Household transmission of SARS-CoV-2 in a rural area in South Africa.

BACKGROUND: Patterns of SARS-CoV-2 spread have varied by geolocation, with differences in seroprevalence between urban and rural areas, and between waves. Household spread of SARS-CoV-2 is a known source of new COVID-19 infections, with rural areas in sub-Saharan Africa being more prone than urban areas to COVID-19 transmission because of limited access to water in some areas, delayed health- seeking behaviour and poor access to care. OBJECTIVES: To explore SARS-CoV-2 infection incidence and transmission in rural households in South Africa (SA). METHODS: We conducted a prospective household cluster investigation between 13 April and 21 July 2021 in the Matjhabeng subdistrict, a rural area in Free State Province, SA. Adults with SARS-CoV-2 confirmed by polymerase chain reaction (PCR) tests (index cases, ICs) and their household contacts (HCs) were enrolled. Household visits conducted at enrolment and on days 7, 14 and 28 included interviewer- administered questionnaires and respiratory and blood sample collection for SARS-CoV-2 PCR and SARS-CoV-2 immunoglobulin G serological testing, respectively. Co-primary cases were HCs with a positive SARS-CoV-2 PCR test at enrolment. The incidence rate (IR), using the Poisson distribution, was HCs with a new positive PCR and/or serological test per 1 000 person-days. Associations between outcomes and HC characteristics were adjusted for intra-cluster correlation using robust standard errors. The secondary infection rate (SIR) was the proportion of new COVID-19 infections among susceptible HCs. RESULTS: Among 23 ICs and 83 HCs enrolled, 10 SARS-CoV-2 incident cases were identified, giving an IR of 5.8 per 1 000 person-days (95% confidence interval (CI) 3.14 - 11.95). Households with a co-primary case had higher IRs than households without a co-primary case (crude IR 14.16 v. 1.75, respectively; p=0.054). HIV infection, obesity and the presence of chronic conditions did not materially alter the crude IR. The SIR was 15.9% (95% CI 7.90 - 29.32). Households with a lower household density (fewer household members per bedroom) had a higher IR (IR 9.58; 95% CI 4.67 - 21.71) than households with a higher density (IR 3.06; 95% CI 1.00 - 12.35). CONCLUSION: We found a high SARS-CoV-2 infection rate among HCs in a rural setting, with 48% of households having a co-primary case at the time of enrolment. Households with co-primary cases were associated with a higher seroprevalence and incidence of SARS-CoV-2. Sociodemographic and health characteristics were not associated with SARS-CoV-2 transmission in this study, and we did not identify any transmission risks inherent to a rural setting.

Usefulness of severity scales for cardiogenic shock in-hospital mortality. Proposal for a new prognostic model.

Cardiogenic shock (CS) is a condition comprising multiple etiologies, which associates high mortality rates. Some scoring systems have been shown to be good predictors of hospital mortality in patients admitted to Critical Care Units (CCU). The main objective of this study is to analyze their usefulness and validity in a cohort of CS patients. METHODS: Observational unicentric study of a cohort of CS patients. SOFA, SAPS II and APACHE II scores were calculated in the first 24 h of CCU admission. RESULTS: 130 patients with CS were included. SOFA, SAPS II and APACHE II scores revealed good discrimination for hospital mortality: (AUC) ROC values (AUC: 0.711, 0.752 and 0.742 respectively; P = .6). Calibration, estimated by the Hosmer-Lemeshow test, was adequate in all cases. Acute coronary syndrome, lactate serum values, SAPS II score and vasoactive inotropic score (VIS) were found to be independent predictors for mortality, upon ICU admission. With these variables, a specific prognostic indicator was developed (SAPS-2-LIVE), which improved predictive capability for mortality in our series (AUC) ROC, 0.825 (95% CI 0.752-0.89). CONCLUSION: In this contemporary CS cohort, the aforementioned scores have been shown to have good predictive ability for hospital mortality. These findings could contribute to a more accurate risk stratification in CS.

Comparison of the periodic slab approach with the finite cluster description of metal-organic interfaces at the example of PTCDA on Ag(110).

We present a comparative study of metal-organic interface properties obtained from dispersion corrected density functional theory calculations based on two different approaches: the periodic slab-supercell technique and cluster models with 32-290 Ag atoms. Fermi smearing and fixing of cluster borders are required to make the cluster calculation feasible and realistic. The considered adsorption structure and energy of a PTCDA molecule on the Ag(110) surface is not well reproduced with clusters containing only two metallic layers. However, all clusters with four layers of silver atoms and sufficient lateral extension reproduce the adsorbate structure within 0.04 Å with respect to the slab-supercell structure and provide adsorption energies of ( -4.45± 0.08 eV) consistent with the slab result of -4.47 eV. Thus, metal-organic adsorbate systems can be realistically represented by properly defined cluster models. © 2018 Wiley Periodicals, Inc.

Short communication: Cytokine profiles from blood mononuclear cells of dairy cows classified with divergent immune response phenotypes.

Genetic selection for enhanced immune response has been shown to decrease disease occurrence in dairy cattle. Cows can be classified as high (H), average, or low responders based on antibody-mediated immune response (AMIR), predominated by type-2 cytokine production, and cell-mediated immune response (CMIR) through estimated breeding values for these traits. The purpose of this study was to identify in vitro tests that correlate with in vivo immune response phenotyping in dairy cattle. Blood mononuclear cells (BMC) isolated from cows classified as H-AMIR and H-CMIR through estimated breeding values for immune response traits were stimulated with concanavalin A (ConA; Sigma Aldrich, St. Louis, MO) and gene expression, cytokine production, and cell proliferation was determined at multiple time points. A repeated measures model, which included the effects of immune response group, parity, and stage of lactation, was used to compare differences between immune response phenotype groups. The H-AMIR cows produced more IL-4 protein than H-CMIR cows at 48 h; however, no difference in gene expression of type-2 transcription factor GATA3 or IL4 was noted. The BMC from H-CMIR cows had increased production of IFN-γ protein at 48, 72, and 96 h compared with H-AMIR animals. Further, H-CMIR cows had increased expression of the IFNG gene at 16, 24, and 48 h post-treatment with ConA, although expression of the type-1 transcription factor gene TBX21 did not differ between immune response groups. Although proliferation of BMC increased from 24 to 72 h after ConA stimulation, no differences were found between the immune response groups. Overall, stimulation of H-AMIR and H-CMIR bovine BMC with ConA resulted in distinct cytokine production profiles according to genetically defined groups. These distinct cytokine profiles could be used to define disease resistance phenotypes in dairy cows according to stimulation in vitro; however, other immune response phenotypes should be assessed.

Minimum oxygen flow needed for vital support during simulated post-cardiorespiratory arrest resuscitation.

According to the ERC and the AHA guidelines, FiO2 should be titrated to achieve an O2Sat ≥ 94%. The aim of this study was to determine the minimum oxygen flow and time needed to reach an FiO2 of 0.32 and 0.80 during post-cardiac arrest care. An experimental analysis was performed that consisted of a simulated post-cardiac arrest situation. Different resuscitators were tested and connected to an artificial lung: Mark IV, SPUR II, Revivator Res-Q, O-TWO. The oxygen flow levels tested were 2, 5, 10 and 15 lpm. Bonferroni and Mann-Whitney U tests were used. An FiO2 of 0.32 or more was obtained using any of the oxygen flow and resuscitators. Only the Mark IV achieved an FiO2 of 0.80 after a minimum of 75s ventilating with 2 or 5 lpm. Clinical and statistical differences (P<.05) were found: at 15 lpm it took 35s to reach an FiO2 of 0.80 or more for Mark IV (85.6 [0.3]) and Revivator (84.3 [1.5]) compared to 50s for SPUR II (87.1 [6.4]); at 2 lpm, all of the devices reached an FiO2 of ≥ 0.32 at 30s(Mark IV (34.8 [1.3]), Revivator (35.7 [1.5]) and SPUR II (34.4 [2.1]), except for O-TWO, which took 35s (36.3 [4.3]). Patients could be ventilated with any of the resuscitators using 2 lpm to obtain an FiO2 of 0.32, although possibly O-TWO would be the last option during the first 60s. In order to reach an FiO2 of 0.80, ventilating with 10 lpm should be sufficient, and preferably using Mark IV or Revivator Res-Q. In conclusion, on observing the results of our study, in any possible scenario, it would be advisable to use Revivator Res-Q or Mark IV rather than O-TWO or SPUR II.

Formation of an Organic/Metal Interface State from a Shockley Resonance.

The emergence of an organic/metal interface state is studied for the interface between 3,4,9,10-perylene-tetracarboxylic acid dianhydride (PTCDA) and Ag(100) by means of time- and angle-resolved two-photon photoemission and density functional theory. The interface state is located in the projected band gap of Ag(100), 2.3 eV above the Fermi level, and has an electron lifetime in the 10 fs range. The interface state has no direct counterpart on the clean Ag(100) surface, which only exhibits a broad resonance at ≈1 eV lower energy. Our results thus show that organic overlayers are able to shift such a resonance substantially and even qualitatively change its character to a distinct electronic state at the interface.

Ecophysiological function of leaf 'windows' in Lithops species - 'Living Stones' that grow underground.

Leaf temperatures were lower when light entry at the leaf tip window was prevented through covering the window with reflective tape, relative to leaf temperatures of plants with leaf tip windows covered with transparent tape. This was true when leaf temperatures were measured with an infrared thermometer, but not with a fine-wire thermocouple. Leaf tip windows of Lithops growing in high-rainfall regions of southern Africa were larger than the windows of plants (numerous individuals of 17 species) growing in areas with less rainfall and, thus, more annual insolation. The results of this study indicate that leaf tip windows of desert plants with an underground growth habit can allow entry of supra-optimal levels of radiant energy, thus most likely inhibiting photosynthetic activity. Consequently, the size of the leaf tip windows correlates inversely with habitat solar irradiance, minimising the probability of photoinhibition, while maximising the absorption of irradiance in cloudy, high-rainfall regions.

The interaction of iron pyrite with oxygen, nitrogen and nitrogen oxides: a first-principles study.

Sulphide materials, in particular MoS(2), have recently received great attention from the surface science community due to their extraordinary catalytic properties. Interestingly, the chemical activity of iron pyrite (FeS(2)) (the most common sulphide mineral on Earth), and in particular its potential for catalytic applications, has not been investigated so thoroughly. In this study, we use density functional theory (DFT) to investigate the surface interactions of fundamental atmospheric components such as oxygen and nitrogen, and we have explored the adsorption and dissociation of nitrogen monoxide (NO) and nitrogen dioxide (NO(2)) on the FeS(2)(100) surface. Our results show that both those environmentally important NO(x) species chemisorb on the surface Fe sites, while the S sites are basically unreactive for all the molecular species considered in this study and even prevent NO(2) adsorption onto one of the non-equivalent Fe-Fe bridge sites of the (1 × 1)-FeS(2)(100) surface. From the calculated high barrier for NO and NO(2) direct dissociation on this surface, we can deduce that both nitrogen oxides species are adsorbed molecularly on pyrite surfaces.

Catastrophizing: a predictor of persistent pain among women with endometriosis at 1 year.

BACKGROUND: Endometriosis is the most common gynecological diagnosis among women with chronic pelvic pain, but the underlying mechanisms of endometriosis-associated chronic pelvic pain remain unclear. Therefore, the objective of this study was to determine the biopsychosocial predictors of pain improvement among women with endometriosis. METHODS: One hundred and fifteen women who presented for treatment of endometriosis-associated chronic pelvic pain at a tertiary referral center at a university-based hospital participated in this prospective observational study of clinical practice. Participants completed questionnaires assessing pain, mental health and catastrophizing at entry and 1 year follow-up. The main outcome measure assessed was the interval change in pain report using the McGill pain 1uestionnaire. RESULT(S): On average, participants experienced a 37.4% reduction in interval pain (P < 0.001). Adjusted for baseline pain, nulliparity (P = 0.002) and catastrophizing (P = 0.04) were associated with decreased probability of interval improvement in pain. Those referred for physical therapy had less interval pain improvement (P = 0.04). However, undergoing hysterectomy was a strong predictor of improvement in pain (P = 0.008). CONCLUSION(S): Our study suggests that chronic pain in endometriosis may be more akin to other idiopathic pain disorders. Specifically, biopsychosocial variables, such as catastrophizing, play an important role in reported severity. Further research on biopsychosocial correlates of chronic pelvic pain in endometriosis is warranted.

Efficacy of spinal manipulation and mobilisation on trunk flexibility and stiffness in horses: a randomised clinical trial.

REASONS FOR PERFORMING THE STUDY: Spinal mobilisation and spinal manipulative therapy (SMT) are being applied to horses; however, there are limited objective measures of their effects on spinal mobility or stiffness in actively ridden horses. OBJECTIVES: To quantify passive spinal movements induced during dorsoventral mobilisation of the trunk and to identify any potential effects of SMT on measures of spinal mobility within the thoracolumbar region in standing horses. We hypothesise that displacement amplitudes will be significantly increased across vertebral levels after SMT, compared to spinal mobilisation only within the control group. METHODS: Passive spinal mobility was assessed in 24 actively ridden mature horses once a week for 3 weeks. Peak vertical displacement, loading and unloading velocities, applied force, stiffness and the frequency of truncal oscillations induced during dorsoventral spinal mobilisation were measured at 5 thoracolumbar sites and compared between treatment (n = 12) and control (n = 12) groups. Each week, outcome parameters were measured pre- and post intervention, 10 min apart. Treatment consisted of manually-applied, high-velocity, low-amplitude (HVLA) thrusts directed at the 5 intervertebral sites. Control horses received no additional intervention. A mixed-effects linear regression model was used to assess the interactive effects of treatment group, vertebral level, week and pre-/post intervention. RESULTS: Post intervention displacement amplitudes of the trunk and applied forces were significantly higher in the SMT group, compared to the control group. A similar trend was found for increased spinal stiffness within the SMT group. Across vertebral levels, SMT induced a 40% increase in displacement, a 20% increase in applied force and a 7% increase in stiffness. CONCLUSIONS AND POTENTIAL RELEVANCE: SMT increased dorsoventral displacement of the trunk, which is indicative of producing increased passive spinal flexibility in actively ridden horses. Further clinical research is needed on the effectiveness of manual therapies in horses with objective measures of back pain, stiffness and poor performance.

Movement of water from old to young leaves in three species of succulents.

A hypothetical adaptive response of succulent plants to drought-stress is the redistribution of water from old to young leaves. We examined the effects of possible movement of water from old to young leaves in three succulent species, Carpobrotus edulis (weak CAM-inducible), Kalanchoe tubiflora (CAM) and Sedum spectabile (possibly a CAM-cycler or CAM-inducible). Old leaves were removed from plants, and photosynthesis, transpiration, f. wt : d. wt ratios, diurnal acid fluctuations, stomatal conductance and internal CO2 concentrations of the remaining young leaves were measured during drought-stress. Comparison was made with plants retaining old leaves. There was no evidence that water moved from old to young leaves during drought-stress as previously hypothesized. Only in drought-stressed plants of K. tubiflora, were photosynthetic and transpiration rates of young leaves greater on shoots with old leaves removed compared with attached. There was a trend in all species for greater fluctuations in acidity in young leaves on shoots that lacked older leaves. For two of the three species studied, the f. wt : d. wt ratios of young leaves were greater under drought-stress, on shoots with old leaves removed than with them attached. Absence of old leaves may reduce competition for water with young leaves, which consequently have higher water content and greater photosynthetic rates.

Yeast desaturases.

The Saccharomyces OLE1 gene encodes the intrinsic membrane-bound Delta-9 fatty acid desaturase. OLE1 expression is regulated at the levels of transcription and mRNA stability by nutrient fatty acids and molecular oxygen. Its transcription is controlled through two distinct promoter elements, the fatty acid response element (FAR) region, and a downstream low-oxygen response element (LORE) that dramatically amplifies FAR-activated expression under hypoxic or cobalt-stimulated growth conditions. Transcription activation through both elements is repressed by unsaturated fatty acids. The half-life of the OLE1 mRNA is also dramatically reduced upon exposure to unsaturated fatty acids. OLE1 expression is governed by two homologous membrane-bound proteins, Spt23p and Mga2p, which activate OLE1 expression through N-terminal polypeptides that are released from the membrane through a ubiquitin-mediated mechanism that involves processing by the 23 S proteosome. Although proteolytic processing of Spt23p can be repressed by polyunsaturated fatty acids, Mga2p processing in normoxic cells appears to be regulated by a different mechanism. Mga2p is essential, however, for the induction of the high levels of expression that are triggered by hypoxia through the LORE promoter element. Surprisingly, Mga2p also plays a critical role in controlling OLE1 mRNA stability, suggesting that there may be a functional linkage between OLE1 transcription and the regulation of OLE1 mRNA stability.

Modification of fatty acids changes the flavor volatiles in tomato leaves.

Expression of the yeast Delta9 desaturase gene in tomato (Lycopersicon esculentum Mill.) resulted in changes in the profiles of fatty acids in tomato leaves. Transgenic leaves displayed a dramatic increase in cis-Delta9 16:1, which only existed in a small quantity in control leaves. Also higher, but not as dramatic, were 18:1 and 16:3 fatty acids. Several fatty acids, viz. 16:0, 18:0, and 18:3 declined in transgenic leaves. Changes in fatty acids were accompanied by changes in certain volatile compounds derived from fatty acids. On a percentage basis, most notable increases (>3-fold) were 1-hydroxy-2-butanone, 1-penten-3-ol, heptanal, 3-hexen-1-ol, 2-octanol,cis-3-hexenal, hexanal and 2-nonenal. Several flavor compounds not known to be biochemically derived from fatty acids, viz. 2-ethyl-furan, 5-ethyl-2-[5H]-furanone, eugenol, and 2-ethylthiophene also showed sharp increases in transgenic leaves.

The membrane proteins, Spt23p and Mga2p, play distinct roles in the activation of Saccharomyces cerevisiae OLE1 gene expression. Fatty acid-mediated regulation of Mga2p activity is independent of its proteolytic processing into a soluble transcription activator.

The Saccharomyces OLE1 gene encodes the Delta-9 fatty acid desaturase, an enzyme that converts saturated fatty acyl-CoAs into cis-Delta-9 unsaturated fatty acids. OLE1 gene expression is regulated by unsaturated fatty acids, which repress transcription and destabilize the OLE1 mRNA. Expression of OLE1 is activated by N-terminal proteolytic fragments of two homologous endoplasmic reticulum membrane proteins, Spt23p and Mga2p. Disruption of either gene does not significantly affect cell growth or fatty acid metabolism; cells that contain null alleles of both genes, however, are unsaturated fatty acid auxotrophs. An analysis of spt23Delta and mga2Delta strains shows that Spt23p and Mga2p differentially activate and regulate OLE1 transcription. In glucose-grown cells, both genes activate transcription to similar levels of activity. Expressed alone, Mga2p induces high levels of OLE1 transcription in cells exposed to cobalt or grown in glycerol-containing medium. Spt23p expressed alone activates OLE1 transcription to levels similar to those in wild type cells. OLE1 expression is strongly repressed by unsaturated fatty acids in spt23Delta or mga2Delta cells, under all growth conditions. To test if OLE1 expression is controlled by fatty acids at the level of membrane proteolysis, soluble N-terminal fragments of Spt23p and Mga2p that lack their membrane-spanning regions (Deltatm) were expressed under the control of their native promoters in spt23Delta;mga2Delta cells. Under those conditions, Mga2pDeltatm acts as a powerful transcription activator that is strongly repressed by unsaturated fatty acids. By comparison, the Spt23pDeltatm polypeptide weakly activates transcription and shows little regulation by unsaturated fatty acids. Co-expression of the two soluble fragments results in activation to levels observed with the Mga2pDeltatm protein alone. The fatty acid repression of transcription under those conditions is attenuated by Spt23Deltatm, however, suggesting that the two proteins may interact to modulate OLE1 gene expression.

MGA2 is involved in the low-oxygen response element-dependent hypoxic induction of genes in Saccharomyces cerevisiae.

Eukaryotes have the ability to respond to changes in oxygen tension by alterations in gene expression. For example, OLE1 expression in Saccharomyces cerevisiae is upregulated under hypoxic conditions. Previous studies have suggested that the pathway regulating OLE1 expression by unsaturated fatty acids may involve Mga2p and Spt23p, two structurally and functionally related proteins. To define the possible roles of each of these genes on hypoxia-induced OLE1 expression, we examined OLE1 expression under normoxia, hypoxia, and cobalt treatment conditions in Deltamga2 or Deltaspt23 deletion strains. The results of OLE1 promoter-lacZ reporter gene and Northern blot analyses showed that hypoxia- and cobalt-induced OLE1 expression was dramatically decreased in a Deltamga2 strain but not in a Deltaspt23 strain. Further analyses using low-oxygen response element (LORE)-CYC1-lacZ fusion reporter assays and electrophoretic mobility shift assays (EMSAs) demonstrated that MGA2 significantly affects the LORE-dependent hypoxic induction pathway of gene expression. When MGA2 was supplied by a plasmid, the LORE-dependent hypoxia-inducible reporter expression was recovered, as was the hypoxia-inducible complex in EMSAs in the S. cerevisiae Deltamga2 strain. Supershift analysis of EMSAs using crude extracts containing mycMga2p indicated that Mga2p is a component of the LORE-binding complex. Another LORE-dependent, hypoxia-inducible gene, ATF1, was similarly affected in the Deltamga2 strain. These results indicate that MGA2 is required for the LORE-dependent hypoxic gene induction in S. cerevisiae.

Identification and characterization of a low oxygen response element involved in the hypoxic induction of a family of Saccharomyces cerevisiae genes. Implications for the conservation of oxygen sensing in eukaryotes.

An organism's ability to respond to changes in oxygen tension depends in large part on alterations in gene expression. The oxygen sensing and signaling mechanisms in eukaryotic cells are not fully understood. To further define these processes, we have studied the Delta9 fatty acid desaturase gene OLE1 in Saccharomyces cerevisiae. We have confirmed previous data showing that the expression of OLE1 mRNA is increased in hypoxia and in the presence of certain transition metals. OLE1 expression was also increased in the presence of the iron chelator 1,10-phenanthroline. A 142-base pair (bp) region 3' to the previously identified fatty acid response element was identified as critical for the induction of OLE1 in response to these stimuli using OLE1 promoter-lacZ reporter constructs. Electromobility shift assays confirmed the presence of an inducible band shift in response to hypoxia and cobalt. Mutational analysis defined the nonameric sequence ACTCAACAA as necessary for transactivation. A 20-base pair oligonucleotide containing this nonamer confers up-regulation by hypoxia and inhibition by unsaturated fatty acids when placed upstream of a heterologous promoter in a lacZ reporter construct. Additional yeast genes were identified which respond to hypoxia and cobalt in a manner similar to OLE1. A number of mammalian genes are also up-regulated by hypoxia, cobalt, nickel, and iron chelators. Hence, the identification of a family of yeast genes regulated in a similar manner has implications for understanding oxygen sensing and signaling in eukaryotes.

Tsc13p is required for fatty acid elongation and localizes to a novel structure at the nuclear-vacuolar interface in Saccharomyces cerevisiae.

The TSC13/YDL015c gene was identified in a screen for suppressors of the calcium sensitivity of csg2Delta mutants that are defective in sphingolipid synthesis. The fatty acid moiety of sphingolipids in Saccharomyces cerevisiae is a very long chain fatty acid (VLCFA) that is synthesized by a microsomal enzyme system that lengthens the palmitate produced by cytosolic fatty acid synthase by two carbon units in each cycle of elongation. The TSC13 gene encodes a protein required for elongation, possibly the enoyl reductase that catalyzes the last step in each cycle of elongation. The tsc13 mutant accumulates high levels of long-chain bases as well as ceramides that harbor fatty acids with chain lengths shorter than 26 carbons. These phenotypes are exacerbated by the deletion of either the ELO2 or ELO3 gene, both of which have previously been shown to be required for VLCFA synthesis. Compromising the synthesis of malonyl coenzyme A (malonyl-CoA) by inactivating acetyl-CoA carboxylase in a tsc13 mutant is lethal, further supporting a role of Tsc13p in VLCFA synthesis. Tsc13p coimmunoprecipitates with Elo2p and Elo3p, suggesting that the elongating proteins are organized in a complex. Tsc13p localizes to the endoplasmic reticulum and is highly enriched in a novel structure marking nuclear-vacuolar junctions.

The Saccharomyces cerevisiae FAT1 gene encodes an acyl-CoA synthetase that is required for maintenance of very long chain fatty acid levels.

The Saccharomyces cerevisiae FAT1 gene appears to encode an acyl-CoA synthetase that is involved in the regulation of very long chain (C20-C26) fatty acids. Fat1p, has homology to a rat peroxisomal very long chain fatty acyl-CoA synthetase. Very long chain acyl-CoA synthetase activity is reduced in strains containing a disrupted FAT1 gene and is increased when FAT1 is expressed in insect cells under control of a baculovirus promoter. Fat1p accounts for approximately 90% of the C24-specific acyl-CoA synthetase activity in glucose-grown cells and approximately 66% of the total activity in cells grown under peroxisomal induction conditions. Localization of functional Fat1p:green fluorescent protein gene fusions and subcellular fractionation of C24 acyl-CoA synthetase activities indicate that the majority of Fat1p is located in internal cellular locations. Disruption of the FAT1 gene results in the accumulation of very long chain fatty acids in the sphingolipid and phospholipid fractions. This includes a 10-fold increase in C24 acids and a 6-fold increase in C22 acids. These abnormal accumulations are further increased by perturbation of very long chain fatty acid synthesis. Overexpression of Elo2p, a component of the fatty acid elongation system, in fat1Delta cells causes C20-C26 levels to rise to approximately 20% of the total fatty acids. These data suggest that Fat1p is involved in the maintenance of cellular very long chain fatty acid levels, apparently by facilitating beta-oxidation of excess intermediate length (C20-C24) species. Although fat1Delta cells were reported to grow poorly in oleic acid-supplemented medium when fatty acid synthase activity is inactivated by cerulenin, fatty acid import is not significantly affected in cells containing disrupted alleles of FAT1 and FAS2 (a subunit of fatty acid synthase). These results suggest that the primary cause of the growth-defective phenotype is a failure to metabolize the incorporated fatty acid rather than a defect in fatty acid transport. Certain fatty acyl-CoA synthetase activities, however, do appear to be essential for bulk fatty acid transport in Saccharomyces. Simultaneous disruption of FAA1 and FAA4, which encode long chain (C14-C18) fatty acyl-CoA synthetases, effectively blocks the import of long chain saturated and unsaturated fatty acids.

MR imaging of epicondylitis.

OBJECTIVE: To systematically evaluate the MR findings in patients with epicondylitis compared with asymptomatic volunteers. DESIGN AND PATIENTS: We imaged 43 elbows: 24 with epicondylitis (22 lateral, 2 medial) diagnosed by clinical examination, and 19 in 16 normal volunteers. MRI was performed at 1.5 T using axial T1-weighted, axial fat-saturated FSE, and coronal or sagittal Fast STIR sequences. Two independent observers evaluated the images for intratendon signal, tendon thickening, periosteal reaction, fluid in the radial head bursa, and anconeus edema. RESULTS: All 24 patients with epicondylitis had increased signal on fat-saturated FSE and Fast STIR images. Twenty-two of these patients had increased intratendon T1 signal, and 19 had tendon thickening. No patient demonstrated fluid in the radial head bursa or periosteal reaction. Only two patients had subtle anconeus edema, while three patients unexpectedly had increased T2 signal within the involved epicondyle. One asymptomatic volunteer (high-performance athlete) had increased T1 and T2 signal with tendon thickening. An additional two asymptomatic volunteers had increased T1 signal only. CONCLUSION: MRI of epicondylitis demonstrates tendon thickening with increased T1 and T2 signal, but these findings may be seen in a small minority of asymptomatic individuals. Anconeus edema, previously demonstrated on MRI in epicondylitis, was only rarely found, and distension of the radial head bursa, surgically described, was not seen. Increased marrow T2 signal within the involved epicondyle is occasionally seen.