacity, by the "guinea pig maximization test". All the aliphatic polyamines caused sensitization. Two of the three cycloaliphatic polyamines sensitized all the animals, while one did not sensitize at all. The aromatic amine sensitized one-fifth of the animals studied. The two polyaminoamides sensitized respectively one-fifth and two-thirds of the animals. Two of the four adducts failed to provoke any reactions. The phenolaccelerated adduct of triethylenetetramine sensitized nearly half of the animals. The adduct of isophorondiamine and epoxy resin sensitized three-fourths of the animals. Judging from the above observations, adducts, when void of aliphatic amines, are probably not potential sensitizers. This may perhaps also be true of the polyaminoamides.


Salpeter-induced dermal changes electron-microscopically indistinguishable from pseudoxanthoma elasticum. Salpeter-induced calcium deposits in ten patients showed a close clinical and histopathological similarity to the lesions of pseudoxanthoma elasticum. By electron microscopy and selected area diffraction analyses of the calcium deposits we find the changes indistinguishable from the changes previously described in involved skin of patients suffering from PXE. Clinically our patients were compared with the fourteen patients with traumatic calcification previously described.


Sensitization capacity of epoxy reactive diluents in the guinea pig. The sensitization capacity of six epoxy reactive diluents was investigated with the "guinea pig maximization test". All the low molecular weight reactive diluents (MW 175 to 360) proved to be sensitizers. One diluent of higher molecular weight (MW 1700) produced no reaction.


Non-thrombocytopenic purpuras.

Non-thrombocytopenic purpuras. 88 cases of microscopical diapedesis of the red blood cells in the superficial blood vessels of the skin were investigated clinically, histologically and immunohistologically. Polymorphonuclear vasculitis (PMNV) was found in 27 cases, 18 of which also demonstrated immunoglobulins (mostly IgM) in the vessel walls. Complement (C3) was found in 16 cases. The cases with PMNV formed a uniform group as regards preceding infections, drug consumption, clinical features, and pathological urinalysis and laboratory data. The second group consisted of 21 cases of lymphocytic perivasculitis with immunoglobulins and/or complement (LP + ig/C3). It clearly differed clinically from the former group in three respects: urinalysis was normal, a slowing of the venous outflow was found in more than half of the cases and the duration of the disease was longer. The biggest group consisted of 40 cases of lymphocytic perivasculitis without detectable immunoglobulins in the vessel walls (LP). In this group the duration of the disease was shortest. The capillary resistance was lower than in the other groups. The results of this investigation indicate that there are at least three pathogenetically differing groups among the clinical purpuric conditions.


Development of gastric dysfunction in dermatitis herpetiformis. In order to study the development of atrophic gastritis, gastric secretory function was examined by a standard pentagastrin test 24 to 78 months after a previous examination (Ex I). The study included 12 patients with dermatitis herpetiformis (DH), 6 patients with functional signs of atrophic gastritis previously, and 8 healthy controls. The surrent examination (Ex II) also included microbiological culture of gastric juice and estimation of gastrin(s), parietal cell and thyroidal antibodies. Most of the controls had increased their maximum acidity and maximum acid output (MAO) between the examinations. This may indicate an altered potency of pentagastrin in recent years. Conversely, 5 of the 6 patients with atrophic gastritis showed a further reduction of maximum acidity and MAO, indicating progressive parietal cell atrophy. In the DH-group, two tendencies were observed: 6/12 patients had an increased MAO at Ex II. They had had lower mean age and higher mean MAO at Ex I, as compared with the remaining 6 patients who had a decreased MAO at Ex II. The latter group more often had parietal cell antibodies.


Food allergy in atopic dermatitis: experimental observations. Results of an investigation of 134 subjects with atopic dermatitis and 29 controls are described. The investigation involved: 1) an allergological enquiry into each patient's medical history, with particular reference to consumption of eggs, milk and fish; 2) a total IgE assay; 3) RAST with egg, milk, cod; 4) a challenge test. Recording of medical histories revealed the existence of clinical sensitivity to egg, milk and cod in respectively 21, 17 and 7 of the 134 subjects with atopic dermatitis, but in none of the controls. In the atopic subjects, RAST demonstrated egg, milk and cod antibodies in 37, 28 and 14 cases respectively, while of the controls only 2 had antibodies--and then in minimal amounts. The maximum incidence of egg and milk antibodies was demonstrated in the initial 3 years of life. Clinical sensitization to cod manifests itself in the form of attacks (urticaria, angi-oedema, vomiting) showing significant correlation between clinical history, results of RAST and the response to challenge, while in the case of sensitivity to eggs and, even less evidently, to milk, the symptoms are more often referred to as "exacerbation of the dermatosis" and the results of RAST, of challenging and the clinical history tend to be contradictory.


Genital occurrence of oral microbiota.

Genital occurrence of oral microbiota. Recent studies indicate that tonsillar gonococcal infection or colonization is fairly common. Carriage rates of about 8% have been found. These studies also indicate that oro-genital contacts are common. Since very little is known about the amount of oral microbiota transmitted to the genitals, we have studied the occurrence of oral streptococci and Neisseria species in urethra and cervix. Among 128 patients attending an STD-clinic we found 10 carriers of oral streptococci, one Streptococcus mitior, four Streptococcus sanguis, one Streptococcus mutans and four Streptococcus salivarius and case of urethritis due to Neisseria menigitidis. Seventy-three of the patients had recently had their genitals exposed to the oral flora of their partners. Despite the heavy contamination with oral microbiota that can be assumed to occur in these cases, there seems to be no colonization of the genitals with oral microbiota.


Trichochromes in red human hair.

Trichochromes in red human hair. The presence of trichochromes B and C in red human hair was confirmed with an analytical procedure which does not give rise to the formation of these compounds as artifacts. The trichochrome precursor 5-S-cysteinyldopa previously demonstrated in red guinea-pig hair was not detected in red human hair.


Subhydroxylated collagen in scleroderma. In sclerodermal skin, the values for proline (Pro), hydroxyproline (Hyp) and hydroxylsine (Hyl) are lower than in normal skin. The molar ratio Hyp to Hyl is lowered. The molar ratio Pro to Hyp was found to be elevated, while that of Pro to Hyl was like that of normal skin. It was concluded that in slceroderma the hydroxylation of Pro to Hyp is incomplete, resulting in an abnormal collagen.


Autoradiographic investigation on benzoyl peroxide treated skin. In vitro autoradiography was used to study the effect of a 5% benzoyl peroxide preparation, or its gel base alone. After three weeks' local application, there was no change in the labelling index of epidermis or follicular infundibulum, while there was a significant reduction in the labelling index of the sebaceous glands. Treatment with the base alone brought about no change.


An ultrastructural study of the epidermis in hyperkeratosis lenticularis perstans. A 60-year-old Japanese man contracted hyperkeratosis lenticularis perstans (Flegel). Biopsy specimens of the keratotic papules on the dorsa of the feet were studied with electron microscopy. An increase in the number of membrane coating granules, keratohyalin granules and tonofilaments was the most prominent feature of the keratinocytes underlying the keratotic papule. However, in the area just above the cone-shaped dermal infiltration, membrane coating granules were absent and keratohyalin granules and tonofilaments were poorly developed.


Bone cysts and spontaneous fractures in two siblings with dyskeratosis follicularis Darier. Two siblings with Darier's disease, multiple bone cysts and recurrent "spontaneous" fractures are described.


Eosinophilic spongiosis in early pemphigus foliaceus. A case of widespread non-specific, itchy exanthema in a 74-year-old female is presented. An early skin biopsy showed eosinophilic spongiosis. Later a dermatitis herpetiformis-like eruption developed. However, immunofluorescence microscopy showed intercellular IgG deposits characteristic of pemphigus. Recognition of eosinophilis spongiosis may be helpful in differentiating early pemphigus from other bullous diseases.


Delayed cold urticaria.

Delayed cold urticaria. Five patients with delayed cold urticaria are described. The urticarial skin response was present between 24 and 72 hours after ice challenge. In two of the patients the cold sensitivity was of clinical relevance. Some of the patients displayed low alpha1-antitrypsin and increased C4 levels in their serum. Our findings may justify the introduction of cold provocation as a routine procedure in the investigation of a patient with chronic urticaria.


Levamisole-induced hypersensitivity.

Levamisole-induced hypersensitivity. A 73-year-old woman with rheumatoid arthritis was treated with Levamisole, 150 mg per day, on 2 days a week. Her arthritis improved, but she developed a severely itching rash, and the treatment was stopped after 6 months. Penicillamine was subsequently given and tolerated without skin complications. 15 months after regular Levamisole was stopped, she was given a single dose of 150 mg which provoked fever of 40 degrees C and rash. Thirteen punch-biopsy specimens were examined by direct immunofluorescence microscopy. During the Levamisole treatment, granular deposits of IgG and C3 were found at the dermal--epidermal junction. Subsequently, the deposits disappeared, but reappeared after Levamisole challenge. The patient's leukocytes were exposed in vitro to Levamisole, and 36% of the total histamine content in the basophils was released. Our results provide further evidence that Levamisole can cause type-I as well as type-III hypersensitivity.


Acne-like eruptions induced by PUVA-treatment. Photochemotherapy with psoralen and long-wave ultraviolet light, so-called PUVA-treatment, is currently being evaluated in many dermatologic departments. Side effects such as nausea, pruritus and erythema are well known. Recently the development of acneiform eruptions was reported in a British patient treated with PUVA (3). We found that 4 out of 80 patients treated in our clinic with 8-methoxy-psoralen according to the usual weight schedule (6) and long-wave ultraviolet irradiation developed perioral dermatitis, in 2 cases, together with acneiform eruptions localised to the forehead.


Intermittent treatment of psoriasis with clobetasol propionate. Clobetasol propionate cream, applied intermittently for 2 weeks, induced a rapid clinical and histological clearing of psoriatic infiltrates. The serum cortisol level was transiently lowered. By maintenance therapy, with the corticosteroid given only one day a week, 8 of 12 patients were kept in remission for an average period of 5 months.


Trichochromes in human malignant melanoma. Decarboxytrichochromes B and C were isolated from a melanoma metastasis in a red-haired man, indicating the presence of trichochromes B and C in the tissue. Trichochromes E and F were not detected.


DNA interstrand crosslinks in normal human skin visualized by electron microscopy. Normal human skin was investigated for DNA interstrand crosslinks by a denaturation electron microscopic method. Skin biopsy materials from the thighs of 8 healthy volunteers were separated into epidermis and dermis. DNA from each layer was isolated, purified and studied in the electron microscope. The presence of DNA interstrand crosslinks in normal human skin was demonstrated. The number of crosslinked DNA molecules and the density of crosslinking within the molecules were determined. Crosslinks were found in 0.8% of the scored molecules and were located almost exclusively in epidermal DNA. The intramolecular density of crosslinking showed a wide range of variation in each volunteer and between the volunteers.


Ultrastructural changes in nuclei and nucleoli in psoriatic epidermis during and after PUVA therapy. Quantitative and qualitative changes of the nuclei and nucleoli in psoriatic epidermis at different intervals during therapy with psoralen and long-wave ultraviolet light (PUVA) were revealed with the electron microscope. Early in the therapy (24 to 96 hr after commencing treatment) changes occur, including irregularity of the nuclear envelope, enlargement of the nuclei and nucleoli, temporary disappearance of fibrillar centres, followed by temporary disappearance of fibrillar centres, followed by temporary increase in nuclear bodies and increasing amounts of heterochromatin. After clearing of lesions, the nuclear and nucleolar structures were identical with those of normal keratinocytes, except that the relative amount of heterochromatin was greater. It is suggested that PUVA therapy brings about inhibition of abnormal keratinization, possibly due to gene repression, and that this may be related to changes in the fibrillar centres of psoriatic keratinocyte nucleoli.


The fluroescent profiles of keratohyalin granules of newborn rat epidermis with a new fluorescent thiol reagent (DACM). A highly specific fluorescent thiol reagent, N-(7-Dimethylamino-4-methyl-coumarinyl)maleimide (DACM) has been used to examine the newborn rat skin in order to elucidate the cystine-rich keratohyalin granules or dense homogeneous deposits. After reduction of the disulphide bond with 2-mercaptoethanol, the periphery of the stratum corneum cells showed a very distinct fluorescence and the cytoplasm of the stratum corneum, granulosum and spinulosum showed moderate fluorescence. No fluorescence was observed, however, over the keratohyalin granules themselves. Intense, spotty fluorescence was observed from the remnants of nuclei and at the periphery of large keratohyalin granules of the stratum granulosum cells, which were assumed to be dense homogeneous deposts according to their distribution pattern. As the intense fluorescence elicited with DACM determines the abundance of cystine content in the material, cystine was found to be very scarce in keratohyalin granules, but abundant in dense homogeneous deposits and at the periphery of the stratum corneum cells.


Plasma-kinin-forming enzyme in human skin: extraction and column chromatographic separation of plasma-kinin-forming enzyme and its inhibitor. The optimal salt concentration for extraction of plasma-kinin-forming enzyme from the human skin was investigated. It was confirmed that a salt concentration higher than 1.5 M in the extraction buffer was required for optimal extraction of this enzyme. An inhibitor of this enzyme was detected in the extract with buffer alone, and the coexistence of this enzyme with the inhibitor in buffer extract was also confirmed by using G-200 gel chromatography.


A leukotactic factor in the stratum corneum of pustulosis palmaris et plantaris: a possible mechanism for the formation of intra-epidermal sterile pustules. Previous studies on the chemotactic properties of soluble substances from psoriatic scales demonstrated the presence of complement-derived chemotactic factors (anaphylatoxins) that seem to play a crucial role in the production of typical psoriasiform tissue changes. The crude extract from the stratum corneum of lesions of pustulosis palmaris et plantaris (PPP) was also noted to contain highly chemotactic components for polymorphonuclear leukocytes (PMN's) as compared with a crude callus extract in the present study. The leukotactic factors contained in the major chemotactic fractions isolated by Sephadex G-75 chromatography revealed in vitro characteristics and in vivo biological activities very similar to those of the leukotactic factor derived from psoriasis scale reported in earlier work, i.e. complement cleavage products (C3a and C5a). On the basis of these findings we have reason to believe that pustule formation in PPP occurs through a mechanism that has a strong resemblance to that in pustular psoriasis. We hypothesize further that similar mechanism may be operative in other dermatoses in which the formation of sterile subcorneal pustules is a characteristic feature.


Absorption of 8-hydroxyquinolines through the human skin. The skin absorption of clioquinol and chloroquinaldol in ointment or cream base was studied in fourteen patients with widespread dermatoses. A serum concentration in the range of 0.3--1.3 microgram/ml using clioquinol and 0.05--0.1 microgram/ml with chloroquinaldol was reached by the second day and persisted throughout the topical treatment. The mean serum half-life of clioquinol and of chloroquinaldol was estimated to 25 and 6 hours, respectively. Local application of zinc oxide ointment after the end of the treatment chelate-bound the skin deposits of clioquinol and lowered the serum half-life to 11 hours. The mean daily urinary excretion of the applied amount of clioquinol and chloroquinaldol was 4.5 and 6%, respectively.


Local absorption of zinc from wounds treated with different concentrations of zinc sulphate. In the present study it was shown in rats that zinc is absorbed from excisional wounds treated with zinc sulphate. Systemic toxic effects were observed in the group treated with 20% zinc sulphate. Local toxic effects were seen in wounds treated with 0.2%, 2% and 20% zinc sulphate. An inhibitory effect of zinc on the migration of granulocytes was suggested on the basis of microscopic observation. In the operated groups which were not treated with zinc and the group treated with 0.02% zinc sulphate a decline was observed in the concentration of zinc in serum. The zinc concentration in serum increased in proportion to the zinc sulphate concentration (0.2%, 2% and 20%) applied to the wounds, while the copper concentration decreased in the groups treated with 2% and 20% zinc sulphate. In all operated groups an increase in zinc and copper concentrations was observed in liver. This was most pronounced in groups treated with higher concentrations of zinc sulphate (0.2%, 2% and 20%). The groups treated with higher concentrations of zinc sulphate also had higher pancreas zinc concentrations than the remaining groups.


Topical nitrogen mustard induced carcinogenesis. Mycosis fungoides is a rare malignant lymphoma that primarily affects the skin. Lymph node and visceral involvement may follow. The disease is chronic in course, displaying numerous clinical variations. Two cases of mycosis fungoides complicated by the development of cutaneous squamous cell carcinoma will be discussed. One of the two patients developed metastatic squamous cell carcinoma from an initial primary cutaneous lesion. Both patients were treated for long periods with topical nitrogen mustard in addition to systemic therapy. The question of topical nitrogen mustard induction of squamous cell carcinoma is raised.


Hunan wart-virus antibodies in patients with genital and skin warts. A comparative follow-up study of the antibody response against human wart-virus was performed, using the immunodiffusion and complement fixation methods on patients with condylomas and skin warts. By the immunodiffusion method, 13% of the patients with skin warts and 3% of the patients with condylomas showed greater than or equal to fourfold increase of antibody titre during the follow-up of 2--35 months. The findings show some typical features of the weak antibody response of a chronic virus infection and suggest a serological overlapping between condyloma viruses and certain group of skin wart-viruses. The antibody prevalence in age-matched controls is shown to be significantly higher than that in the wart or condyloma patients' initial serum samples. This is indicative of the protective function of antibodies against warts and condylomas. Also the analysis of the history of warts in patients with condylomas suggests that protection against condylomas can be acquired from previous warts, evidently by immunological mechanisms. In a control group of medical students, human wart-virus antibodies were frequently (52%) found in subjects without any history of warts. This finding supports the view that human wart-virus can frequently induce latent or subclinical infections in human beings.


Micropapular sarcoidal facial eruption in a child: Gianotti-type perioral dermatitis. The case is described of a boy aged 13 years reported with a diffuse facial micropapular eruption which had a granulomatous histology. The diagnosis is consistent with perioral dermatitis of the Gianotti type (4, 5). It is postulated that bubble gum was the cause of the condition, the most likely ingredient being the essential oils in the gum, which may actually be selectively absorbed by the follicle to produce a perifollicular granulomatous reaction. In this article we have added yet another possible cause for perioral dermatitis.


Acne treatment with oral zinc and vitamin A: effects on the serum levels of zinc and retinol binding protein (RBP). The serum levels of zinc, vitamin A and retinol binding protein (RBP) were studied in 75 acne patients before and during oral treatment with zinc, vitamin A or placebo. In the zinc-treated patients an increase in the mean serum zinc level was seen after 2 weeks, when also the first clinical improvement occurred. After 4 weeks the zinc level had increased by about 30% and no further significant increase was observed during 3 months of treatment. In 33 healthy subjects there was an increase of 14% after 4 weeks of zinc therapy. Vitamin A and placebo induced no significant changes in the serum zinc status. Prior to therapy the serum levels of vitamin A and RBP were lower in the acne patients than in the controls. Zinc + vitamin A treatment raised the serum RBP value to normal after 4 weeks. In patients given vitamin A alone, a probable increase in RBP was achieved. Zinc and placebo treatment did not change the serum level of RBP.


Oral zinc in acne vulgaris: a clinical and methodological study. In a double-blind investigation of 54 patients suffering from acne vulgaris, the effect of 0.6 g of oral zinc sulphate daily versus placebo was studied. During the active treatment period of 6 weeks, the acne improved by about one-third, as rated with a score system. Clinical comparison with placebo showed the result of zinc sulphate therapy to be slightly, but statistically significantly better. Various methods for the clinical evaluation of acne were studied. Counting of acne lesions was found to be a more precise method than assessment according to the opinion of the patients and the assessment of colour photographs. The correlation between the results obtained by lesion counting and evaluation by photographs was low, as was also the reproducibility of photographic assessment.


Treatment of acne with cyproterone acetate and ethinyl estradiol. Tablets containing 2 mg cyproterone acetate and 0.05 mg ethinyl estradiol in a calendar package of 21 days were used as an oral contraceptive to treat acne. The series comprised 20 patients. The women were 18--43 years of age, and all had acne which had previously been resistant to therapy. The treatment was continued for 6 months. Serum testosterone, 17-OHCS, 17-KS, serum ALAT and gamma-GT were recorded prior to the treatment at 3 and 6 months. Ten patients responded well to the treatment, 5 responded moderately well, 3 experienced no change, and 2 became worse. The serum testosterone level fell during the therapy and the ALAT level rose, though only one pathological ALAT value was recorded.


Systemic miconazole treatment of a patient with chronic granulomatous mucocutaneous candidiasis. Miconazole given intravenously and orally was evaluated in the treatment of a patient with drug-resistant, chronic, widespread horny candida lesions. Marked clinical improvement occurred after 4 weeks of the first course of treatment, in which 400 mg of miconazole per day was given intravenously. After 3 months no signs of the disease were apparent. The patient remained free of symptoms for 3 months, after which the lesions recurred and a second course of treatment, with 200 mg of miconazole per day intravenously, was started. A prompt clinical clearing was seen within 3 weeks, after which the miconazole (1 000 mg per day) was given orally. During oral administration of the drug a limited degree of recurrence of horny patches occurred. Systemic miconazole seems to be the drug of choice in the treatment of systemic chronic mucocutaneous candidiasis.


Axillary hyperhidrosis: local treatment with aluminum chloride hexahydrate 25% in absolute ethanol. Eleven women (group I) with axillary hyperhidrosis were treated ad modum Shelley & Hurley with local application of aluminium chloride hexahydrate 25% in absolute ethanol and plastic foil occlusion during two successive nights once a week. The patients were observed for 24 weeks. Another 12 women (group II) were treated for up to 12 weeks with aluminium chloride hexahydrate 25% in absolute ethanol but without using plastic foil occlusion. We have attempted to determine the effect of the treatment by sweat measurements, leaving the left axilla untreated during the initial 1--2 weeks as a control. An immediate reduction in sweat production of the treated axilla was found in both groups. The reduction increased during the first weeks and was maintained thereafter. The degree of sweat reduction was the same during standardized work and during rest (group II). At the final controls all patients in group I had discontinued the occlusion and had individualized the treatment, most of them using local application 1--2 times weekly. Two patients in group I had to stop the treatment because of unbearable itching. The other patients found the treatment completely satisfactory. All patients had experienced itching and smarting when starting the treatment. Often these discomforts were temporary but they seem to require active support by close medical control during the initital period of treatment. In contrast to previous investigations, effective treatment was achieved both with and without occlusion.


Phototoxic reaction to long-term low-dose administration of chlorpromazine in mice. The mouse tail in vivo technique for the study of acute drug phototoxicity was tested in experiments involving prolonged administration of chlorpromazine and ultraviolet light. A phototoxic reaction was induced with lower combined doses of drug and radiation than was the case in single exposures.


Demonstration of hepatitis B-surface antigen in liver biopsies. A comparative investigation of immunoperoxidase and orcein staining on identical sections of formalin fixed, paraffin embedded tissue. Staining of 1000 consecutive liver biopsies with orcein showed positive reaction in the cytoplasm of ground-glass hepatocytes in 18 of the biopsies. On new sections of the 18 orcein positive biopsies immunoperoxidase staining was performed in order to demonstrate hepatitis B-surface antigen (HBsAg). After destaining the same sections were stained with orcein. All biopsies showing positive orcein staining showed positive immunoperoxidase reaction. The number of positive cells was in biopsies with less than 20 positive cells per mm2 biopsy larger using the immumoperoxidase staining than with orcein staining. Further the staining contrast was more pronounced. Immunoperoxidase staining thus seems more sensitive than orcein staining for demonstrating HBsAg in liver tissues. Orcein stains HBsAg in liver tissue even though the antigen determinants are blocked by antibodies.


Comparison of anoxic changes in isolated rat cardiac myocytes in suspension and in histological sections. Cardiac myocytes from adult rat were isolated by heart perfusion in the presence of collagenase and incubated in the absence and presence of oxygen. As a result of anoxia, there was a gradual increase in plasma membrane permeability, noted as an increase in succinate-stimulated oxygen uptake, a decrease in trypan blue exclusion frequency, a leakage of cytosolic lactate dehydrogenase activity and an increased proportion of swollen, irregularly contracting myocytes. The contractions of the damaged myocytes resembled the known non-physiologic contractions of the heart, i.e. extrasystoles, arrhythmia, fibrillation or block. After different periods of time of anoxia myocyte pellets were fixed in formalin, sectioned and examined by light microscopy. The appearances of the myocytes in suspension were compared with those in paraffin-embedded sections. Special attention was given to the hematoxylin basic fuchsin picric acid stain and it was noted that the basic fuchsin was taken up by contracted or damaged myocytes, which according to their morphology in suspension revealed irregular contractions, but not by undamaged or necrotic myocytes.


Flow cytometric DNA analysis in fine needle aspiration biopsies from patients with prostatic lesions. Diagnostic value and relation to clinical stages. Transrectal fine needle aspiration biopsy was carried out in 59 patients with cancer of the prostate, benign prostatic hypertrophy and prostatis for cytomorphological examination and flow cytometric DNA analysis. One major single peak in the DNA histogram was observed in patients with benign hyperplasia, prostatitis and in some patients with carcinoma. A DNA histogram with a second or third peak was always compatible with carcinoma and was found both in patients with carcinoma in stage I--II and stage III--IV. It is suggested that patients with hyperploid cancers in stage I--II could benefit from early treatment.


Concentration and distribution of heavy metals in nasal mucosa of nickel-exposed workers and of controls, studied with atomic absorption spectrophotometric analysis and with Timm's sulphide silver method. Biopsy specimens from nasal mucosa of 30 nickel-exposed individuals and 6 controls were analysed by atomic absorption spectrophotometry to determine the content of nickel, copper, cobalt, zinc and iron. Timm's sulphide silver staining method was used for visualizing heavy metals in cryostat sections of biopsy material from each individual. The purpose of the investigation was to study the sulphide silver staining pattern in nasal mucosa of nickel-exposed individuals and controls, and to establish whether variations in heavy metal concentrations, especially nickel, affect the histochemical pattern in the mucosa. No consistent differences were found in the histochemical pattern between biopsies with high and low concentrations of nickel or any of the other metals. Some difference in epithelial types between specimens from the nickel-exposed group and the control group was seen, but the staining pattern was quite similar for corresponding epithelial types in the two groups. Two nasal carcinomas from nickel workers were virtually unstained with the sulphide silver staining method.


Immunochemical studies on Staphylococcus aureus plasma membrane. 2. Antigenic properties. Cytoplasma membrane and lipoteichoic acid (LTA) isolated from S. aureus Cowan I were examined serologically. LTA contains both alpha- and beta-glucosyl substituents at glycerol and most probably ester-linked alanine as well, all being antigenic determinants. In addition to LTA, the membrane contains a glycoprotein exhibiting antigenic determinant(s) in both the protein and sugar moieties.


Precipitating antibodies against Neisseria meningitidis in normal sera and their possible origin. Crossed immunoelectrophoresis was used to study precipitating antibodies against Neisseria meningitidis in sera from 214 normal persons and in human gammaglobulin. A polyspecific meningococcal antigen preparation and a corresponding rabbit antiserum constituted the reference system. All the sera contained one to five precipitins against N. meningitidis which could be identified and quantified by means of the reference system. The precipitin score, which expresses the number and titre of precipitins in each serum, increased with age. More than 25 precipitins were found in the gammaglobulin preparation, including antibodies against the groupspecific polysaccharides A, B and C. Absorption of three of the five normally occurring precipitins (nos. 4, 19, 22) with antigens from various bacteria showed that precipitins nos. 4 and 19 could be completely absorbed by antigens from other Neisseria species, but not by antigens from other genera. This indicates that these two precipitins may have been induced by non-pathogenic Neisseria species or by meningococci. The finding of protective antibodies in gammaglobulin might indicate the possibility of its use in prophylaxis or therapy meningococcal infections.


The DNA content and nuclear size in normal, dysplastic and carcinomatous laryngeal epithelium. A spectrophotometric study. The aim of the present study was to obtain a more objective evaluation of nuclear hyperchromasia and polymorphism in laryngeal epithelium. The method is based on a photometric assay of nuclear size and DNA content in Feulgen stained slides. The DNA content of hyperchromatic nuclei should exceed the mean for nuclei of normal epithelium by more than twice the standard deviation. In comparison with lymphocytes (2N) the DNA content in hyperchromatic nuclei corresponds to 3.3 N. The mean DNA contents of normal, dysplastic and carcinomatous laryngeal nuclei were 69, 71 and 118 A.U. The mean nuclear area for normal, dysplastic and carcinomatous epithelial cells were 48, 41 and 73 micrometer2. The higher the degree of atypia displayed by the cells the greater was the variability of the DNA content and the nuclear size. The investigation shows that the method may provide a more objective basis for evaluating hyperchromasia and polymorphism.


Further evidence for the involvement of microtubules in the proximo-distal intra-axonal transport of acetylcholine and related enzymes in rat sciatic nerve. The two mitotic inhibitors colchicine (COL) and podofyllotoxin (POD) and their respective isomers, lumiCOL and picropodofyllin (picPOD) were tested for their effect on the intraaxonal transport (AXT) of acetylcholine (ACh) and the cholinergic enzymes in rat sciatic nerve. The mitotic inhibitors and their isomers were dissolved in saline+10% ethanol (COL and lumiCOL) or dimethylformamid (DMFA) (POD and picPOD) and injected (3-5microliter) subepineurally into the sciatic nerve. As controls the vehicle (saline +10% ethanol or DMFA) alone were injected into some rats. 2 h later a crush operation was performed 15 mm distal to the site of injection. The accumulation of ACh or the two enzymes, ACh-esterase (AChE) and cholineacetyltransferase (CAT), in the nerve segment proximal to the crush (12 h before death) was used as a measure of the AXT. COL and POD were very effective in inhibiting AXT of all 3 substances, while their isomers, lumiCOL and picPOD, were essentially without effect on AXT in equimolar concentrations (0.1 M). The effects on AXT of the 4 test substances thus appear related to their affinity to bind to tubulin, which is several orders of magnitude higher for COL and POD than for their isomers. The results further support the view that intact microtubules are essential for AXT of both membrane-bound (ACh E) and soluble (CAT) enzymes, as well as of ACh in rat motor nerves.


Effects of pressure on fast axoplasmic flow. An in vitro study in the vagus nerve of rabbits. By using a radially directed stream of fluid it is possible to compress a section of a nerve in vitro by known pressures without interrupting the supply of oxygen and nutrients to the nerve. Motor neurons of the vagus nerves of rabbits labelled with 3H-leucine were used at room temperature. When a nerve was subjected to 20 mmHg pressure fast axoplasmic flow was not altered, but at 30 mmHg there was a slight but consistent inhibition, which was even more marked at 60 mmHg and still more at 90 mmHg. The pressure induced block of axoplasmic flow was reversible when 60 mmHg pressure was applied for as long as 4 h, when the nerves were compressed for 15 h there was reversibility in only 5 out of 8 cases.


Ventricular extrasystoles and intracellular electrolytes in hypokalemic patients before and after correction of the hypokalemia. Fifty-four initially hypokalemic patients, 43 of whom were on diuretic treatment, were given potassium supplementation until they showed a repeatedly normal serum potassium level. Muscle specimens obtained by percutaneous biopsy revealed that there were no concomitant increases in muscle potassium content, nor in intracellular potassium concentration, except in the very small group (6 patients) with a muscle magnesium content of greater than or equal to 3.95 mmol/100 g fat free dry solids (FFDS) and an initially lower muscle potassium content (less than or equal to 39.9 mmol/100 g FFDS). ECG, registered for 3 hours on a portable ECG tape recorder before and after correction of the serum potassium level, showed no change in the frequency of ventricular ectopic beats.


Whipple's disease. Clinical and histopathological changes during treatment with sulphamethoxazole-trimethoprim. A case of Whipple's disease involving the small intestine, bone marrow and peripheral lymph nodes diagnosed one year after the initial arthritic symptoms by peroral jejunal biopsy and rapidly responding to treatment with sulphamethoxazole-trimethoprim, is described. The effectiveness of therapy was monitored by clinical evaluation of the patient's condition, repeated intestinal biopsies, determination of body weight, faecal lipid excretion and haematological values. Clear-cut clinical remission was induced promptly; and after nine months' treatment, the patient was in perfect health. The drug was well tolerated and folate deficiency did not occur. After four months of antibiotic therapy, bacterial capsular remnants could still be demonstrated in the bone marrow aspirate. The characteristic small bowel mucosal PAS-positive macrophages, visualized in the jejunal biopsy specimens, persisted in reduced number after nine months of treatment.


Serum haptoglobulin changes in patients with severe isolated head injury. From a series of 74 patients with severe head injuries and massive brain lesions, we measured the changes in haptoglobulin in 33 cases. On average, the globulin was, after a short drop in the first two days, increased about three times towards the seventh day, and was still above normal after ten days. Seen together with the total alpha-2-globulin increase after head injury this rise seems mainly to be caused by the haptoglobulin changes. Haptoglobulin was found to be a good indicator of the extent of brain tissue damage. It cannot on its own be used for prediction of survival chances.


Immunochemistry of bovine serum albumin. Two fragments were isolated from BSA one was derived from the first terminal third of the molecule and the second from the last third of the molecule. Each fragment inhibited the reaction of BSA-anti BSA by 90% or better. An immunoabsorbent of each bound 90% of anti BSA. Each fragment bound two antibody molecules per mole of fragment. These results are explained by the concept that BSA contains repeating identical or similar antigenic determinants. Conformational non identity of various batches of BSA was revealed by reactivity of the disulfide bonds at the neutral transition. Trypsin was found to cleave GSA, PSA, and HSA to yield an immunochemically reactive fragment. At least in the case of HSA, the fragment exhibited all the immunochemical reactivity of the native protein.


Analysis of a complex antigenic site on horse cytochrome c. Of the antigenic determinants so far identified for cytochrome c, only one involves more than a single amino acid substitution between the immunogen and host proteins. Both a threonine at position 89 and a glutamic acid at position 92 control one of the three antigenic sites identified in horse cytochrome c, as expressed in rabbits. Three antibody subpopulations, all directed against this region of the molecule, were isolated from the serum of a single rabbit by adsorption on a series of insolubilized cytochromes c. Antibody fluorescence quenching titrations with a variety of cytochromes c were used to confirm the identification of the antigenic determinant and to examine the subtle differences in the specificities of the three subpopulations. The determinant in the region of Residues 89-92 is affected by amino acid substitutions at positions 88 and 96. Since all these residues are in an alpha-helix the farthest distance between them is only 12 A and therefore, the Residues 88-96 can all be accommodated in the antibody binding site. The ability to identify and describe the antigenic determinant, as well as separate subpopulations directed against this site, demonstrates the resolution possible using a series of homologous protein antigens.


T-lymphocyte activation by immunogenic determinants. Synthetic antigens have been of great value in elucidating the relationships between antigen structure and lymphocyte activation. The compound RAT behaves as a monofunctional antigen in guinea pigs and mice, inducing T-lymphocyte responses without appreciable circulating antibody, although the ABA-specific B cell population is expanded by immunization with the monovalent molecule. On the other hand, bifunctional antigens composed of one RAT moiety serving as a carrier and a second chemical group, either identical to or different from RAT, serving as a hapten, induced antibody responses. In such responses, T cell specificity was always directed against the RAT component. Using symmetrical bifunctional antigens with rigid or flexible spacers between the two determinants, marked differences in structural requirements for cell triggering, assessed by antigen-induced lymphocyte proliferation, and for cell cooperation, determined by antibody formation, were found. Rigidly spaced bifunctional antigens serve admirably for cooperation but poorly for T cell activation, underscoring the advantage of two-point binding for the latter.


Immunochemical studies on the tobacco mosaic virus protein. The decapeptide having the amino acid sequence Thr-Thr-Ala-Glu-Thr-Leu-Asp-Ala-Thr-Arg has been shown to be a major antigenic determinant of the tobacco mosaic virus protein in rabbits, mice and guinea pigs. The antigenic specificity of the decapeptide is attributed to its C-terminal tripeptide Ala-Thr-Arg. Although this tripeptide has no demonstrable binding with antibodies to the protein, its N-octanoylated derivative exhibits specific binding with antibodies as well as the capacity to elicit delayed skin reactions in guinea pigs immunized with the protein. The latter results suggest that both B cells and T cells have antigen receptors of identical specificities. Although all mouse strains tested responded equally to TMVP, with the production of anti-protein, the response to the decapeptide was shown to be correlated (albeit not absolutely) with Ig allotype Iga exhibiting generally high responsiveness while Igb exhibiting generally low responsiveness. The low responsiveness could not be attributed to suppression of the secondary immune response.


First consequences of the determination of the entire antigenic structure of sperm-whale myoglobin. By using the antigenic structure of sperm-whale Mb as a model we have established that the antigenicity of its sites is independent of any sequence identities between the injected myoglobin and the Mb of the immunized animal. Furthermore, the ability to produce in rabbits autoantibodies to rabbit Mb and the successful extrapolation of the antigenic structure of sperm-whale Mb to human hemoglobin strongly demonstrated that the antigenicity of certain parts of a protein molecule is primarily dependent on the uniqueness of their conformational locations.


Peptides and autoimmune disease.

Peptides and autoimmune disease. The use of derived and synthetic peptides has contributed greatly to our understanding of encephalitogenic determinants in the basic protein molecule. Peptides derived from BP by use of trypsin, pepsin, cathepsin D (brain and liver) and BNPS-skatole have proven most useful. Synthetic peptides have served to define the disease-inducing determinants with precision. A remarkable feature of these studies is that different antigenic determinants serve as encephalitogenic sites in different species. The encephalitogenic sites comprise short peptide domains of the BP polypeptide chain, only 8 residues (rat), 9 residues (guinea pig), and 10 residues (rabbit) in length. In view of the requirement for both haptenic and carrier specificity of an immunogenic molecule, it is impressive that these peptides themselves elicit the autoimmune disease, EAE. While less active than BP on a molar basis, they are nonetheless potent encephalitogens, producing clinical signs in rats and guinea pigs at less than 1 microgram dose. The data indicate that for most animal species (guinea pig, rat, monkey) there appears to be only one major encephalitogenic determinant, an unusual finding in view of the number of antigenic determinants for cell-mediated immunity existing in the BP molecule. Possibly a combination of genetic and anatomical factors may account for this phenomenon. A relationship may exist between multiple sclerosis and EAE as shown by peptide studies; lymphocytes are found in MS patients during exacerbation sensitized to the same region of BP active in the monkey. The major encephalitogenic sites are: Guinea Pig (9) Phe-Ser-Trp-Gly-Ala-Glu-Gly-Gln-Lys(Arg); Rabbit (10) Thr-Thr-His-Tyr-Gly-Ser-Leu-Pro-Gln-Lys; Rat (8) Ser-Gln-Arg-Ser-Gln-Asp-Glu-Asn; Monkey (14) Phe-Lys-Leu-Gly-Gly-Arg-Asp-Ser-Arg-Ser-Gly-Ser-Pro-Hser.


Recognition of lysozyme by lymphocyte subsets. Extensive studies with antisera from a variety of animals have failed to detect any cross-reactivity between egg white lysozyme and its reduced, S-carboxymethylated (CM-) derivative. In contrast, a number of studies addressing the specificity of T lymphocytes have revealed that these two forms of lysozyme cross-react rather extensively at that level. Preliminary attempts to eliminate this latter cross-reactivity by further denaturation and/or chemical modification have so far proven unsuccessful. In a second line of experimentation the response to CM-lysozyme of mice which are genetically unresponsive to native lysozyme was assessed and found deficient. The implications of these findings are discussed.


Genetic control of the T-lymphocyte proliferative response to cytochrome c. Cytochromes c have been used as antigens in a murine T-lymphocyte proliferation assay in order to characterize the nature of determinants whose recognition is under immune response (Ir) gene control. The cytochromes are advantagous as antigens because 1) they have well-characterized primary and tertiary structures, 2) they are antigenically simple, differing from mouse cytochrome c at only a small number of amino acid residues, and 3) there exist a large number of evolutionary variants which can be used to locate antigenic sites by cross-stimulation. In the present studies, the T-lymphocyte proliferative response to pigeon cytochrome c was shown to be under the control of two complementing major histocompatibility (MHC)-linked Ir genes in mice of the H-2a and H-2k haplotypes. Mice of the H-2b, H-2d, H-2p, H-2q, H-2s, and H-2u haplotypes were low or nonresponders. Complementation was demonstrated by showing that an F1 hybrid between two nonresponder recombinant strains, B10.A(4R) and B10.A(5R), could respond to pigeon cytochrome c. The determinant on the cytochrome recognized in this immune response was located to the C-terminal portion of the molecule around residues 89 and/or 100. This was shown by the failure of closely related cytochromes from the Pekin duck and chicken to cross-stimulate T lymphocytes immune to pigeon cytochrome; position 89 and 100 carry the only residues different from those in mouse cytochrome c that are unique to pigeon cytochrome among the three bird cytochromes tested. This localization was further substantiated by demonstrating that the cyanogen bromide cleavage-fragment (residues 81-104) from pigeon cytochrome, but not the same fragment from Pekin duck cytochrome, was as good a stimulant of T cells immune to the whole molecule as the intact cytochrome. These results identify the immunogenic site on the molecule as one which differs from mouse cytochrome c by only one or two amino-acid residues. Thus, T-cell immune responses, which are under MHC-linked Ir gene control, are as capable as antibody responses of recognizing subtle differences in protein structure. However, the ability of T cells to respond equally well to stimulation with polypeptide fragments or with the whole molecule suggests either that T-cell recognition involves certain differences from B cell recognition or that in some cases the fragments possess a similar spatial structure to that of the corresponding segment in the native protein.


The precise and entire antigenic structure of lysozyme: implications of surface-simulation synthesis and the molecular features of protein antigenic sites. Intensive research in the author's laboratory over a 10-year period has now culminated in the precise determination of the entire antigenic structure of native hen egg-white lysozyme. The protein carries three antigenic sites. Each site is made up of spatially adjacent surface residues that are not in direct peptide linkage. The residues of each site describe an imaginary line which circumscribes part of the surface topography of the protein and act functionally towards the antibody as if they are in direct peptide bond linkage. The reactivity of each site is fully satisfied by an appropriate surface-simulation synthetic peptide, and the three synthetic sites account for the full immunochemical reactivity of the native protein. Each site is subject to conformational restrictions and exhibits directionality which is a function of side chain orientations. The antigenic sites of myoglobin and lysozyme are compared. It is proposed that antigenic sites of the type found in myoglobin are called "continuous sites", while antigenic sites of the type seen in lysozyme are defined as "discontinuous sites".


Genetic control of the immune response to insulin: its dependence upon a macrophage mediated selection of distinct antigenic sites. The immune response to insulin, in both mouse and guinea pig , is under control of H-linked immune response genes. When immunized with either pork or beef insulin to CFA, both strain 2 and 13 guinea pigs respond by antigen-specific lymphocyte proliferation and synthesis of specific antibody. The specificity of the elicited antibodies are indistinguishable between these inbred strains. By contrast, strain 2 T cells recognize a distinct region of the A chain alpha loop consisting of amino acids residues 8, 9 and 10, while strain 13 T cells see an as yet undefined region of the B chain. H2b (A chain alpha loop responder) and H2d (B chain responder) mice similarly discriminate which area of the molecule are recognized by their T lymphocytes. The function of the Ir gene, in both the guinea pig and mouse appears to be an intramolecular selection of discrete regions within the antigen for recognition by the T cell. The data presented suggest that this function operates at the level of the macrophage.


Evidence for complement activation following the oral administration of cow's urine concoction in rats. Oral administration of cow's urine concoction in rats causes the activation of the third complement component in the serum. The product of this activation has some histame-releasing effects and causes a characteristic acute fall in neutrophil and monocyte counts in the peripheral blood which is reversed within four hours. The importance of these observations to the areas of the world where cow's urine concoction is used for the treatment of childhood convulsion is discussed.


Histochemical and immunohistochemical study of diffuse large-cell lymphomas. Ninety diffuse large-cell lymphomas (diffuse histiocytic lymphoma) were subclassified into B-cell, T-cell, and histiocytic types according to their enzyme histochemical and immunohistochemical characteristics. The B-cell type was characterized by presence of intracellular monoclonal immunoglobulin; negative or weakly positive diffuse acid phosphatase activity; and an occasional focal nodular pattern or preceding nodular lymphoma. The T-cell type was characterized by moderate, focal acid phosphatase activity; convoluted nuclear structure; and frequent preceding cutaneous manifestations. The histiocytic type was characterized by strong nonspecific esterases and diffuse acid phosphatase activity and presence of lysozyme and phagocytic activity. Most of the lesions (74 cases) were of the B-cell type. This group was further subdivided into follicular center cell type and B-cell immunoblastic sarcoma, according to the stage of cellular transformation. Preliminary clinical correlation suggests that the histiocytic type is most resistant to treatment. B-cell immunoblastic sarcomas were much more aggressive than neoplasms of the follicular center cell type.


Rapid detection of ringed sideroblasts with bromchlorphenol blue. A test for demonstrating ringed sideroblasts was developed, using the dye bromchlorphenol blue. The test is based upon the ability of the dye to form a colored water-insoluble precipitate with iron. Because it is rapid and permits localization of ferritin-containing cytoplasmic inclusions, the reaction may be useful in the assessment of sideroblastic anemias.


Prenatal diagnosis of congenital nephrosis in 23 high-risk families. The efficacy of maternal serum and amniotic fluid alpha-fetoprotein (AFP) estimation for the prenatal detection of congenital nephrosis was assessed in 23 pregnancies of couples with a previously affected child. At 15 to 18 weeks' gestation, amniotic fluid AFP concentration was elevated in seven of 23 cases, and maternal serum AFP level in five of these. Legal abortion was carried out at 18 to 19 weeks in all those cases where he amniotic fluid AFP concentration was abnormally high, and in all cases the fetus was found to be affected. The diagnosis of intrauterine congenital nephrosis was obvious by electron microscopic examination of the fetal kidney, but not by light microscopy. The child was born without congenital nephrosis in all 16 cases where amniotic fluid AFP level was normal, and in 16 of 18 cases (89%) where maternal serum AFP concentration was normal. Thus, the amniotic fluid AFP assay is more reliable and is recommended whenever congenital nephrosis is suspected on the basis of family history.


Developmental and psychoeducational sequelae of chronic otitis media. The developmental, psychological, and educational sequelae of chronic otitis media occurring during the first three years of life were investigated in a selected group of children with educational difficulties. One group of children had a history of chronic and severe otitis media, requiring myringotomy after 3 years of age. The control group had infrequent and relatively mild episodes of otitis media and no surgical intervention. Children with chronic and severe otitis media manifested substantial delays in speech and language, auditory processing deficits, disturbances in auditory-visual integration, reading disorders, and poor spelling skills. Early medical and educational intervention for children with a high-risk profile of language delay and severe chronic otitis media is suggested as a possible strategy for preventing the development of a specific learning disability.


Isoprinosine in classical acute viral hepatitis. A prospective double-blind controlled study was performed on the effects of isoprinosine in 81 patients with clinical and histological features of classical acute viral hepatitis. Appetite was assessed daily. Serum alanine aminotransferase, aspartic aminotransferase, and bilirubin levels were assessed weekly. Changes in biochemical parameters, number of deaths, number of patients showing rise of aminotransferase levels despite treatment, and duration required for appetite to return to normal in each group were compared. Observation was continued for 4 weeks. Contrary to preliminary reports, no therapeutic benefit was demonstrated in any of these parameters. We concluded that isoprinosine was ineffective in the acute phase of classical acute viral hepatitis.


Significance of meconium in midtrimester diagnostic amniocentesis. The importance of meconium passage as a sign of the third-trimester fetus in distress is well known. The significance of similar observations in the midtrimester fetus is much less certain. The increasing use of amniocentesis during the middle trimester for the diagnosis of genetic disease makes ascertainment of such data important. Ten instances of meconium-stained amniotic fluid in the last 514 amniocenteses performed at our institution for antenatal genetic diagnosis are reported. Seven pregnancies have ended at term with normal deliveries. Three pregnancies have terminated with fetal death. Meconium staining may be a sign of impending fetal death when passed during the second trimester. When accompanied by alpha-fetoprotein elevation two of three pregnancies terminated with fetal death.


Cultured Plasmodium falciparum used as antigen in a malaria indirect fluorescent antibody test. Human sera obtained from persons infected with Plasmodium falciparum were tested by a standard indirect fluorescent antibody (IFA) technique using antigen obtained from long term in vitro cultures of two strains of P. falciparum, and antibody in high titer was reproducibly detected. Sera from uninfected persons had undetectable or very low titers of antibody. The use of cultured parasites offers a convenient, stable source of antigens from different P. falciparum strains without requiring their adaptation to primates. Differences observed in IFA titers obtained by reacting immune serum with two different P. falciparum strains suggests the need for further evaluation of strain specificity.


Binding of antigen to surface Ig on isolated rat mast cells. This study concerns observations on isolated rat mast cells sensitized to specific antigen. Specific binding of antigen to the surface immunoglobulins of these cells could be demonstrated by immunofluorescence technique as well as by radiolabelled antigen binding and the degree of binding parallelled the allergic reaction as judged by histamine release. Exposure of mast cells to antigen at both low and high antigen concentrations did not change the distribution of the surface immunoglobulins. Furthermore, neither capping not shedding could be induced, even by excessive antigen stimulation. The amount of antigen molecules bound to surface Ig was linearly correlated to the allergic histamine release. When mast cells were sensitized to two antigens of different molecular weight, the cells showed the highest sensitivity to the antigen having the highest molecular weight. These results indicate that both the amount of antigen and the molecular weight of  the antigen bound to surface Ig are important factors in the allergic histamine release from mast cells, whereas surface Ig redistribution is not.


[Light microscopic and histochemical results on the innervation of Brunner's glands in the duodenum of laboratory animals (author's transl)]. Brunner's glands of cats and guinea-pigs for instance are staffed with a well formed nerve plexus of cholinergic and adrenergic axons. Corresponding to numerous neurophysiologic results the excretory glandular cells and the enterochromaffine cells of Brunner's glands are subjected to a differentiated adreno-cholinergic influence.


PAS-lead hematoxylin as a stain for small-granule endocrine cell populations in the lungs, other pharyngeal derivatives and the gut. Epithelial cells of the several subtypes that comprise the small-granule cell population of the respiratory system are little studied, partly because adequate silver, monoamine fluorescence and other specific light microscopical preparations have been more difficult to obtain than in the gut and other organs possessing diffuse endocrine systems. Periodic acid-Schiff (PAS) in combination with MacConaill-Solcia's lead hematoxylin has in our hands proven dependable for routine staining of serial 2-micrometer glycol methacrylate sections used in mapping the distributions of these cells along the airway. In lungs of mice, hamsters, kittens, and fetal rabbits, typical small-granule cells stain weakly or not at all with lead hematoxylin alone, hence are easily overlooked. PAS adds to the cytoplasm a diffuse magenta coloration; and because it is diastase-resistant, less brilliant than that of mucus but more so than bronchiolar cell secretions, and finer textured than lysosomal staining of other cells present, the effect is to highlight small-granule cells whether solitary or in clusters. Additional PAS staining of basement membranes and lead hematoxylin staining of cilia enhance the combined stain's resolving power. In thyroid gland, parafollicular cells stand out boldly against follicular elements; in small intestine, hematoxylin-positive endocrine cells are well differentiated from absorptive, mucous, and Paneth cells that surround them. Using a complementary monoamine fluorescence technique on plastic sections of lungs from control and 5-hydroxytryptophan-pretreated animals prior to staining, we can show that fluorescent epithelial cells are identical with those stained by PAS-lead hematoxylin.


Genetic polymorphism of the vitamin D binding protein and another post-albumin protein in horse serum. Horizontal polyacrylamide gel electrophoreses, on 10% separation gel, of horse serum revealed polymorphism of the vitamin D binding protein (Gc protein) and another post-albumin protein (Pa). Family data supported the hypothesis that Gc and Pa types were controlled by autosomal codominant alleles. For both Gc and Pa proteins, the homozygous types showed a single fraction while the heterozygous type had two fractions. Pa types were found to be identical to the post-albumin types reported earlier by starch gel electrophoresis. Two Gc alleles, GcF and GcS, and three Pa alleles, Pa D, Pa F and Pa S, were observed in samples from Swedish (four breeds), Lipizzaner and Arab horses. The frequency of the more common allele at the two loci, i.e. GcF and PaF, ranged from 0.72-0.93 and from 0.58-0.99, respectively, in the different breeds studied. Plasma samples showed an extra protein fraction near the GcS fraction and thus were found unsuitable for Gc typing.


Standardization of allergen extracts.

Standardization of allergen extracts. A review of clinical and experimental data pertaining to the standardization of allergenic extracts is presented. Various problems facing practicing allergists are outlined: potency and stability of extracts the reactivity of the patient, in vitro and in vivo methods for assessing potency. Practical recommendations for allergists concerning allergenic extracts and their use are enumerated.


Serologic examination of the Westphal-type lipopolysaccharides of Pasteurella multocida. The serologic specificities of the Westphal-type lipopolysaccharides from 16 serotypes of Pasteurella multocida were compared with those reported for the heat-stable typing antigens in the gel diffusion precipitin test. Like the heat-stable typing antigen, the Westphal-type lipopolysaccharide from each of the serotypes reacted only with its homologous antiserum in 14 of the 16 serotypes. The lipopolysaccharide from type strains representing serotypes 2 and 5 reacted with either of the corresponding typing sera, as did field isolates of these serotypes to a lesser extent. Although differences among the properties of these antigens were minor, the lipopolysaccharide appeared to be a major component of the heat-stable antigen responsible for the type specificity.


Relief of malignant obstructive jaundice by percutaneous insertion of a permanent prosthesis in the biliary tree. Twelve patients with malignant obstruction of the biliary tree were treated by dilating the lesion percutaneously and inserting an internal large-bore teflon prosthesis in place bridging the the stricture. All 12 patients had unresectable neoplasms. The procedure was devised because existing modes of palliation using surgical techniques are associated with significant mortality or mobidity. There are also many problems with nonsurgical catheter drainage. Decompression was achieved in all 12 patients as shown radiographically by passage of contrast into the duodenum. Disappearance of pruitus was achieved in seven of seven patients, and in 10 of 12 disappearance of jaundice (bilirubin, before prosthesis, 18.4 +/- 4.5 mg/dl [mean +/- 1 SD], bilirubin 1 month after prosthesis, 1.8 +/- 0.6 mg/dl [mean +/- 1 SD], P less than 0.001) with improvement of general clinical status was achieved. Percutaneous placement of a permanent biliary tract prosthesis is safe and effective for the palliative decompression of malignant biliary tract obstruction.


[New fluorescence method for staining of human spermatozoa]. 300 human ejaculates from a andrological group of patients were stained for visualisation of the spermatozoal morphology with the new fluorochrome DAPI (4',6-diamidino-2-phenylindol) and examined by fluorescence microscopy. DAPI has proven to be stable, since the preparations do not fade even after long storage periods. It can be utilized in small amounts and has a special affinity for the spermatozoon nucleus, which it stains light yellow, in contrast to the acrosomal parts which are fluorescent green. The method is surprisingly simple and can be carried out in a few minutes. A further advantage of the DAPI procedure lies in the additional staining of leucocytes and microorganisms, so that inflammatory processes of the male accessory organs can be diagnosed as well.


Palliative dilation for dysphagia in esophageal carcinoma. We retrospectively reviewed our experience with palliative dilation for dysphagia in esophageal carcinoma. During a 3-year period 26 patients with squamous-cell carcinoma of the esophagus underwent peroral esophageal dilation for relief of dysphagia. Twenty-four were able to resume a soft or regular diet after dilation. This improvement was accomplished with low morbidity and no mortality. Dilations were done without additional risk in patients with malignant tracheoesophageal fistulae and in patients undergoing radiation therapy. We conclude that esophageal dilation can be done safely and effectively in patients with squamous-cell carcinoma of the esophagus. Palliative dilation can significantly improve the quality of life for these patients and should be considered an important part of their management plan.


[Development of follicular and parafollicular adenomas in the thyroid of rats treated with thiamazole (author's transl)]. Hypothyroidism is induced in rats treated with thiamazole, an antithyroid drug. If this phase lasts long enough, the follicular cells develop adenomas. Within the same period, the number of parafollicular or C cells increases threefold on an average, but without producing corresponding adenomas, the formation of which is inhibited. When treatment is stopped, a second phase appears during which thyroid function reverts to normal. The previously observed inhibition disappears, and, after a period of latency, the hyperplastic C cells develop parafollicular adenomas. The type of tumors to be found in treated rats (either of them, or both simultaneously) is determined by respective durations of the two phases. The endocrine mechanisms of these phenomena are discussed.


[Problems in detection of mycoplasma contamination in cell cultures (author's transl)]. A total of 135 cell lines was examined for mycoplasma contamination using two techniques: isolation and specific DNA-staining with DAPI or "Hoechst 33258". The two techniques showed similar results in 64% of the cases of cell contamination while the remainder was detected only by one or the other techniques: 12,82% by the staining technique and 23% by the isolation technique, which shows that the 2 techniques are complementary. The staining technique is very quick, easy to execute, very sensitive, and should be the method of choice to detect contamination when the mycoplasma do not grow in acellular media.


Effect of enzymatic adenylylation on dihydrostreptomycin accumulation in Escherichia coli carrying an R-factor: model explaining aminoglycoside resistance by inactivating mechanisms. Strains of Escherichia coli carrying R-factor R71(a), which codes for a streptomycin-spectinomycin adenylyltransferase, have elevated levels of resistance to dihydrostreptomycin (DHS) compared with isogenic R(-) bacteria. DHS accumulated by whole cells and spheroplasts of R(+) bacteria is lower than that observed for R(-) strains, a result of the absence of the second and more rapid of the two energy-dependent phases of DHS uptake seen in susceptible E. coli. A mutant of R(+)E. coli with reduced DHS resistance has been shown to have reduced levels of streptomycin-spectinomycin adenylyltransferase activity as well as enhanced drug accumulation. Actively accumulated DHS was recovered from R(+) cells as the adenylylated derivative. Neither was inactivated antibiotic detected in culture filtrates, nor was actively accumulated drug lost from R(+) cells under normal conditions. The cellular distribution of actively accumulated DHS in R(+) and R(-) cells was found to be the same. Membranes isolated from these cells retained only a small fraction ( approximately 1%) of the total cell-associated drug. The R(+) derivative of a mutant with defective energy transduction (E. coli NR-70) and reduced ability to transport aminoglycosides has a significantly higher minimal inhibitory concentration of DHS than its R(+) parent (strain 7). Streptomycin-spectinomycin adenylyltransferase activity, from comparisons of K(m) values and total activities of enzyme, was the same in both strains. The enzyme has been localized to the exterior surface of the bacterial inner membrane, although isolated membranes lacked detectable enzyme activity. The preceding observations are consistent with the proposal that the level of R71(a)-mediated DHS resistance is the outcome of competition between the rate of adenylylation and the rate of the first energy-dependent phase of DHS transport. When the rate of adenylylation exceeds the first energy-dependent phase, adenylylated DHS is accumulated, apparently in a manner identical to the accumulation of DHS. Unlike DHS, adenylylated DHS does not interact with ribosomes, and, consequently, there is a failure to initiate ribosomally dependent sequelae such as the second energy-dependent phase of accumulation, inhibition of protein synthesis, and/or misreading of mRNA.


Iontophoretic application of adenine arabinoside monophosphate to herpes simplex virus type 1-infected hairless mouse skin. Several antiviral agents were applied topically or by iontophoresis to hairless mouse skin inoculated with herpes simplex virus type 1 (HSV-1), and the chemotherapeutic effectiveness was evaluated. Topical application of iododeoxyuridine, arabinoside A, and adenine arabinoside monophosphate (ara-AMP) moderately decreased the average lesion score, number of mice with paralysis, and number of mice dying in HSV-1-infected animals. Also, the mean survival time was moderately prolonged by topical application of those antiviral agents. When ara-AMP was applied by cathodal (-) iontophoresis to the HSV-1-infected skin, the average lesion score, number of mice with paralysis, and number of mice dying were greatly decreased. Furthermore, the mean survival time of mice was highly increased by cathodal (-) iontophoresis of ara-AMP. The therapeutic efficacy of ara-AMP iontophoresis was much superior to the topical application of iododeoxyuridine, arabinoside A, and ara-AMP. These data suggest that ara-AMP iontophoresis would be the method of choice for the management of HSV-1 skin lesions in hairless mice.


Histochemistry of the dermal ground substance of initial psoriatic lesions. 30 cases of initial psoriasis were studied. Pinhead-sized lesions, recently appeared, were excised and studied with histochemical methods for acid mucopolysaccharides. In all specimens a well defined reduction of the material stained with colloidal iron, with alcian blue with critical electrolyte concentrations and with toluidine blue was observed in the upper dermis, in particular in the papillary dermis in the center of the lesions, in correspondence of the squirting papillae. The more evident reduction of the staining with alcian blue with 0.1 M and 0.2 M MgCl2 suggests that the changes of the dermal ground substance are predominantly due to an alteration of the hyaluronic acid.


Extrahepatic biliary obstruction by metastatic colon carcinoma. Extrahepatic biliary obstruction can be caused by cancer metastatic from the colon to the lymph nodes adjacent to the bile duct. This report details our experience with eight such cases treated at the Massachusetts General Hospital in the last seven years. The interval between resection of the primary tumor and appearance of jaundice averaged 13 months. The location of the obstruction, preferably defined preoperatively by cholangiography, was low on the common duct in three cases and high in the porta hepatis in five. Relief of biliary obstruction was accomplished by biliary-enteric bypass (four cases), internal biliary stenting by permanent indwelling tube (two cases), or by portal irradiation (two cases). In addition to palliating the symptoms of obstructive jaundice, the period of comfortable survival appears to have been extended: the bypassed patients lived 13-38 months. Erosion of tumor into the duodenum, with resulting gastrointestinal hemorrhage, was an additional problem in three patients. Our overall experience illustrates the value of distinguishing this subgroup of patients from the larger number whose jaundice results from extensive liver metastases, and of treating aggressively those with extrahepatic biliary obstruction.


Cellular organization of locus coeruleus in the rat as studied by Golgi method. The locus coeruleus (LC) of rats at different ages was studied in sections stained by the Golgi-rapid method. The LC is composed of two types of neurons, medium-sized (20mu x 35mu) and small cells (10mu x 15mu). The medium-sized cells are mainly fusiform in shape and occupy the dorso-caudal part of the nucleus, while pyramidal or multipolar cells frequently occur in the ventral part. Both types of cells bear a moderate to large number of spines on the surface of their soma, which form the main criterion for identifying the LC neurons. Though they radiate several relatively short dendrites in different directions, which bifurcate once or twice, the medio-ventral ramification is prevailing, extending beyond the limit of cells to the pontine central gray adjacent to the LC. The latter region might constitute the principal receptive sites of LC for inputs from various origins. Axons of LC neurons take their initial course either latero-rostral- or latero-caudalwards, while some axons divide dichotomously rostral- and caudalwards. Fine axon collaterals are frequently demonstrated within the LC and sometimes seem to contact with dentrites of LC neurons. Besides this, two kinds of afferents of unknown origins are observed between LC neurons. At least three kinds of small oval neurons are occasionally impregnated, though their axonal distributions cannot be demonstrated.


Experimental demonstration of the cell types in the adenohypophysis of the gobiid fish, Rhinogobius brunneus. The hypophysis of the common freshwater goby, Rhinogobius brunneus, accompanying gonadal maturation, was studied light microscopically to determine the cell types by the aid of the administration of antithyroidal and antiadrenocortical drugs. It is peculiar in that the hypophysis is entirely buried in the hypothalamus, and that the entire adenohypophysis is covered with a thin layer of the neurohypophysis. Two types of cells, azocarminophil prolactin cells and PbH-positive adrenocorticotrophs, are discernible in the rostral pars distalis. The proximal pars distalis consists of three types of cells: weakly AF positive thyrotrophs, strongly AF positive gonadotrophs and orangenophil somatotrophs. In spite of the application of several stainings, only one cell type, the PbH-positive cell, is demonstrated in the pars intermedia.


Differential staining of collagens type I, II and III by Sirius Red and polarization microscopy. Organs of fish, amphibian, reptile, bird and mammals when stained by Sirius Red and studied with polarization microscopy present different colors in regions where collagens I, II and III have been described. Collagen type I presented a yellow, orange or red color while collagen type III appeared green. Collagen type II, present in cartilage and chondrosarcoma showed a variable color according to the tissue and the species. Its color and morphology however always permitted its clear distinction from collagens type I and type III.