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Biomedical subjects

B Alexander

Publications and source records attributed to B Alexander.

At least 19 recordsLinked to original sources

The effect of 6-dimethylaminopurine (6-DMAP) and cycloheximide (CHX) on the development and chromosomal complement of sheep parthenogenetic and nuclear transfer embryos.

The effects of activation by 6-dimethylaminopurine (6-DMAP) and cycloheximide (CHX) on the development and chromosomal complement of sheep parthenogenetic and SCNT embryos were investigated. The results revealed that the blastocyst development of parthenogenetic embryos was significantly higher (P < 0.05) in 6-DMAP activated oocytes, compared to those activated with CHX (21.0 +/- 0.9 vs. 14.9 +/- 0.5, respectively). In contrast, the blastocyst frequencies did not significantly differ (P > 0.05) between the two activation treatment groups for SCNT embryos. The 6-DMAP or CHX treatment did not result in any significant difference in the blastocyst total cell number in either parthenote or SCNT embryos. The chromosomal analysis revealed that all the parthenogenetic embryos (100.0%) derived from 6-DMAP treatment, were chromosomally abnormal whereas in CHX-treated embryos, it was significantly lowered (93.6%, P < 0.05). Conversely, the proportions of chromosomally abnormal SCNT embryos did not significantly differ (P > 0.05) among the 6-DMAP and CHX- treated embryo groups (60.0% vs. 56.2%, respectively). This study demonstrated that oocyte activation agents such as DMAP and CHX have differing effects on meiotic or mitotic nuclei. The study also highlighted the feasibility of using bovine X and Y chromosome specific painting probes in sheep embryos.

Adenine↗

The impact of chromosomal alteration on embryo development.

Chromosome alterations, such as those affecting telomere erosion, predictably occur with each cell division, others, which involve changes to the expression and replication of the X-chromosome occur at particular stages of development, while those that involve loss or gain of chromosomes occur in a random and so far unpredictable manner. The production of embryos in vitro and by somatic cell nuclear transfer (SCNT) has been associated with altered expression of marker genes on the X-chromosome and an increased incidence of chromosomally abnormal cells during early development. In the case of SCNT embryos chromosome abnormalities may be associated with the nuclear donor cell. Telomere rebuilding subsequent to SCNT appears to vary according to species and type of donor cell used. It is speculated that the rate of telomere erosion and incidence of chromosome abnormalities affects developmental potential of early embryos and may be potential predictors of developmental outcome.

Animals↗

Analysis of the sex pheromone extract of individual male Lutzomyia longipalpis sandflies from six regions in Brazil.

Although the phlebotomine sandfly Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) is generally accepted to be a species complex, it is unclear how many members there are, how they are related and which are the main vectors of leishmaniasis. The vectorial capacity of each sibling species is likely to differ, thus a means of identifying the most important vector species is of critical importance to the epidemiology and control of this debilitating disease in South and Central America. In Brazil four chemotypes have been distinguished by sex pheromone analysis. In this study the sex pheromone extracts of L. longipalpis from six regions of Brazil were analysed in detail. Samples included the sympatric 1-spot, 2-spot and intermediate spot morphotypes from Sobral, Ceará State. The results strongly suggest that members of the complex that produce different sex pheromones are reproductively isolated, thus strengthening the argument that the different chemotypes represent true sibling species. The study also found significant differences in morphology and the amounts of sex pheromone produced by members of each chemotype from different parts of Brazil, which suggests population substructuring that has not previously been recognized. Evidence of a fifth chemotype in Brazil is also presented.

Animals↗

Risk factors for work related violence in a health care organization.

OBJECTIVE: Identify the exposure effects of job family, patient contact, and supervisor support on physical and non-physical work related violence. DESIGN: Cross sectional study of employees in a Midwest health care organization, utilizing a specially designed mailed questionnaire and employer secondary data. SUBJECTS: Respondents included 1751 current and former employees (42% response rate). RESULTS: Physical and non-physical violence was experienced by 127 (7.2%) and 536 (30.6%) of the respondents, respectively. Multivariate analyses of physical violence identified increased odds for patient care assistants (odds ratio (OR) 2.5, 95% confidence interval (CI) 1. 1 to 6.1) and decreased odds for clerical workers (OR 0.1, 95% CI 0.03 to 0.5). Adjusted for job family, increased odds of physical violence were identified for moderate (OR 5.9, 95% CI 2.1 to 16.0) and high (OR 7.8, 95% CI 2.9 to 20.8) patient contact. Similar trends were identified for non-physical violence (OR 1.4, 95% CI 1.1 to 2.0 and OR 1.7, 95% CI 1.3 to 2.3). Increased supervisor support decreased the odds of both physical (OR 0.7, 95% CI 0.6 to 0.95) and non-physical violence (OR 0.5, 95% CI 0.4 to 0.6), adjusting for job family and demographic characteristics. CONCLUSIONS: Increased odds of physical violence were identified for the job family of nurses, even when adjusted for patient contact. Increased patient contact resulted in increased physical and non-physical violence, independent of job family, while supervisor support resulted in decreased odds of physical and non-physical violence.

Cross-Sectional Studies↗

Control of phlebotomine sandflies.

Phlebotomine sandflies (Diptera: Psychodidae) transmit many zoonotic diseases (arboviruses, bartonelloses and especially leishmaniases) of importance to human health in at least 80 countries. Measures used to control adult sandflies (Lutzomyia and Phlebotomus) include the use of insecticides (mostly pyrethroids) for residual spraying of dwellings and animal shelters, space-spraying, insecticide-treated nets, impregnated dog-collars and personal protection through application of repellents/insecticides to skin or fabrics. Because the breeding-sites of sandflies are generally unknown, control measures that act specifically against immatures are not feasible, although the effectiveness of a few biological and chemical agents has been demonstrated in laboratory evaluations. Reports of insecticide-resistance refer to only three sandfly species (P. papatasi, P. argentipes and S. shorttii) against DDT in one country (India), although there are reports of DDT-tolerance in several countries. Current knowledge of sandfly susceptibility to various insecticides is summarized. Constraints and advantages of different compounds, formulations and delivery methods for sandfly control under different environmental conditions are discussed.

Animals↗

Location and function of intrahepatic shunts in anaesthetised rats.

BACKGROUND: In the present study we determined the proportion of shunt flow due to patent intrahepatic portal systemic shunts in the normal rat liver and its relationship with microsphere induced portal hypertension. METHODS: Systemic and hepatic haemodynamics were measured continuously before, during, and after intraportal injection of 15 micro m diameter microspheres in anaesthetised male Wistar rats. Functional hepatic blood flow and intrahepatic shunt flow were determined by the use of constant intraportal infusion of sorbitol and simultaneous measurements in the portal vein, hepatic vein, and carotid artery. The percentage of large shunts of diameter >15 micro m were estimated by intraportal injection of (51)Cr labelled 15 micro m diameter microspheres. RESULTS: Hepatic sorbitol uptake was 97.9 (0.5)% in normal control rats, with functional hepatic blood flow equalling total hepatic blood flow (2.52 (0.23) ml/min/100 g body weight). Microsphere injection decreased sorbitol uptake to 12.8 (4.3)% and further to 4.1 (0.7)% when followed by hepatic arterial ligation. In the latter two groups, intrahepatic shunt flow (1.46 (0.15) and 1.16 (0.19) ml/min/100 g body weight, respectively) was not significantly different from portal venous flow (1.36 (0.20) and 1.20 (0.20) ml/min/100 g body weight, respectively). Portal venous flow remained at 70% of basal values and portal venous pressure only increased by 50% from baseline. (51)Cr labelled microsphere shunt fraction through large shunts (>15 micro m) was less than 1.0%. CONCLUSION: The site of confluence between the hepatic artery and portal vein is in zone II. Intrahepatic shunts originate in presinusoidal regions in zone I in the normal liver and, when activated by intraportal injection of microspheres, divert 70% of the total portal blood flow away from zone III and thereby reduce acute increases in portal venous pressure.

Animals↗

Some Psychodidae (Diptera) from Atlantic forest in south-eastern Brazil, with descriptions of Trichomyia dolichopogon sp. nov. and Trichomyia riodocensis sp. nov.

The family Psychodidae includes the medically important phlebotomine sand flies and four other subfamilies that have been little studied in the Neotropics. The authors here report the results of a trip to collect psychodids in Parque Estadual do Rio Doce, a national park in Minas Gerais that contains one of the largest surviving areas of Atlantic Forest in Brazil. Collections made by Malaise and CDC light traps as well as from diurnal resting sites included 15 species of Psychodinae and Trichomyiinae, among them Trichomyia dolichopogon sp. nov., T. riodocensis sp. nov. and 13 other species new to science but represented by females only. Twelve species of phlebotomine sand flies (Lutzomyia spp.) were also collected. Collections from an undisturbed area of the park were much richer faunistically than those from an area that was destroyed by fire in 1967 but had since regenerated, suggesting that recovery after environmental disturbances of this type may be prolonged. This pattern was not seen for phlebotomine sand flies, whose greater abundance and species richness in the disturbed section of the park may be related to their dependence on small mammal hosts, known to be more diverse in this type of habitat.

Animals↗

Sulfur and oxygen isotope analysis of sulfate at micromole levels using a pyrolysis technique in a continuous flow system.

The discovery of a mass-independent isotopic composition (delta17O = (delta17O - 0.512 * delta18O) no equal to 0) in aerosol sulfate and the identification of its origin (aqueous-phase oxidation by 03 and H2O2) have renewed interest in measuring the oxygen isotopic content of sulfate. In this paper, we present a new method to measure both delta17O and delta18O in SO4, with the possibility of sulfur isotope analysis on the same sample. The technique takes advantage of the easy pyrolysis of Ag2SO4 to SO2, O2, and Ag metal in a continuous flow system. Because the technique is not quantitative in oxygen (yield approximately 45% for O2), a calibration is needed. Correction factors of +0.87 and +0.44% were obtained for delta18O and delta17O, respectively. A technique to convert micromole levels of sulfate in any form to silver sulfate is described. To reach this goal, a solid electrolyte (Nafion membrane) is used in an electrolysis apparatus. Reproducibilities for micromole sample sizes are (1sigma) 0.5, 0.3, and 0.1% for delta18O, delta17O, and delta17O, respectively. No memory effects or isotopic exchange during the treatment of the sample is observed. The main advantages of this new method over the existing ones are no fluorinating agent is needed, both oxygen and sulfur isotopes can be measured on the same sample, only very small amounts of sulfate are needed (down to 100 microg (1 micromol)), it is relatively fast and inexpensive, and the possibility exists to couple this technique to an on-line analysis.

Journal Article↗

Functional and compositional studies of arteries stored in University of Wisconsin solution compared with Krebs-Bülbring buffer.

Endothelium-dependent and -independent vasorelaxation in ring segments of rabbit thoracic aorta is reduced and noradrenaline-induced vasoconstriction unaltered after prolonged storage in University of Wisconsin solution (UW) compared to arteries stored in extracellular-type solutions such as Krebs-Bülbring buffer (KBB). The aims of the present study were to determine whether angiotensin-II-induced vasoconstriction, alterations in myosin light chains, protein synthetic capacity, and subcellular structures are altered after 8 days of UW storage at 4 degrees C. The present study showed reduced contractility to angiotensin II, following 8 days of cold storage in UW, that was not reversed in the presence of a nitric oxide synthase inhibitor, N(G)-nitro-l-arginine methyl ester (100 microM). Measurements of contractile protein ratios in the same tissues after cold storage in UW or KBB did not show any significant alterations in smooth muscle myosin light chains or protein synthetic capacity (reflected by total RNA). It is concluded that reductions in vasoconstriction in UW-stored tissue are unlikely to be due to increased release of nitric oxide nor reduced availability of myosin light chains for phosphorylation and vasoconstriction.

Angiotensin II↗

Hepatic arterial perfusion regulates portal venous flow between hepatic sinusoids and intrahepatic shunts in the normal rat liver in vitro.

Intrahepatic shunts regulate portal venous pressure during periods of acute portal hypertension when the transhepatic portal resistance is momentarily increased in the normal rat liver in vivo. Hepatic arterial inflow may also increase the transhepatic portal resistance and activate intrahepatic shunts. In the present study, the transhepatic portal resistance and the activity of intra-hepatic shunts were measured in vitro and the point of confluence between the hepatic artery and portal vein in the rat determined. Livers of male Sprague-Dawley rats were single-pass, dual-perfused in vitro. Total cessation or diversion of the hepatic arterial inflow to the portal venous vasculature, whilst maintaining total hepatic perfusate flow, decreased intrasinusoidal pressure, increased transhepatic portal venous resistance and opened the portal venous intrahepatic shunts in a manner similar to intraportal injection of 15-microm diameter microspheres. Injections of the microspheres into the hepatic arterial circulation increased hepatic arterial pressure dramatically, consistent with complete occlusion of the arterial vasculature. The intrahepatic shunts are located at a pre-sinusoidal level because no increases were detected in hepatic arterial pressure following intraportal injection of microspheres. The hepatic arterial vasculature, unlike the portal supply, does not possess a collateral shunt circulation and coalesces with the portal vein at an intrasinusoidal location

Animals↗

Reciprocal regulation of endothelial substrate adhesion and barrier function.

OBJECTIVE: To examine how cell-substrate adhesion is regulated during barrier changes produced by exposure to inflammatory mediators. METHODS: Lung microvascular endothelial monolayers were treated with test agents +/- blockers, and barrier was measured by transendothelial resistance; cell-substrate adhesion was assessed by surface area conservation after trypsin treatment of monolayers. Protein phosphorylation and distribution were assayed by immunoblotting and fluorescent microscopy, respectively. RESULTS: H2O2, histamine, bradykinin, and thrombin, decreased endothelial barrier function, and enhanced adhesion to the substratum. H2O2 enhanced cell adhesion to the substrate in a concentration (0-1 mM)- and time (0-60 minutes)-dependent fashion. This effect of H2O2 reversed within 120 minutes of removal of H2O2 and was blocked by the mean arterial pressure (MAP) kinase inhibitor, PD98059 and by chelating cytoplasmic Ca2+ but not PKC or PKG inhibition. H2O2 also stimulated tyrosine phosphorylation of several proteins and increased the association of the focal adhesive proteins paxillin, talin, and vinculin with the cytoskeleton and may promote localization of these proteins to junctions. CONCLUSIONS: Our data indicate that inflammatory mediators reduce cell-cell contact, contributing to reduced solute barrier and simultaneously enhanced substrate binding, which may be reciprocal events in barrier regulation in vitro and in vivo.

Animals↗

Nitric oxide modulates acetylcholine-induced vasodilatation in the hepatic arterial vasculature of the dual-perfused rat liver.

The role of nitric oxide in the modulation of hepatic arterial vascular reactivity was investigated in an isolated dual-perfused rat liver preparation. Twelve male Wistar rats (200-250 g) were anaesthetized with sodium pentobarbitone (60 mg kg-1 i.p.). The livers were then excised and perfused in vitro through hepatic arterial and portal venous cannulae at constant flow rates. Concentration-dependent dose-response curves to acetylcholine (10(-8)-10(-5) M), sodium nitroprusside (10-6(-5) x 10(-4) M), and adenosine triphosphate (ATP) (10(-8)-10(-5) M) in the hepatic artery were constructed after the tone was raised by addition of methoxamine (3 micorM L(-1)). Acetylcholine-induced vasodilatation in the hepatic artery was significantly attenuated with inhibition of nitric oxide synthase by using NG-nitro-L-arginine methyl ester (30 microM), Emax = 51.7 +/- 2.8 vs. 32.5 +/- 3.1 mmHg, before vs. after NG-nitro-L-arginine methyl ester, respectively. ATP-induced hepatic arterial vasoconstriction which was significantly enhanced with L-NAME, Emax = 94.0 +/- 9.3 vs. 127.0 +/- 8.0 mmHg, before vs. after NG-nitro-L-arginine methyl ester, respectively. Sodium nitroprusside-induced hepatic arterial vasodilatation remained unchanged with NG-nitro-L-arginine methyl ester, Emax = 57.0 +/- 3.4 vs. 57.0 +/- 4.1, before vs. after NG-nitro-L-arginine methyl ester, respectively. The data from the present study suggest that acetylcholine-induced vasodilatation in the intrahepatic arterial vasculature of the rat liver is at least, in part, mediated by the release of nitric oxide. In addition, ATP-induced hepatic arterial vasoconstriction is also modulated by the release of nitric oxide (*P < 0.05, Student's paired t-test).

Acetylcholine↗

First report of Lutzomyia (Nyssomyia) umbratilis Ward & Frahia, 1977 outside of Amazonian Region, in Recife, State of Pernambuco, Brazil (Diptera: Psychodidae: Phlebotominae).

Lutzomyia umbratilis, a known vector of Leishmania guyanensis in the north of Amazon basin, has been exclusively found in the Amazonian region. Here we report for the first time the occurrence of this species in northeastern Brazil. The epidemiological importance of the occurrence of this species in the Atlantic Forest is commented.

Animals↗

Mortality among radiologic technologists in the united states (1926-1997). 2(nd) Follow up.

PURPOSE: To evaluate risk for all-cause and cause-specific mortality in a large, primarily female (73%) cohort of radiologic technologists.METHODS: The study consists of 145,915 radiation technologists, certified in the American Registry of Radiologic Technologists (1926-1982) and followed through 1997. Causes of death were obtained from death certificates or, more recently, through NDI Plus. Standardized Mortality Ratios (SMR) were computed and tests of homogeneity were performed to detect differences in mortality among causes. Poisson models were used to estimate risks using an internal comparison group.RESULTS: Significantly low SMRs were observed for all causes (0.76), all cancers (0.82), and diseases of circulatory system (0.69). Compared to U.S. women, the risk for breast cancer mortality bordered around unity (SMR 1.01, 95% CI 0.94-1.09). However, relative to all other cancers, breast cancer mortality was significantly increased (RSMR 1.24, p < 0.01). Elevated risk for breast cancer was associated with certification before 1940 (SMR 1.55, 95% CI 1.24-1.91), and duration of certification of 20-29 (SMR 1.21, 95% CI 1.06-1.37) and 30+ years (SMR 1.77, 95% CI 1.54-2.02). A 35% increase in leukemia risk was evident for women certified for a duration of 20-29 years and a 36% increase among women certified for 30+ years. Poisson analysis revealed a significant increase in breast cancer risk with increasing number of years certified among women first certified before 1940 (p < 0.001) and during 1940-49 (p = 0.05) compared to women first certified in 1950 or later.CONCLUSIONS: Preliminary findings of this study suggest increased breast cancer risk associated with occupational radiation exposures prior to 1950 and with long-term cumulative exposures. However, potential confounding by reproductive and other risk factors needs to be evaluated.

Journal Article↗

Trends in reproductive, smoking, and other chronic disease risk factors by birth cohort in a large occupational study.

PURPOSE: To illustrate the value of using large cohort studies to identify birth cohort trends in several chronic disease risk factors.METHODS: In collaboration with the American Registry of Radiologic Technologists (ARRT) and the University of Minnesota, the National Cancer Institute (NCI) initiated a cohort study of radiologic technologists who were certified by ARRT for at least two years between 1926 and 1982. Over 90,000 technologists (nearly four-fifths female) from all 50 states responded to a mailed questionnaire on reproductive, medical, work, and lifestyle factors. Ten, mostly five-year, birth cohorts, from before 1920 through 1960 and later, were evaluated.RESULTS: In this population, the mean height of both men and women generally rose in each subsequent birth cohort. The proportion of men who smoked before age 18 fell among those born since the late 1920s. In contrast, the proportion of women smoking before age 18 rose among those born since the early 1950s, reaching 14.2% among those born in 1960 and later. The mean age at menarche fell, until leveling off at 12.5, among those born after 1940. Recent birth cohorts (since 1950) show among the highest mean ages at birth of first child (>26 yeras), highest rates of nulliparity at age 25 (>/=63%), and lowest mean parity levels (</=1.7), compared with earlier birth cohorts. The proportion of women ever using oral contraceptives rose steadily by birth cohort, reaching 86 percent in the most recent birth cohort.CONCLUSIONS: Some of the trends observed may help explain historic chronic disease incidence patterns and predict future disease burdens.

Journal Article↗

HIV-1 Tat increases endothelial solute permeability through tyrosine kinase and mitogen-activated protein kinase-dependent pathways.

OBJECTIVE: HIV-1 infection is associated with alterations of several vascular endothelial functions including adhesion molecule expression, growth, and vascular permeability. The bases of these errors are not known, but might involve secretion of the HIV-1 derived transcription factor 'Tat-1'. This study investigated Tat-1 mediated endothelial barrier changes and second message regulation of this phenomenon. METHODS: We exposed human umbilical vein endothelial cell monolayers to Tat-1 (0-150 ng/ml) for up to 48 h and measured resulting changes in monolayer permeability. We also investigated the role of tyrosine and mitogen activated protein (MAP) kinases, and protein kinase G using the pharmacological blockers genistein, PD98059 and KT5823 respectively. RESULTS: Tat-1 significantly reduced monolayer barrier and increased albumin permeability within 24 h. Tat-1 also stimulated tyrosine phosphorylation of multiple endothelial proteins, disorganized junctional phosphotyrosine staining and increased the number of these immunostaining structures. The increased permeability produced by Tat-1 was blocked by genistein and PD98059, but not by KT5823. Genistein and PD98059 pretreatment also prevented the changes in phosphotyrosine immunostaining produced by Tat-1 and blocked phosphorylation of several proteins including MAP kinase. CONCLUSION: These results suggest that HIV may dysregulate endothelial barrier through the effects of Tat-1. These blocker experiments suggest that the effects of Tat are transcription/translation-dependent. These data demonstrate that Tat increases endothelial albumin permeability in vitro through tyrosine kinase and MAP kinase, but not protein kinase G pathways.

Blotting, Western↗

A quantitative evaluation of the permeability of the blood brain barrier of portacaval shunted rats.

The integrity of the blood-brain barrier (BBB) was measured in male Sprague Dawley rats subjected to 16 weeks of portacaval shunting (PCS), the optimal time required for the cerebral changes to develop, by using an in situ brain perfusion technique. The penetration of a vascular space marker 14C mannitol, and labelled amino acids 3H-phenylalanine or 3H-glutamate were measured in brain and cerebrospinal fluid (CSF) using an in situ brain perfusion technique, over 2 or 20 minutes. The patency of the surgical shunt was confirmed by measurement of significantly increased plasma ammonia (131.5 +/- 14.8 micromol x l(-1)) and AST (159.5 +/- 19.9 IU x l(-1)) concentrations compared to controls 39.9 +/- 3.7*, and 82.5 +/- 6.6* respectively. Brain and CSF 14C-mannitol space (ml x 100g(-1)), was not increased by PCS where brain space was 1.31 +/- 0.27 mL x 100g(-1) compared to control 1.19 +/- 0.49 mL x 100g(-1), and CSF was 0.14 +/- 0.06 mL x 100g(-1) compared to control 0.15 +/- 0.05 (PCS n=10, control n=8). The uptake for 3H-glutamate, which is required for cerebral ammonia detoxification, was also unchanged in both brain and CSF. However, brain uptake of 3H-phenylalanine was significantly reduced from 871 +/- 80 microL x min(-1) x g(-1) to 356 +/- 154* microl x min(-1) x g(-1) (n=4), although there was no change in CSF uptake. These data suggest that there is no generalized breakdown of the blood-brain or blood-CSF barriers during PCS as assessed by mannitol penetration. The reduction in phenylalanine uptake into the brain may help stabilize high cerebral aromatic amino acid levels. *P<0.05, Two-tailed, Student's unpaired t-test.

Acetates↗

Leukemic cells from murine myeloid leukemia display an intrinsic ability for autonomous proliferation.

OBJECTIVE: Human acute myeloid leukemia (AML) cells can proliferate in vitro in the absence of added growth factors when cultured at high cell density. Autocrine growth factor production is a postulated mechanism of autonomous growth. We sought to examine this using murine AML cells. MATERIALS AND METHODS: We have utilized a Moloney murine leukemia virus (M-MuLV) model of AML to investigate the nature of autonomous in vitro growth of myeloid leukemic cells. RESULTS: Like human AML, M-MuLV-induced myeloid leukemic cells displayed autonomous growth in unstimulated high cell density cultures. However, replating of individual, primary, growth factor autonomous colonies of leukemic cells demonstrated the presence of clonogenic cells capable of autonomous growth when cultured at low cell density. In addition, there was heterogeneity in the progeny of these cells: both factor-dependent leukemic cells and cells autonomous of exogenous factor were observed. CONCLUSION: We propose that clonogenic cells capable of autonomous growth at low cell density represent leukemic progenitors while the majority of leukemic cells derived from these "autonomous" leukemic cells are factor-dependent.

Acute Disease↗