Search

Loss and degradation of grasslands in the Great Plains region have resulted in major declines in abundance of grassland bird species. To ensure future viability of grassland bird populations, it is crucial to evaluate specific effects of environmental factors among species to determine drivers of population decline and develop effective conservation strategies. We used threshold models to quantify the effects of land cover and weather changes in "lesser prairie‐chicken" and "greater prairie‐chicken" (Tympanuchus pallidicinctus and T. cupido, respectively), northern bobwhites (Colinus virginianus), and ring‐necked pheasants (Phasianus colchicus). We demonstrated a novel approach for estimating landscape conditions needed to optimize abundance across multiple species at a variety of spatial scales. Abundance of all four species was highest following wet summers and dry winters. Prairie chicken and ring‐necked pheasant abundance was highest following cool winters, while northern bobwhite abundance was highest following warm winters. Greater prairie chicken and northern bobwhite abundance was also highest following cooler summers. Optimal abundance of each species occurred in landscapes that represented a grassland and cropland mosaic, though prairie chicken abundance was optimized in landscapes with more grassland and less edge habitat than northern bobwhites and ring‐necked pheasants. Because these effects differed among species, managing for an optimal landscape for multiple species may not be the optimal scenario for any one species., We quantified the effects of land cover and weather changes in Galliformes in Kansas. We found evidence of nonlinear, threshold effects of land cover change in abundance of all four focal species, suggesting that the presence of both grassland and cropland is important in the landscape to simultaneously maintain these populations.

The inflammatory response plays an important role in the pathophysiology of multiple injuries. This study examines the effects of severe trauma and inflammatory response on markers of neuronal damage. A retrospective analysis of prospectively collected data in 445 trauma patients (Injury Severity Score (ISS) &ge, 16) is provided. Levels of neuronal biomarkers (calcium-binding Protein B (S100b), Enolase2 (NSE), glial fibrillary acidic protein (GFAP)) and Interleukins (IL-6, IL-10) in severely injured patients (with polytrauma (PT)) without traumatic brain injury (TBI) or with severe TBI (PT+TBI) and patients with isolated TBI (isTBI) were measured upon arrival until day 5. S100b, NSE, GFAP levels showed a time-dependent decrease in all cohorts. Their expression was higher after multiple injuries (p = 0.038) comparing isTBI. Positive correlation of marker level after concomitant TBI and isTBI (p = 0.001) was noted, while marker expression after PT appears to be independent. Highest levels of IL-6 and -10 were associated to PT und lowest to isTBI (p &lt, 0.001). In all groups pro-inflammatory response (IL-6/-10 ratio) peaked on day 2 and at a lower level on day 4. Severe TBI modulates kinetic profile of inflammatory response by reducing interleukin expression following trauma. Potential markers for neuronal damage have a limited diagnostic value after severe trauma because undifferentiated increase.

Background: Anatomic landmarks located on the proximal femur have only recently been defined, and there is a lack of radiographic guidelines for their locations presented in the literature. With the confident identification of these landmarks, radiographs could provide more assistance in preoperative evaluations, intraoperative guidance, and postoperative assessments. Purpose: To quantify the radiographic locations of endoscopic landmarks of the proximal femur. Study Design: Descriptive laboratory study. Methods: Ten cadaveric specimens were dissected, and radio-opaque hardware was placed for each landmark of interest. Radiographs were obtained and measurements recorded in anteroposterior (AP) and Dunn 45° views. Results: In the AP view, the gluteus medius insertion was located a mean 12.9 ± 2.4 mm and 34.7 ± 5.1 mm from the piriformis fossa and vastus tubercle, respectively. The piriformis fossa was a mean 14.8 ± 5.9 mm and 4.9 ± 1.9 mm from the anterior and posterior tips of the greater trochanter, respectively. The anterior and posterior tips of the greater trochanter were a mean 14.8 ± 5.1 mm from each other. In the Dunn 45° view, the piriformis fossa was a mean 13.3 ± 2.0 mm, and the vastus tubercle was a mean 21.5 ± 6.0 mm, from the gluteus medius insertion. Moreover, the vastus tubercle was a mean 33.5 ± 6.4 mm from the anterior tip of the greater trochanter and 31.6 ± 8.5 mm from the posterior tip of the greater trochanter. Conclusion: In spite of the variation in cadaveric sizes, quantitative descriptions of endoscopic landmarks were reproducible in clinical views. Clinical Relevance: A detailed understanding of how the described landmarks present radiographically is relevant to preoperative planning, intraoperative evaluations, and postoperative assessments.

Patients with hereditary or acquired haemophilia A may develop inhibitory factor VIII (FVIII) antibodies. These disrupt FVIII activity predominantly by preventing the formation of the tenase complex, leading to a serious bleeding disorder. Antibodies without inhibiting activity, however, can also be found when screening patients with haemophilia A under FVIII supplementation. Therefore, the detection of only these allo- or autoantibodies from plasma is not sufficient. Rather, the characterization of the antibody-induced effects on the coagulation cascade should be considered due to its great diagnostic importance. Currently, inhibitory activities are detected by the functional Bethesda assay, which directly measures the delay in clotting time by the patient plasma. However, this assay does not provide information on the cause of the inhibition. Here, we report the development of a surface plasmon resonance (SPR) biosensor that has the potential to integrate both quantitative and functional information on patient antibody characteristics in one measurement. Recombinant FVIII protein was immobilized on the sensor surface to detect antibodies from patient plasma. The interaction of the FIX- and FXa-clotting proteins with the formed anti-FVIII/FVIII complex could be detected subsequently within the same SPR measurement cycle. Inhibitory antibodies led to the prevention of these interactions. Thus, discrimination between the clinically relevant inhibitory and non-inhibitory antibodies was enabled. In a group of 16 patients with inhibitory antibodies (both ELISA- and Bethesda-positive), 5 patients with non-inhibitory antibodies (ELISA-positive but Bethesda-negative) and 12 healthy controls, diagnostic sensitivity and specificity data of 100% for the FIX interaction were achieved using this biomimetic biosensor approach. The new method allows for detection and quantification, as well as for evaluation of inhibitory activity of allo- and autoantibodies, using small sample volume and short analysis time.

BACKGROUND The parallelization of clinically relevant antigens in a microarray format is of growing importance due to the ability to measure multiple antigen-antibody interactions. With the development of a microarray for the detection of antiphospholipid antibodies we focussed on one important autoimmune disease that is still diagnostically challenging. Reasons are the heterogeneity of the autoantibodies and the unspecific clinical symptoms. METHODS For the covalent immobilization of antigenic structures, glass transducers were coated with 11-aminoundecyltrimethoxysilane (11-AUTMS). In total 35 antiphospholipid syndrome (APS) patients, six patients with lupus erythematosus and 24 healthy controls were investigated on a microarray format using polarized imaging reflectometric interference spectroscopy. RESULTS The novel surface modification based on the short derivative 11-AUTMS resulted in a selective biosensor allowing a clear differentiation of patient and control samples. It combined proteinogenic as well as phospholipid-derived antigens, namely β2-glycoprotein I (β2-GPI), prothrombin, cardiolipin (CL) and a β2-GPI/CL complex. With optimized regeneration conditions, up to 20 consecutive measurements could be performed on one chip. Sensitivity was determined to be 0.800-0.929, specificity was between 0.733 and 0.969, depending on the respective antigen. CONCLUSIONS Multiplexed determination of serological parameters has a great potential. We have shown that our biosensor is capable of detecting four different APS relevant antibodies in parallel exhibiting a sensitivity and specificity comparable to existing ELISA methods.

Prescribed fire is a commonly used as a follow-up procedure to mechanical top growth removal methods such as mowing and roller chopping, but the effects of fire on spinescence and tannin content of shrub sprouts produced after mechanical top growth removal are unknown. Following mowing, (1) height, spinescence, and tannin content in sprouts produced after burning; (2) nutrient and fiber contents in sprouts of the 3 study species; and (3) utilization of sprouts of each species in burned and unburned plots were determined in each of blackbrush acacia (Acacia rigidula Benth.), honey mesquite (Prosopis glandulosa Torr.), and spiny hackberry (Celtis pallida Torr.). Averaged across sampling periods, burned blackbrush acacia and honey mesquite had 54% and 94%, respectively, shorter thorns than unburned plants. Burned and unburned spiny hackberry plants had similar thorn lengths. Averaged across species, sprouts of burned plants had similar tannin levels as unburned plants 6 and 12 weeks after burning. S...

We hypothesized that top growth removal results in long-term increases in antiherbivore defenses of blackbrush acacia ( Acacia rigidula ), honey mesquite ( Prosopis glandulosa ), and spiny hackberry ( Celtis pallida ). In 1998, we determined chemical composition and thorn characteristics of leaves and twigs from plants roller chopped once in 1989, from plants roller chopped in 1989 and 1995, and from untreated plants. Honey mesquite had greater crude and digestible protein than untreated plants, and spiny hackberry had lower acid detergent fiber and acid detergent lignin than untreated plants 3 years after roller chopping in 1989 and 1995. Thorns were longer and denser in blackbrush acacia 9 years after one roller chopping treatment. Thorn densities were greater in all three species after two treatments. Long-term increases in thorn length and density after roller chopping may reduce value of the shrubs as browse for herbivores.

We used caged and uncaged plants to compare shoot growth, biomass, spinescence, nutrient, and tannin content of blackbrush acacia (Acacia rigidula Benth.), honey mesquite (Prosopis glandulosa Torr.), and spiny hackberry (Celtis pallida Torr.) 6 and 12 weeks after mowing, and estimated white-tailed deer (Odocoileus virginianus Raf.) use of sprouts of each species stripped of spines and unstripped. Blackbrush acacia grew slowly and had more thorns, protein-precipitating tannins, and fiber than honey mesquite or spiny hackberry. Spiny hackberry, a shrub adapted to fertile sites, grew slower and was more physically defended than honey mesquite but lower in tannins than blackbrush acacia, and was the most palatable to deer of the three species. Thorn removal did not alter palatability of the three species to deer.

Serbia’s defeat of the Austro-Hungarian invasion of 1914 counts among the major upset victories of the last century. This article, which examines Austro-Hungarian tactical performance in Serbia in that campaign, offers unique insights into the causes of the Habsburg debacle. Analysis of the battlefield performance of the Austro-Hungarian 21st division, which fought and lost the first Habsburg engagement in August 1914, reveals much about the operational shortcomings of Vienna’s field forces that summer and autumn; in particular, this article details the impact of the controversial nationalities issue on Habsburg battlefield effectiveness, offering a revisionist perspective about the realities and liabilities of a multinational army.

The simultaneous quantification of β-fructofuranosidase (invertase, EC 3.2.1.26) and glucan 1,4-α-glucosidase amyloglucosidase, EC 3.2.1.3) activities in an one-step assay is reported. The approach is based on chemometric evaluation of the mid-IR spectra of the reaction mixture, measured with a sequential injection (SI) system with FTIR spectroscopic detection. Two partial least squares (PLS) calibration models were established automatically. The first model was calculated from reaction mixtures after the action of standard mixtures of both enzymes. The other was built up with mixtures of the substrates (maltose, sucrose) and products (fructose, glucose) without any enzymatic reaction. The establishment of the second model was more rapid as no time for an enzymatic reaction is necessary. Both models gave comparable results for the prediction of mixtures containing 0–2.0 U/ml of each enzyme, with mean errors between 0.08 and 0.05 U/ml.

A flow injection analysis (FIA) system with fourier transform infrared (FTIR) spectroscopic detection for the simultaneous determination of amyloglucosidase (exo-glucan 1,4-α-glucohydrolase, EC 3.2.1.3) and α-amylase (endo-glucan 1,4-α-glucanohydrolase, EC 3.2.1.1) activities in aqueous solution is proposed. Starch hydrolysis catalyzed by both enzymes was monitored in the mid-IR range by recording IR-spectra of the assay solutions before and after hydrolysis. The intensities of the resulting difference spectra were directly related to the enzyme activities. Furthermore, the different reaction pathways of the two enzymes under investigation produced distinct spectral changes resulting in difference spectra characteristic for each enzyme. Using partial least squares (PLS) regression it was possible to simultaneously determine both enzyme activities covering a range of 38–150 U/l (630–2500 nkat/l) for amyloglucosidase and a range of 500–3750 U/l (8300–62 000 nkat/l) for α-amylase. Absorption due to the sample matrix was successfully eliminated by calculation of the difference spectra. The developed method was also successfully applied to the analysis of spiked fermentation broth samples yielding mean deviations of 2.6% and 10.8% for amyloglucosidase and α-amylase, respectively.

31P magnetic resonance spectroscopy (MRS) was used to study an open therapeutic trial of coenzyme Q10 (CoQ) in mitochondrial encephalomyopathies. Eight patients were treated with 150 mg CoQ per day for 6 months. 31P MRS spectra of calf muscle were recorded at rest, during exercise and in the immediate postexercise recovery period. Although there was an improvement of the mean ratio of phosphocreatine (PCr) to inorganic phosphate during the post-exercise recovery period after 3 months of treatment, this finding was mainly due to a single therapy responder and did not reflect a beneficial effect on the whole group. Improved repletion of PCr persisted after 6 months of therapy. Our study identified a single responder to this therapy, whose response could not be predicted on the basis of clinical, biochemical or molecular data. These findings suggest that therapeutic trials of CoQ should be performed under close metabolic monitoring in order both to identify responders for subsequent long-term treatment and to evaluate possible mechanisms of this supportive therapy.

ERGIC-53 is a lectin-type membrane protein that continuously recycles between the ER, ER-Golgi intermediate compartment (ERGIC) and the cis-Golgi. To identify the targeting signals that mediate this recycling, N-glycosylated and myc-tagged variants of ERGIC-53 were constructed. By monitoring endoglycosidase H resistance, we measured the loss from the ER-ERGIC-cis-Golgi cycle of ERGIC-53. A domain exchange approach with the plasma membrane reporter protein CD4 showed that the transmembrane and the lumenal domains are not sufficient, while the cytoplasmic domain of ERGIC-53 is required and sufficient for pre-medial-Golgi localization. However, the ERGIC-53 cytoplasmic domain on CD4 lead to increased ER-staining by immunofluorescence microscopy indicating that this domain alone cannot provide for unbiased recycling through the ER-ERGIC-cis-Golgi compartments. Complete progress through the ER-ERGIC-cis-Golgi recycling pathway requires the cytoplasmic domain acting together with the lumenal domain of ERGIC-53. Dissection of the cytoplasmic domain revealed a COOH-terminal di-lysine ER-retrieval signal, KKFF, and an RSQQE targeting determinant adjacent to the transmembrane domain. Surprisingly, the two COOH-terminal phenylalanines influence the targeting. They reduce the ER-retrieval capacity of the di-lysine signal and modulate the RSQQE determinant.

In the present study, we have extended earlier taxonomic, biochemical and experimental investigations to characterize two species of Taenia from carnivores in Kenya by use of the sequences of a variable domain (D1) of nuclear ribosomal DNA and the cytochrome c oxidase subunit 1 and NADH dehydrogenase 1 genes of mitochondrial DNA. Emphasis was placed on the characterization of Taenia madoquae from the silver-backed jackal (Canis mesomelas) and Taenia regis from the lion (Panthera leo), given the previous absence of any DNA sequence data for them, and on assessing their genetic relationships with socioeconomically important taeniids. The study showed that T. regis was genetically most closely related to T. hydatigena, and T. madoquae to T. serialis, T. multiceps or T. saginata. The present findings provide a stimulus for future work on the systematic relationships and epidemiology of lesser-known taeniid cestodes in Africa and other continents, employing mitochondrial sequence data sets.

Ternidens deminutus (Strongylida) is a parasitic nematode infecting non-human and human primates in parts of Africa, Asia and the Pacific islands. The present study genetically characterized T. deminutus and defined genetic markers in nuclear ribosomal DNA (rDNA) as a basis for developing molecular-diagnostic tools. The sequences of the second internal transcribed spacer (ITS-2) of rDNA were determined for adult specimens of T. deminutus (Nematoda: Strongylida: Oesophagostominae) from the Olive baboon and the Mona monkey. The length and G+C content of the ITS-2 sequences was 216 bp and approximately 43%, respectively. While there was no sequence variation among individual T. deminutus specimens from the baboon, 6 (2.8%) nucleotide differences were detected in the ITS-2 between the parasite from baboon and that of the Mona monkey, which is similar to the difference (3.2%) between 2 other species of Oesophagostominae (Oesophagostomum bifurcum and O. stephanostomum) from non-human primates, suggesting significant population variation or the existence of cryptic (i.e. hidden) species within T. deminutus . Pairwise comparisons of the ITS-2 sequences of the 2 operational taxonomic units of T. deminutus with previously published ITS-2 sequences for selected members of the subfamilies Oesophagostominae and Chabertiinae indicated that species from primates (including those representing the subgenera Conoweberia and Ihleia) are closely related, in accordance with previous morphological studies. The sequence differences (27-48.3%) in the ITS-2 between the 2 taxonomic units of T. deminutus and hookworms (superfamily Ancylostomatoidea) enabled their identification and delineation by polymerase chain reaction (PCR)-based mutation scanning. The genetic markers in the ITS-2 provide a foundation for improved, PCR-based diagnosis of T. deminutus infections and for investigating the life-cycle, transmission patterns and ecology of this parasite.

Isopseudodicentric chromosome 18 is very rare and results in a combination of partial trisomy and partial monosomy of chromosome 18. We report here a hypotrophic newborn with a lateral cleft lip and palate and multiple craniofacial dysmorphisms, a combined heart defect, unilateral hypoplasia of the kidney, bilateral aplasia of thumbs, and generalized contractures. Cytogenetic analysis revealed an isopseudodicentric chromosome 18 with breakpoint in 18q (46,XX,psu idic(18)(pter --> q22.1::q22.1 --> pter)). The isopseudodicentric chromosome 18 was observed in 100% of blood lymphocytes and umbilical cord fibroblasts, thus indicating a non-mosaic finding of the isopseudodicentric chromosome in the child. An elongated derivative chromosome 18 had also been found prenatally in amniotic cells. In contrast, a terminal deletion (18q-) was detected in placental cell cultures. The breakpoint was mapped to a 0.9 Mb region on 18q22.1 (located 64.8-65.7 Mb from the telomere of the p-arm) by a novel quantitative PCR approach with SYBR green detection. The results indicate an identical breakpoint of the isopseudodicentric chromosome 18 in the child and the 18q- chromosome in the placenta. To our knowledge this is the first report that a fetus carrying an isopseudodicentric chromosome 18 with breakpoint in 18q (46,XX,psu idic(18)(pter --> q22.1::q22.1 --> pter)) in non-mosaic form can be viable, but is associated with severe congenital malformations of the child.

ERGIC-53 (former designation, p53) is a 53-kDa nonglycosylated, dimeric, and hexameric type I membrane protein that has been established as a marker protein for a tubulovesicular intermediate compartment in which protein transport from the endoplasmic reticulum to the Golgi apparatus is blocked at 15 degrees C. Although ERGIC-53 is not a resident protein of the rough endoplasmic reticulum its cDNA sequence carries a double lysine endoplasmic reticulum retention motif at the cytoplasmically exposed COOH terminus. Here we report that overexpression of ERGIC-53 in COS cells saturates its intracellular retention system leading to the appearance of ERGIC-53 at the cell surface. Cell surface ERGIC-53 is efficiently endocytosed by a mechanism that is disturbed when the two critical lysines of the endoplasmic reticulum retention motif are replaced by serines. The results suggest a mechanistic similarity of pre-Golgi retention by the double lysine motif and lysine-based endocytosis.

In approximately 5% of cancer patients undergoing radiotherapy, this treatment has to be interrupted because of an acute reaction of normal tissues. To test the possibility of predicting this type of reaction, the micronucleus assay was used to determine radiosensitivities of peripheral blood lymphocytes of 15 patients with severe acute reaction of normal tissue, 15 patients without this reaction and 15 healthy donors. Whole-blood cultures were irradiated with X-rays (4 Gy, 1.08 Gy/min) and treated with cytochalasin B. The micronuclei scores observed in irradiated cells were corrected for the scores in unirradiated cells. Intra-individual and interindividual variations in micronuclei scores were analysed in samples from healthy donors, and highly significant interindividual differences were found (P0.001). Scores of cells not irradiated in vitro were higher for cancer patients before radiotherapy than for healthy donors (P0.001), and those for cancer patients after radiotherapy were higher than for patients before radiotherapy (P0.001). Average micronuclei scores induced by in vitro irradiation were significantly higher in samples from cancer patients compared with those from healthy donors (P0.01). Moreover, all subgroups of cancer patients included individuals with very high levels of micronuclei after in vitro irradiation. There was, however, no relationship between the micronuclei scores and the occurrence of severe acute reactions in normal tissues.

Three patients with chronic progressive external ophthalmoplegia of adult-onset, generalized muscle atrophy and myalgia are described. Two patients fulfilled the histological criteria for centronuclear myopathy, the third those for fiber-type disproportion. Additionally, typical ragged red fibers were found in all muscle specimens, and several muscle fibers were cytochrome c oxidase negative. NADH and succinate dehydrogenase stains showed increased subsarcolemmal accumulation of mitochondria. To determine whether these findings are coincidental or whether they indicated an additional mitochondrial disorder, all patients were investigated using biochemical analysis of the respiratory chain, molecular genetics, magnetic resonance spectroscopy of quadriceps muscle and ergometry. These tests suggested an additional mitochondrial dysfunction. Mitochondrial dysfunction seems to be more common in this group of myopathies than previously estimated, and may be of importance in the pathogenesis of these disorders.

The transmembrane protein CD16 (Fc gamma RIII) is detected on activated macrophages, natural killer (NK) cells and a small subset of T lymphocytes. From CD3-CD56+ CD16+bright NK cells and CD3+ CD56+ CD16+dim non-major histocompatibility complex (MHC)-restricted cytotoxic T lymphocyte (CTL) clones were generated reflecting the stable, but different, CD16 expression of the respective peripheral blood subpopulations. To compare the role of CD16 on NK cells and non-MHC-restricted CTL, Fc gamma RIII activation and its mechanisms were investigated using monoclonal antibodies (mAb). Cross-linking of CD16 induced Ca2+ influx in CD16+bright NK clones. In contrast, there was no Ca2+ mobilization after CD16 activation in CD16+dim CTL, which revealed a good response to cross-linking of CD3 antigen. Pretreatment with CD16 mAb alone or cross-linked CD16 mAb did not block the CD3 response of CD16+dim CTL. Again, CD16 cross-linking induced more interferon-gamma transcription in NK cell clones than in non-MHC-restricted CTL clones. Also a higher tumor necrosis factor-alpha production of NK clones after CD16 cross-linking compared to CD16+dim CTL could be observed. These data suggest that after CD16 activation CD16+dim CTL and CD16+bright NK cells use different second messengers. In addition, signal transduction via CD3 and CD16 appears to function independently in CD16+dim non-MHC-restricted CTL.

Interaction of human Fc receptors with their ligands or monoclonal antibodies (mAB) has been shown to trigger the release of cytokines, induce phosphoinositide turnover, and increase cytotoxicity [1, 2]. The Fc receptor with low affinity for IgG (FcγRIII, CD16) mediating antibody-dependent cellular cytotoxicity (ADCC) has been described in peripheral blood on monocytes, polymorphonuclear leukocytes (PMN), and natural killer (NK) cells. As a result of two different genes, FcγRIII is expressed as a glycosyl-phosphoinositol anchored form on PMN and as a transmembranously anchored glycoprotein on monocytes and NK cells [3, 4]. In addition to CD3− CD16+bright NK lymphocytes also a small T cell subset expressing CD16 antigen at lower density has been observed [5, 6]. These T lymphocytes, mainly of a T cell receptor (TCR) γδ+ CD4− CD8− phenotype, mediate non-MHC-restricted cytotoxicity and ADCC [7, 8]. The role of CD16 antigen in signal transduction on TCR αβ+ lymphocytes with coexpression of the FcγRIII in comparison to NK cells has not been studied yet.

The objective of the study was to evaluate sperm nuclear shape in sex-sorted and non-sorted semen using Fourier Harmonic Amplitudes (FHA). Frozen semen was obtained from a single commercially available source. Mature bulls (n = 15) from the same breed with an average age of 4.42 ± 2.08 years were collected and the semen was either frozen or X-chromosome sorted using a flow cytometer and then frozen. Frozen straws were transported to the lab and analyzed for FHA. Briefly, straws were thawed and cells were incubated with 1.6 μm Hoechst 33342. Cells were then washed, fixed, dried to a slide and analyzed for nuclear head shape. Harmonic amplitudes 0 to 5 (HA0–HA5), derived from FHA, were previously shown to be an accurate, objective, and repeatable measure of sperm nuclear shape. HA0 describes the overall nuclear size of the sperm, whereas HA1 describes the anterior head, HA2 the length of the sperm along the longitudinal axis, and HA3 to 5 the distal, post-nuclear curvature of the sperm head. Each unit of semen was evaluated for motility and FHA. There was a significant decrease in motility in the sorted group (77 ± 1% v. 54 ± 3%; P < 0.0001). Multivariate ANOVA showed that there were differences between the sorted and non-sorted groups in HA1 to 4 (P < 0.02). Harmonic amplitude means ± SD (microns) for sorted and non-sorted treatments are as follows: HA1 (0.117 ± 0.003 v. 0.109 ± 0.003), HA2 (1.087 ± 0.005 v. 1.063 ± 0.005), HA3 (0.139 ± 0.003 v. 0.130 ± 0.003), and HA4 (0.201 ± 0.004 v. 0.191 ± 0.004), respectively. The nuclear shape of X-sorted sperm is longer and more pinched in both the anterior and posterior head. Interestingly there was no difference in HA0 (P = 0.119) indicating that the overall size of the sperm head is not affected by the sorting process. The differences in harmonic amplitudes may be due to the size and a restricted location of the X v. Y chromosome in the sperm nucleus.

Pretreatment with deuterium oxide (D2O) has been shown to protect mice against lethal effects of X-rays. In contrast, X-irradiation of cultured mammalian cells in D2O-containing medium has previously been reported to result in increased cell killing. Therefore, the effects of preincubation in medium containing 20% D2O on radiosensitivity were tested, using cells of a heat-sensitive cell-cycle mutant (21-Tb) of the murine mastocytoma P 815-X2. The mutant cells proliferate at 33 degrees C and are arrested in G1 phase in a state of reversible proliferative quiescence at 39.5 degrees C. Prior to irradiation with single X-ray doses of 0-10 Gy, the cells were cultured in normal or D2O-containing medium, either for 96 h at 33 degrees C ('proliferating cells'), or for 72 h at 33 degrees C followed by 24 h at 39.5 degrees C ('arrested cells'). After X-irradiation the cells were resuspended in normal medium, and cell survival was determined by the capacity of cells to form colonies in fibrin gels. Preincubation in medium containing 20% D2O resulted in a radioprotective effect on both proliferating and arrested cells, particularly at the higher X-ray doses. This radioprotection was manifested as a decreased slope of the semilogarithmic survival curves, whereas pretreatment with D2O had no significant effect on postirradiation repair as judged from Dq values. These results support the interpretation that the increase in postirradiation survival may be attributed to incorporation of deuterium into cellular metabolites during the period of preincubation.

Palatability of shrub sprouts to white-tailed deer (Odocoileus virginianus Raf.) differs among species, which causes selective browsing and can shift shrub species composition to dominance by less palatable species. The hypothesis was tested that differences in palatability of new sprouts among shrub species following roller chopping small (4 ha) patches within a shrubland matrix would result in a shift in shrub species composition within the patches toward less palatable species. Relative density and relative canopy cover of all woody species in plots 9 years after 1 roller chopping treatment, in plots 3 years after 2 roller chopping treatments, and in untreated plots were estimated. Relative density of blackbrush acacia (Acacia rigidula Benth.) was 3 times greater and relative canopy cover was 12 times greater 9 years after the first roller chopping treatment compared to untreated plots, but relative density and relative canopy cover of blackbrush acacia in roller chopped plots were similar to relative density and relative canopy cover in untreated plots 3 years after the second roller chopping treatment. Relative canopy cover of spiny hackberry (Celtis pallida Torr.) in plots roller chopped in 1989 and 1995 was higher than in untreated plots. Relative density and canopy cover of all other species were similar between roller chopped and untreated plots. Shrub community composition 9 years after 1 roller chopping treatment or 3 years after 2 roller chopping treatments in the subtropical thornscrub communities in southern Texas did not shift toward greater dominance of less palatable species. DOI:10.2458/azu_jrm_v56i6_schindler

TO THE EDITOR: Spontaneous bacterial peritonitis (SBP) is a common complication in patients with ascites. It is estimated that 8–27% of patients with cirrhosis develop this complication (1). The organisms involved in SBP are usually gastrointestinal pathogens. Escherichia coli, Streptococci, and Klebsiella account for approximately 80% of the cases, but various other organisms such as Enterococcus, Enterobacter, and Clostridium species have been known to cause SBP (1). Although different species of Streptococcus have been isolated from ascitic fluid, Streptococcus bovis has only been reported in a few cases. We report a case of spontaneous bacterial peritonitis due to this organism.

Cell suspensions were prepared from normal and regenerating liver of adult rats by perfusion with a calcium-chelating agent (EGTA), collagenase and hyaluronidase, and the cells were incubated in culture medium. In cultures prepared from regenerating liver at 20 h after partial hepatectomy, 23 ± 4% of parenchymal cells initially incorporated [3H]TdR. This incorporation was shown to reflect semiconservative DNA replication. At least some parenchymal cells were able to complete their DNA synthesis and to progress through G2 and mitosis. Numbers of hepatocytes in mitosis increased up to 12 h of culture. On the other hand, no entry of hepatocytes into the S period was detectable in cultures prepared from normal or regenerating liver.

In cultures of a murine mastocytoma, endogenous synthesis of thymidine phosphates, as determined by the incorporation of [3H]deoxyuridine into DNA, was reduced within 15 min to less than 3% of control values by the addition of amethopterin (10 µM) in combination with hypoxanthine and glycine. If [3H]thymidine and unlabeled thymidine were added simultaneously with amethopterin, the increase with time of radioactivity in cellular DNA was linear at least between 30 and 90 min, while radioactivity in the acid-soluble nucleotide fraction remained constant during this time interval, indicating that intracellular thymidine nucleotides had the same specific activity as exogenously supplied [3H]thymidine. This permitted calculation of the amount of thymidine incorporated per hour into DNA of 106 cells. In conjunction with the base composition of mouse DNA, these results were used to calculate rates of DNA synthesis. Cell proliferation rate, cell cycle time, and the duration of the S period were not affected to any appreciable extent by the addition of amethopterin and thymidine. Rates of DNA synthesis, as derived from thymidine incorporation rates, were in good agreement with those derived from the measured mean DNA content of exponentially multiplying cells and rates of cell proliferation.

In order to study the influence of the colloid osmotic pressure on albumin and total liver protein synthesis, rat hepatocytes were isolated by collagenase perfusion and incubated in Krebs-Ringer-buffer for 4 h. The colloid osmotic pressure produced by different bovine serum albumin (BSA) or dextran 60 concentrations varied from 3 to 80 mm Hg. A physiological colloid osmotic pressure of 20 mm Hg was obtained with 5.7 g BSA or 3.7 g dextran 60 per 100 ml of buffer. Albumin synthesis was measured by Laurell rocket immunoelectrophoresis. Total liver protein and total secretory protein synthesis were determined by the measurement of 1-14C-leucine incorporation. Albumin synthesis was not primarily regulated by the colloid osmotic pressure as was demonstrated by a lack of inhibition after addition of BSA. There was no significant influence of the oncotic pressure on the incorporation of 14C-leucine into total liver proteins. The incorporation into total secretory proteins was inhibited by an increasing colloid osmotic pressure, mediated either by BSA or dextran, suggesting an inhibition of the secretion of plasma proteins other than albumin.

Normal murine peritoneal mast cells were fused to serum-deprived, non-proliferating cells of a cultured subline (41-SB-4) of the P-815 murine mastocytoma. Upon reincubation in medium containing 10% horse serum for 48 h, mono- and binuclear 41-SB-4 cells reentered S phase of the cell cycle, while mast cell X 41-SB-4 heterokaryons as well as mono- and binuclear mast cells remained in proliferative quiescence, indicating dominant expression of the quiescent state of mast cells. The quiescent state of normal mast cells thus resembles that of cold-sensitive (cs) mutant cells (21-F) of the undifferentiated P-815 mastocytoma: at the non-permissive temperature of 33 degrees C, the 21-F cells were found to enter a state of quiescence which is characterized by its dominant expression in heterokaryons and by morphological differentiation with the formation of metachromatically staining granules similar to those of mast cells. This suggests that the cellular control mechanisms involved in entry into proliferative quiescence and in morphological differentiation of cs 21-F cells may be analogous to those of normal mast cells and/or their precursors.

Cultures in vitro of a murine mastocytoma were incubated in a series of semisynthetic media. In order to produce specific nutritional deficiencies media were used in which one amino acid ( l -leucine, l -histidine or l -tryptophan, respectively) was present in optimal and various suboptimal concentrations. Appropriate reduction of the concentration of l -leucine or l -histidine resulted in a decrease of cell multiplication and in an increase of the relative number of dead cells, whereas omission of l -tryptophan had no appreciable effect on cell proliferation during the observation period of 96 h. After incubation of cells in these media during 4 days, the mean cellular histamine and 5-hydroxytryptamine contents as well as the incorporation of 35 SO 4 into the protein-bound heparin fraction were determined. The mean cellular content of histamine or 5-hydroxytryptamine was reduced in media containing suboptimal concentrations of the corresponding precursor amino acid ( l -histidine or l -tryptophan, respectively). In contrast, appropriate reduction of the l -leucine level in the culture medium resulted in an increase of the mean cellular 5-hydroxytryptamine content and an increased 35 SO 4 incorporation.

1. In menschlichen Gewebekulturen (Tumorzellstamm KB) gezuchtete GelbfiebervirusstAmme (Asibi, 17D) wurden auf ihre InfektiositAt, PathogenitAt und immunisierende Potenz in Versuchen an Affen gepruft. 2. Die Dauerzuchtung des pantropen Asibistammes fuhrte zu Varianten von betrAchtlich verminderter Viscerotropie und unverAnderter neurotroper QualitAt. Der Grad der Mitigierung war annAhernd derselbe wie bei den modifizierten KulturstAmmen 17E und 17AT vonTheiler. 3. Die Dauerpassage von 17D-StAmmen im menschlichen Gewebe bewirkte keine Reaktivierung viscerotroper Eigenschaften, sondern einen hochgradigen Verlust der Neurotropie, wie ein solcher bei andern SubstAmmen des 17D-Virus bisher wohl nicht beobachtet wurde.

Resume Des conjugues azoproteiques, prepares par couplage d'agents cytostatiques (amethopterine et un derive methylhydrazine) au fibrinogene et a l'albumine exercent, in vitro, une activite antitumorale, determinee sur les cellules du mastocytome murin P- 815 -X 2 . Certains ont montre une activite egale ou inferieure a celle de l'agent cytotoxique seul. D'autres se sont reveles plus actifs que le medicament libre. Ces resultats sont discutes selon les differentes acquisitions concernant la permeabilite des cellules tumorales aux proteines plasmatiques.

Office balance, as applied to the toll telephone network, can be divided into two major divisions, those of through and terminal balance. Through balance concerns the connection of one 4-wire intertoll trunk with another, switched on a 2-wire basis. Terminal balance is required for the interconnection of an intertoll trunk with a toll connecting trunk to the subscriber's central office. Because of the extent of private networks presently being engineered, it is necessary to balance the interconnection of calls within these networks. This paper is concerned with the basic principles of through and terminal balance, but primarily centers on terminal balance in the private and DDD networks, because that is the area of the majority of engineering problems.

In this brief review, the development of modern quantitative methods for the cultivation of mammalien cellsin vitro is described, and present knowledge of their nutritional requirements is presented. In addition, some applications of these techniques in biochemistry, pharmacology, virology, genetics, and radiobiology are discussed.

Frettchen uberleben eine sicher todliche Infektion mit einem virulenten Staupevirus, wenn ihnen gleichzeitig die hundertfache Menge eines avirulenten Staupevirus injiziert wird. Erfolgt die virulente Infektion 24 Stunden vor der Injektion der hundertfachen Dosis des avirulenten Virus, so kann eine deutliche Verlangerung der Inkubationszeit beobachtet werden, Wurde die tausendfache Dosis des avirulenten Staupevirus 24 Stunden nach der virulenten Infektion verabfolgt, so uberlebte von zwei Tieren eines ohne Krankheitserscheinungen, das andere verendete nach typischem Krankheitsverlauf.

131Iododeoxyuridine (131IDU) was injected into normal and partially hepatectomized rats, and the specificity of incorporation of this thymidine analogue into liver DNA was determined 2, 24 and 48 hr following intramuscular injection. At 2 and 24 hr after 131DU injection, a major proportion of radioactivity in the liver was in the acid-soluble fraction, whereas 48 hr after injection the label in the acid-soluble fraction had decreased considerably. In liver obtained 2 hr after injection of 131IDU, only 1.8–16.6% of the total radioactivity were in DNA. If, however, the tissue was subjected to formalin fixation, the acid-soluble label was extracted selectively, and of the remaining radioactivity 64–88% was in DNA. Therefore, the radioactivity that is not extracted by formalin may be used as a measure of DNA synthesis at the time of injection of 131IDU, thus obviating time-consuming biochemical fractionation procedures.

Cultures of a murine mast cell tumor were partially synchronized by reversible inhibition of DNA synthesis with amethopterin (in combination with hypoxanthine and glycine) during different time intervals and subsequent addition of thymidine. In these cultures, the duration of the G2 period was measured (a) by observing the time-lag between addition of thymidine and resumption of mitotic activity, (b) by evaluating the time-lag between the addition of 3 H-thymidine and the appearance of labeled mitoses. Inhibition of DNA synthesis did not significantly change the median duration of the subsequent G2 period. It is, therefore, concluded that the G2-specific processes cannot begin before DNA synthesis has been completed, and that a triggering mechanism may be responsible for the transition of a cell from the S into the G2 period.

In cultures of murine neoplastic mast cells, the duration of different phases of the division cycle (G1, S, G2, and mitosis [M]) was determined under optimal and several well-defined suboptimal growth conditions. Two methods of evaluation were applied to the same culture system: first, the relative number of G1, S, G2, and M cells was determined by pulse labeling of samples with thymidine-3H and subsequent radioautography in conjunction with a microfluorometric technique permitting rapid measurements of cellular DNA content; second, after pulse labeling with thymidine-3H, the variations with time of the mitotic labeling index were analyzed. Suboptimal culture conditions were obtained by reducing the concentration of single essential medium components (leucine, glucose, or serum) or by the addition of specific metabolic inhibitors (actinomycin D, amethopterin). Growth-limiting culture conditions resulted in increased generation times. Even under control conditions, the cell number doubling time exceeded the generation time, and this difference was more pronounced in suboptimal media. Under most of the suboptimal conditions tested, the increase in generation time was attributable primarily to an extended duration of the G1 phase. Under certain growth-limiting conditions, however, other phases were also prolonged. In addition, the variabilities of the generation time and of certain cell cycle phases were increased under suboptimal culture conditions. Results obtained by the two methods of evaluation were, in general, in good agreement with each other. Some differences were, however, observed and interpreted in terms of cell death and/or asymmetric frequency distributions of cell cycle parameters.

Partially synchronous cultures of a murine mastocytoma were prepared by centrifugation of cells in isotonic sucrose gradients and reincubation of slowly sedimenting early interphase cells under steady state conditions. Cell multiplication in these cultures occurred in a stepwise manner with a periodicity of approx. 8 h. In aliquots that were withdrawn from synchronous cultures at 1.5 h intervals, 3 H-thymidine incorporated into DNA by intact cells and DNA polymerase activities in cell homogenates were determined. Whereas synchronous growth of the cell population was reflected by pronounced fluctuations of 3 H-thymidine incorporation into DNA of intact cells, variations with time of DNA polymerase activity, using native or denatured DNA as primer, were much smaller and parelleled those of cellular protein content. Maxima and minima of 3 H-thymidine incorporation by intact cells preceded those of DNA polymerase activity and of cellular protein content by 1.5 to 3 h. It is concluded that total DNA polymerase activity as measured in disrupted cells does not determine changes in rate of DNA synthesis in the cell culture system studied.

A method of preparing synchronous cell cultures is presented which is based on a selection of postmitotic cells by density gradient centrifugation. Gradients were prepared by appropriate mixing of normal culture medium with medium in which NaCl was replaced by sucrose at a concentration providing for isotonicity. The cells sedimenting most slowly (2–5% of the original cell number) were collected, suspended in sucrose-free medium and cultured under conditions of an optimal steady state. Cell viability, as determined by the capacity of cells to form macroscopically visible colonies in a semisolid medium, was not altered by this treatment. Rhythmic variations of mitotic activity were observed, with a first maximum at 8 h after collection of postmitotic cells. The percentage of cells in the S period was determined at regular time intervals by pulse labeling with 3H-thymidine. A high proportion of DNA-synthesizing cells was found during low mitotic activity, while a low proportion of cells in S phase was observed at the time of maximum mitotic activity.

A heat-sensitive (hs, arrested at 39.5 degrees C, multiplying at 33 degrees C) and a cold-sensitive (cs, arrested at 33 degrees C, multiplying at 39.5 degrees C) cell cycle variant were isolated from an undifferentiated P-815 murine mastocytoma line. At the respective nonpermissive temperature, both the hs and the cs variant cells were reversibly arrested with a DNA content, typical of G1 phase. The cells of two cs variant subclones, when exposed to the nonpermissive temperature of 33 degrees C, formed metachromatically staining granules with an ultrastructure resembling that of mature mast cells. In addition, the cellular 5-hydroxytryptamine content underwent a marked increase, and the cells responded to compound 48/80 by degranulation as described for normal mast cells. On the other hand, in cells of two hs variant subclones, essentially no mast cell granules were detectable at either 33 or 39.5 degrees C. As previously reported, the cs cell cycle variant phenotype is expressed dominantly in heterokaryons obtained by fusing cs with wild-type cells, whereas hs cell cycle variant cells, similar to other hs mutants, were found to behave recessively under these conditions. Thus the state of proliferative quiescence induced in the cs cells at 33 degrees C is qualitatively different from the state of cell cycle arrest observed in hs cells at 39.5 degrees C and may represent a model for proliferative quiescence of differentiated cells in the intact organism.

The parameters of the cell cycle are analyzed in terms of the stochastic theory of cell proliferation for a murine mastocytoma line. The cells were grown in suspension culture under steady-state conditions in a chemostat. Initial estimates of the parameters from synchronous growth indicate that agreement of the data with the model is obtained only if the model is modified to include an initial proliferating fraction of less than 100%, and a cell loss continuing throughout the course of the experiment. The analysis verifies that the modified theory adequately describes the data, and that similar parameters are obtained from both desynchronization and percent labeled mitosis experiments. The average cycle time from 10 desynchronization experiments was 8.24 ± 0.52 h with a cellular standard deviation of 1.28 ± 0.18. The combined parameter obtained by dividing the cellular standard deviation by the cycle time is shown to be a useful measure of biological variability well defined over many different experiments. The rate constant for cell loss is about 0.009 which gives an 8% cell loss per cycle. The cell loss is sufficient to account for the apparent deficit in initially proliferating cells. The initial distribution of the synchronous cells is qualitatively examined and is found to be peaked late in G1 or early in S.

A heat-sensitive (hs, arrested at 39.5 degrees C, termed 21-Ta) and a cold-sensitive (cs, arrested at 33 degrees C, termed 21-Fb) clonal cell cycle variant were isolated from the same clone of the P-815 murine mastocytoma line. At the respective nonpermissive temperatures, both the hs and the cs variant were reversibly arrested in G1 phase, and numbers of cells forming colonies upon reincubation at the permissive temperature remained nearly constant for at least 6 days. Cells arrested in G1 by incubation at the respective nonpermissive temperatures were fused to cells of another P-815 clone (31-S) that had been arrested by serum deprivation. Upon reincubation in medium containing 10% serum for 48 h at 39.5 degrees C, 21-Ta x 31-S heterokaryons, similar to 31-S x 31-S homokaryons, entered the S phase, whereas at 33 degrees C, 21-Fb x 31-S heterokaryons, similar to 21-Fb x 21-Fb homokaryons, remained arrested in G1, indicating a recessive expression of the hs and a dominant expression of the cs phenotype.

Es wird vom Auftreten einiger Hepatozoon-Infektionen in ostafrikanischen, wildlebenden Saugetierarten berichtet, die bisher als Wirte fur die Parasiten dieser Gattung weniger bekannt sind. In einer 1965/66 planmasig getesteten Serie von 310 kleinen Saugetieren in 31 Arten aus der Gegend von Tororo in Ost-Uganda wurde nur eine solitare Infektion in Mastomys natalensis bemerkt. Die Gameten im Peripherblut und die Schizozysten in der Milz sind beschrieben und abgebildet. Ferner wird von einer Massenuntersuchung des Blutes von 77 freilebenden Carnivoren (Caniden, Feliden und Viverriden) aus dem Serengeti-Nationalpark in Tanzania berichtet, die in den Jahren 1966/67 durchgefuhrt wurde. Es konnten etwa in der Halfte der 56 untersuchten Massailowen (Panthera leo massaicus) intraleukozytar gelagerte Gameten einer Hepatozoon-Art gesehen werden, die sich von allen anderen bisher bekannten Formen durch ihre Grose auszeichneten. In der Tupfelhyane (Crocuta crocuta) kommt etwa in dem gleichen Prozentsatz eine andere Hepatozoon-Art vor. Sie ist deutlich kleiner und variations-freudiger. Vom Lowenparasiten werden selektiv die Monozyten des Peripherblutes parasitiert, der in Hyanen liegt oft auch in gelapptkernigen Granulozyten.

During the last 15 years, cultures of cells in vitro, because of the intro­ duction of quantitative methods of high precision, have become a valuable tool for studies of the effects of certain classes of pharmacological agents. Cell cultures offer the possibility of observing the effects of drugs on cells in an artificial environment, without interference by nervous and hutnoral factors present in the intact organism. Furthermore, because during prepa­ ration of primary cultures the cells are released from the three-dimensional framework of tissue architecture, the influence of intercellular contact is greatly decreased or completely abolished. Cell culture systems thus permit an analysis of the responses of isolated cells to pharmacological agents. In this review, various applications of cell culture methods to the analysis of the mode of action of drugs are discussed. The action of certain drugs is characterized by a high specificity, and inhibition of a single process within the cultured cells is observed. Such specific inhibitors have become exten­ sively used in analyzing the role of various processes in cellular activities. This "use of pharmacology in cell culture" will, however, not be considered. After a primary culture of dispersed cells has been prepared from a piece of tissue by mechanical disruption or by the application of a suitable enzyme such as trypsin, in most cases only a small proportion of the cell population is capable of proliferation. Furthermore, under conditions in vitro the cells usually undergo marked changes of their metabolic activities. Even a primary culture, therefore, cannot be regarded as representative of the tissue of origin. Under cell culture conditions, in general a selection of cell types capable of continued proliferation takes place. In contrast to the situation in the intact organism, cell multiplication in vitro is not limited by growth-regulating mechanisms. Cells in prolonged culture, therefore, are, in some respects, comparable to mUltiplying microorganisms. Genetically pure cell populations may be obtained by culture of isolated cells. Normal human cells in culture usually retain their diploid chromosome set. They appear, however, to be limited in their proliferative capacity to a number of -ap­ proximately 60 cell divisions and then undergo degenerative changes which

The relative incorporation into RNA and DNA of 3 H-5-uridine, 3 H-cytidine and 3 H-6-thymidine was determined in brain, liver, kidney and spleen of the mouse as well as in neoplastic cell cultures. In systems with a high proliferative activity (spleen, cell cultures), the specificity of 3 H-5-uridine as a precursor of RNA was higher than that of 3 H-cytidine, although not as high as the specificity of 3 H-thymidine as a precursor of DNA. In organs with a low proliferative activity (brain, liver and kidney), the results based on a biochemical analysis did not permit any conclusions as to the distribution of label from 3 H-5-uridine and 3 H-cytidine between RNA and DNA of the small proportion of DNA-synthesizing cells.