Morphine and phenytoin binding to plasma proteins in renal and hepatic failure. The binding of morphine and phenytoin to plasma proteins was examined in healthy subjects and in patients with renal and hepatic failure. In the uremic patients without hepatic failure, morphine binding was dependent on the concentration of total serum proteins and albumin, but not the severity of renal failure as measured by creatinine clearance. Binding of phenytoin, however, was dependent on the degree of renal failure and albumin concentration, but not on total serum protein concentration. Renal transplant in 1 patient restored the binding of both drugs to a value within the normal range. The combination of hypoalbuminemia and hyperbilirubinemia resulted in the greatest impairment to binding for both drugs. It is concluded that patients with uremia, jaundice, hypoalbuminemia, particularly in combination, are sensitive to usual clinical doses of morphine, at least in part, because of decreased binding to plasma proteins.


Comparative effects of tolamolol and propranolol on cardiac and peripheral circulatory function in patients with coronary artery disease. In a clinical study comparing the cardiocirculatroy effects of intravenous tolamolol to those of propranolol, tolamolol, 16 mg, induced similar reduction in resting heart rate as 8 mg propranolol in 16 coronary patients. Tolamolol did not disturb cardiac pump performance and exerted less negative inotropic action than propranolol as assessed by mechanical contractility indices. Myocardial beta-one chronotropic and inotropic stimulation by exogenous epinephrine was blocked equally by tolamolol and propranolol. Tolamolol exerted less systemic vascular beta-two blockade than propranolol as assessed by the peripheral resistance and vasopressor responses to epinephrine infusion. Tolamolction than propranolol and is thereby suitable for careful extension of beta blockade therapy to certain patients with pulmonary  and ventricular dysfunction.


Fenmetozole in acute alcholol intoxication in man. Forty healthy adult male volunteers were studied to determine the efficacy of fenmetozole to antagonize the effects of acute alcholol intoxication. Twenty subjects receive placebo and 20 fenmetozole in dosage of 100 mg and 200 mg in a double-blind paradigm. Pretreatment with fenmetozole failed to antogonize or attenuate cognitive, perceptual, motor and affective changes associated with acute alchol intoxication.


A review and discussion of goals in community dentistry. The author discusses the need for clearly stated goals in community dentistry, the choice of goals, and the chances to achieve them. Goals in community health planning should be based on the values, needs and resources of the community in question. When analyzing goals a distinction should be made between dental health and dental care. Some goals for dental care, e.g. equity of access, may concern social justice but do not necessarily lead to better dental health. It is generally agreed that good dental health for a whole community cannot be achieved by restorative treatment alone; good dental health habits are considered to be a better way to promote dental health. Existing research indicates that there are chances to attain an acceptable level of dental health if the community effort is systematically planned and executed. There are indications that a reduction in the prevalence of dental disease can be brought about by new and effective means in the future. Long-term planning should be flexible enough to respond to new opportunities.


Cerebral Paragonimiasis.

Cerebral Paragonimiasis. The first case of cerebral paragonimiasis was reported by Otani in Japan in 1887. This was nine years after Kerbert's discovery of the fluke in the lungs of Bengal tigers and seven years after a human pulmonary infection by the fluke was demonstrated by Baelz and Manson. The first case was a 26-year-old man who had been suffering from cough and hemosputum for one year. The patient developed convulsive seizures with subsequent coma and died. The postmortem examination showed cystic lesions in the right frontal and occipital lobes. An adult fluke was found in the occipital lesion and another was seen in a gross specimen of normal brain tissue around the affected occipital lobe. Two years after Otani's discovery, at autopsy a 29-year-old man with a history of Jacksonian seizure was reported as having cerebral paragonimiasis. Some time later, however, it was confirmed that the case was actually cerebral schistosomiasis japonica.  Subsequently, cases of cerebral paragonimiasis were reported. However, the majority of these cases were not confirmed histologically. It was pointed out that some of these early cases were probably not Paragonimus infection. After World War II, reviews as well as case reports were published. Recently, investigations have been reported from Korea, with a clinicla study on 62 cases of cerebral paragonimiasis seen at the Neurology Department of the National Medical Center, Seoul, between 1958 and 1964. In 1971 Higashi described a statistical study on 105 cases of cerebral paragonimiasis that had been treated surgically in Japan.


Angiostrongyliasis cantonensis (eosinophilic meningitis, Alicata's disease). Angiostrongyliasis is an infectious disease caused by nematode parasites of the genus Angiostrongylus. The rat lung worm Angiostrongylus cantonensis, primarily a parasite of rodents, is largely responsible for human cases of eosinophilic meningitis, or meningoencerphalitis, which occurs on many Pacific islands and in Southeast Asia. The disorder, which frequently occurs in epidemic extent, is caused by invasion of the central nervous system by developing larvae of the parasite. The infection is most frequently due to ingestion of food containing the infective, third-state, larvae. Meningitic and ocular forms of the disease have been recognized. The disease has been described or referred to under a variety of synonyms. The terms eosinophilic meningitis, eosinophilic meningoencephalitis, and epidemic eosinophilic meningitis were first used to describe the disease before its etiology was known. These terms, however, lack specificity, because the eosinophilic meningitic syndrome may accompany many other parasitic as well as nonparasitic diseases of the central nervous system. Nevertheless, they are still being widely used, since in most cases only the epidemiology of the disease points to the etiologic role of A. cantonensis. Direct clinical or laboratory evidence of the etiologic agent is usually not established because reliable tests are not yet available. The term angiostrongylosis, or angiostrongyliasis, if used without the adjective, also may give rise to confusion, since the same term is applied to the pulmonary infection of dogs by A. vasorum and might be used for infection by any other Angiostrongylus species. Even the term cerebral or ocular angiostrongyliasis may prove in the future to have similar disadvantages. The scientifically correct term angiostrongyliasis cantonensis has been used recently. It is sufficiently specific and formed in analogy to the names of other parasitic diseases of man, like schistosomiasis japonica, schistosomiasis mansoni, schistosomiasis haematobia. For the murine infection, as well as for the disease produced experimentally in animals, the term angiostrongylosis cantonensis ought to be reserved because of the preferential use by veterinarians of the ending osis for designation of pathological changes produced in animals by parasites. After the recent discovery of A. costaricensis, another rat parasite causing human disease in Costa Rica, it becomes necessary to distinguish between angiostrongyliasis cantonensis (eosinophilic meningitis) and angiostrongyliasis costaricensis (intra-abdominal eosinophilic granulomatosis). A potential disadvantage of these terms may be encountered in case of a systemic revision or reclassification of the parasite. This has happened at intervals. A. cantonensis was named Pulmonema cantonensis by its discoverer and later was described under the name Haemostrongylus ratti.


Cerebral Malaria.

Cerebral Malaria. Cerebral malaria is an acute diffuse encephalopathy associated only with Plasmodium falciparum. It is probably a consequence of the rapid proliferation of the parasites in the body of man in relation to red cell invasion, and results in stagnation of blood flow in cerebralcapillaries with thromobotic occlusion of large numbers of cerebral capillaries. The subsequent cerebral pathology is cerebral infarction with haemorrhage and cerebral oedema. The wide prevalence of P. falciparum in highly endemic areas results in daily challenges to patients from several infected mosquitoes. It is thus important to understand the characteristics of P. falciparum, since this is one of the most important protozoan parasites of man and severe infection from it constitutes one of the few real clinical emergencies in tropical medicine. One of the more important aspects of the practice of medicine in the tropics is to establish a good understanding of the pattern of medical practice in that area. This applies to malaria as well as to other diseases. The neophyte might be somewhat surprised to learn, for example that an experienced colleague who lives in a holoendemic malarious area such as West Africa, sees no cerebral malaria. But the explanation is simple when the doctor concerned has a practice which involves treating adults only. Cerebral malaria is rare in adults, because in highly endemic areas, by the age of 1 year most of the infants in a group under study have already experienced their first falciparum infection. By the time they reach adult life, they have a solid immunity against severe falciparum infections. In fact, "clinical malaria" could occur in such a group under only two circumstances: 1) in pregnancy, a patent infection with P. falciparum might develop, probably due to an IgG drain across the placenta to the foetus;2) in an individual who has constantly taken antimalarials and who may have an immunity at such a low level that when antimalarial therapy is interrupted, clinical malaria might ensue. The above examples emphasise the paramount importance of the clinician dealing with malaria having some insight into the complex immunity processes operative in the human host; these have been reviewed by McGregor.


Patterns of Amyotrophic Lateral Sclerosis and Parkinsonism-Dementia on Guam. The striking concentration of amyotrophic lateral sclerosis (ALS) and parkinsonismdementia (PD) among the Chamorro people of Guam has attracted attention since the early 1950's. Extensive studies conducted over the past 20 years have not revealed the causes of these diseases and the reasons for their remarkable concentrations on Guam. Approximately 1 in 10 Guamanian deaths over age 25 is from ALS, and another 1 in 10 is from PD. ALS still occurs approximately 50 times more frequently on Guam than in the continental United States. Advances in virologic and biochemical techniques, and long years of study, appear to be leading us to a point where some of the problems will be resolved. We shall attempt to review the pertinent background information and summarize the data and observations up to the present time.


Viral-antibody complexes in canine adenovirus type 1 (CAV-1)ocular lesions: leukocyte chemotaxis and enzyme release. Canine adenovirus-type 1 (CAV-1)-antibody complexes caused severe anterior uveitis with corneal edema ("blue eye") when injected into the anterior chamber of normal dogs. The response of the anterior uvea to such immune complexes (IC) was similar to the spontaneously occurring disease. In the presence of complement (C'), IC caused release of neutrophile chemotactic factors. Following phagocytosis of IC-C' leukocytes released lysosomal enzymes, as indicated by the presence of acid phosphatase in the surrounding medium. Membrane bound viral aggregates, presumably IC, were common in neutrophiles and in macrophages that had infiltrated the anterior chamber of opaque eyes that occurred after intravenous (IV) inoculation with attenuated CAV-1. These data were incorporated into a postulated scheme for the pathogenesis of CAV-1 uveitis with corneal edema.


The planning, operation, and function of a clinical laboratory in a teaching hospital. Shortly after World War I, laboratories began to grow in number and complexity in the hospitals throughout the United States. Need dictated the funding and therefore, expansion of these laboratories. In general, very little overall planning was introduced into the development of these laboratories but rather empiricism and reaction to pressure dictated the day-to-day, week-by-week development. Since a great deal of the teaching at that time was done on a one-by-one basis with laboratory learners, and since much of the laboratory work was done by physicians and used directly upon the patient, there was very little need for considering the laboratory as a more global resource for the teaching institution. As the science of medicine advanced and as the diversity and complexity of laboratory occupations increased, it was necessary to place more stress upon careful planning and operation of the clinical laboratory in the teaching institution. The laboratory was approached not only as a training area in its own right but as an adjunct to the undergraduate and postgraduate medical education occurring in the teaching institution. It became paramount then to consider the detail planning necessary to bring about a structural and functional entity that could respond to these more global teaching needs. It was imperative that the laboratory itself respond to its role and responsibility in the tripartite function of a teaching institution -- research, education, and service. In any good planning environment, effort must be expended towards setting goals and objectives; analytical methods applied to making assumptions and establishing premises. Alternate means for achieving goals and objectives were developed. Energy was expended in making forecasts and projecting results. Alternate means were sorted out and choices were made. Implementation guidelines were designed and an evaluation and feedback mechanism was instituted. In more recent years, we have attempted to apply these steps in planning to the development of laboratories and to the installation of operational procedures which would allow the clinical laboratory to truly function as a teaching unit and as a vital resource for the total training program of the institution in which it is housed.


Physiological changes in blood volume.

Physiological changes in blood volume. Measurement of blood volume is an important clinical tool in establishing a diagnosis as in polycythemia vera, in assessing the true significance of a low blood count in a patient with splenomegaly, and in evaluating a bleeding patient. In theory, blood volume measurements should be of great value in treating acute blood loss, but this is often too time-consuming to be practicable. The main value of blood volume studies so far has been in clinical research where it is essential for correct interpretation of the peripheral blood measurements. Estimates using the hematocrit (packed cell volume) could be misleading since this measurement does not take into account total blood volume and changes in the plasma volume. A variety of isotopes that tag the red blood cell have been used to measure total blood volume and red cell mass. Most commonly used at present is autologous 51-Cr labeled red cells, thus eliminating the risk of hepatitis in transfusion to the subjects. Radio labeled albumin and other plasma proteins were used to measure plasma volume. A large molecular protein is ideal to eliminate overestimation from extravascular diffusion of labeled material for accurate determination of plasma volume. Physiologic changes in the newborn, in pregnant women, during exercise, bed rest, and altitude in the nonpathologic state are discussed. Studies in the normal subject's adaptation in different physiologic conditions have revealed a variety of factors that regulate blood volume.


Prodigiosin-like pigments.

Prodigiosin-like pigments. Prodigiosin, the bright red tripyrrole pigment from Serratia marcescens, has also been identified in Pseudomonas magnesiorubra, Vibrio psychroerythrus, and two Gram-negative rod-shaped mesophilic marine bacteria not members of the genus Serratia. Prodigiosin is sometimes bound to proteins; thus, extracts may require acid treatment before isolation of the pigment. Higher homologs of prodigiosin have been detected by mass spectroscopy. A mutant strain of S. marcescens produced nor-prodigiosin, in which the methoxy group of prodigiosin is replaced by a hydroxy group. Another mutant strain produced a blue tetrapyrrole pigment whose structure is a dimer of prodigiosin's rings A and B. Three novel biosynthetic analogs of prodigiosin have been obtained using a colorless mutant which does make rings A and B but not ring C and which can couple rings A and B with some added monopyrroles similar to ring C. The structures of three prodiginine (prodigiosin-like) pigments from streptomyces have been elucidated. All have the methoxytripyrrole aromatic nucleus of prodigiosin and all have an 11 carbon aliphatic side chain attached at carbon 2 of ring C. In two of the pigments the side chain is also linked to another carbon of ring C. The earlier literature about prodiginine pigments from actinomycetes has been interpreted and evaluated in light of the most recent findings. The structure elucidation of six prodiginine pigments from Actinomadurae (Nocardiae) has been completed. Only one, undecylprodiginine, is the same as from a streptomycete. For three of the six pigments, nine carbon side chains are observed and in four of them the side chain is attached to carbon 5 of ring A as well as carbon 2 of ring C so that a large ring is formed which includes the three pyrrole moieties. A section on identification summarized useful methods and presents information with which any known prodiginine pigment can be identified. The final step in the biosynthesis of prodigiosin was known to be the coupling of methoxybipyrrolecarboxaldehyde (rings A and B) with methylpentylpyrrole (ring C). Recent work using 13C-labeled precursors and Fourier transform 13C nuclear magnetic resonance has shown the pattern of incorporation for acetate, proline, glycine, serine alanine, and methionine into prodigiosin. Each pyrrole ring is constructed in a different way. Two of the streptomyces pigments have also been investigated; the pattern of incorporation is similar to that for prodigiosin. The biological activities of some prodiginine pigments are summarized. All show activity against several Gram-positive bacteria; some have anti-malarial activity. Prodigiosin has been tested clinically against coccidioidomycosis.


The therapeutic activity of diftalone in rheumatoid arthritis: crossover comparison with indomethacin. A multi-centre double-blind crossover study was carried out in 70 hospitalised patients with rheumatoid arthritis to compare the activity of diftalone, a new anti-inflammatory agen, with that of indomethacin. Patients received either 750 mg. diftalone or 100 mg. indomethacin daily for 2 weeks and were then crossed over to the alternative treatment for a further 2 weeks. Routine clinical and laboratory assessments of disease activity were carried out before the start of the trial and after each treatment period. The results indicate that both drugs produced a favourable response and that there was no statistically significant difference between them in the parameters measure, except for the erythrocyte sedimentation rate which fell only after the administration of diftalone. Patient and physician assessments on the course of the disease were in general agreement and were favourable to the same extent for both treatments. Both products were well tolerated and there were few reports of side-effects.


A controlled trial of bromhexine ('Bisolvon') in out-patients with chronic bronchitis. Bromhexine, 16 mg. 3-times daily, was compared with placebo in a double-blind crossover trial in 41 out-patients with chronic bronchitis and irreversible airways obstruction, and who were considered to be in a steady state. Each treatment was given for 3 weeks with a week of placebo in between; in addition, all patients took oxytetracycline, 500 mg. twice daily, starting 1 week before the trial and continuing throughout the duration of it. All patients completed the trial but the results from 5 were excluded because during the first 3 weeks they developed an influenza-like illness accompanied by a decrease in FEV1,and peak expiratory flow rate. Results from the remaining 36 patients showed a statistically significant reduction in phlegm stickiness (p smaller than 0.05) as judged by the patient, and a significant improvement in overall clinical state (p smaller than 0.05) as judged by the physician during bromhexine treatment, but no significant change in symptoms like cough, chest tightness, ease of breathing or sputum volume, peak expiratory flow rate and FEV1 etc. Five patients reported 6 side-effects, 3 with bromhexine, 1 with placebo and 1 with both treatments.


Pizotifen (BC 105) in migraine prophylaxis. Pizotifen, an antaminic, was investigated in a double-blind placebo controlled trial to evaluate its efficacy in migraine prophylaxis. It was found to be and effective prophylactic in a proportion of migraine patients, reducing the frequency as well as the severity of headaches. Some patients showed weight gain. Several patients have continued treatment for up to 2 years with good control of their migraine and a reduction in the amount of medicament required for the acute attack.


Clinical trial of a low dose combined oral contraceptive ('Ovranette'). A combination of 150 mug. d-norgestrel with 30 mug. ethinyl oestradiol was administered to 120 women for a total of 767 cycles. No pregnancies occurred and cycle control was good, 99% of the cycles being within the range of 28 plus or minus 3 days. Breakthrough bleeding occurred in 7.2% of the cycles, spotting in 8.9% and amenorphoea in 0.8%. Side-effects were minimal.


Six-day clotrimazole therapy in vaginal candidosis. Two hundred and two patients with symptoms, signs and mycological evidence of Candida vaginitis were entered in a single-blind trial comparing 6-day clotrimazole therapy with a control group receiving 6 days' nystatin therapy. Adequate data was collected on 69 clotrimazole and 72 nystatin treated patients for analysis. Judged by both the severity of symptoms and signs and the number of patients with negative mycological findings, 6-day clotrimazole therapy produced statistically significantly better results than obtained in the control group. When reviewed 4 weeks after starting therapy, 93% of the clotrimazole treated patinets had culturally negative results for Candida compared with 74% of the patients treated with nystatin. It is concluded that 6-day clotrimazole therapy is effective in the treatment of Candida vaginitis.


Evaluation of pindolol in the treatment of hypertension. Twenty hypertensive patients were treated for 15 weeks with pindolol, a new anti-hypertensive agent. Blood pressure while resting and after standing was reduced in all patients whether previously treated with an antihypertensive agent or not. All patients completed the study and no side-effects were reported. Laboratory tests showed no abnormalities and pindolol was very well tolerated in all patients. The dose of other antihypertensive agents was reduced in 7 patients.


Lampbrush chromosomes in spermatocytes of Chironomus. In the primary spermatocytes of Chironomus pallidivittatus lampbrush chromosomes in pachytene and diplotene stages can be isolated by using a spreading technique.--In the earliest analyzable stages of the pachytene the lampbrush loops, although different in size, are identically structured, consisting of spiralized fibers covered with a thin matrix. These loops contain an average DNA length of 33 mum. They are connected by axial fibers of 6 nm in diameter at intervals of 0.15--0.07 mum without any interposing chromomeres. The axis is double-stranded.--On the basis of the DNA distribution patterns it is supposed that the loops contain those chromatid regions which are included in the bands of the polytene chromosomes of the same species. The lack of a typical RNP transcript in the loops and the finding that 98% of the total DNA is found in the loops lead to the conception that the lampbrush organization in these spermatocytes is primarily involved in the orderly course of crossing-over and in the preparation for the metaphase contraction of the chromatids.


The proteins of polytene chromosomes of Drosophila hydei. It is reported that chromatin can be prepared from highly purified polytene nuclei from the salivary glands of third instar larvae of Drosophila hydei; such chromatin differs from that of diploid nuclei mainly by deficiencies in certain nonhistone chromosomal proteins. It is suggested that these proteins are important components of constitutive heterochromatin, which is severely underrepresented in polytene chromosomes. Chromosome morphology, including the pattern of induced puffs, is maintained throughout the mass isolation of glands and sucrose gradient purification of nuclei, as indicated by studies on temperature-shocked and control larvae. No significant alteration in the chromosomal proteins following puff induction by heat shock could be detected on analysis of the isolated protein fractions by disc gel electrophoresis. More sensitive techniques must be developed to study the apparent rearrangement or accumulation of protein at puff sites, and to elucidate the role of this protein in gene activation.


Accuracy of actuarial and clinical predictions for length of stay and unauthorized absence. Multivariate-equation predictions of length of hospital stay and probability of unauthorized absence were compared with similar predictions made by clincians and actual outcome for 167 patients drawn from a random sample of admissions during a two-month period at five state hospitals. The computer proved to be about as accurate as clinicians in predicting length of stay. Neither computer nor clinician was able to usefully predict unauthorized absence because of its infrquent occurrance. Use and improvement of such actuarial predictions within the Missouri automated data base were discussed.


[Postanastomotic renal-artery stenosis after renal transplantation]. A particular kind of renal-artery stenosis was observed in three of 60 renal transplantations performed between 1969 and 1974. It was always 1.0-1.5 cm distal to the anastomosis and, as histological examination demonstrated, was definitely due to vessel injury after removal of the transplant. To avoid it the proximal arterial stump should be cut off before transplantation. The postanastomotic region must be dissected free during the operation and an extended graft venoplasty or sleeve resection performed.


[Ectopic mammary gland on the back].

[Ectopic mammary gland on the back]. A supernumerary, fully developed mammary gland between both shoulder blades above the vertebral column was discovered in a 32-year-old woman when it caused symptoms. It was removed by operation. Histological examination revealed fibrous mastopathy. The only abnormal radiological feature was a spina bifida of T6 at the site of the ectopic breast.


Cyclic variations in the increased responsiveness of the pituitary to luteinizing hormone-releasing hormone (LHRH) induced by LHRH. Following a single iv injection of 50 ng of LH-releasing hormone (LHRH), a small increase in plasma LH titers was observed and this response was greater on proestrus and estrus than in other stages of the cycle. When a second injection of a similar dose of the neurohormone was given one hour later, the response to the second injection was markedly enhanced on proestrus but only slightly increased at other stages of the cycle. The response to LHRH in androgenized females was similar to that observed on proestrus, and, as on proestrus, an enhanced response to a second injection occurred. Only small increases in FSH followed these injections and there was no increase in responsiveness to the second injection in any of the groups. It is postulated that rising estrogen titers can bring about a priming action of LHRH which enhances the response of the gland to a second injection on proestrus. The priming action of LHRH may magnify the response of the pituitary to endogenous decapeptide release to further increase the release of LH on the afternoon of proestrus.


Ovarian steroids directly alter luteinizing hormone (LH) release by pituitary homografts. The direct role of estradiol and progesterone in sensitizing the adenohypophysis to the releasing action of LHRH (luteinizing hormone-releasing hormone) has been examined in rats bearing pituitary homografts. Groups (10-14/group) of virgin female rats (CDF) strain) were hypophysectomized, ovariectomized, and pituitaries obtained from long-term ovariectomized donors were implanted under the kidney capsule. The animals were treated on the sixth day after transplantation with sesame oil vehicle, estradiol (1.6 mug/100 g BW), progesterone (0.6 mg/100 g BW), or estradiol plus progesterone. This injection regimen was repeated 12 h later. Ninety minutes after the steroid injection on day 7 half of the animals in each group received LHRH (ip) and half received saline. Basal levels of LH in the steroid-treated groups not injected with LHRH were, in general, elevated as compared to the oil-injected group. Following LHRH, blood samples were collected by cardiac puncture under light ether anesthesia at 5, 15, 30, 60, and 90 min postinjection. Animals receiving LHRH responded with significant elevations of LH both in the presence and absence of steroids while saline was without effect. However, the magnitude of the LH peak was found to be significantly blunted by estrogen treatment. Moreover, the peak LH response to LHRH occurred 30 min following injection in all groups except the progesterone primed animals. In this group the peak LH was delayed by 30 min. These results suggest that estradiol directly inhibits the release of LH at the level of the pituitary and that the ovarian steroids, estradiol and progesterone probably interact to determine the time of optimal pituitary sensitivity to LHRH.


Localization of luteinizing hormone-releasing hormone in the preoptic area and hypothalamus of the rat using radioimmunoassay. To determine the localization of luteinizing hormone-releasing hormone (LHRH), five brains from adult male rats were serially sectioned in a cryostat at - 10 C in either the frontal, horizontal or sagittal planes. Acetic acid-ethanol extracts of each section were assayed for LHRH using radioimmunoassay (RIA) and in some cases using bioassay as well. Approximately 0.2 ng LHRH was concentrated in medial basal preoptic (MB-PO) tissue overlying the rostral portion of the optic chiasm. This LHRH appears to be associated with the organum vasculosum of the lamina terminalis and/or adjacent neural tissue. Uniform, low levels of LHRH were detected in hypothalamic tissue between the preoptic area (POA) and arcuate-median eminence (ARC-ME) region. In the ARC-ME region 2.7 ng of LHRH were concentrated primarily in the median eminence. The lateral distribution of LHRH in the ARC-ME region extended beyond the median eminence into tissue corresponding to the lateral aspect of the ventromedial nucleus. Concomitant bioassay and RIA determinations of LHRH were highly correlated. Of the sections bioassayed, only those sections containing LHRH released FSH. These results confirm the presence of LHRH in the POA and in the rostral hypothalamus of the rat brain. The possible significance of LHRH in the POA for the regulation of LH release is discussed.


Effect of alloxan on permeability and hexose transport in rat pancreatic islets. The in vitro effect of alloxan exposure on the permeability of collagenase isolated rat pancreatic islets to sucrose, D-mannitol, and L-glucose has been investigated. Determination of changes in cell volume with a non-wash double label isotope procedure indicates that alloxan treatment exerts no measurable effect on permeability to sucrose, D-mannitol, or L-glucose as compared to nonalloxan-treated islets. In addition, neither prior exposure nor the concomitant presence of alloxan alters the rate of D-glucose or 3-0-methyl-D-glucose transport into rat pancreatic islets. It is concluded that the in vitro effect of alloxan on abolishing glucose-induced insulin release in isolated rat pancreatic islets does not appear to be the result of permeability changes to small organic molecules or alteration in the transport of D-glucose into the beta-cell.


Biological aspects of cytochrome P450 and associated hydroxylation reactions. A review is made of the distribution and functions of cytochrome P450 in various tissues. "Normal" levels of the enzyme are tabulated and factors inducing higher levels are discussed. Evidence suggestive of the existence of different species of the enzyme is discussed, as is the hyothesis that genetically defective (or absent) isoenzymes of cytochrome P450 can provide an explanation for the clinical findings caused by the adrenogenital syndrome.


Some reflections on clinical research.

Some reflections on clinical research. The place of clinical research in the veterinary profession is discussed against the author's personal experiences and in the context of how research workers, clinicians, teachers and veterinary students might be brought into a more cohesive unit through the development of a Faculty of clinical research and experimental medicine. It is argued that students should receive training in research and teaching and that  efforts should be made to break down the attitude of "them" and "us" which tends to separate the clinicians and academics.


Surgical repair of cleft palate in the horse. Surgical repair of a cleft palate was carried out in three horses. Mandibular symphisotomy allowed adequate exposure of the defect. The first subject, a young foal died from inhalation pneumonia but the other two made satisfactory recoveries. The problems of closure of the lip and symphysis are discussed.


Maintenance of fertility in the horse including artificial insemination. A high fertility  rate depends on many different factors and is always related to inheritance and enviorment. The successful feritly control system in the German Thoroughbred breeding industry shows that fertility can be increased by good management and  veterinary supervision. The  insemination of horses with frozen semen is discussed. Replacement of natural service by A.I. with frozen semen is not generally  accepted in horsebreeding, as the conditions are entirely different from cattle breeding. However, there are several ways in which A.I. can be assistance in stud management.


Structure and function of the genome of coliphage T5. Transcription in vitro of the "nicked" and "nick-free" T5+ DNA. Purified T5+ DNA in nicked form and after repair of the specific single-strand interruptions with DNA ligase was used in transcription studies using Escherichia coli RNA polymerase (holoenzyme) in the presence and absence of E. coli termination protein rho. The transcriptional products were analyzed with respect to their size distribution and the sequences transcribed from the different templates. The results indicate that the single-strand breaks in the DNA of bacteriophage T5, though in genetically defined positions, do not have any specific effect on transcription in vitro. Furthermore, the E. coli rho protein, although it depresses net RNA synthesis and reduces the average molecular weight of the transcripts, seems to act in a non-specific way in this system.


Glucose catabolite repression in Escherichia coli K12 mutants defective in methyl-alpha-d-glucoside transport. 1. Two spontaneous Escherichia coli K12 mutants resistant to glucose catabolite repression were isolated using minimal agar plates with methyl alpha-D-glucoside. Mutants grow well on glucose and mannitol. 2. Glucose does not inhibit the inducible enzyme synthesis in isolated mutants: mutant cell (in contrast to parent cells) produce high levels of beta-galactosidase and L-tryptophanase under the conditions of glucose catabolite repression. 3. The isolated mutants are negative in methyl-alpha-D-glucoside transport; glucose uptake is not severely damaged. But the mutants (named tgl, transport of glucose) retained the ability to phosphorylate methyl alpha-D-glucoside in vitro at the expense of phosphoenolpyruvate. 4. The tgl mutation is cotransduced with purB and pyrC markers, i.e. locates near 24 min of the E. coli chromosome map. 5. It is thought that E. coli cells possess two glucose transport systems. The first one is represented by the glucose-specific enzyme II of the phosphoenolpyruvate-dependent phosphotransferase system. The second glucose transport system (coded for tgl gene) functions as permease and possesses high affinity to methyl alpha-D-glucoside. The integrity of glucose permease determine the sensitivity of the cell to glucose catabolite repression.


Nuclease digestion of synthetase x tRNA complexes. Phenylalanyl-tRNA and seryl-tRNA synthetase protect strongly though not completely their cognate tRNAs against nuclease attack, as had been shown previously. In an investigation of the mechanism of protection it was demonstrated that the low susceptibility of phenylalanyl-tRNA-synthetase x tRNA-Phe complexes to nucleases is due to free tRNA present in equilibrium with synthetase. The equilibrium can be shifted by an excess of synthetase or by dilution of the complex. It therefore appears that synthetase competes with the nuclease for free tRNA. Degradation of the complex is low, however, because under the conditions of partial digestion the synthetase has a greater affinity for the tRNA than does the nuclease. Fragmented tRNAs, as they are formed during partial nuclease digestion, bind to synthetase to different degrees. tRNA-Phe with a lesion in the dihydrouridine loop binds very poorly whereas a nick in the anticodon loop reduces the strength of binding to a much lesser extent. In a systematic study of the stoichiometry of protection it was confirmed that under standard conditions one phenylalanyl-tRNA synthetase protects one tRNA-Phe and one seryl-tRNA synthetase two tRNA-Ser molecules against nuclease attack. Under certain conditions, however, (concentration of the complex higher than 10 mu-M, or alternately in buffers of low ionic strength) it is observed that phenylalanyl-tRNA synthetase binds up to 1.6 molecules tRNA-Phe. In the serine system, these special conditions do not affect the binding properties of seryl-tRNA synthetase.


Studies on bacteriophage M13 DNA. 2. The gene order of the M13 genome. The double-stranded replicative form DNA of bacteriophage M13 was cleaved into 13 specific fragments by the restriction endonuclease from Haemophilus aphirophilus. The individual DNA fragments from wild-type replicative form molecules were then annealed to circular, single-stranded DNAs of phage M13, bearing amber mutations as genetic markers. When such DNA hybrids infected competent Escherichia coli cells, only those duplexes which were genetically heterozygous gave rise to wild-type phages in the progeny. In this way, the genetic markers carried on the individual DNA fragments could be determined. In addition, marker rescue in each gene was obtained with the 10 specific fragments of M13 replicative form DNA, produced by cleavage with the restriction endonuclease from Haemophilus aegyptius. From these results and the enzyme cleavage maps of both types of restriction fragments a distribution of genetic markers along the physical map could be obtained, which allowed an arrangement of M13 genes into a genetic map. Evidence is presented that the gene order of M13 is: IV-(I,VI)-III-VIII-VII-V-II with II and IV being contiguous on the circular map.


The effect of tRNA derivatives bound with natural or synthetic mRNA on the interaction of Escherichia coli ribosomes with colicin E3. Ribosomal binding complexes directed by poly(U) or T4 mRNA were formed with aminoacyl-tRNA or its derivatives bound to predominantly the P or A binding site. The defined binding complexes were reacted with colicin E3 and the reaction was assessed by the ability of the complexes to proceed with polypeptide synthesis. The results indicated that only one of the four complexes tested was completely resistant to colicin E3-induced inactivation: that of Phe-tRNA bound in the presence of poly(U) to the A-site. The poly(U) directed complex of AcPhe-tRNA and the T4-mRNA-directed complex at the A-site appeared slightly resistant, while the T4 mRNA initiation complex was inactivated by colicin E3 in a manner similar to non-complexed ribosomes. Colicin E3 added to ribosomes after protein synthesis had been initiated affected the subsequent polymerization in a manner corresponding to the response of the binding complexes. Thus, poly(U)-translating ribosomes were less affected than ribosomes translating the viral mRNA. The vulnerability of natural-mRNA-directed binding complexes to inactivation by colicin E3 is in accord with the mode of inactivation by the colicin in vivo.


Comparative effect of heparin on RNA synthesis of isolated rat-liver nucleoli and purified RNA polymerase A. The polyanion heparin has been employed to study the interaction of rat liver DNA-dependent RNA polymerase A and its template under various conditions. Heparin very efficiently inhibits polymerase molecules, which are not bound to DNA or are associated with the template in a loose, i.e. non-specific fashion. Purified nucleoli, isloated from rat liver nuclei, contain RNA polymerase A in abundant quantities of which only a portion is bound in a transcriptional complex. Excess enzyme, which is contained in the nucleolus in a quasi free form, can be transferred to an exogenously added template and can be completely inhibited by the prior addition of heparin. The enzyme contained in a transcriptional complex, however, initiated in vivo and completing these RNA chains in vitro, is fully resistant to heparin. In contrast to these results it has been found that RNA polymerase A extracted from nuclei and purified by various chromatographic steps does not form heparin-resistant complexes, even after the enzyme has been bound to the DNA template. Moreover it has been found that purified RNA polymerase A transcribes truly native DNA extremely poorly, indicating that the enzyme is highly deficient in the act of initiation on duplex DNA. It is therefore questionable whether the interaction of the purified enzyme and isolated DNA represents binding to true initiation complexes as is observed in the intact nucleolus.


Madopar versus sinemet. A clinical study on their effectiveness. Fifteen well-documented Parkinson patients treated with Madopar were switched to Sinemet for 3 months. The results were measured with an objective test procedure and compared. The effects of Madopar and Sinemet were equal provided th amount of levodopa was the same. There was one drop-out, because of ventricular extrasystoles. Attention is paid to the occurrence of involuntary hyperkinesias and the on-off-effect.


Gaseous persufflation during machine perfusion of human kidneys before transplantation. Massive air embolisation (persufflation) of four machine-perfused human kidneys occurred during preservation. This lasted for 60-120 min and caused increasing perfusion resistance. Pressure and flow returned to normal within 2 h after reperfusion. All kidneys showed immediate function after transplantation. Experiments with dog kidneys revealed closure of cortical vessels and increased kidney temperature during persufflation. Cortical flow became normal after 1 h of reperfusion.


Hemopoietic colony forming units in fresh and cryopreserved peripheral blood cells of canines and man. Colony forming units (CFUa) were assayed in the peripheral blood and separated mononuclear cells of canines and man.  Fresh and cryopreserved samples were studied.  By plating 3 x 10-5 canine buffy coat cells or 3 x 10-6 human buffy coat cells between 33 plus or minus 6.6 and 38 plus or minus 1.0 colonies were observed.  A high degree of reproducibility was shown for duplicate plates and on repeated testing.  Cryopreservation for a one-month period resulted in a minimal recovery of 80 per cent CFUa for canine cells and 96 per cent for human cells.  In vivo correlation between peripheral blood CFUa and marrow repopulation was assessed indogs following supralethal whole body irradiation.  Prompt repopulation of the marrow was observed and this correlated well with the CFUa assay following infusion of stored buffy coat cells.  It was concluded that the peripheral blood of both man and dog have significant numbers of cells with CFUa capabilities that may be potentially useful for marrow grafting purposes.


Factors which influence development of macrophage-layer and spleen colonies in mice. Irradiated mice infused with bone marrow cells developed colonies both in the spleen and on the macrophage layer of the cellulose acetate membrane which had been inserted into their peritoneal cavity. Factors influencing the ratio of spleen to macrophage-layer colonies were examined. The ratio of the colonies depended on the condition of the recipients rather than the condition of the injected cells provided there was no marked histoincompatibility between donors and recipients.


Treatment of canine malignancies by 1200 R total body irradiation and autologous marrow grafts. Twenty-five dogs with malignant lymphoma (L) and 18 dogs with solid, nonhematologic tumors (ST) were treated with 1200 R total body irradiation (TBI). Rescue from the otherwise lethal hemopoietic toxicity by infusion of autologous marrow aspirated before TBI was attempted, and survival, response to TBI, and immune reactivity post-grafting were determined. Eight L dogs survived more than 14 days post TBI and marrow grafting, and 12 out of 19 evaluable dogs showed a decrease of 75 per cent or more in clinically detectable tumor. There was no evident relationship between clinical status or marrow status before TBI and survival of more than 14 days or tumor response to TBI. Seven of the 8 survivors ultimately developed recurrent tumor. Eight ST dogs survived more than 14 days. Only 4 of 14 evaluable ST dogs showed significant clinical response of their tumor to TBI. Humoral and cellular immune reactivity were significantly impaired during the 10-week period following TBI and marrow grafting in all dogs studied. These results indicate that therapy in addition to lethal doses of TBI is necessary to cure spontaneous L or to significantly affect ST in dogs. They also provide baseline data which are necessary to assess the immunotherapeutic effectiveness of allogeneic marrow grafts.


Bone marrow regeneration after local injury: a review. This paper is focused on a mechanically depleted medullary cavity as an experimental model for analysis of marrow regenerative programs. The reconstitution of marrow in an evacuated cavity is basically a local phenomenon in respect to the stimulus for regeneration and the origin of the responsible cells. The nature of the triggering stimulus is unknown, but it is probably related to disruption of the continuity of the marrow stroma and endosteum. The initiating cells appear to be independent lines of mesenchymal and hematopoietic stem cells bound to bone, most likely within the endosteum and haversian system. The mesenchymal cells form the characteristic marrow stroma. Hemic cell regeneration can occur without immigrant hematopoietic stem cells, although such cells are known to contribute to later stages of repopulation. The formation and resorption of trabecular bone appears to be intimately related to the development of a sinusoidal matrix, perhaps by serving as a callus or supporting lattice and perhaps by providing a mechanism for distribution of stromal progenitors. Hematopoiesis is initiated in sites of active bone resorptive. The interplay of events consequent to marrow removal is strikingly similar to that seen with heterotopic marrow implants. Because stromal stem cells, unlike hematopoietic stem cells, do not migrate from distant sites, marrow stroma is the limiting factor in recovery from localized injury. Stromal stem cells are fairly radiosensitive but are not as sensitive as hematopoietic stem cells. The apparent radioresistance of stromal elements in an intact marrow seems to be due to their very low turnover rate. Latent radiation damage can be readily unmasked by conditions that promote their proliferation. This no doubt accounts for the radiosensitivity of stroma in an evacuated femur or heterotopic implant in contrast to its continued functional integrity with similar irradiation of in situ marrow. Even in an intact marrow, however, exposures in the 1000 rad range can lead to slowly evolving hypocellularity associated with diminished blood flow. With higher doses, aplasia of the irradiated site becomes progressively more generalized. It remains to be seen whether this limiting condition is due to the loss of specific regulatory functions or stromal components or merely reflects sinusoidal damage.


Augmentation of marrow growth by thymocytes separated by discontinuous albumin density-gradient centrifugation. Mouse thymocytes were separated by discontinuous albumin density-gradient centrifugation. The ability of B6 thymocytes from the gradient fractions to increase the number of spleen nodules formed by B6 marrow cells in B6D2F1 recipients was compared with that of similar numbers of B6 thymocytes from an unfractionated cell suspension. Thymocytes from any of the gradient fractions were not more effective than thymocytes from an unfractionated suspension in augmenting marrow-cell growth, indicating that the presumptive 'effective cell type' in this system cannot be separated from the total thymocyte population by its buoyant density properties.


Inhibitors of the adhesiveness of enteropathogenic E. coli. The entero-pathogenic strain of E. coli 0125: K 70 was able to adhere to washed guinea-pig erythrocytes and to cause their agglutination. Electron microscopy revealed this strain to be rich in fimbriae. They were able to cause hemagglutination by themselves, generally at protein concentration of 10 to 100 mug per ml. D-mannose and alpha-methylmannoside were able to inhibit hemagglutination by the whole bacteria or their isolated fimbriae at concentrations of 0.003 to 0.012 mg/ml. L-mannose, D-lactose, D-glucose, N-acetylglucosamine, D-galactose, N-acetyl-D-galactosamine, D-fucose, did not exert any inhibiting effect at concentrations as high as 50 mg/ml. N-acetyl-neuraminic acid was also uneffective at concentration as high as 9 mg/ml.


[B-lymphocytes subpopulation. Rapid identification with a histoimmunological method (author's transl)]. After extraction of tissues (spleen, thymus, blood) B cell bearing surface immunoglobulins were specifically labelled with Fab-peroxidase conjugate in light and electron microscopy. Positive and negative labelled lymphocyte were counted. This procedure allowed us to quantitate B and T cell in different diseases in peripheral blood and within lesions. It may represent a new immunopathological approach.


Effect of excision repair system on antibacterial and mutagenic activity of daunomycin and other intercalating agents in Salmonella typhimurium. Daunomycin and adriamycin, are more mutagenic and antibacterial for a strain of Salmonella typhimurium defective for the uvrB gene than for its uvr+ counterpart. Other intercalating agents, as some acridine dyes, affect equally the two bacterial strains.


Does pentagastrin supress secretin induced pepsin secretion in Heidenhain pouch dogs? Secretin-stimulated pepsin secretion from Heidenhain pouches was significantly depressed by concomitant pentagastrin. Pentagastrin by itself was without effect on pouch pepsin. Methacholine, on the other hand, did not antagonize secretin-stimulated pouch pepsin.


Are there somatostatin-containing nerves in the rat gut? Immunohistochemical evidence for a new type of peripheral nerves. Antibodies to somatostatin, a recently isolated hypothalamic peptide inhibiting growth hormone release, were used in immunohistochemical studies on the gastrointestinal tract. Somatostatin containing cells in the stomach, and somatostatin-containing nerves in the small and large intestine, could be demonstrated. These findings give evidence of a new type of nerve in the gut.


U.S. organized family planning programs in FY 1974. Most organized family planning programs combine the delivery of contraceptive services with a wide range of preventive health services. An estimated 3.3 million women, 97 percent of all who received family planning services in organized programs in FY 1974, received one or more medical services. At least 2.7 mission women received each of the 'core' medical services recommended by DHEW -- pap smear, blood pressure and pelvic and breast examinations. About 2.4 million women were tested for VD; 2.1 million women received urinalyses and 1.9 million received blood tests. Other services provided included pregnancy testing, infertility workup, and rubella and sickle cell screening.


Calcium antagonists and islet function. I. Inhibition of insulin release by verapamil. Verapamil is a potent calcium antagonist known to inhibit excitation-contraction coupling in both myocardium and myometrium. Its effect upon glucose- and sulfonylurea-induced insulin release was investigated in the isolated perfused rat pancreas. After twenty-five minutes' pretreatment and at concentrations ranging between 0.8 and 8.1 muM, verapamil caused a dose-related inhibition of glucose-induced insulin release during both the early and late phase of the secretory process. At a concentration of 0.8 muM, the degree of inhibition was more marked when the exposure time to verapamil prior to stimulation with glucose was increased to sixty minutes. Verapamil also inhibited gliclazide-induced insulin release. Infusion of verapamil during the late phase of the secretory response to glucose demonstrated that the inhibition of insulin release was an immediate and reversible phenomenon. The inhibitory effect of verapamil was enhanced at a subnormal calcium concentration and reduced at a high calcium concentration. These findings are consistent with the well-known calcium dependency of both glucose- and sulfonylurea-induced insulin release and suggest that verapamil might be a promising tool for further studies on the interactions between cations and secretagogues in the beta-cell secretory process.


Basal and postprotein insulin and glucagon levels during a high and low carbohydrate intake and their relationships to plasma triglycerides. The effects of acute and chronic differences in the carbohydrate content of the diet on plasma insulin, glucagon, insulin-glucagon molar ratio (I/G), and triglycerides were studied. Acute effects were studied by varying the carbohydrate content of a single test meal, while chronic effects were determined by varying the carbohydrate content of the diet for a week. A test meal containing 0.6 gm of gelatin per kilogram plus 0.6 gm. per kilogram of glucose resulted in much higher levels of insulin and I/G (p smaller than 0.005), lower glucagon levels (p smaller than 0.05), and slightly higher triglycerides (N.S.) than did a meal of 1.2 gm, per kilogram of gelatin alone. One week of a 12 gm. carbohydrate, 2870-calorie diet lowered insulin (p smaller than 0.001), I/G (p smaller than 0.05), and triglycerides (p smaller than 0.001) and increased glucagon (N.S.), whereas a 390-gm. carbohydrate, 2784-calorie intake significantly increased insulin, I/G, and triglycerides (p smaller than 0.005) and lowered glucagon (p smaller than 0.02) within two days; even greater changes in hormones were observed on a 510-gm. carbohydrate intake. Of those patients in whom a high carbohydrate intake induced a triglyceride rise of at least 40 mg. per deciliter, a significant correlation between the change in I/G and the change in triglycerides was noted (r equals 0.85; p smaller than 0.01). The results are compatible with but do not prove the proposal that pancreatic alpha and beta cells play a mediating role in carbohydrate induction of hyperlipidemia.


Paradoxical glucagon response after stimulation with glucose and arginine in isolated pancreatic sand rat islets. Isolated pancreatic islets of normoglycemic sand rats do not respond to 2.5 mM glucose with an enhanced glucagon secretion, which could be observed in normal Wistar rats. Arginine stimulates glucagon release in the presence of 2.5 mM glucose in Wistar rats as well as in sand rats. The secretion pattern is not caused by insulin deficiency since sand rat islets are characterized by an increased insulin secretion rate in vitro. This paradoxical glucagon secretion is not caused by a changed glucagon content but might be related to this species which is able to develop a diabetic syndrome spontaneously.


An apparent abnormaltiy of the B-cell microtubular system in spiny mice (Acomys cahirinus). The pancreatic B-cell contains microtubules, which are thought to participate in the process of insulin release. In order to disclose a possible abnormaltiy of this B-cell microtubular system in animals with islet dysfunction, isolated islets from normal rats and mice, as well as from diabetic mutant mice (DBM mice) and from spiny mice (Acomys cahirinus) were incubated in the presence of vincristine, which causes the precipitation of the microtubular protein into paracrystalline deposits. Ultrastructural examination of the islets indicated that the volume-density of vincristine-induced deposits was markedly reduced in B-cells of spiny mice, when compared to that found in normal rats and mice and DBM mice. Exposure of the islets from spiny mice to a high glucose concentration, concomittantly to vincristine, caused a further reduction in vincristine-induced crystals content of the B-cell. It is speculated that an impairment of the B-cell microtubular system may account for the deficiency of insulin release found in spiny mice.


[Carcinoma of the uterine cervix and herpes simplex virus type 2. Investigations on the causal relationship (author's transl)]. Epidemiological serological studies have shown that women with carcinoma of the uterine cervix show a higher incidence of Herpes Simplex Virus Type 2 (HSV 2) than controls. The attempt to demonstrate the genomic DNS of the HSV 2 in cells of carcinoma of the cervix by experiments of hybridization is reported. The investigations were carried out directly on material from carcinoma of the cervix and also on cell cultures from tumor material. The majority of the cultures grew fibroblasts but some cultures grew cells with the characteristics of malignant cells. The hybridization of DNS from portions of carcinoma or of DNS from cell cultures with complementary HSV 2 radioactive RNS showed in no case HSV-2-DNS in a quantity of one or one half genomic equivalent per cell. These investigations suggest that a casual relationship between carcinoma of the uterine cervix and an infection with HSV 2 is unlikely. However, it cannot be excluded that carcinoma of the uterine cervix has different quantitative proportions than the Burkitt-lymphoma and that only a fraction of the genomic material of the virus (smaller than 1/10) is incorporated into the malignant cell.


Age-related changes in the bone marrow and spleen of SAS/4 mice. The total number of nucleated cells in the bone marrow of SAS/4 mice increase some twofold between 1 and 24 months of age but when related to body weight remains essentially constant over a wide range of ages. The concentration of CFU-S in femoral marrow is also constant with age and since other bones containing marrow appear, at least in young mice, to have the same CFU-S concentration as the femur it is concluded that the CFU-S compartment size of the whole bone marrow is independent of age when expressed on a body weight basis, In contrast, both the absolute number and the concentration of exogenous CFU-S in the spleen decline markedly in old mice. Smilary there is a decline in the number of endogenous colony-forming cells and the spleens of 24-month-old mice seem virtually devoid of such colonies. Not only were older mice less capable of supporting the growth of endogenous colonies, but their spleens also appear to provide a poorer environment for exogenous colony growth when compared with growth in younger recipient spleens.


Serum antibodies in human pancreatic disease. An immunofluorescent study of sera from patients with various pancreatic diseases and from controls was carried out using human pancreas as substrate. A reproducible pattern of immunofluorscence, consisting of a coarse granular appearance in the cytoplasm of acinar cells of the pancreas and independent of the ABO blood group system was found. This occurred in 21 of 61 patients with acute pancreatitis; it was not found in other pancreatic diseases, and occurred in only four of 170 control subjects consisting of mixed hospital patients and healthy adults. Preliminary absorption experiments suggest that the  antigen is located in the microsomal fraction of pancreatic homogenates and may be organ-specific. The relationship of positive antibody tests to aetiological factors is discussed.


The promoters and bidirectional transcription in the immC region of P 22 and L phages. It is proposed that the transcription of the c-2 repressor gene initiates at two promoters, pre and prm, similarly to the transcription of the c-I gene of lambda phage. In the P22 phage, these promoters are defined by mutations of c54 and K5.


Penicillinamidohydrolase in Escherichia coli. I. Substrate specificity. Substrate specificity of the bacterial penicillinamidohydrolase (penicillinacylase, EC 3.5.1.11) from Escherichia coli was determined by measuring initial rates of enzyme hydrolysis of different substrates within zero order kinetics. Some N-phenylacetyl derivatives of amino acids and amides of phenylacetic acid and phenoxyacetic acid of different substituted amides of these acids or amides, structurally and chemically similar to these compounds, served as substrates. Significant differences in ratios of initial rates of the enzyme hydrolysis of different substrates were found using a toluenized suspension of bacterial cells or a crude enzyme preparation, in spite of the fact that the enzyme is localized between the cell wall and cytoplasmic membrane, in the so-called periplasmic space. N-phenylacetyl derivatives are the most rapidly hydrolyzed substrates. Beta-phenylpropionamide and 4-phenylbutyramide were not utilized as substrates. The substrate specificity of the enzyme is discussed with respect to a possible use of certain colourless compounds as substrates, hydrolysis of which yields chromophor products suitable for a simple and rapid assay of the enzyme activity.


Growth of Cryptococcus neoformans in UV-irradiated excreta of pigeons. UV irradiation of pigeon droppings resulted in an increased concentration of some inhibitors (peroxides) of growth of Cryptococcus neofarmans. This may be, in addition to the direct germicidal action of sunshine, another cause of the rare occurrence of this fungus in pigeon droppings on unsheltered sites in natural habitats.


The in vitro and in vivo effects of carrageenin on humoral and cellular factors of natural resistance. The inhibitory effect of carrageenin, a sulfated algal polygalactose, on humoral factors of natural resistance is discussed. In dependence on dose and time, the influence of non-specific and specific bacteriolysis, on the in vitro and in vivo opsonic activity and on the course of infection was studied.


Novel approaches to the mode of action of colicins. According to the theory of Fredericq (1949) and Nomura (1964), colicins are attached by specific receptor sites in the cell walls of sensitive bacteria, which mediate their inhibitive effects. During last years, a great variety of experimental data have been accumulated, some of which cannot be easily interpreted in terms of this theory. There exist considerable discrepancies concerning the chemical nature and molecular weight of isolated receptors. The attachment of a colicin onto its receptor need not be irreversible. The inhibition of numerous membrane-associated functions in colicin-tolerant mutants suggests their pleiotropic deletion nature. The difference between colicin resistance and colicin tolerance does not seem to be clear-cut. Cells of stable L-forms of protoplast type, completely devoid of their walls, retain in most cases the same patterns of sensitivity to colicins as rods of the same strains. Experimental changes in the relationship between the cell wall and the cytoplasmic membrane decrease colicin sensitivity of the cells. Colicin E3 has been found to be a specific endoribonuclease, able to cleave a terminal fragment from the 16 S rRNA also in isolated ribosomes in vitro: not only in ribosomes from sensitive bacteria, but also in those from resistant ones and from eukaryotic cells. A destabilization of the DNA helix was induced by colicin E2 in vitro as in vivo. It seems that there exist two distinct types of colicin receptors with different functions: those in the cell wall, and those in the cytoplasmic membrane. Only the contact of colicins with the latter ones is biologically effective and starts both stages of their inhibitive effect: the reversible and the irreversible ones.


Developing a utilization review model for community mental health centers. During their struggles to develop a utilization review model for a community mental health center, members of an interdisciplinary committee identified several issues they believe are important in establishing such a model. They include comprehensiveness of participation and of areas for review (the review committee should represent all disciplines and programs, and should be concerned with any aspect of center functioning), a problem-review approach in which subcommittees carry out documented studies of issues or problems, and specific provision for feedback and implementation of the results.


Adsorption in vitro to Escherichia coli of antibodies and other proteins in bovine serum and colostrum and its effects on the production of E. coli agglutinins. IgGl, IgG2, IgA and IgM from bovine serum and colostrum are adsorbed by Escherichia coli in vitro; lactoferrin is also adsorbed from colostrum and alpha2 macroglobulin from serum. The colostral adsorbed proteins on E. coli appear to reduce formation of agglutinins when the treated bacteria are injected into rabbits and guinea-pigs. Assay of the concentration of proteins dissociated from colostrum-treated cells showed equal amounts of secretory IgA AND IgGl, half that amount of IgG2, and traces of IgM and lactoferrin. Dissociation of proteins from serum-treated E. coli yielded equal amounts of IgGl and IgG2, traces of IgA and an alpha2 macroglobulin, but no IgM.


Promotion of secondary anti-DNP antibody production in mice by type III pneumococcal polysaccharide (SIII) and dinitrophenylated rabbit antibody to SIII. Type III pneumococcal polysaccharide (SIII) is able markedly to increase the adoptive IgG ANTI-DNP antibody response of B cells primed to DNP-flagellin and stimulated with DNP conjugated to the heterologous carrier, rabbit globulin, provided the latter has anti-SIII activity. The stimulatory effect is apparently accessory cell-dependent as well as being unequivocally T cell-dependent. Although no positive evidence is available, the possibility exists that non-specific T-cell activation is involved in the stimulating effect of anti-SIII plus SIII.


Splenic mediated erythrocyte cytotoxicity in malaria. Cell-mediated cytotoxicity in virulent rodent malaria has been demonstrated in vitro, whereby splenic cells effected specific lysis of 51Cr-labelled erythrocytes from parasitized animals. More than one cellular cytolytic effector system appeared to be operative in the mouse. One effector system involved splenic macrophages, from normal or immune animals, which were increasingly cytotoxic to target cells in the presence of antibody. A second effector system involved nylonpurified immune spleen cells which were significantly more cytotoxic than similary prepared normal spleen cells in the presence of immune serum. Although antibody alone was not cytolytic, the data strengthen the concept that immune spleen cells and antibody can interact in a co-operative fashion to mediate cytotoxic reactions in malaria.


Experimental aspergillosis in rats infected via intraperitoneal and subcutaneous routes. Normal adult rats infected via the subcutaneous (s.c.) route with viable spores of Aspergillus fumigatus develop serum antibodies, measured by passive haemagglutination and passive cutaneous anaphylaxis (PCA), to extracts of the fungus. No significant histological abnormality was detected but s.c. inoculation in cortisone-treated animals induced the formation of granulomas frequently found in lymph nodes including those of the mesentery and paratracheal group. These granulomas were associated with the presence of both spores and fungal hyphae. Intraperitoneal inoculation of A. fumigatus spores occasionally produced lesions containing hyphae in the liver, spleen and mesenteric lymph nodes. Lesions were more frequent and extensive in cortisone-treated rats where again they were most prominent in the mesenteric lymph nodes. None of the regimes produced fatal hyphal aspergillosis. Both normal and cortisone-treated rats developed serum agglutinins but not reagins following intraperitoneal injection of spores.


Incorporation of stainless steel, titanium and Vitallium in bone. Operative trauma, implant instability and corrosion are recognized as aetiological factors in the production of the soft-tissue capsule often present round a metal implant in bone. In the present experimental study, the isolated effect of corrosion was evaluated under standardized experimental conditions when operative trauma was minimal and implant movement was negligible. The reaction of bone to stainless steel, titanium and Vitallium was studied by histological and biomechanical methods, and compared to that provoked by copper. No soft-tissue capsule was found round the modern materials, but copper gave rise to a marked tissue reaction. It is concluded that a traumatic operative techniques and implant immobility are more effective for stable incorporation of metallic implants in bone than the nature of the implant itself, provided that one of the modern, low-corrosive metals or alloys is used.


Osseous lesion simulating a bone tumour due to an unsuspected fragment of wood in the foot. A case is reported in which an osteolytic lesion in the third metatarsal bone was proved at operation to be the result of an injury by a splinter of wood which had remained embedded in the tissues. A short review of the relevant literature is appended.


In vitro lymphocyte response to purified protein derivative, BCG and Mycobacterium leprae in a population not exposed to leprosy. Lymphocytes from 14 BCG-vaccinated donors, seven tuberculin positive and seven tuberculin negative by skin testing, were stimulated in vitro with four mycobacterial antigens, purified protein derivative (PPD), PPD/BCG, whole BCG bacilli, and whole Mycobacterium leprae and also with Candida antigen and phytohemagglutinin. The response was measured by incorporation of (3)H-labeled thymidine. The response to PPD, PPD/BCG, and BCG was found to correlate with the result of skin testing with turberculin. The turberculin-positive group also responded more strongly to M. leprae, whereas the two groups did not differ significantly in their response to Candida antigen or phytohemagglutinin. These findings indicate a certain degree of cross-reactivity between BCG and M. leprae. The use of the lymphocyte transformation test to measure antigenic cross-reactivity is discussed.


In vivo behavior of revertants from Stahylococcus aureus L-phase variants compared with the parent strain. The in vivo behavior of revertants of Staphylococcus aureus L-phase variants were studied in mice. The Giorgio strain of S. aureus was induced into L-phase variants by methicillin. After 13 serial passages in methicillin agar and six passages in antibiotic-free agar, reversion occurred. Six revertants, all phage untypable, coagulase negative, and mannitol negative, were inoculated intravenously into Swiss albino mice in comparison with the parent strain. Inoculation of 10(8) colony-forming units resulted in 38% mortality at 28 days for the Giorgio group and 0 to 2% for the revertants. The geometric mean titer of staphylococci in the kidneys was 3.5 x 10(5) for the Giorgio group and 0.5 x 10(0) to 1.0 x 10(2) for the revertants. Serial colony counts of blood, lungs, spleen, liver, and kidneys in mice inoculated with 10(7) colony-forming units showed faster clearance and failure of the revertant to colonize the kidneys. It is concluded that the in vivo behavior of the L-phase revertants was vastly different from that of the parent Giorgio strain.


Bacteremia in suckling rabbits after oral challenge with Vibrio parahaemolyticus. Vibrio parahaemolyticus, a halophilic marine enteropathogen, produces diarrhea in man after ingestion of contaminated seafood. Only strains capable of producing beta-hemolysis (Kanagawa-positive strains) are enteropathogenic. Yet the majority of marine isolants are nonhemolytic and Kanagawa negative. Studies were initiated in suckling rabbits in an attempt to elucidate pathogenetic mechanisms. Fasting animals were infected orally with Kanagawa-positive and Kanagawa-negative strains of V. parahaemolyticus, V. cholerae, two enteropathogenic strains of Escherichia coli, Shigella flexneri, and strains of salmonellae. Seven hours postchallenge, cardiac blood, liver, and spleen cultures were obtained. V. parahaemolyticus strains failed to induce intestinal fluid accumulation during this study period. Bacteremia occurred in animals challenged with Kanagawa-positive V. parahaemolyticus, S. flexneri, and salmonella strains only. Animal passage increased the ability of V. parahaemolyticus to cause bacteremia. Liver and spleen cultures were positive in approximately 50% of animals challenged with the Kanagawa-positive strains of V. parphaemolyticus. The ability of this organism to penetrate the intestinal epithelium of suckling rabbits may represent a specific property that plays a role in pathogenesis.


Lymphocyte transformation in syphilis: an in vitro correlate of immune suppression in vivo? Suppression of cellular immunity during primary and secondary infection may explain, in part, the unusual clinical evolution of syphilis. We have previously shown that lymphocytes from normal subjects undergo blastic transformation when exposed in vitro to Treponema refringens. This response was suppressed in patients with syphilis. the suppression being unrelated to serum factors. In the present paper we studied lymphocyte response in vitro to T. refringens, T. reiter, and T. pallidum as well as to monilia and trychophytins. The response to these antigens was suppressed in patients with syphilis although the response to phytohemagglutinin. pokeweed mitogen, and streptolysin was normal. These data support the hypothesis that human infection with T. pallidum is followed by a complex interaction between cellular and humoral immunity, the former being suppressed in primary and secondary stages.


Role of immunoglobulins in protection against Shigella-induced keratoconjunctives. Various immunoglobulin preparations were tested for their ability to protect guinea pig eyes from infection by a virulent strain of Shigella flexneri. Secretory immunoglobulin A was effective in delaying or preventing keratoconjunctivitis in eight guinea pigs when it was used to precoat the organism, and was also placed in the eye with the inoculum. Neither immunoglobulin G nor immunoglobulin M gave any protection when used in the same way. Protection by secretory immunoglobulin A appeared to be related to the antishigella antibody content of the immunoglobulin since a low-titered preparation gave less protection than a higher-titered one.


Altered phospholipid metabolism in Escherichia coli accompanying killing by disrupted granulocytes. The effect of bactericidal concentrations of disrupted rabbit granulocytes and of partially purified granulocyte fractions on phospholipid metabolism by Escherichia coli has been investigated. Previous studies in this laboratory have shown that, during and after killing of E. coli by granulocytes, bacterial macromolecular synthesis continues. Similarly, despite almost complete loss of viability within 15 min, incorporation of [1-(14)C]palmitate, [2-(14)C]glycerol, and [1-(14)C]acetate into E. coli phospholipids, in the presence of granulocyte preparations, remains the same as in control E. coli populations for at least 1 h. Incorporation of [1-(14)C]oleate into E. coli phospholipids is actually stimulated during the first 60 min of incubation in the presence of granulocyte preparations (more than twofold at 30 min and 40% at 60 min). With all labeled lipid precursors, bactericidal granulocyte preparations cause a relative increase in the labeling of E. coli cardiolipin, with a corresponding drop in labeled phosphatidyl-glycerol. Labeled lyso-compounds accumulate in the presence of granulocyte preparations when [1-(14)C]palmitate, but not when [1-(14)C]oleate is the labeled precursor. Since oleate occurs mainly in the 2-acyl position of E. coli phospholipids, whereas at least 50% of palmitate occurs in the 1 position, it appears that a phospholipase A(2) acts on the E. coli phospholipids. These various effects are also seen when E. coli are exposed to highly purified granulocyte preparations that possess potent bactericidal and phospholipase A(2) activities. We speculate that this phospholipase A(2) in the granulocyte preparations stimulates oleate but not palmitate incorporation by initiating increased turnover of the fatty acid in the 2-acyl position of E. coli phospholipids, causing formation of 1-acyl lyso-compounds likely to be preferentially reacylated with unsaturated fatty acids.


Isolation and purification of Treponema pallidum from syphilitic lesions in rabbits. Treponema pallidum were extracted from testicular syphilomas of corticosteroid-treated rabbits and purified by differential centrifugation. The steroid therapy allowed a longer holding time for infected rabbits, which produced greater treponeme yields, averaging 1.58 x 10(10) treponemes per rabbit. The treatment, which also diminished cellular infiltration and increased the extracellular mucoid material in lesions, produced much cleaner suspensions than preparations from nontreated animals. Most of the treponemes in the purified suspensions were still motile, and none carried demonstrable host immunoglobulin. The preparations were free of recognizable host tissue debris and they contained, on the average, 1.9 x 10(-7) mug of protein per treponeme.


Simplified assay for concanavalin A-dependent bacterial agglutination by using cell surface mutants. Two cell surface mutants of Escherichia coli isolated through supersensitivity to sodium dodecyl sulfate were found to be agglutinated by concanavalin A in the intact rod form. By using these strains, the assay method for the concanavalin A-dependent agglutination was greatly simplified since neither spheroplast formation nor protease treatment is needed. This rod cell agglutination was characterized and applied for a semiquantitative determination of the amount of bound concanavalin A.


Yersinia pestis: correlation of ultrastructures and immunological status. Ultrastructural identification and localization of the fraction 1 "envelope" antigen in the plague bacillus Yersinia pestis were the primary objectives of this brief study. The antigenicity of extra-cellular material between the bacilli in undisturbed cultured colonies and that of the pathogen per se were measured and correlated by means of the semi quantitative complement fixation method after incubation for 72 h at 37 C. When the amount of extracellular substance in wild-type T1 (virulent) bacteria was compared by electron microscopy with that in avirulent strains of Y. pestis, with and without passage through guinea pigs, we found that the material of interest was greatly attenuated or even absent in colonies that had not been passed through animals, whereas passage markedly augmented production of the material. We also explored the requirement for larger quantities of Ca(2+) and Mg(2+) in the culture media and discovered that without these ions production of the extracellular material was limited. These observations support the hypothesis that this extracellular substance between cultured Y. pestis bacilli of various strains represents the source of the fraction 1 envelope antigen.


In vitro N-nitrosodimethylamine formation by some bacteria. It was found that human pathogenic bacteria and some species of intestinal bacteria in rats have the ability to catalyze the formation of N-nitrosodimethylamine from dimethylamine and nitrate in vitro.


Genetic instability of M protein and serum opacity factor of group A streptocci: evidence suggesting extrachromosomal control. The M antigen, a primary determinant of virulence in group A streptococci that is expressed biologically as resistance to phagocytosis, is known to undergo a variety of phenotypic changes both in vivo and in vitro. These changes are nonrandom and can occur at a high frequency. Using the previously described relationship between the serum opacity reaction (associated with certain strains) and the presence of the M antigen, the phenotypic instability of the M antigen was analyzed. The results support the conclusion that M protein synthesis and the serum opacity reaction are directly or indirectly controlled by the same gene or by genes which are linked and can segregate as a unit. Moreover, growth conditions and the curing agents rifampin and ethidium bromide had a discernible influence on the segregation of clones unable to exhibit serum opacity factor and to resist phagocytosis by human leukocytes. Serial transfer of stationary-phase cultures of four strains of group A streptococci significantly increased the number of colonies negative for the serum opacity reaction and the M antigen. For two of four strains both ethidium bromide and rifampin also increased the segregation of colonies with this phenotype. In light of these experiments and the necessary controls, the possible influence of plasmids or bacteriophage in regulating M protein synthesis is discussed.


Factors affecting filamentation in Candida albicans: changes in respiratory activity of Candida albicans during filamentation. Glucose metabolism and respiration of Candida albicans were compared under conditions which permitted either maximal filamentous or maximal yeast growth. Changes in metabolism were monitored by comparing the quantities of ethanol produced, CO2 evolved, and oxygen consumed. Filamenting cultures produced more ethanol and less CO2 than yeasts, with oxygen consumption in the former concomitantly slower than that of the latter. Studies involving cofactors and inhibitors associated with electron transport imply that a transfer of electrons away from flavoprotein is required for maintenance of yeast morphology. Conditions consistent with a buildup of reduced flavoprotein, however, favored filament formation. These changes were expressed metabolically as a shift from an aerobic to a fermentative metabolism. The results presented are consistent with hypotheses correlating filament production with changes in carbohydrate metabolism and an interruption of electron transfer within the cell.


Cellular streptolysin S-related hemolysins of group A Streptococcus C203S. Group A streptococci strain C203S, grown in heart infusion broth with 0.3% maltose, produce two cellular hemolysins related to extracellular streptolysin S (SLS). Enzymatic lysis of the streptococci by group C streptococcal phage-associated lysin results in release of low titer, labile hemolysin, which can be stabilized by ribonucleic acid (RNA)-core (RNA preparation from yeast). This labile hemolysin can be detected only after the higher titer cellular streptolysin O is removed by erythrocyte membranes or inactivated by N-ethylmaleimide. The other cellular SLS-related hemolysin is released in a latent state (potential hemolysin) which can be activated to high-titer hemolysin by sonication with RNA-core. The titer of such activated hemolysin depends upon the intensity of sonic energy, duration of sonication, and amount of RNA-core. RNA obtained from the streptococci is far less effective than RNA-core. When the cocci are disrupted by sonication or grinding, potential hemolysin and/or activated form may be released, depending upon the conditions employed. The potential hemolysin material is large and heterogeneous; activation appears to involve, in part, disaggregation or fragmentation. Labile hemolysin, potential hemolysin, and the activated form of potential hemolysin can all be converted to hemolysin having the same hemolytic and physical properties as RNA-core SLS, suggesting that all have the same hemolytic moiety. The presence of glucose in heart infusion broth prevents formation of both potential hemolysin and RNA-core SLS by log-phase cells, whereas addition of glucose to a culture in heart infusion broth with 0.3% maltose stops accumulation of potential hemolysin but does not affect continuation of RNA-core SLS release. These results suggest that potential hemolysin is a cellular precursor to RNA-core SLS.


Use of the K88 antigen for in vivo bacterial competition with porcine strains of enteropathogenic Escherichia coli. Infant mice were used to measure the amount of fluid accumulation (enterosorption) in the intestinal tract after oral inoculation of a porcine strain of enteropathogenic Escherichia coli (K88-+, Ent-+). Significant reduction in the amount of fluid found in the intestinal tract was observed if the mice were first inoculated with a K88-possessing, non-enterotoxigenic strain of E. coli. The protection provided is thought to be due to specific competition for attachment sites on cells of the small intestine.


Competition between Chlamydia psittaci and L cells for host isoleucine pools: a limiting factor in chlamydial multiplication. L cells (mouse fibroblasts) supported the multiplication of the obligately intracellular parasitic bacterium Chlamydia psittaci (strain 6BC) when incubated in fresh growth medium (medium 199 + 5% fetal calf serum). When incubated in the medium supernatant from a 24-h-old culture of uninfected L cells (24-h used medium), uninfected cells did not divide and infected cells did not provide an adequate environment for the multiplication of C. psittaci, which persisted in a noninfectious latent state within the host cells. The failure of both L cells and chlamydiae to divide resulted from an overall reduction in the rate of protein synthesis by both host and parasite brought about by an insufficiency of the essential amino acid isoleucine in 24-h used medium. The concentration of isoleucine required to activate minimal growth of C. psittaci also minimally stimulated uninfected L cells to divide. The addition of cycloheximide to 24-h used medium also activated the latent chlamydial infection because it stimulated the incorporation of host protein-derived isoleucine into chlamydial protein. The results suggest that the chlamydial parasite and the L-cell host compete for the isoleucine in the soluble pool of the host cell and that the parasite is capable of sequestering isoleucine for its own biosynthetic needs only when the concentration of isoleucine in the host pool rises above the level required to maintain the hose in the stationary state. Extrapolation of the results obtained with the L cell-C. psittaci model system to natural latent chlamydial infections is discussed.


Prolonging morbidity in rabid dogs by intrathecal injection of attenuated rabies vaccine. Dogs vaccinated intrathecally with attenuated rabies vaccine developed antibodies that were detected in the cerebrospinal fluid, blood, and brain; dogs similarly vaccinated but with an inactivated vaccine developed no antibodies in the brain. When the attenuated vaccine was administered to rabid dogs, a prolongation of the morbidity period was noted and, in some dogs, recovery from the disease. Rhesus monkeys died when administered any of the available attenuated vaccines intrathecally, and further studies with that species could not be undertaken.


Quantitation of human peripheral blood T and B lymphocytes. Centrifugation of heparinized peripheral blood on Ficoll-Hypaque under defined conditions is a most common method of lymphocyte purification in clinical studies. A loss of 25-30% of lymphocytes may occur during this procedure due to incomplete recovery of lymphocytes from the plasma layer after the gradient centrifugation and/or to poor recovery during the washing process. The loss is not selective since T and B cells are lost in the same proportions as determied by several different methods for T and B cell identification. Quantitation of T and B lymphocytes in terms of total number/mm-3 of peripheral blood is necessary in order to determine deficiencies in either of these two cell populations. The percentage values are inadequate in clinical evaluations. Thus, patients with chronic lymphocytic leukemia show elevated absolute number of T cells absolute numbers of T and B cells as well as percentages ought to be reported in all clinical studies.


In vitro lymphocyte transformation in the development of experimental allergic thyroiditis. In vitro lymphocyte transformation to homologous thyroid antigen has been studied in cultures of lymph node cells obtained from 97 guinea pigs at weekly intervals after a single injection of homologous thyroid extract in complete Freund's adjuvant, using a method for measurement of 3-H-thymidine incorporation. Lymph node cell cultures from animals 1 week after sensitization showed a significant increase in thymidine incorporation after the addition of antigen. Two weeks after sensitization the stimulation was maximal, and then a lower but significant response continued during the next 5 weeks. There was no correlation between lymphocyte transformation and the level of circulating antibodies throughout the post-sensitization period examined. The early lymphocyte transformation contrasted with the later development of macrophage migration inhibitory factor activity.


Human, rat and mouse liver-mediated mutagenicity of vinyl chloride in S. typhimurium strains. Exposure of S. typhimurium strains TA 1530, TA 1535 and G-46 to vinyl chloride increased the number of His+ revertants/plate 16, 12 or 5 times over the spontaneous mutation rate. After 6 h of exposure to vinyl chloride, the mutagenic response for TA 1530 strain was enhanced 7-, 4- or 5-fold when fortified postmitochondrial liver fractions from humans, rats or mice were added. The enzyme-mediated vinyl chloride mutagenicity was dependent on an NADPH generating system and the enzyme activity was localized in a liver microsomal fraction; 9,000 times g liver supernatant was three times more active than microsomes, while liver cytosol or alcohol dehydrogenase did not affect the mutagenicity. Phenobarbitone pretreatment of rats and mice increased the mutagenic response by up to 15-40 percent as compared to untreated controls. The relative mutagenic activities of VCM, taking the value from mouse liver as 100, for TA 1530 strain mediated by 9,000 times g tissue fractions were: rat liver, 80; mouse and rat kidney, 20 and 16; mouse and rat lung, less than 7; human liver (from four biopsy specimens) 170, 64, 70 and 46. Chloroacetaldehyde and chloroacetic acid, a urinary metabolite of VCM, showed toxic effects, while chloroethanol was weakly mutagenic for TA 1530 strain.


[Aminoglycoside antibiotics from clinical viewpoint]. Indications, side effects, dosage and several pharmacokinetic properties of gentamicin, tobramycin, sisomicin, kanamycin, amikacin, and streptomycin are delt with in the first part. Hereafter, some aspects are briefly mentioned concerning combinations of antibiotics with aminoglycosides, as well as indications for the evaluation and analysis of the aminoglycoside concentration in the serum.


[Aminopenicillins from microbiological viewpoint]. In this report it was tried to emphasize several microbiological aspects of the amino-penicillin. Hereby it turns out that the activity differences of all the substances are very similar on microbiological part. A break of the cross-resistance does not happen with all the amino-penicillins. With regard to the appearance of resistant strains, it can be told that the conditions still are advantageous on non-hospitalized patients in the totality. In the clinic a high part of resistant strains are found on different grounds.


Aminopenicillins - clinical pharmacology and use in disease states. Ampicillin introduced in 1961 has been administered in the treatment of diverse infections by both oral and parenteral means. Oral infections of the upper airways such as otitis media, bronchitis, and pneumonia have responded with high success rates since the microorganisms involved have remained sensitive to ampicillin. Similarly, out-patient urinary tract infections caused by Escherichia coli, Proteus mirabilis, and enterococci are cured. Typhoid fever may yet be treated with ampicillin, but shigellosis has become refractory with the development of resistant strains. Ampicillin has assumed a prominent role in the treatment of gonorrhoea. Parenteral ampicillin is still a mainstay of the treatment of Hemophilus meningitis, but the recent appearance of ampicillin resistant strains may become a serious problem. A number of derivatives and analogues of ampicillin have been developed. Among the compounds, hetacillin, metampicillin and pivampicillin which hydrolyze in the body to yield ampicillin, only pivampicillin appears to offer advantage over the parent compound. Blood levels are twice those of a comparable dose of ampicillin. However, more comparisons with ampicillin in clinical situations are needed. The other analogues of ampicillin are epicillin, cyclacillin and amoxicillin. Epicillin has no superiority to ampicillin, and the cyclacillin data do not show clear superiority over ampicillin in spite of initially high blood levels, since the compound is less active and so rapidly cleared from the body. Amoxicillin, on the other hand, has been shown to have it vitro activity equal to ampicillin and to produce higher blood levels for a longer period of time. Clinical studies have substantiated efficacy in treatment of otitis media, pharyngitis, bronchitis, pneumonitis, and urinary tract infections at doses half those of ampicillin. It has been effective in gonorrhoea and typhoid, but not in shigellosis. It would seem that to date only pivampicillin and amoxicillin, particularly the later, should be considered as replacements of ampicillin in oral therapy.


[Cephalosporin antibiotics from microbiologic viewpoint. A comparison of antibacterial and pharmacokinetic properties]. With the necessity not only of differentiating between effectiveness and non-effectiveness, as in the earlier phases of chemotherapy, but also of detecting graduated differences in effect quantitatively, animal experiment and clinical trial have been shown to be no longer adequate. In a correlating comparison of antibacterial activity with the drug level in vivo as the two decisive basic elements of chemotherapeutic effect, new and interesting possibilities arise for the evaluation of antibiotics. In a comparative evaluation of nine different cephalosporin antibiotics, not only the objective antibacterial and pharmacokinetic properties are taken into consideration, but also the dosage recommendations of the manufacturers as subjective factors. The result is that the cephalosporins in the treatment of gram-positive infections (except enterococci) are almost equivalent. In the gram-negative field, there are marked differences which produce a different species-dependent therapeutic value for the various preparations. A fundamentally superior "universal" cephalosporin for all bacterial infections does not exist.


[Cephalosporins from the clinical viewpoint]. Cephalosporines are discussed as viewed by the clinician. For the clinician it is difficult to choose among the offer of manifold preparations the most promising. Antibacterial activity, pharmacologial properties and possible side-effects are shown and discussed in all Cephalosporines to make the clinic's choice easier.


Peptide synthesis with carbodiimide.

Peptide synthesis with carbodiimide. An isotope dilution assay for racemization during acid couplings is described and applied to carbodiimide-mediated synthesis. Using the coupling of t-BOC-L-phenylalamine with glycine derivatives as a model, racemization in solution falls in the 0.01 to  0.1% range and it is a function of reagent concentrations. A typical coupling using the solid phase method shows 0.03% racemate.


[Mutagenicity testing of irradiated laboratory animal feed in the 'host-mediated' assay with Salmonella typhimurium G 46]. Food irradiated with 10 MeV-electrons (dose: 4-5 Mrad) was tested for mutagenic effects, using the host-mediated assay. Irradiated 'Altomin' (a pelleted laboratory diet) was fed to mice for 9 to 44 days. After the 30th day of feeding, the animals additionally received irradiated glucose-solution instead of drinking water. Results did not indicate any mutagenic activity of the irradiated diet on the histidine-auxotroph strain of Salmonella typhimurium used in the assay.


The lung-colony assay: extension to the Lewis lung tumour and the B16 melanoma--radiosensitivity of B16 melanoma cells. Experiments are described which demonstrate that a lung-colony assay can be used to study the viability of unknown cell populations from the B16 Melanoma or the Lewis Lung Tumour. It is shown that the number of lung colonies formed can be increased by the addition of plastic microspheres to the injected cell suspension or by pre-irradiating the lungs of the recipient mice. The colony technique has been used to isolate melanotic and amelanotic cell-lines from the B16 Melanoma which were found to have different growth-rates. In vitro radiation survival curves for B16 Melanoma cells have also been established, and these have parameters in the usual range for mammalian cells.