Stimulation of WI-38 cell cycle transit: effect of serum concentration and cell density.

Flow microfluorometry has been used to characterize the effects of serum concentration and cell density on the initiation of cell cycle transit of stationary phase (G0) human diploid fibroblasts (strain WI-38). The concentration of serum used to stimulate these cultures had no effect on the time cells began appearing in S (the DNA synthetic period), nor on the synchrony with which they moved around the cell cycle. However, as the serum concentration increased, the fraction of the stationary phase population released from G0 increased. Cell density modulated the ability of serum to stimulate cell cycle traverse. For example, at a cell density of 1.81 X 10(4) cells/cm2, 78% of the population was sensitive to serum stimulation; whereas, when the density was increased to 7.25 X 10(4) cells/cm2, only 27% of the population could be stimulated. This effect of cell density on the serum response is not simply the result of changing the ratio of serum concentration to cell density, but appears to reflect a true modulation of the population's sensitivity to serum stimulation. These results are consistent with the interpretation that the primary action of serum is to determine the transition of cells from a non-cycling G0 state to a cycling state and that cell density determines the proportion of the population capable of undergoing this transition.


Control of normal differentiation of myeloid leukemic cells. XI. Induction of a specific requirement for cell viability and growth during the differentiation of myeloid leukemic cells.

Normal hematopoetic cells require the presence of a protein (MGI) in the appropriate conditioned medium (CM) for cell viability and growth and for differentiation to mature macrophages and granulocytes. Clones of myeloid leukemic cells have been established in culture (D+ clones) which require CM with this protein for differentiation, but not for cell viability and growth. It has been shown that these leukemic cells can be induced by CM to again require, like normal cells, the presence of CM for cell viability and growth. Induction of this requirement, which will be referred to as RVG, occurred before the D+ cells differentiated to mature granulocytes. Clones of myeloid leukemic cells (D- clones) that could not be induced to differentiate to mature cells, did not show the induction of RVG. The steroid hormones prednisolone and dexamethasone can induce some, but not all the changes associated with differentiation of D+ cells, Incubation with these steroids did not result in the induction of a requirement for these steroids for cell growth and viability. Studies with CM from different sources have shown, that all batches that induced RVG also induced differentiation of D+ cells and that both activities were inhibited after treating the CM with trypsin. It is suggested that the same protein (MGI) may be involved in both activities. Incubation of D+ cells with CM resulted in an increase in agglutinability by concanavalin A and this increase was maintained even in the absence of CM. This suggests, that the induction of RVG in D+ myeloid leukemic cells is associated with a change in the cell surface membrane.


Enhanced synthesis and extracellular accumulation of hyaluronic acid during stimulation of quiescent human fibroblasts by mouse epidermal growth factor.

The effect of mouse epidermal growth factor (mEGF) on the synthesis of glycosaminoglycans and glycoproteins by human fibroblasts has been studied. The addition of physiological concentrations (10(-9)M) of mEGF to quiescent cultures preincubated in the absence of serum was found to elicit an increased incorporation of 3H-glucosamine into the glycosaminoglycans and glycoproteins of both the cellular and extracellular fractions. Although the growth response to the factor, as measured by DNA replication, was minimal under these conditions as compared with the effect of serum, the mEGF-induced incorporation of glucosamine into these cellular constituents and into the extracellular glycoproteins was comparable to that elicited by serum shift-up. Serum, however, caused a significantly larger incorporation of glucoasimine into extracellular, acid-soluble glycosaminoglycans, which were shown to contain hyaluronic acid as the major component. As previously demonstrated, the growth response to mEGF can be enhanced several fold by an mEGF-binding arginine esterase, which is normally associated with the factor in vivo, and by ascorbate. The esterase was found to increase markedly the mEGF-induced incorporation of glucosamine into extracellular hyaluronic acid, while the addition of ascorbic acid did not significantly alter glucosamine incorporation.


Inhibition of sulfate transport in Ehrlich ascites tumor cells by 4-acetamido-4'-isothiocyano-stilbene-2,2'-disulfonic acid(SITS).

The effects of the nonpenetrating amino reactive reagnet 4-acetamido-4'-isothiocyano-stilbene-2-2'-dilsulfonic acid (SITS) on anion transport (sulfate, chloride, and inorganic phosphate) were investigated in Ehrlich ascites tumor cells. Short time exposure to SITS produces a reversible inhibition (92%) of sulfate transport. The kinetics of interaction suggest that reversibly bound SITS competitively inhibits sulfate transport, Ki = 3 X 10(-6)M. Incubation of tumor cells with SITS (1 X 10(-4)M) for longer periods of time results in a time dependent irreversible inhibition of sulfate transport which obeys first order kinetics. The rate coefficient for the inactivation process is 0.040 min-1. The kinetics of irreversible inhibition is best explained by the irreversible binding of SITS to the sulfate transport site, and therefore makes SITS a potentially useful probe for the quantiation of these sites in the tumor cell. The lack of effect of irreversibly bound SITS on either chloride or inorganic phosphate transport points to a specificity in the interaction of SITS with the tumor cell membrane, as well as indicating that an alternate pathway exists for the movement of these anions across the membrane.


Regulation of lipid synthesis from acetate in diploid fibroblast cultures -- variation with passage level and stage of cell growth.

Regulation of lipid synthesis from acetate in human diploid fibroblast cultures has been studied at various passage levels and at different stages of cell growth. When cultures were transferred to lipid free medium, a stimulation of [14C]acetate incorporation into lipid occurred within three to six hours after removal of exogenous lipid. In early passage cultures, this stimulation was observed whether cells were transferred to protein-free medium or medium supplemented with delipidized serum protein. However, in late passage cultures the presence of delipidized serum protein was required for the stimulation of lipid synthesis. When logarithmically dividing and stationary phase cultures were compared, the cultures in log phase showed stimulation of acetate incorporation into lipid in the presence or absence of delipidized serum protein, whereas in the stationary cultures the delipidized serum protein was required. When cultures were partially synchronized by a thymidine block, stimulation of acetate incorporation into lipid in the blocked cells only occurred in the presence of delipidized serum protein; in released cells stimulation occurred in protein free medium. When inhibition of lipid synthesis from acetate was compared in young vs. old or dividing vs. stationary cultures, however, no differences were observed. The data indicate the response of diploid fibroblast cultres to change in exogenous lipid is dependent on passage level and state of growth.


Endosymbiosis in amoebae: recently established endosymbionts have become required cytoplasmic components.

A strain of large, free-living amoeba that became dependent on bacterial endosymbionts which had infected the amoebae initially as intracellular parasites, was studied by micrurgy and electron microscopy. The results show that the infected host cells require the presence of live endosymbionts for their survival.Thus, the nucleus of an infected amoeba can form a viable cell with the cytoplasm of a noninfected amoeba only when live endosymbionts are present. The endosymbiotic bacteria are not digested by the host amoebae and are not themselves used as nutritional supplement. While the host amoebae are dependent specifically on the endosymbionts, the latter can live inside amoebae of different strains, indicating that their dependence on the host cells is not yet strain specific.


Argon laser microirradiation of mitochondria in rat myocardial cells in tissue culture. VII. fibrillation in ventricle and auricle cells.

Rat myocardial cells in vitro were irradiated in individual mitochondria with an argon ion laser microbeam. The contractile respone termed fibrillation in single and multicellular groups of both ventricle and auricle cells were compared. Specific correlations were made between fibrillation duration, the number of cells in the group, and the number of times the cells had fibrillated. Correlations were also made for the number of laser shots needed to induce fibrillation and the number of cells in the group. Another set of correlations were made between the pre-irradiation beat frequency and the beat frequency following recovery. Several differences and similarities of the above parameters were detected between auricle and ventricle cells. A comparison of the morphology and ultrastructure of auricle and ventricle cells also revealed significant differences.


Electron-microscope observations on cell nuclei in various tissues of a teleost fish: the nucleolus-associated monolayer of chromatin structural units.

Observations on stain uptake by thin sections through condensed interphase chromosomes in cells from epithelial and muscle tissue in kidney and intestine, and also in fibroblasts, show a distribution into DNA-rich and DNA-poor phases similar to that already described in cells from the connective tissue blood. In all the nuclei the nucleolus, when adjacent to the nuclear envelope, is separated from the inner membrane by a monolayer of chromatin structural units, similar to the monolayer enclosed on both sides by nuclear envelope, previously described in a wide variety of organisms. The data provide further support for the hypothesis that the condensed interphase chromosomes in eukaryotes are characterized by essentially similar structural units folded to form similar patterns. This hypothesis, regarding the higher order units, is consistent with data of others which show that histones and DNA fold to form similar repeating subunits in chromatin, irrespective of the base sequence in the DNA and the origin of the histones.


Ultrastructural site variations in mouse epidermal organization.

Mouse dorsal, ear, tail and foot epidermis are compared according to their tissue architecture and cell kinetics. Cell proliferation is expected in terms of the daily volume of keratin replaced. The stratum corneum may be organized into vertical columns of squames, which may have minimal overlap as in dorsum and ear, or maximal overlap as in tail. Individual areas are adapted to their function both in squame fine structure and rate of cell replacement. The surface keratin loss/replacement rate is at its highest in foot and tail, and lowest in ear and dorsum. Observations on hairless mouse dorsum are also included.


The blepharoplast of Marsilea: its de novo formation and spindle association.

Blepharoplast formation and function has been investigated by light and electron microscopy in developing microspores of the water fern Marsilea vestita. When single-celled microspores are hydrated they undergo a series of 9 cell divisions producing 7 sterile cells plus 32 spermatids. The blepharoplast, a densely staining, spherical inclusion (0-5-1-0 mum in diameter) interpenetrated by numerous lightly stained channels, is a precursor organelle for the production of the 100-150 basal bodies of the motile sperm. During development it arises twice in the dividing spermatogenous cells; first in telophase of the second to last or 7th division, and again in telophase of the next to last or 8th division. The blepharoplast first forms on the distal side of the nucleus during telophase of the 7th division but soon degenerates as the cell enters prophase and metaphase of the 8th division. The blepharoplast first forms on the distal side of the nucleus during telophase of the 7th division but soon degenerates as the cell enters prophase and metaphase of the 8th division. During telophase of the 8th division the blepharoplast again arises in the cytoplasm near a small indentation of the nuclear envelope. The forming blepharoplast consists of a sphere of lightly stained flocculent material within which 2 plaques separated by 40-50 nm arise. Each plaque possesses 2 dense layers (20 nm) separated by a light layer (10 nm). During subsequent development the distal layer of each plaque becomes progressively more densely stained. Light-staining channels appear within the accumulating dense material a-d 2 hemispherical blepharoplasts emerge. The blepharoplasts become spherical through continued growth and finally separate from each other as they move to the spindle poles during prophase of the last or 9th division. The blepharoplast appears to act as a microtubule organizing centre during formation of the spindle apparatus in prophase of the 9th division. While it remains at the pole throughout mitosis it does not continue to serve as the focal point for the spindle tubules. During metaphase-anaphase of the 9th division the blepharoplast swells, and the channels at its surface begin transforming into procentrioles and finally basal bodies. The results support the contention that basal bodies in Marsilea arise de novo, since no preexisting template such as a centriolar pinwheel is observed and sine the intermediates which initially occur are structurally dissimilar from a procentriole.


Ultrastructural and enzymic modulation of HeLa cells induced by sodium butyrate and the effects of cytochalasin B and colcemid.

Sodium butyrate causes HeLa cells to assume an elongated and jagged shape. Ultrastructurally this change is associated with the formation of bundles of microfilaments. Desmosomes were present between adjacent cells. No increase in microtubules was observed in the butyrate-treated cells. Butyrate induces an increase in the activity of 2 membrane-bound enzymes, alkaline phosphatase and 5'-nucleotidase; however, the activity of a third membrane enzyme, acetylcholine esterase, is reduced. The activities of the several other enzymes with different subcellular localizations are not significantly increased. Colcemid and cytochalasin B prevent or reverse the butyrate-mediated change in HeLa cell morphology and also partially inhibit the induction of alkaline phosphatase activity in these cells. The effect of cytochalasin B on alkaline phosphatase induction may be caused by a reduction in protein synthesis produced by this fungal metabolite.


Pollen-wall proteins: quantitative cytochemistry of the origins of intine and exine enzymes in Brassica oleracea.

Simultaneous coupling methods for detection of acid phosphatase and non-specific esterase produce a coloured reaction product that is quantitatively related to enzyme content in freeze-sectioned Brassica pollen and tapetal cells. The intine-located acid phosphatase has 2 periods of synthesis: the first in late vacuolate period, associated with the completion of deposition of the intine polysaccharides; the second during pollen maturation, apparently reflecting cytoplasmic synthesis, Esterase activity accumulates in the tapetal cells until dissolution at early maturation period, when there is a dramatic rise in pollen-wall esterase activity, reflecting the transfer from tapetum to exine cavities. These quantitative studies confirm the gametophytic and sporophytic origins of the intine and exine proteins.


Identification of Propionibacterium acnes and related organisms by precipitin tests with trichloroacetic acid extracts.

The serological identification of Propionibacterium acnes, P. granulosum, and P. avidum, using trichloroacetic acid extracts, is described. With antisera prepared against reference strains, the method has been tested on 142 strains recently isolated from human skin. All except two of the strains could be identified serologically, and there was excellent agreement between the serological results and the fermentation pattern of the strains. Two serological types of P. acnes and two of P. avidum were identified, but only one of P. granulosum.


Occurrence of Trichophyton megninii in Ontario. Identification with a simple cultural procedure.

Five strains of Trichophyton migninii were isolated in Toronto from four people. One patient probably was infected in Toronto since he had not visited the European endemic area. These were the first isolates made in Ontario; a requirement for L-histidine was determined. A cultural differentiation of T. migninii and T. rubrum is described. Attention is directed to the similarity of the metabolism for dextrose in Bromocresol purple milk dextrose agar by T. megninii and T. mentagrophytes which differs from T. rubrum.


Plaque assay of rabies virus on porcine kidney cell monolayers.

The ERA strain of rabies virus was adapted to growth on monolayers of a porcine kidney cell line (PK-2A). A reproducible plaque assay system was subsequently developed, which appears to be satisfactory for conducting plaque neutralization tests.


Photostatic copying of immunoelectrophoretic patterns with the use of color print filters.

Immunoelectrophoretograms can be replicated inexpensively, rapidly, and to scale by the use of color print filters on a photostatic copy machine. By this method additional copies are readily available for patient charts, permanent record, and any other use.


Comparison of urethral swabs, urine, and urinary sediment for the isolation of Chlamydia.

In 70 male patients with urethritis, Chlamydia was recovered from 18 urethral swabs (26%), 5 urinary sediments (7%), and 3 urine specimens (4%). All Chlamydia isolates were recovered from urethral swabs.


Retention of preservative levels of formaldehyde in desiccated biological products.

Concentrations ranging from 8 to 100% of the preservative level of formalin (37% formaldehyde solution) were retained by desiccated biologics, with most products retaining about 50% regardless of the amount originally present.


Retention of preservative levels of phenol in desiccated biological products.

Preservative levels of phenol retained by desiccated biological samples ranged from 3 to 73%, with highly proteinaceous samples retaining the most phenol.


Incidence of bacteremia in adults without infection.

To determine the incidence of bacteremia in healthy adults, blood cultures were obtained from 240 patients who had no demonstrable foci of infection. Five patients (2.1%) had positive blood cultures. Staphylococcus epidermidis was isolated from four patients and Alcaligenes faecalis from one. In each of these patients, the organism isolated probably represented contamination rather than bacteremia.


Clinical laboratory evaluation of the fifty-milliliter vacutainer blood culture tube.

The efficacy of the 50-ml vacutainer culture tube (Becton-Dickinson) for recovery of microorganisms from blood cultures was compared with laboratory-prepared blood culture media. The isolation of Bacteroides sp. was significantly higher in the 50-ml vacutainer culture tube whereas that of nonfermenting gram-negative rods and yeasts was higher in the in-house media. There was no significant difference in the recovery of all other organisms.


Study on the usefulness of hypertonic culture media.

Specimens from 300 patients were studied using five to nine aerobic and anaerobic culture media, including five that were hypertonic, Groups studied included fever of unknown origin, suspected endocarditis, endocarditis during therapy, bacteremia during therapy, abscess and cellulitis, presumed infectious arthritis, renal transplantation during rejection, collagen disease, sarcoidosis, lymphoma, and colitis. Isolates in hypertonic media were reverted to parent form by agar passage. In only 5% of these selected cases were organisms found in hypertonic, but not conventional, media that appeared on the basis of repeated isolation and/or serological studies to come from the patient. Nine of the 16 appeared to be of major significance. The two groups in which use of highly enriched, hypertonic media seemed most helpful were suspected endocarditis and undefined meningitis with negative cultures using standard media. The most effective of the hypertonic media used was 0.3 M sucrose in brain heart infusion with 20% horse serum. In most instances, the organism grew only in the hypertonic sucrose, and in most cases it appeared in conventional rather than aberrant form. Hypertonic media, especially 0.3 M sucrose, are of substantial helpin a small number of carefully selected cases.


Septicemia due to Neisseria lactamica.

Neisseria lactamica was isolated from the blood of a pediatric patient who had signs of septicemia and otitis media. Organisms morphologically resembling Neisseria, as well as gram-positive cocci, were seen on a Gram stain of fluid from the middle ear. It is hypothesized that the N. lactamica septicemia was secondary to infection of the middle ear by this organism.


Critical analysis of hypertonic medium and agitation in detection of bacteremia.

Over 18,000 clinical specimens collected in Vacutainer tubes with sodium polyanethol sulfonate were inoculated into modified Columbia broth (MCB) with and without 10% sucrose. The effects of venting and shaking on recovery were studied. The volume of the blood had a definite effect on the recovery rate. When inoculum size was held constant, recovery of aerobic and facultative organisms was maximal in vented and shaken bottles; the presence of sucrose had no demonstrable effect, recovery of anaerobes was maximal using an unvented bottle incubated under stationary conditions; a significantly greater recovery of facultatives and a marginally greater recovery of anaerobes was obtained with the hypertonic formulation. We conclude that a hypertonic formulation of MCB offers no advantage in the recovery of anaerobes but is of value in the recovery of facultatives and anaerobes. It is recommended that blood cultures be routinely inoculated into isotonic MCB and then vented and shaken for at least 4 hours, and hypertonic MCB incubated without venting or shaking.


Method for phage typing group A type 49 streptococci.

A method of phage subtyping group A type 49 streptococci is described. The method is similar to that used for phage typing staphylococci, except that lysates obtained by induction with mitomycin C rather than propagated stock phages were used. Five type 49 strains were used as phage donors. Seventy-two strains of type 49 streptococci isolated from 10 worldwide sources were examined by this method. Among these strains, five distinct subtypes (I through V) could be distinguished on the basis of their lytic patterns. Only a few of the type 49 strains could not be classified into one of these phage subtypes (6% using 100 X routine test dilution). Strains from a single source were generally homogeneous with respect to their phage subtype. The method proved useful in discriminating between type 49 strains isolated from different geographical sources and from the same place in different years. Studies in progress suggest that it may be useful for subtyping other strains of special epidemiological interest, such as strains of other serological types associated with nephritis.


Bacteriocin (klebocin) sensitivity typing of klebsiella.

High-titer preparations of Klebsiella bacteriocins (klebocins) were obtained by using mitomycin to induce standard strains of Klebsiella. Of 296 clinical isolates of Klebsiella, 67% could be typed on the basis of their sensitivity to klebocins. The method proposed in this paper affords a standard basis for the further development of klebocin typing as a suitable procedure for hospital laboratories concerned with epidemiological investigations of hospital-associated infections. Evidence is also provided to show that high-titered klebocin typing can be used in conjunction with biochemical typing to provide a sensitive epidemiological marker for Klebsiella.


Primary isolation of Mycobacterium avium complex-serotype 6 on blood agar.

Mycobacterium avium complex-serotype 6 was isolated in pure culture on blood agar plates from inocula taken from the heart blood, lungs, liver, kidneys, and spleen of a naturally infected captive female opossum (Didelphis marsupialis virginiana). Repeat cultures from stored tissues and transfer of colonies from original blood agar plates revealed that the mycobacterium grew on tryptose, brilliant green, eosin-methylene blue. Sabouraud glucose, and mycobiotic agar plates and in Fletcher leptospira medium. The cultural, biochemical, and serological characteristics of the test isolate were compared with other mycobacteria. This is the first report to describe the primary isolation of a serotype from the M. avium complex from an animal species on blood agar or in Fletcher broth. In addition, this is the second documented report describing the isolation and identification of a mycobacterial species from the American opossum.


Caprylate-thallous agar medium for selectively isolating Serratia and its utility in the clinical laboratory.

A defined agar medium (hereinafter designated caprylate-thallous [CT5 agar) containing 0.01% yeast extract, 0.1% caprylic (n-octanoic) acid, and 0.025% thallous sulfate is highly selective for all Serratia species and effectively discriminates against most non-Serratia strains likely to be in the same habitats. The selectivity of CT agar is demonstrated by the very high efficiency of colony formation (mean, 80.7% of that on a nonselective complex medium) on CT agar by known Serratia strains and the very low efficiency of colony formation (close to zero) on CT agar by bacterial strains known not to be Serratia. The utility of this medium in actual clinical laboratory practice is demonstrated by the more rapid and higher recovery of Serratia on this selective medium as compared to conventional procedures of in-tandem runs of 513 consecutive urine, feces, and sputum specimens. Pigmented and nonpigmented Serratia strains deliberately added to fecal specimens can be selectively and quantitatively recovered on CT agar. CT agar compares favorably with, or in some cases is an improvement over, other selective media which have been recommended for isolating Serratia. This selective CT agar medium could be quite useful in ecological surveys, especially those related to hospital-acquired infections.


Recent experience with the complement fixation test in the laboratory diagnosis of rickettsial diseases in the United States.

Sera from patients suspected of having rickettsial infections were tested in the complement fixation test with antigens prepared from the rickettsiae of Rocky Mountain spotted fever (SF), rickettsial pox (RP), murine typhus, epidemic typhus, and from Rickettsia canada (RC). Eight units of antigen were used in all cases and two units in man. Only those patients with antibody titers of 1:16 or higher were included in the study. Largely on the basis of comparative titers, the patients were divided into two groups: 102 with SF and 35 with infections by one of the members of the typhus group. The antibody titers were higher with SF antigen than RP antigen in 72% of the SF patients, and in only two SF patients was the RP titer higher, and then by only one tube (twofold dilution). There seemed little advantage in including the RP antigen in the battery of rickettsial antigens. Cross-reaction with at least one of the typhus antigens was observed in the sera from 64% of the SF patients. It was extensive enough to be confusing (within one tube) in 17% with eight units of antigen, but the differentiation was more distinct with two units of antigen. The cross-reaction with typhus antigens was as frequent in children with SF as it was in adults; thus, it is unlikely that these cross-reactions resulted from previous typhus vaccination. The serological differentiation between murine typhus and epidemic typhus was frequently difficult, but the epidemiological background was distinct. Five patients had higher titers to RC antigen, and four of these may possibly have had RC infections.


Rapid method for the detection and identification of mycolic acids in aerobic actinomycetes and related bacteria.

A rapid method for the identification of lipids characteristic of the genera Corynebacterium, Mycobacterium, Nocardia, and the "rhodochrous group" has been developed. Modifications of previously described methods make this procedure suitable for use in the clinical laboratory. Thin-layer chromatography is used to demonstrate the presence of the lipid characteristic of Nocardia spp. (type A) in some corynebacteria, nocardias, and members of the "rhodochrous group." Precipitation in ether and ethanol is used to demonstrate the presence of mycobacterial mycolic acids. Since this procedure can be carried out in less than 2 days and the lipids are extracted from the same batch of cells grown for diaminopimelic acid and whole-cell sugar analyses, it can readily be added to the battery of tests performed in reference laboratories that deal with aerobic actinomycetes and related bacteria.


Use of Mueller-Hinton agar as amylase testing medium.

Readily available Mueller-Hinton agar was used to detect starch hydrolysis by Streptococcus bovis.


Electron microscopic autoradiographic studies of gliogenesis in rat optic nerve. I. Cell proliferation.

Electron microscopy and 3H-thymidine autoradiographic techniques were used to study the fine structure of proliferating cells in developing rat optic nerve. Before the closure of the optic canal almost all of the cells incorporating radioactive thymidine are ventricular cells, but after closure (16 days of gestation) the vast majority are differentiating astroblasts or oligodendroblasts. Labeled astroblasts show a range in their degree of differentiation; some cells lack 90 A cytoplasmic filaments while others have glial filaments and abundant cytoplasmic organelles. In contrast to astroblasts, all of the labeled oligodendroblasts are in the early stages of differentiation. The proliferation of oligodendroblasts starts at five days postnatal, approximately a day or two before the onset of myelination.During myelinogenesis a few of the labeled oligodendroblasts show presumptive connections to myelin sheaths. Microglial cells do not appear to play a major role in gliogenesis since they form less than 2% of all the labeled cells. The results of this study indicate that astroblasts and oligodendroblasts, rather than undifferentiated glioblasts, are the major source of macroglia. The finding that proliferating glia are in the processof differentiation agrees with recent studies which show that differentiated cells can divide.


Electron microscopic autoradiographic studies of gliogenesis in rat optic nerve. II. Time of origin.

The time of origin for astrocytes and oligodendrocytes in rat optic nerve was studied by 3H-thymidine autoradiographic techniques similar to those used in dating the time of origin for neurons. This study shows that astrocytes are formed throughout late embryonic and all of postnatal development, while oligodendrocytes are generated only during the postnatal period. A few astroglia undergo their final cell division as early as 15.5 days of gestation, but most astrocytes are not generated until the first week of postnatal development. Although the final cell division for more than half of the astrocytes takes place before the end of the first postnatal week, fully mature, fibrous astrocytes are not observed in electron micrographs until after 14 days of age. This time lag implies that the differentiation of these early generated cells takes place gradually over a 2-to 3-week interval. Oligodendroglia begin their final division a day or two before the onset of myelination (6-7 days postnatal), but the vast majority are produced during the period of myelinogenesis. After almost all of the axons have been myelinated, oligodendrocytes are still being generated in small numbers. These late forming cells are generally less differentiated in appearance than those formed earlier; this suggests that the degree of differentiated of oligodendrocytes may be dependent upon the number of axons available for myelination. As with astrocytes, oligodendrocytes show a lag of about two weeks from the time of final cell division until they transform into morphologically differentiated cells. In transverse sections of the optic nerve heavily labeled neuroglia are randomly distributed, indicating there are no temporal-radial gradients for the individual cell types. This observation taken together with the other information obtained from the present and the previous study (Stoff et al., '76) strongly suggest that the factors controlling gliogenesis are different from those governing neuronogenesis.


The influence of phenobarbital on biotransformation of 25-hydroxycholecalciferol.

Altered vitamin D metabolism has been implicated as a cause of anticonvulsant-induced osteomalacia. Previous studies have demonstrated accelerated biotransformation of vitamin D3 to 25-hydroxycholecalciferol (25-OHD3). However, it is not known whether the 25-OHD3 is metabolized to 1,25-dihydroxycholecalciferol (1,25-(OH)2D3), the tissue-active metabolite of vitamin D3. This study using rats was undertaken to study the influence of phenobarbital on the biotransformation of 25-OHD3 to 1,25-(OH)2D3. The disappearance rate of 25-OHD3 was virtually the same in the pheno-barbital-treated group (re = 0.0615 pmole/min) and the control group (re - 0.0549 pmole/min). Similarly, the appearance rate of 1,25-(OH)2D3 was virtually the same in the treated group (ra = 0.0133 pmole/min) and the control group (ra = 0.0134 pmole/min). These data suggest that phenobarbital does not affect the biotransformation of 25-OHD3 to 1,25-(OH)2D3. The implication of this observation is that altered vitamin D metabolism does not account for phenobarbital-induced osteomalacia.


The pharmacologic effect of aerosolized terbutaline sulfate in exercise-induced bronchospasm.

Bronchospasm can be induced in asthmatics when exercised according to a multistage branching treadmill protocol that allows them to achieve 80 per cent of maximal age-predicted heart rate. This degree of exercise is usually achievable and allows inducible bronchospasm to occur. This present study was undertaken to investigate the effect of terbutaline sulfate aerosol in exercise-induced broncho-spasm. Asthmatics were exercised to 80 per cent of their maximal heart rate, and FEV1.0 and MMEFR were assessed while standing using a Jones Pulmonar II waterless spirometer at 5, 15, and 30 minutes after exercise. After a standard rest period defined by a return to baseline of FEV1.0, MMEFR, and heart rate for 30 minutes, the subjects were administered either 0.50 mg aerosolized terbutaline sulfate or placebo and then exercised again. The pulmonary function parameters were again recorded after this exercise. Preterbutaline and postplacebo exercise resulted in a significant reduction in FEV1.0 and MMEFR, while after treatment with terbutaline not only did bronchospasm not occur but bronchodilation occurred (P less than 0.01). Inhaled terbutaline appeared to normalize the exercise tolerance of the asthmatics and restore physiologic pulmonary airway conductance according to the parameters of FEV1.0 and MMEFR.


The effect of cycloserine on pyridoxine-dependent metabolism in tuberculosis.

Measurements were made of urinary tryptophan metabolites of 13 tuberculosis patients in order to reveal characteristics of pyridoxine-dependent metabolism before and during cycloserine treatment. The abnormally high level of xanthurenic acid excretion in untreated patients suggests a decreased availability of pyridoxal phosphate related to the disease process. Although plasma cycloserine levels were kept high once therapy began, xanthurenic acid excretion before and after tryptophan load became progressively more normal as symptoms diminished. This observation suggests that the convulsions which may sometimes accompany cycloserine administration are not due to a direct pyridoxine antagonism by the drug. Throughout the study, no significant changes in 5-hydroxyindoleacetic acid excretion were observed. Presumably, the metabolic pathway of serotonin is unaffected by tryptophan loading, cycloserine administration, or the apparent pyridoxine depletion associated with tuberculosis.


Recovery from deafferentation by cricket interneurons after reinnervation by their peripheral field.

The physiological and morphological recovery of cricket (Acheta domesticus) abdominal giant interneurons following varying periods of deafferentation and subsequent regeneration was examined. The principal afferent input to two identified interneurons was removed by surgically ablating an abdominal sensory appendage, the cercus. Deafferentation restricts the growth of the dendrites of the medial giant interneuron. Reinnervation by the peripheral sensory field leads to recovery of the dendrite length which is dependent on the time allowed for recovery. The response properties of the reinnervated neurons never completely recovers irregardless of how short the period of deafferentation. Reinnervated neurons respond more weakly to standard tones than do control neurons. This is due in part to faciliation of an inhibitory synaptic input which is activated by the control cercus. The results suggest that the balance between the excitatory and the inhibitory synaptic inputs to these interneurons is irrevocably altered by brief periods of deafferentation early in life.


Cells of origin of entorhinal cortical afferents to the hippocampus and fascia dentata of the rat.

The pathway from the entorhinal cortical region to the hippocampal formation has previously been shown to be comprised of two sub-systems, one of which projects predominantly to the ipsilateral fascia dentata and regio inferior of the hippocampus proper, and a second which projects bilaterally to regio superior. The goal of the present investigation was to determine if these two pathways might originate from different cell populations within the entorhinal area. The cells of origin of these entorhinal pathways were identified by retrograde labeling with horseradish peroxidase (HRP). Injections which labeled the entorhinal terminal fields in both the fascia dentata and regio superior resulted in the retrograde labeling of two populations of cells in the entorhinal area. Ipsilateral to the injection, HRP reaction product was found in the cells of layer II (predominantly stellate cells) and the cells of layer III (predominantly pyramidal cells). Contralateral to the injections, however, the reaction product was found almost exclusively in the cells of layer III. With selective injections of the entorhinal terminal field in regio superior, only the cells of layer III were labeled, but these were labeled bilaterally. Selective injection of the entorhinal terminal field in the fascia dentata, however, resulted in the labeling of cells of layer II, but not of layer III, and these cells of layer II were labeled almost exclusively ipsilaterally. A very small number of labeled cells in layer II were, however, found contralateral to the injection as well. No labeled cells were found either in the presubiculum or parasubiculum following injections of the hippocampal formation. These cell populations were found capable of retrograde transport of HRP, however, since cells in both presubiculum and parasubiculum were labeled following HRP injections into the contralateral entorhinal area. These results suggest that the projections to the fascia dentata originate from the cells of layer II, while the projections to regio superior originate from the cells of layer III of the entorhinal region proper. The very slight crossed projection from the entorhinal area to the contralateral area dentata probably originates from the small population of cells in layer II which are labeled following HRP injections in the contralateral area dentata.


Method factors associated with assessment of child behavior: fact or artifact?

A sample of 64 phobic children between the ages of 6 and 16, with their general population matched controls, were rated by parents on the Louisville Behavior Check List and Louisville Fear survey and by School Behavior check list. The analysis of ratings by factor analysis revealed three instrument factors while a multiple discriminant analysis elicited a multidimensional phobic trait that crossed observers, situations, and instruments. The results are discussed in terms of the implications for multivariant analysis of behavior change as well as for classification of psychopathological disorders of childhood. The thesis is advanced that instrument factors elicited by factor analysis in psychotherapy change studies are statistical artifacts and that a multiple discriminant analysis is a more appropriate technique for the study of change as well as for classification.


The effects of biasing information on behavioral observations and rating scales.

The effects of biasing information on behavioral observations and rating scales were studied. Forty-one undergraduate students trained in making reliable behavioral observations were given differential expectations concerning the activity level of a target child. They then viewed videotape recordings of that child and tallied frequency counts of six behavioral categories simultaneously. In addition, subjects completed postexperimental rating scales composed of specific, identifiable behaviors in regard to the target child. Results indicated that, for the most part, neither the behavioral observations nor the rating scales were significantly affected by the biasing information. It is suggested that rating scales constructed of items as discrete and readily identifiable as those of behavioral observation measures may prove resistant to biasing effects.


The cheating and intention of a partner as determinants of evaluative decisions among juvenile offenders.

Male juvenile offenders were asked to evaluate the work of an experimental partner (confederate) who completed a work task four times. The work partner cheated on two trials and did not cheat on two trials while working for a charity (one cheat and one noncheat trial) or for selfish gain (one cheat and one noncheat trial). It was found that subjects rated the work done on cheating trials lower than work done on noncheating trials. In addition, subjects shared less reward money on cheating trials relative to noncheating trials. However, the intentions of the conferate, defined as the reward recipient, failed to affect subjects' judgments. These results were described as supporting previous studies which have indicated that delinquent fail to account of others' intentions when making moral judgments.


An experiment in social engineering in serving the families of predelinquents.

One hundred and two predelinquent adolescents were randomly assigned to one of two conditions: an experimental condition in which behavioral contracting services were offered both at home and in school or a control condition in which clients were told that they could not be accomodated by project therapists and were informed about the possibility of their receiving treatment in other programs in the community. Most of those who were assigned to the control condition did not avail themselves of services in these other programs. Results indicate that, relative to the controls, the behaviorally treated youth scored small but statistically significant improvement relative to the controls on five measures: ratings of school behavior by the person who originally referred them for service, by their teachers, by their mothers, and by their fathers, and ratings of their parent-child interactions as measured by their mothers. Gains were also reported in mother's ratings of their children's behavior at home. The failure of significant gains to be made in two measures of school performance--grades and attendance--is explained by the failure of the treatment techniques to over-come an age-related deterioration in these areas found among most children in the participating schools. The failure of fathers to find significant improvement in their relationships with the referred adolescents is viewed as a consequence of unanticipated changes in the intrafamilial balance of power resulting from contracting services. Finally, a subsidiary analysis showed that the contracting service appeared to offer the greatest gain to the demographically defined sub-groups of youths who are among the population-at-risk in many juvenile courts.


A motivational environment for behaviorally deviant junior high school students.

A contingency management program was established in a junior high school to better manage and educate students having histories of severe misconduct. School administration selected 46 eighth-graders having multiple suspensions for misbehavior. Students (N = 32) in two of the schools remained in traditional programs, serving as controls, whereas students (N = 14) in the third school participated in a token reinforcement program. Reinforcers provided in the afternoon were contingent upon achievement and discipline during morning academic periods. Home-based reinforcers were established to support school behavior. Compared with the control group, significant reductions in negative school behavior as well as greater increases in academic achievement were obtained for the treatment group, thus supporting the efficacy of contingency management for adolescents school misbehavior.


Time-out with children. Effects of an explanation and brief parent training on child and parent behaviors.

The purpose of the present study was twofold: (a) to determine the effect of an explanation prior to or after time-out on child compliance and on child disruptive behavior during time-out and (b) to determine the effect of brief parent training in time-out on child and parent behaviors. Thirty-two mother-child pairs served as subjects and were assigned to one of the following four groups: control, time-out only, explanation prior to time-out, or explanation following time-out. Each mother-child pair was observed for one session under pretraining, training, and posttraining conditions. The results indicated that time-out significantly increased compliance but the addition of an explanation did not further alter the effectiveness of time-out. Training in the use of time-out decreased the incidence of maternal interruptions but did affect maternal responses that were not trained. Finally, following brief time-out training for noncompliance, the mothers used the procedure only 50% of the time following noncompliance.


Overselectivity, developmental level, and overtraining in autistic and normal children.

Thirteen autistic children were compared to 13 normal children matched to them in mental age, on performance of a visual discrimination task. Form, color, and size were relevant and redundant cues. The groups did not differ significantly in mean trials to reach criterion or in breadth of learning, and both groups increased their breadth of learning after 50 trials of overtraining. Form was preferred to color and size by both autistic and normal children. Within each group, rank on mental age was highly correlated with rank in breadth of learning. Verbal and nonverbal autistic children did not differ in breadth of learning or in dimensional preference. Even nonverbal autistics equaled the performance of their normal controls. Our results suggest that overselective attention is better understood as part of a general developmental lag in cognition in autistic children than as a specific deficit underlying psychotic behavior.


Behavior problems of Anglo and Mexican-American children.

The present investigation compared Anglo (N = 1,999) and Mexican-American children (N = 192) on the four factors of the Behavior Problem Checklist (BPCL). Teachers provided general information and BPCL ratings. The Anglo children manifested more problems on three of the factors than did the Mexican-American children. Significant interactions of grade, sex, and social class with ethnic group were revealed for two of the factors.


Incidence of exercise-induced asthma in children.

The incidence of exercise-induced asthma (EIA) was studied in 134 asthmatic and 102 nonasthmatic atopic children and compared to that in 56 nonatopic children. Pulmonary function tests measuring forced vital capacity (FVC) and 1-sec forced expiratory volume (FEV1) were performed on each child prior to and serially for 20 min following free running exercise. The incidences of EIA among the asthmatic and atopic nonasthmatic children were 63% and 41%, respectively. This phenomenon is widespread among allergic children and cannot be accurately predicted from the history. A simple and easily performed outpatient procedure is described for the diagnosis of EIA.


Diagnosis of food allergy by the radioallergosorbent test.

The radioallergosorbent test (RAST) was positive in 52.5% of 200 sera representing 200 food hypersensitivities from 108 patients with a history of definite immediate-type reactions to foods. Corresponding prick test was performed for 170 of the sera. The latter test was positive in 70%, the RAST was positive in 52%, and iether prick test or RAST was positive in 74%. It is concluded that the RAST is positive less frequently than the prick test in the diagnosis of immediate-type food allergy in clinically sensitive patients, but that the performance of both tests increases slightly the possibility of confirming the diagnosis. However, the RAST is useful for further evaluating positive prick tests with foods that do not correlate with clinical hypersensitivity.


Reactivation of tuberculosis: a problem of aging.

Six examples demonstrate the reactivation of pulmonary tuberculosis in elderly patients. In 3 cases the tuberculosis became reactivated 25, 47 and 51 years respectively after the initial diagnosis. In aged patients with pulmonary symptoms and abnormal x-ray findings, the presence of reactivated tuberculosis should always be considered. Sometimes steroid or x-ray therapy for other disease may be the reactivating factor.


Femorotibial bypass in the elderly for revascularization of the severely ischemic lower extremity.

In the elderly, revascularization of a severely ischemic lower extremity with occlusion of both femoral and popliteal arteries often can be achieved by surgical construction of a distal bypass to the tibial or peroneal arteries. An aggressive diagnostic and therapeutic approach is necessary, in an attempt to prevent recourse to primary amputation. Femoroperoneal or femorotibial bypass can be performed safely and is recommended in elderly patients with advanced ischemia of a lower extremity with absolute indications for surgical intervention, e.g., gangrene, gangrenous ulceration or rest pain. A significant number of limbs can be salvaged by this method. Although the mortality rate in the older age groups is predictably higher, the overall rate for this operation compares favorably with that for primary amputation.


Physician's attitudes toward the ill aged and nursing homes.

A survey of physicians in private practice (exclusive of pediatricians) was conducted in a medium sized suburban city in the New York metropolitan area, to determine whether physicians' attitudes toward the ill aged and nursing homes were predictors of the quality of medical care available to area nursing home patients. Questionnaires were circulated to 302 practitioners. Of the 28 percent who responded, 32 percent were psychiatrists, 15 percent primary care physicians and 8 percent orthopedists. Physicians felt competent to manage the ill aged, although 50 percent had had no significant degree of exposure to geriatric medicine in their medical education, and 70 percent of the primary care group had had none. Primary care and older physicians were more likely to treat patients in nursing homes. Almost 40 percent viewed the nursing home as a place to die. Although 85 percent studied that physicians should be involved in the nursing home displacement process, only 21 percent believed that they continued to be in charge of their patients after placement. The findings demonstrate generalized medical disinterest in the care of ill aged patients in institutions. The persons responsible for awarding government grants and those involved in planning medical school curricula should pay more attention to the needs of the chronically ill aged.


Effect of size on the function of three day hospitals: the case for the small unit.

The work load of three day hospitals attached to active departments of Geriatric Medicine was studied over a one-year period. A significant difference in the functional efficiency of the three units was noted. The smaller 12-place and 15-place units provided effective short-term rehabilitation, whereas the 28-place unit did not. Size seemed to be the major factor in the success or failure of a given unit. This study throws serious doubts upon the wisdom of a policy of building large day hospitals with all their accompanying problems of staffing and transport. An effective unit for out-patient rehabilitation can be provided with 10 to 15 places per day if the staff members are chiefly remedial therapists. In the districts studied, a provision of 0.5 place per 1,000 of the population aged 65 or older would have been sufficient to meet the perceived need for rehabilitation. However, this would not have been sufficient to satisfy the requirement for out-patient investigation and medical treatment on a day basis.


Use of the Holter electrocardiographic monitor in the diagnosis of transient ischemic attacks.

The diagnosis of transient ischemic attacks usually is based on an accurate history and physical examination (with the finding of carotid bruits), a complete neurologic examination, a brain scan, an encephalogram and an angiogram. Recently the Holter technique for continuous 24-hour electrocardiographic monitoring has been used as a diagnostic aid in these problem cases. Severe bradycardia, sick sinus syndrome, tachy-brady syndrome, recurrent tachycardia and heart block have been found to be the cause of the central-nervous-system symptoms in aged atherosclerotic patients. Correction with anti-arrhythmia drugs and pacemakers, with or without carotid endarterectomy, has been highly successful. Data are presented on 204 patients tested by Holter monitoring over a 2.5-year period in a small community hospital; 6 cases are illustrated in detail. The Holter monitor is a valuable tool in evaluating the patients with cerebrovascular insufficiency.


Behavioral change in the chronicity pattern of dementia in the institutional geriatric resident.

To determine whether behavioral change as a result of environmental stimulation can be reported in individualized terms for the aged with organic brain syndrome or senile dementia, a program of multimodal group interactive therapy was started in an extended care facility. During a 1.5-year period, the study included 108 residents, whose average age was 84 years. The technique used was a combination of modified Reality Orientation, Remotivation, Activity and Milieu therapies. Thirteen groups residing in different wings of the facility were treated for 55 sessions by 11 activity specialists and 2 elderly volunteers under the supervision of the consulting clinical psychologist. Data were collected on emerging behavioral changes with respect to awareness, attention, verbalized ideation, voice quality, speech quantity, sociability, self-care, quality of affect, energy level, expressed somatic concern and physical movement. Of the initial 108 residents, 80 had chronic brain syndrome of varying severity or senile dementia. During treatment, 35 of these 80 residents (44 percent) were improved, 17 (20 percent) were unchanged, 14 (18 percent) became worse, and 14 (18 percent) remained unrated. The results warrant a redifinition of the psychotherapeutic approach. It should include factors other than the severity and chronicity of disease in determining the prognosis for behavioral change in geriatric institutional residents.


Inheritance of 6-phosphogluconate dehydrogenase variants in Culex pipiens quinquefasciatus Say.

The formal genetics of two variants of a 6-phosphogluconate dehydrogenase locus of the adult mosquito, Culex pipiens quinquefasciatus, was analyzed by starch gel electrophoresis. These two alleles, PgdF and PgdS are codominant, and autosomally inherited, and appear to be linked to the Pgm locus by 27 crossover units in linkage group III.


Selection effects on a linked neutral locus.

Selection changes the frequency of alleles at a linked locus as well as at those under selection if the population is not in linkage equilibrium. The magnitude of this frequency change depends on the tightness of the linkage, the selection intensity, and the deviation from linkage equilibrium. Allowing a population to mate randomly without selection brings the population closer to linkage equilibrium. This decreases the effect of selection on allelic frequencies at a linked neutral locus. However, if linkage is very tight it can take many generations to make a large difference in the effect of the linked locus. The loss due to undesirable changes in allelic frequencies at linked loci when the population is not in linkage equilibrium must be weighed against the time and effort saved by beginning intense selection for the primary trait in an early generation. Effects of selection intensity, linkage intensity, and delayed selection on changes in allelic frequency at a neutral linked locus are demonstrated.


Inheritance of limber legs in Jersey cattle.

Pedigree information from 105 cases of limber leg Jersey calves indicate that this semilethal abnormality is inherited as a simple autosomal recessive. The increased incidence of this abnormality in recent years suggests a rise in the gene frequency, probably due to the popularity and widespread use of a carrier bull Favorite Commando, his daughter Marlu Milady, and many of their descendants. Steps to control this abnormality have been taken by the American Jersey Cattle Club.


A further study of the inheritance of racing performance in thoroughbred horses.

In an analysis of handicap ratings of 1158 3-year-old thoroughbred racehorses, estimates were obtained for the effects of sex differences, the correlation between mates, the effectiveness of selection in males and females, and the heritability of racing performance. The results agreed closely with those of a previous study, and indicated a heritability of about 0.35 to 0.40, some assortative mating, and highly effective selection for performance.


Immunoelectronmicroscopic localization of IgA in skin of patients with dermatitis herpetiformis.

Ultrastructural localization of in vivo-bound IgA in the skin of patients with dermatitis herpetiformis was determined by using a modified peroxidase-antiperoxidase multistep method. Three types of reaction product deposition are seen. The most common type of reaction product deposition, that which is identified by direct immunofluorescence as the speckled type of IgA deposit, shows up as clumps and yarnlike fibrils. The second type of IgA deposition, which is a linear band by direct immunofluorescence, appears to be associated with anchoring fibrils. The third of IgA deposition, which is also linear by immunofluorescence, is confined to the lamina lucida.


Ultrastructural localization of immunoglobulins in skin of patients with dermatitis herpetiformis.

A multistep immunocytochemical method utilizing horseradish peroxidase as an immunologically bound marker was used to detect and localize IgA deposits in skin of patients with dermatitis herpetiformis at the ultrastructural level. IgA was found in the upper papillary dermis forming irregular aggregates in seemingly haphazard distribution. These aggregates were associated with microfibrillar bundles and with the microfibrillar component of the elastic tissue. IgA was also detected on anchoring fibrils, but showed no topical relationship to the basal lamina which was always spared. This finding indicates that basal lamina components do not serve as target sites for the immunologic reaction occurring in dermatitis herpetiformis. The selective affinity of IgA deposits to microbfibrillar bundles may be relevant to the hypothesis that the skin pathology in dermatitis herpetiformis is caused by circulating gluten-antigluten complexes, trapped in the skin by reticulin-bound antireticulin antibodies which cross-react with gluten.


Black bands on a starch gel stained for serum esterases in red grouse.

From 36 crosses of red grouse (Lagopus l. scoticus), 136 offspring were coded for a new locus (Rg). The three possible phenotypes were probably determined by two codominant alleles at an autosomal locus.


Prolonged ultraviolet light-induced erythema and the cutaneous carcinoma phenotype.

A considerable amount of evidence exists in support of the role of ultraviolet radiation as a major etiologic factor in human skin cancer, both melanoma and carcinoma types. On the basis of epidemiologic studies a phenotype has been described which helps to identify the persons who are more susceptible to skin cancer. In an attempt to further define this population, patients with cutaneous carcinoma and a normal control group were exposed to artificial ultraviolet light (UVL) and the erythema and tanning responses of each group were measured over a 21-day period. UVL-induced erythema was prolonged in a significantly higher percentage of patients with skin cancer than in control patients, lasting two to three weeks after single exposures to 6 and 8 times the patient's minimal erythema dose. The presence of prolonged erythema correlated with this history of previous skin cancer but did not correlate with other established risk factors for cutaneous carcinoma, i.e., fair skin, light hair and light eyes, easy sunburning and poor tanning, and Celtic ancestry. Prolonged erythema following UVL radiation may therefore represent an additional risk factor and help to identify the skin cancer-susceptible population.


Chromosome complexity in the Indian long-tailed tree mouse.

The chromosomes of the Indian long-tailed tree mouse, Vandeleuria oleracea nilagirica, have been studied in both sexes and the diploid number established to be 2n = 28 with a simple XX/XY sex chromosome mechanism. The sex chromosome polymorphism encountered in different taxa is discussed in the light of the present findings.


Randomized trial of a hexachlorophene preparation and P-300 bacteriostatic soaps.

Twenty-six patients undergoing chemotherapy for acute leukemia on a protected environment-prophylactic antibiotic program were randomly assigned to bathe with either P-300 or a 3% hexachlorophene preparation. Both agents substantially reduced the microbial flora of the skin, but the latter preparation was slightly more effective. The bacteriostatic compounds in both preparations were absorbed through the skin but no toxic effects were observed.


8-methoxypsoralen and UVA promote sister-chromatid exchanges.

8-Methoxypsoralen (8-MOP) and long-wave UV light (UVA 365 nm) are now being use to treat vitiligo and psoriasis. Cultured human lymphocytes exposed to these agents in vitro show an increased infrequency of sister chromatid exchanges, related to 8-MOP-DNA photoadducts. Although these data raise questions regarding the biologic consequences of this therapeutic regimen, it is unknown whether 8-MOP and UVA cause mutations or extracutaneous somatic cell recombinants in vivo. Exchanges may represent cellular repair of DNA damage.


The variation of recurrence risks with penetrance for isolated cases of autosomal dominant conditions.

The relationship between the penetrance and the recurrence risk for sibs of isolated cases of autosomal dominant conditions with incomplete penetrance is presented.


Renal glycosuria due to gentamicin in rabbits.

Gentamicin sulfate was given to rabbits for four weeks in two dosage regimens, either 4 or 8 mg/kg subcutaneously twice a day. Two of the seven animals in each group regained nearly normal renal function after a transient rise in the level of serum creatinine, but the remainder developed severe renal failure leading to the death of all but one of these rabbits. None of the animals showed significant hyperglycemia, buy glycosuria appeared in all and was significantly correlated with the extent of renal damage. These findings demonstrate gentamicin-induced derangement of proximal tubular function and thus amplify the previously demonstrated histologic changes due to the drug. When the drug was administered twice daily, there was marked nephrotoxicity in rabbits given dosages of gentamicin only slightly higher than those employed clinically.


Hepatitis B surface antigen in environmental samples from hemodialysis units.

For assessment of environmental contamination with hepatitis B surface antigen (HBS Ag), the presence of blood and of HBS Ag was determined in samples from two completely separated hemodialysis units. Samples from mucosal and skin surfaces and samples of saliva from patients were also investigated. The surfaces of both units were divided into epidemiological categories to determine the distribution of environmental contamination with HBS Ag. HBS Ag was determined by radioimmunoassay. Oral samples from all patients seropositive for HBS Ag were psoitive for HBS Ag. The rectum and the skin were positive in three instances. The presence of blood did not coincide with HBS Ag. In the unit in which HBS Ag-positive patients were treated, 4.3% of the total samples were HBS Ag-positive. Of the samples that were free of blood, 1.5% were HBS Ag-positive, whereas 18.5% of the blood-contaminated samples contained the antigen. HBS Ag-positive samples were taken from surfaces in the immediate environment of HBS Ag-positive patients, as well as from surfaces with which patients had no direct physical contact. The data lead to the conclusion that spilled blood and hemodialysis equipment are not the only sources of contamination. This fact implies that HBS Ag-positive patient undergoing hemodialysis should be completely separated from HBS Ag-negative patients.


Coagulopathy in experimental sepsis with Streptococcus pneumoniae in rabbits: effect of drug therapy and splenectomy.

The cause of death in bacteremia due to Streptococcus pneumoniae remains unclear. The role of intravascular coagulation and splenectomy was investigated in rabbits with lethal pneumococcal bacteremia. The staphylococcal clumping titer in serum, a measure of fibrin degradation products, increased early and persisted until death. This titer correlated with the level of bacteremia. The partial thromboplastin time and platelet-rich plasma clotting time also increased as the disease worsened. However, the prothrombin time remained normal. 125I-labeled fibrinogen was cleared normally from the plasma of infected rabbits, whether intact or splenectomized. Similarly, the concentration of fibrogen in plasma remained normal, even though the level of fibrin degradation products increased, and no difference in these parameters was noted between intact and splenectomized rabbits. Fibrin deposition could not be detected in any of the organs studied. Neither the level of fibrin degradation products nor survival was affected by treatment with hydrocortisone, hexadimethrine, cytochrome c, carboxypeptidase B, epsilon-aminocaproic acid, or heparin. These data suggest that intravascular coagulation occurs in this experimental infection prior to the onset of shock but probably plays only a minor role in lethality.


Significance of diuresis-provoked bacteriuria.

Of 52 clinically healthy, ambulant elderly subjects with urine that was sterile or contained less than 0.2 X 10(5) bacteria/ml before diureisi, 11 (21.2%) responded to oral water loading and intravenous treatment with furosemide with a bacterial excretion rate of greater than 2.4 X 10(5)/min. Subjects with bacteriuria had decreased blood pressure, rate of creatinine clearance, maximal level of urinary creatinine, and maximal urinary flow. Twenty-nine of the nonbacteriuric and eight of the subjects who developed bacteriuria after diuresis were retested one to two years later. No subject had received antibacterial agents in the interim. The infections of four of the original subjects who had developed bacteriuria had cleared up and showed functional improvement. The remaining four subjects who had developed bacteriuria during the first study and three subjects who had been nonbacteriuric during the first study had concentrations of bacteria ranging from 0.05 X 10(5) to 1.80 X 10(5)/ml before diuresis in the second study, and excreted increasing numbers of bacteria (from 0.12 X 10(5) to 9.0 X 10(5)/min) in response to the diuretic loading. Two of the subjects who were bacteriuric in both surveys were infected with different bacteria during the two studies. The data show the high frequency of occult bacteriuria in ostensibly healthy elderly patients that is revealed only by forced diuresis, the detectable functional deficits, the improvement after clearing of the infections, and the rapid turnover between infected and clear periods.


Studies on opacity factor production by pharyngeal isolates of group A streptococci.

Production of opacity factor (OF) by 841 pharyngeal isolates of group A streptococci from schoolchildren in Casper, Wyoming was studied along with T- and M-typing of the streptococcal strains. The majority (89%) of M-typable strains were OF-negative in contrast to M-nontypable isolates, 45% of which were OF-positive. There was no apparent relation between the production of OF by the M-nontypable isolates and the type of infection (symptomatic vs. asymptomatic), degree of positivity of initial culture, antistreptolysin O response, and failure to eradicate the organisms by treatment with antibiotics. However, seasonal shifts in the prevalence of M-nontypable OF-positive strains provided evidence of change in streptococcal ecology, such a shift would not have been detected by T-typing alone. Thus, OF production is an additional epidemiologic marker that is helpful in differentiating M-nontypable group A strains that bear common T-antigens.


Intravenous injection of gentamicin and tobramycin without impairment of hearing.

The aminoglycoside antibiotics gentamicin and tobramycin were given to six healthy volunteers by intravenous injection. A dose of 80 mg was injected within 1 min. The curves relating serum concentration to time did not differ significantly for the two drugs. The maximal serum concentrations of drug exceeded 10 mug/ml on seven of 12 occasions. Despite these high concentrations, loss of hearing was not significant at any frequency (1,000-8,000 Hz) up to one month after injection. It is suggested that the aminoglycoside antibiotics penetrate into the inner ear lymph very slowly and that the concentration at that site is relatively unaffected by transient high concentrations of antibiotic in the blood plasma.


Pharmacokinetics of intravenously administered tobramycin in normal volunteers and in renal-impaired and hemodialyzed patients.

The pharmacokinetics of intravenously administered tobramycin, an aminoglycoside antibiotic, were studied in 16 volunteers with normal renal function and 10 patients with different degrees of renal impairment. In all subjects, the disappearance of tobramycin from serum followed first-order kinetics, and the elimination rate constant decreased proportionally with increasing degree of renal impairment. The half-life average 82 min in normal subjects (endogenous creatinine clearance, greater than 80 ml/1.73 m2 per min) and was 33.7 hr in a totally anephric subject. Linear relations were defined between the half-life of tobramycin in serum and the reciprocal of the endogenous creatinine clearance and between the half-life of the drug and the concentration of serum creatinine. These relations may be used for dosage adjustment in patients with renal impairment. In hemodialyzed patients, the half-life decreased six- to ninefold during dialysis for 6 hr. The extent of tobramycin elimination by hemodialysis may be predicted from relations established between the elimination of tobramycin and that of creatinine and blood urea nitrogen. Dosage schedules must be compatible with the pharmacologic and pharmacokinetic properties of the antibiotic.


Administration of tobramycin and gentamicin by the intravenous route every 6 hr in patients with normal renal function.

The feasibility and safety of administration of tobramycin and gentamicin in every 6 hr rather than every 8 hr was studied in 18 patients with normal renal function. Eleven patients received tobramycin in a dose of 1.0-3.2 mg/kg every 6 hr, and seven received gentamicin in a dose of 1.4-2.0 mg/kg every 6 hr. The mean peak serum concentrations, excluding the levels in the three children given tobramycin, were 5.5 and 5.8 mug/ml in each group, respectively, after maximal dosage; trough levels 2 hr before the next dose were less than or equal to 1.8 mug/ml in both groups of patients. Neither drug accumulated in the serum when the dosage regimen was maintained for three to 19 days. There was no evidence of nephrotoxicity or ototoxicity. Administration of tobramycin and gentamicin every 6 hr seems to be a reasonable program for patients with normal renal function, pending further studies on safety.


Pharmacokinetics of tobramycin in rats with reduced renal parenchyma and in experimental shock.

Ten days after four-fifths nephrectomy in rats (four-fifths of the renal parenchyma removed), the total and renal elimination constants of tobramycin were lowered by one-half. The biological half-life of tobramycin doubled. These results suggest that soon after nephrectomy the residual nephrons are able to increase the rate of excretion of tobramycin. The increase in mass of the remaining renal tissue between day 10 and day 80 after nephrectomy had no effect on the biological half-life, on the total and renal elimination constants, and on the renal clearance of tobramycin. During endotoxin shock and burn shock, concentrations of tobramycin in serum decreased at a slower rate. The highest serum concentrations of tobramycin were observed in rats with shock due to burns.


Comparison of urinary excretion of tobramycin and gentamicin in adults.

The urinary excretion of tobramycin and gentamicin was measured after intramuscular injections of 1 mg of each drug/kg in six normal volunteers. The antibiotics were assayed by the adenylyltransferase enzymatic method, which enables urine concentrations to be estimated accurately without dependence on urinary pH. The levels of the antibiotics in urine varied with the urine volume. The concentration of gentamicin was greater than 113 mug/ml and that of tobramycin was greater than 94 mug/ml 1 hr after administration. Urinary excretion of both antibiotics was rapid; 63.5% of the gentamicin and 74% of the tobramycin was excreted with 24 hr, with a large proportion being excreted by 4 hr after injection. There was a striking uniformity of individual values, and the method offers considerable advantages over existing microbiologic assays.


Single daily doses of tobramycin in therapy of urinary tract infections.

Results of therapy with tobramycin for infections caused by susceptible pathogens have been impressive. The usual multiple dose regimen is aimed at maintenance of the serum concentration of tobramycin above the minimal inhibitory concentration of the causative pathogen throughout the course of therapy. Evidence is accumulating, however, that the urinary concentration of antibiotics may be of greater importance than the serum concentration in the treatment of urinary tract infections. This concept has been supported by results of studies with nitrofurantoin, naladixic acid, tetracycline, and, more recently, gentamicin. The present study of 90 patients with urinary tract infections treated with a single intramuscular daily dose of tobramycin resulted in round-the-clock urinary levels of tobramycin that exceeded the minimal inhibitory concentration of the offending pathogen. The clinical and bacteriologic response to therapy was excellent, and no oto- or nephrotoxicity occurred. This regimen frequently allows for therapy on an outpatient basis, fewer painful injections for the patient, and the saving of nursing time and expensive supplies.


Comparison of tobramycin and gentamicin in the treatment of complicated urinary tract infections.

The efficacy and safety of intramuscular administration of either of the two aminoglycoside antibiotics tobramycin and gentamicin every 8 hr for seven days were compared in a prospective, randomized study of 75 elderly male patients with complicated urinary tract infections. All bacteria were sensitive to both antibiotics by the disk diffusion method. The two groups of patients (each receiving one of the two drugs) were comparable in terms of infecting microorganisms and underlying pathology of the urinary tract (mostly prostatic hyperplasia or carcinoma and urethral strictures). No patients had indwelling catheters. All patients had normal renal function (serum creatinine, less than or equal to 1.5 mg/100 ml and/or blood urea nitrogen, less than or equal to 25 mg/100 ml). In both groups 64% of the patients were cured of the infection, with cure defined as a negative urine culture result seven days after discontinuation of therapy. A statistically but not clinically significant increase in the level of serum creatinine in both groups of patients was noted after treatment.


Laboratory and clinical evaluation of tobramycin, a new aminoglycoside antibiotic.

The in vitro susceptibility to tobramycin of 1,466 bacterial isolates, including several genera and strains of common pathogenic gram-negative bacilli and Staphylococcus aureus, was determined by conventional serial dilution methods. Tobramycin exhibited marked in vitro antibacterial activity; almost all of the isolates tested were inhibited by less than or equal to 2.5 mug of the drug/ml. The antibiotic was used in the treatment of several types of infection in 29 patients with normal renal function. Observations were made on the therapeutic and secondary effects of the drug and on possible toxicity to the eighth cranial nerve. Clinical therapeutic results were satisfactory, The hearing of one patient was impaired, but the relation of this effect to tobramycin therapy is uncertain. Serum levels of the antibiotic were measured in 10 patients to investigate whether the drug accumulated during repeated administration. Mean serum levels of tobramycin 1 hr after intramuscular injection of 1.0 mg/kg were between 2.5 and 3.0 mug/ml on days 1, 3, and 6 of therapy. No accumulation of the antibiotic was observed after repeated administration.


Tobramycin in pediatric use.

The new aminoglycoside antibiotic, tobramycin, was used for treatment of gram-negative and staphylococcal infection in 38 neonates, infants, and children in a pediatric surgical unit. Levels of drug in serum after administration by intramuscular, intravenous, and intraperitoneal routes were monitored, and control of infections was generally good within the therapeutic range of 2.0-10.0 mug/ml with a standard dosage regimen of approximately 5 mg/kg per day. Impairment of renal function and concurrent lincomycin therapy were important factors causing variation of levels in serum outside this range. Levels of tobramycin in cerebrospinal fluid after intraventricular instillation varied greatly in one patient, but were satisfactory in another. Clinical and bacteriological assessment of results indicated only two failures of treatment, although infection with a known resistant organism supervened in three cases. Screening for renal, hepatic, and hematological toxicity revealed only one case of transient and reversible renal impairment. Response to tobramycin therapy was generally rapid and satisfactory in a group of young patients with moderately severe infections, many of which were complicated by the presence of a congenital anomaly.


Tobramycin and cephalothin sodium in treatment of infected patients with acute leukemia.

Tobramycin has been shown to be safe and effective when given intravenously in combination antibiotic therapy for infected patients with acute leukemia or other profound pancytopenic states. Tobramycin in combination with one or more antibiotics (usually cephalothin) produced satisfactory clinical results in 21 of 30 episodes of fever. Toxic effects were minimal and were limited to occasional mild thrombophlebitis.


Tobramycin and cephalothin for treatment of suspected sepsis in neutropenic children with cancer.

The safety and efficacy of tobramycin and cephalothin in treatment of suspected sepsis were studied in neutropenic children with various malignancies. Twenty episodes of suspected sepsis in 19 febrile children with cancer were treated with parenteral tobramycin and cephalothin; the duration of therapy ranged from one to 80 days. In 14 of the 20 episodes of suspected sepsis, a favorable clinical response was achieved within five days after initiation of antibiotic therapy. These episodes included a urinary tract infection with Proteus mirabilis and sepsis due to Escherichia coli. In four of the additional six episodes, clinical deterioration was though to be caused by the underlying malignancies. Two episodes included a case of E. coli spesis that ended fatally and a nosocomial infection with Eikenella corrodens. Results of this study suggest that combination therapy with tobramycin and cephalothin is safe and efficacious in treatment of suspected sepsis in febrile children with malignancies and neutropenia.


Clinical use of tobramycin in patients with surgical infections due to gram-negative bacilli.

Twenty-three patients with acute peritonitis, intraabdominal abscess, wound infections (including infection of serious burns), sepsis accompanying renal transplantation, or urinary tract infection associated with cancer were treated with various dosage regimens of tobramycin, and their clinical responses were analysed. For the conditions studied, the optimal regimen was 3 mg/kg per day in three divided doses. No clinical effectiveness was noted in this study for doses of greater than or equal to 4 mg/kg per day.


Clinical evaluation of tobramycin in respiratory and systemic infections in immunodepressed and normal patients.

Twelve patients with acute or chronic pneumonia due mainly to gram-negative bacilli, two patients with pseudomonas endocarditis, and two patients with seratia sepsis were treated with 80-160 mg of tobramycin in two daily doses. Fourteen infected patients with underlying leukemia or lymphoma received this dose of tobramycin combined with cefazolin or penicillin. Most respiratory infections were cured or markedly improved. with eradication or significant reduction in the number of infecting organisms. One case of pseudomonas endocarditis and both cases of serratia sepsis were also cured. Combined treatment with tobramycin and beta-lactam antibiotics resulted in clinical and bacteriological improvement in 50% of systemic immunodepressed patients with sepsis and/or pneumonia.


Tobramycin therapy of infections due to Pseudomonas aeruginosa in patients with cystic fibrosis: effect of dosage and concentration of antibiotic in sputum.

Established respiratory infections with mucoid Pseudomonas aeruginosa in patients suffering from cystic fibrosis were treated with conventional as well as larger doses of tobramycin. The infection was eradicated in four of the 17 patients treated, but the duration of follow-up study of one patient was short. It appeared that treatment was most successful in those patients in whom the highest peak concentrations of tobramycin in sputum were obtained. No side effects were noted, even when tobramycin was given in large doses for two weeks.


Pharmacokinetics and ototoxicity of gentamicin, tobramycin, and amikacin.

The pharmacokinetics of gentamicin, tobramycin, and amikacin in inner ear fluids, serum, cerebrospinal fluid, and the compartments of the eye were studied and compared in guinea pigs. The concentrations of antibiotic were determined by microbiologic methods and were confirmed by the use of 14C-labeled gentamicin. Retention was clearly demonstrated in perilymph, in which the half-lives of gentamicin, tobramycin, and amikacin were 12, 11, and 10 hr, respectively. The concentrations of drug in perilymph were symmetrical and were many times higher than the concentrations of antibiotic in the brain. There was no difference between the concentration of drug in endolymph and that in perilymph. A linear relation between concentrations in the perilymph and the dosage of gentamicin was ascertained. Long-term treatment did not influence the pharmacokinetics of the three antibiotics in the inner ear. However, increased levels of drug in the inner ear in animals with uremia and in some animals with otitis media explained the increased ototoxicity that occurs in treatment of these two conditions. Suboccipital puncture and diuresis did not change the concentrations of aminoglycoside antibiotics in the inner ear. Antibiotics applied locally in the middle ear had high degrees of ototoxicity.


Tobramycin adenylyltransferase: a new aminoglycoside-inactivating enzyme from Staphylococcus epidermidis.

Certain strains of Staphylococcus epidermidis resistant to the aminoglycoside antibiotics were shown to contain an enzyme that inactivates the kanamycins, neomycins, butirosins, paromomycin, gentamicin A, amikacin, and tobramycin by adenylylation. Tobramycin adenylyltransferase, as this enzyme is called, was found to be optimally active at pH 5.5. With paromomycin or neomycin B and C as substrates, however, two pH values (5.5 and 9.0) for optimal activity were observed. The enzyme requires Mg++ for activity and is stabilized significantly by dithiothreitol. It is probable that the 4'-hydroxyl group of ring I of the antibiotics is adenylylated. Those aminoglycosides that are not substrates for the enzyme lack a hydroxyl group in the corresponding position.


Emergence of gentamicin-resistant bacteria: experience with tobramycin therapy of infections due to gentamicin-resistant organisms.

A computerized system for testing and surveillance of bacterial susceptibility to antibiotics was used in monitoring the emergence of gentamicin-resistant strains of aerobic and facultative gram-negative bacilli at Massachusetts General Hospital since the release of gentamicin for clinical use in 1971. During the period studied, there was a significant increase in the prevalence of gentamicin-resistant bacteria, particularly among Pseudomonas, Acinetobacter (Herellea), and Proteus and, more recently, among Enterobacter and Klebsiella. Most gentamicin-resistant strains of Pseudomonas aeruginosa and Acinetobacter calcoaceticus var. anitratum (Herellea varginicola) retained susceptibility to tobramycin. Of the other gentamicin-resistant organisms, most were also resistant to tobramycin. Twelve patients with infections caused by gentamicin-resistant organisms were treated with tobramycin. All 12 patients were seriously ill, and all but one had failed to respond to previous therapy with gentamicin. Nine patients responded favorably to tobramycin, and six were cured. P. aeruginosa and A. calcoaceticus var. anitratum were most frequently the infecting organisms in these patients. Patients received tobramycin for three to 42 days; no significant drug-related toxicity was noted. These results emphasize the increasing clinical importance of gentamicin-resistant bacteria and suggest that tobramycin may be effective for treatment of some, but not all, infections caused by gentamicin-resistant bacteria.


Evaluation of tobramycin in severe urinary tract infection.

Twenty-five patients with severe urinary tract infections were treated with 3 mg of tobramycin/kg per day (given in two doses). Susceptible organisms were Escherichia coli, Proteus, Klebsiella, Pseudomonas, Streptococcus, and enterococcus. Clincal conditions in which treatment produced excellent bacteriological results included a wide range of urological disorders; the most common were pyelonephritis, cystitis, and epididymo-orchitis, Three patients had septicemia, and 12 had an infection that was the result of urinary tract obstruction requiring surgery.


Effects of electric-shock delivery on schedule-induced water intake: delay of shock, shock intensity, and body-weight loss.

In each of four experiments, schedule-induced water intake in the rat was studied under fixed-time 40-sec food delivery. Experiments I and II studied the temporal relationship between response-independent electric-shock delivery and licking. Shock was delivered under a variable-time 60-sec schedule. A lick-dependent delay was imposed so that licking and shock delivery were systematically separated in time by a minimum of 1 to 15 sec. Over a wide range of shock intensities the data failed to reveal a consistent delay-of-shock effect. Similar shock intensities led to similar reduction of water intake at each delay of shock interval. Experiments III and IV studied the effects of body-weight loss on water intake during independent shock delivery. In Experiment III, shock was delivered under variable-time 60-sec with a minimum separation between shock and licking of 5 sec. In Experiment IV, shock was delivered under variable-time 180-sec. The minimum separation between shock and licking was 10 sec. In each study, the resistance of water intake to suppression by shock delivery increased as the degree of body-weight loss increased. Schedule-induced water intake was affected more by shock when the animal was maintained at 90% of free-feeding weight than at 70%.


A national assessment of propoxyphene in postmortem medicolegal investigation, 1972-1975.

Eighteen medical examiners, coroners, and forensic science laboratories and offices, representing a total jurisdictional population of 52.6 million, were visited during November 1975, and more than 1200 cases occurring in the four years from 1972 through 31 July 1975 were evaluated for inclusion in the study. The sites were distributed across the United States and Ontario, Canada, and included urban and rural, states, city, and county jurisdictions. Scientific data and circumstantial information was gathered consistently for each case and site by means of five questionnaires. Finally, 1022 cases were compiled and examined, and the data were analyzed to form the body of data from which this report is written. The following conclusions may be drawn. 1. The number of deaths involving propoxyphene is increasing each year, and at a faster rate than total drug deaths. The absolute numbers and rate are different in urban and rural areas, but the frequency reached 6.0 deaths per million population in 1974. Deaths attributed to suicide as well as those determined to be accidental deaths and undetermined have increased. 2. Approximately 66% of all the cases studied had the word propoxyphene included in the cause of death statement on the death certificate. Approximately 46% of the cases were classified as suicide (64% of them female and 36% male), 26% as accidents, and 21% as undetermined. 3. The deceased were mainly middle-class, Caucasian, urban dwellers, with male and female evenly distributed. Their ages were from 20 to 50 years, with few outside this range. Female ages were uniformly distributed, but males in their early twenties were very prominent. This is different from the U.S. population age distribution, which is currently dominated by teenagers. Propoxyphene does not appear to be a pediatric problem, as seen in the study. 4. The deceased were not part of the illegal drug abuse population and had no particular propensity for the use of heroin or narcotics, but rather they were a particular medical population of those who misuse prescription drugs and alcohol. 5. The deceased did have a marked tendency to hypochondria, chronic minor illnesses and emotional problems. Some 43% had recent medical histories, and 82% had a documented psychiatric history which often included (51%) self-destructuve behavior such as suicide attempts. Almost all received a wide range of prescription drugs, particularly tranquilizers, which they often misused in the sense of self-medicating, multiple drug ingestion, and combining alcohol with their medication. Approximately 34% had a history of misusing some drug, and 20% could be defined as abusers in that they were prone to excessive use of their medications; 44% had diazepam available to them, and 17% were either problem drinkers or alcoholics.


A precipitin-inhibition test on denatured bloodstains for the determination of human origin.

A new method has been developed for determination of the species of origin of denatured bloodstains. Antiserum with known titer is incubated with the denatured bloodstain. If the bloodstain has the same origin as the antiserum, a substantial amount of antibody will be absorbed by the stain, resulting in an attenuation of the antiserum titer. By reacting the supernatant with a known serum by using immunodiffusion it is possible to detect any appreciable attenuation of the antiserum titer. We have been able to determine the species of origin of various denatured bloodstains using this inhibition method.


Detection of amphetamine and methamphetamine-type materials in pharmaceutical and biological fluids by fluorometric labeling.

A rapid and sensitive method for detecting amphetamine and methamphetamine in drug preparations and biological fluids has been developed. Amphetamine and methamphetamine in pharmaceutical and clandestine drug preparations can be easily screened from other contaminating drugs and readily identified by their fluorescence, with subsequent separation accomplished by TLC. The same general procedure can also be used to detect amphetamine and methamphetamine in human urine at concentrations of 0.1 mug/ml.


Several unusual cases of child abuse.

All childhood deaths which occurred in New Mexico during 1974 and 1975 were reviewed. Nine fatal instances of abuse were identified representing the entire spectrum of physical abuse: neglect, abuse in a single episode of injury, repetitive abuse, or sexual abuse. Several cases are summarized. These are unusual either in the distribution of pathologic findings or in the problems encountered in court presentation.


Short-range ammunition--a possible anti-hijacking device.

Short-range ammunition was developed for use by law enforcement personnel in congested, enclosed areas and primarily as a hijacking deterrent in commercial airliners. This ammunition was expected to reduce the risk of injury to innocent bystanders and prevent damage to the aircraft walls. Experimentally, short-range ammunition is effective in its wounding capacity at close ranges, and it does not exit from the body. Due to the particular structure of the projectile, injuries due to ricochet are averted. At ranges of 6 ft (1.8 m), the risk of damage to the full thickness of an aircraft wall does exist. Short-range ammunition deserves serious consideration as an alternative to regular ammunition for use in specialized, close-quartered situations, such as hijacking.


Studies of the spatial distribution of firearms discharge residues.

Fingerprint lifting tape has been applied to the lifting of firearms discharge residue from various surfaces. Lifting efficiencies from coarse fabric ranged from 30 to 70%. Use of the technique has been demonstrated in identifying a bullet hole and in measuring residue patterns on floor and firing hand surfaces. In hand lifts it has been shown that autoradiography alone does not uniquely distinguish the hand of someone who has fired a revolver from the hand of someone who has not. Neutron activation analysis of lifts from selected hand areas may, however, provide such a distinction.


Programmed energy dispersive X-ray analysis of top coats of automotive paint.

Elemental, qualitative, and semiquantitative analyses of 1974 U.S. automotive paints by a Finnigan Model 900F Energy Dispersive X-ray (EDX) Spectrometer are examined. This EDX analytical system incorporates a minicomputer to receive, store, and present data. The paint samples were received from the National Bureau of Standards (NBS) Law Enforcement Standards Laboratory as a collection representative of the top coats used on all U.S. manufactured automobiles in 1974. Aspects considered include EDX system stability, homogeneity of similarly catalogued samples from different NBS collections, categorization of paints by predominant constituents, and comparison of data on paint standards that were similar. Automated analysis represents an important application of forensic science to the characterization of paint samples and provides for the nondestructive analysis of a variety of materials in a large range of sample sizes and concentrations.


Examination of paints by trace element analysis.

Trace element analysis using neutron activation analysis is effective and valuable, particularly in the examination of white household paint. Although physical appearance and resin composition are generally similar in these paint samples, trace element composition provides an effective way of distinguishing among them. In the case of automobile paint samples, NAA serves as an important additional technique for discrimination. The technique is important when sample sizes are very small. The technique developed takes a few minutes for sample preparation, a few hours of irradiation time (during which the examiner's presence is not required), and then a few minutes for counting and obtaining quantitative multielement concentration patterns. A technician can easily handle 30 to 50 samples per day.


Forensic odontology in missing persons cases.

Forensic odontology can provide a service early in cases of missing persons where foul play is suspected. The combined effort of the professional police investigator and the odontologist prior to the discovery of an unidentified remains allows for an efficient, accurate identification procedure. The forensic dental interview can provide a working format or starting point in developing a planned interrogation procedure concerning dental information. In such a manner, the planned interview can enhance the investigator's efforts in missing persons cases and other cases requiring dental identification.