| 2021 | Novel Ampeloviruses Infecting Cassava in Central Africa and the South-West Indian Ocean Islands. | Viruses | Cassava is one of the most important staple crops in Africa and its production is seriously damaged by viral diseases. In this study, we identify for the first time and characterize the genome organization of novel ampeloviruses infecting cassava plants in diverse geographical locations using three high-throughput sequencing protocols [Virion-Associated Nucleotide Acid (VANA), dsRNA and total RNA], and we provide a first analysis of the diversity of these agents and of the evolutionary forces acting on them. Thirteen new isolates were characterized in field-grown cassava plants from the Democratic Republic of Congo (DR Congo), Madagascar, Mayotte, and Reunion islands. The analysis of the sequences of the corresponding contigs (ranging between 10,417 and 13,752 nucleotides in length) revealed seven open reading frames. The replication-associated polyproteins have three expected functional domains: methyltransferase, helicase, and RNA-dependent RNA polymerase (). Additional open reading frames code for a small transmembrane protein, a heat-shock protein 70 homolog (), a heat shock protein 90 homolog (), and a major and a minor coat protein ( and respectively). Defective genomic variants were also identified in some cassava accessions originating from Madagascar and Reunion. The isolates were found to belong to two species tentatively named Manihot esculenta-associated virus 1 and 2 (MEaV-1 and MEaV-2). Phylogenetic analyses showed that MEaV-1 and MEaV-2 belong to the genus , in particular to its subgroup II. MEaV-1 was found in all of the countries of study, while MEaV-2 was only detected in Madagascar and Mayotte. Recombination analysis provided evidence of intraspecies recombination occurring between the isolates from Madagascar and Mayotte. No clear association with visual symptoms in the cassava host could be identified. |
| 2021 | Sodium R-lipoate and enzymatically-modified isoquercitrin suppressed IgE-independent anaphylactic reactions and stress-induced gastric ulceration in mice. | Int Immunopharmacol | Anaphylaxis is a life-threatening allergic reaction, for which the worldwide prevalence is rapidly increasing. The currently used synthetic antiallergic drugs have a high tendency to cause adverse effects, like gastric ulcers, in long-term use. Therefore, a great deal of attention has been given to develop new safer and more effective antiallergic agents from natural compounds that are chemically/enzymatically-modified. Here, we evaluated/compared the efficacy of two different doses (50 and 100 mg/kg body weight "b.w", given orally) of sodium R-lipoate (NaRLA) and enzymatically-modified isoquercitrin (EMIQ) in alleviating both local/systemic non-immunological anaphylactic reactions and stress-induced gastric ulceration in mice, in comparison with sulfasalazine (SSZ) as a reference drug. The results indicated that the pre-treatment of animals with NaRLA or EMIQ (especially at 100 mg/kg b.w) completely succeeded, as SSZ, in alleviating the hind paw edema induced by either histamine or compound 48/80 (Cpd 48/80). Furthermore, NaRLA and EMIQ prevented the mast cell degranulation and anaphylactic shock caused by Cpd 48/80 (in a dose-dependent manner) and reduced significantly (P < 0.001) the histamine release from the mouse peritoneal mast cells, like SSZ. Moreover, their use was associated with alleviating both gastric histopathological and biochemical alterations in the water-restraint stress (WRS) mice model towards the control values. They also decreased the percentage of degranulated mesenteric mast cells in the WRS mice model. In conclusion, our findings provide possibility that both NaRLA and EMIQ may serve as an effective therapeutic agents for mast cells-dependent anaphylactic reactions without risks of inducing gastric ulcers. |
| 2021 | Cell Population Data (CPD) for Early Recognition of Sepsis and Septic Shock in Children: A Pilot Study. | Front Pediatr | Innovative Cell Population Data (CPD) have been used as early biomarkers for diagnosing sepsis in adults. We assessed the usefulness of CPD in pediatric patients with sepsis/septic shock, in terms of early recognition and outcome prediction. We revised 54 patients (0-15 y) admitted to our Pediatric Intensive Care Unit (PICU) for sepsis/septic shock during a 4-year period. Twenty-eight patients were excluded, 26 septic patients were enrolled (G1). Forty children admitted for elective surgery served as controls (G2). Data on five selected CPD parameters, namely neutrophils fluorescence intensity (NE-SFL), monocytes cells complexity (MO-X), monocytes fluorescence intensity (MO-Y), monocytes complexity and width of dispersion of events measured (MO-WX), and monocytes cells size and width dispersion (MO-WZ), were obtained at time of PICU admission () by a hematological analyzer (Sysmex XN 9000®). As the primary outcome we evaluated the relevance of CPD for diagnosing sepsis/septic shock on PICU admission. Furthermore, we investigated if CPD at were correlated with C-reactive protein (CRP), patient survival, or complicated sepsis course. On PICU admission (), NE-SFL, MO-WX, and MO-Y were higher in sepsis/septic shock patients compared to controls. NE-SFL values were correlated with CRP values in G1 patients ( = 0.83). None of the five CPD parameters was correlated with survival or complicated sepsis course. We found higher values of NE-SFL, MO-WX, and MO-Y in children with sepsis/septic shock upon PICU admission. These parameters may be a promising adjunct for early sepsis diagnosis in pediatric populations. Larger, prospective studies are needed to confirm our preliminary observations. |
| 2021 | Ubr1-mediated ubiquitylation orchestrates asexual development, polar growth, and virulence-related cellular events in Beauveria bassiana. | Appl Microbiol Biotechnol | The E3 ubiquitin ligase Ubr1 is a core player in yeast ubiquitylation and protein quality control required for cellular events including proteasomal degradation and gene activity but has been rarely explored in filamentous fungi. We show here an essentiality of orthologous Ubr1-mediated ubiquitylation for the activation of central developmental pathway (CPD) and the CPD-controlled cellular events in Beauveria bassiana, a filamentous fungal insect pathogen that undergoes an asexual cycle in vitro or in vivo. As a result of ubr1 disruption, intracellular free ubiquitin accumulation increased by 1.4-fold, indicating an impaired ability for the disruptant to transfer ubiquitin to target proteins. Consequently, the disruptant was compromised in polar growth featured with curved or hook-like germ tubes and abnormally branched hyphae, leading to impeded propagation of aberrant hyphal bodies in infected insect hemocoel and attenuated virulence. In the mutant, sharply repressed expression of three CDP activator genes (brlA, abaA, and wetA) correlated well with severe defects in aerial conidiation and submerged blastospore (hyphal body) production in insect hemolymph or a mimicking medium. Moreover, the disruptant was sensitive to cell wall perturbation or lysing and showed increased catalase activity and resistance to hydrogen peroxide despite null response to high osmolarity or heat shock. Most of the examined genes involved in polar growth and cell wall integrity were down-regulated in the disruptant. These findings uncover that the Ubr1-mediated ubiquitylation orchestrates polar growth and the CDP-regulated asexual cycle in vitro and in vivo in B. bassiana. KEY POINTS: • Ubr1 is an E3 ubiquitin ligase essential for ubiquitylation in Beauveria bassiana. • Ubr1-mediated ubiquitylation is required for activation of central development pathway. • Ubr1 orchestrates polar growth and asexual cycle in vitro and in vivo. |
| 2020 | Reviewing the value of leukocytes cell population data (CPD) in the management of sepsis. | Ann Transl Med | Sepsis is a medical emergency that describes the body's systemic immune response to an infection and can lead to end-stage organic dysfunction and death. Despite the advances in understanding the pathophysiology of this syndrome and therapies, sepsis remains one of the leading causes of morbidity and mortality in critically ill patients. Early diagnosis and rapid intervention are essential to improve outcomes, which inspired the concept "golden hour," during which the correction of shock and organic dysfunction can improve the patients' outcomes. But the initial presentation of sepsis is often nonspecific and its severity is difficult to assess. Anomalies in temperature, heart and respiratory rates and leukocyte counts are manifestations of systemic inflammatory response syndrome (SIRS). Diagnosis, management and follow-up of patients with sepsis remains a challenge, and diverse biomarkers have been proposed for the timely diagnosis and prognosis of septic patients: lactic acid, procalcitonin (PCT), C-reactive protein, immature granulocytes. The host's initial response to infection is a humoral, cellular and neuroendocrine reaction to infection, and leukocytes interact with endothelial cells. The new generation of hematological analyzers incorporates technological innovations allowing to expand the information derived from the complete blood count: new leukocyte derived parameters are emerging as potentially useful markers in different clinical situations. Additional research parameters cell population data (CPD), characterizing different leukocyte populations have become available, and preliminary observations suggest their utility in the diagnosis of sepsis. This review emphasizes the value of CPD, reported by modern cellular counters for early recognition of sepsis, and therefore the potential improvement in patient outcomes. |
| 2016 | Clinical significance of cell population data (CPD) on Sysmex XN-9000 in septic patients with our without liver impairment. | Ann Transl Med | This study evaluated the clinical significance of cell population data (CPD) parameters obtained on Sysmex XN-9000 in septic patients admitted to intensive care unit (ICU) and stratified according to liver function.The study population consisted in 84 patients, 44 of whom did not develop sepsis (NS), whereas the remaining 40 developed sepsis (SE) (n=24) or septic shock (SS) (n=16). Two hundred ostensibly healthy blood donors [healthy subjects (HS)], undergoing routine blood testing before a regular blood donation, were studied.Except for neutrophils and lymphocytes cell size (NE-FCS and LY-Z), all other CPD values were significantly different in ICU patients compared to HS. Neutrophils and monocytes fluorescence intensity (NE-SFL and MO-X) values were significantly higher in SS compared to sepsis and not develop sepsis patients. The value of many parameters was also different according to liver function. Overall, MO-X and neutrophils fluorescence intensity (NE-SFL) exhibited the best performance for diagnosing sepsis in all patients (AUC, 0.75 and 0.72), as well as in those with (AUC, 0.95 and 0.89) or without (AUC, 0.72 for both) liver impairment. These parameters were also significantly correlated with Sequential Organ Failure Assessment (SOFA) score.This study suggested that some novel CPD parameters (namely NE-SFL and MO-X) may provide useful information for diagnosis and management of sepsis. |
| 2016 | Pain. Part 7: Trigeminal Neuralgia. | Dent Update | Trigeminal neuralgia (TN) is also known as 'tic douloureux' (in French, 'painful twitch'). It is a rare chronic facial pain syndrome, characterized by severe, brief, stabbing, 'electric shock-like 'recurrent pain attacks felt in one or more divisions of trigeminal nerve innervation areas. So intense is the elicited pain that TN has a significant effect on a sufferer's quality of life, rendering many patients unable to consider a future with the ongoing threat of recurrent pain. The aim of this article is to discuss the diagnosis and management of this disabling facial pain condition. CPD/Clinical Relevance: As general medical practitioners may struggle differentiating TN from toothache, primary care dentists have an important role in excluding odontogenic cause of pain, diagnosing TN and referring patients to a facial pain clinic for further investigations and multidisciplinary team management. |
| 2013 | Candidate gene expression associated with geographical variation in cryoprotective dehydration of Megaphorura arctica. | J Insect Physiol | A number of small and permeable invertebrates survive subzero temperatures by cryoprotective dehydration (CPD) in which animals readily lose water to equilibrate body fluid melting points with surrounding temperature thereby avoiding the risk of freezing. Population studies are useful for detecting evolutionary climatic adaptation by comparing populations from locations differing in climatic characteristics. To identify the existence of adaptive variation for important physiological mechanisms underlying the CPD capacity we investigated the gene expression profile of five candidate genes as well as water content and cryoprotectant concentrations in five natural populations from diverse climatic origins. Our results show that Arctic populations, originating from an area with severe winter conditions (Svalbard), respond differently than the populations coming from more benign conditions (Mainland Norway). The Svalbard populations lost water and accumulated trehalose faster in response to cold exposure at -6 °C. The gene expression results suggests that the Svalbard populations experience less cellular perturbation and has a lesser need for molecular chaperones (hsp70) during CPD, but handles the stress by early and rapid induction of cryoprotectant producing enzymes (tps) and oxidative stress scavengers (sod) and possibly also membrane modifications (fad). Thus, these traits relate to the severity of the climate adapted to and are likely markers of their adaptive history. |
| 2011 | Inositol hexakisphosphate-dependent processing of Clostridium sordellii lethal toxin and Clostridium novyi alpha-toxin. | J Biol Chem | Clostridium sordellii lethal toxin and Clostridium novyi α-toxin, which are virulence factors involved in the toxic shock and gas gangrene syndromes, are members of the family of clostridial glucosylating toxins. The toxins inactivate Rho/Ras proteins by glucosylation or attachment of GlcNAc (α-toxin). Here, we studied the activation of the autoproteolytic processing of the toxins by inositol hexakisphosphate (InsP(6)) and compared it with the processing of Clostridium difficile toxin B. In the presence of low concentrations of InsP(6) (<1 μM), toxin fragments consisting of the N-terminal glucosyltransferase (or GlcNAc-transferase) domains and the cysteine protease domains (CPDs) of C. sordellii lethal toxin, C. novyi α-toxin, and C. difficile toxin B were autocatalytically processed. The cleavage sites of lethal toxin (Leu-543) and α-toxin (Leu-548) and the catalytic cysteine residues (Cys-698 of lethal toxin and Cys-707 of α-toxin) were identified. Affinity of the CPDs for binding InsP(6) was determined by isothermal titration calorimetry. In contrast to full-length toxin B and α-toxin, autocatalytic cleavage and InsP(6) binding of full-length lethal toxin depended on low pH (pH 5) conditions. The data indicate that C. sordellii lethal toxin and C. novyi α-toxin are InsP(6)-dependently processed. However, full-length lethal toxin, but not its short toxin fragments consisting of the glucosyltransferase domain and the CPD, requires a pH-sensitive conformational change to allow binding of InsP(6) and subsequent processing of the toxin. |
| 2010 | Comparison of ex vivo and in vitro human fibroblast ageing models. | Mech Ageing Dev | Several studies have analyzed modulation of gene expression during physiological ageing with interesting, but often contradictory results, depending on the model used. In the present report we compare age-related metabolic and synthetic parameters in human dermal fibroblasts (HDF) isolated from young and old subjects (ex vivo ageing model) and cultured from early up to late cumulative population doublings (CPD) (in vitro ageing model) in order to distinguish changes induced in vivo by the aged environment and maintained in vitro, from those associated with cell senescence and progressive CPD. Results demonstrate that fibroblasts from aged donors, already at early CPD, exhibit an impaired redox balance, highlighting the importance of this parameter during ageing, even in the presence of standard environmental conditions, which are considered optimal for cell growth. By contrast, several proteins, as those related to heat shock response, or involved in endoplasmic reticulum and membrane trafficking, appeared differentially expressed only during in vitro ageing, suggesting that, at high CPD, the whole cell machinery becomes permanently altered. Finally, given the importance of the elastic component for a long-lasting connective tissue structural and functional compliance, this study focuses also on elastin and fibulin-5 synthesis and deposition, demonstrating a close relationship between fibulin-5 and ageing. |
| 2010 | Severe, short-duration (0-3 min) heat shocks (50-52 degrees C) inhibit the repair of DNA damage. | Int J Hyperthermia | The goal of this study was to determine whether short-duration (15 s-3 min) high-temperature (50 degrees C) heat shocks inhibit the repair of DNA damage.Cultured HeLa cells were used. DNA damage was measured after UV exposure or X-irradiation. Three methods were used to measure DNA damage: alkaline comet assay with the endonuclease, UVDE, for single strand breaks and UV photoproducts, antibodies specific for cyclo-pyrimidine dimers (CPD) or for 6-4 photoproducts (64PP), and the appearance-resolution of gamma-H2AX foci for DNA double strand breaks.Heat shocks of 15-30 s at 50 degrees C inhibited repair of DNA damage after UV exposure or X-irradiation detected by the alkaline comet assay (after UV) or by persistence of gamma-H2AX foci (after X-rays). The phosphorylation of histone, H2AX, induced by 1 or 4 Gy of X-rays was inhibited in a time-dependent manner after 15-45 s at 52 degrees C. When the excision of UV-induced PP was measured, heat shocks of more than 60 s at 50 degrees C were required to show measurable inhibition.Severe (50 degrees C) short-duration (15 s or greater) heat shocks inhibit repair of UV-induced DNA damage. The ability to detect the inhibitory effects of very short, 15-60 s, heat shocks was assay dependent. The comet assay could detect repair inhibition after a 15-s heat shock. Detection of DNA damage by specific antibodies could only detect repair inhibition after 1-3-min heat shocks. Using the gamma-H2AX foci method 30 s at 50 degrees C induced a significant delay in the repair of DNA damage after 1 Gy of X-rays. |
| 2009 | Continuous peritoneal dialysis compared with daily hemodialysis in patients with acute kidney injury. | Perit Dial Int | In some parts of the world, peritoneal dialysis is widely used for renal replacement therapy (RRT) in acute kidney injury (AKI), despite concerns about its inadequacy. It has been replaced in recent years by hemodialysis and, most recently, by continuous venovenous therapies. We performed a prospective study to determine the effect of continuous peritoneal dialysis (CPD), as compared with daily hemodialysis (dHD), on survival among patients with AKI.A total of 120 patients with acute tubular necrosis (ATN) were assigned to receive CPD or dHD in a tertiary-care university hospital. The primary endpoint was hospital survival rate; renal function recovery and metabolic, acid-base, and fluid controls were secondary endpoints.Of the 120 patients, 60 were treated with CPD (G1) and 60 with dHD (G2). The two groups were similar at the start of RRT with respect to age (64.2 +/- 19.8 years vs 62.5 +/- 21.2 years), sex (men: 72% vs 66%), sepsis (42% vs 47%), shock (61% vs 63%), severity of AKI [Acute Tubular Necrosis Individual Severity Score (ATNISS): 0.68 +/- 0.2 vs 0.66 +/- 0.22; Acute Physiology and Chronic Health Evaluation (APACHE) II: 26.9 +/- 8.9 vs 24.1 +/- 8.2], pre-dialysis blood urea nitrogen [BUN (116.4 +/- 33.6 mg/dL vs 112.6 +/- 36.8 mg/dL)], and creatinine (5.85 +/- 1.9 mg/dL vs 5.95 +/- 1.4 mg/dL). In G1, weekly delivered Kt/V was 3.59 +/- 0.61, and in G2, it was 4.76 +/- 0.65 (p < 0.01). The two groups were similar in metabolic and acid-base control (after 4 sessions, BUN < 55 mg/dL: 46 +/- 18.7 mg/dL vs 52 +/- 18.2 mg/dL; pH: 7.41 vs 7.38; bicarbonate: 22.8 +/- 8.9 mEq/L vs 22.2 +/- 7.1 mEq/L). Duration of therapy was longer in G2 (5.5 days vs 7.5 days; p = 0.02). Despite the delivery of different dialysis methods and doses, the survival rate did not differ between the groups (58% in G1 vs 52% in G2), and recovery of renal function was similar (28% vs 26%).High doses of CPD provided appropriate metabolic and pH control, with a rate of survival and recovery of renal function similar to that seen with dHD. Therefore, CPD can be considered an alternative to other forms of RRT in AKI. |
| Change of excitability in brainstem and cortical visual processing in migraine exhibiting allodynia. | Headache | Clinical and neurophysiological manifestations of information processing associated with central sensitization are little known. Allodynic migraine (AM) can be caused by the sensitization of trigeminal neuron, but no study has reported on AM between attacks using blink reflex (BR) and pattern-reversal visual evoked potentials (PVEPs).We explored the characteristics of AM between attacks associated with central sensitization using BR and PVEP.We recruited 13 patients with interictal AM and 15 patients with nonallodynic migraine (NA), and 30 healthy subjects (HS). BRs were obtained using paired pulses delivered at the interstimulus interval (ISI) of 150, 300, and 500 ms. The ratio of the area in the R2 of the second to R2 of the first shock was measured for each ISI. PVEP were recorded with 2 spatial frequencies (0.5 and 4.0 cpd) and 2 low and high contrasts (29% and 98%, respectively). Amplitudes of P100 were measured.For BR, there were no significant differences in the ratio of the area of the R2 between the sides of stimulation, and the sides of headache. AM patients had less suppression of the R2 at the ISI of 150 and 300 ms when compared with the NA patients and HS. For PVEP, at 0.5, there were significant differences of amplitude between AM patients and HS, and between NA patients and HS in low and high contrast. At 4.0 cpd, there were significant differences of amplitude between AM patients and HS in low contrast, and between AM patients and HS, and NA patients and HS in high contrast. In AM patients, there was a significant difference of amplitude ratio between 0.5 and 4.0 cpd. Conclusions.-Our BR and PVEP study showed that migraine patients exhibiting allodynia may show central sensitization of brainstem trigeminal neuron and have contrast modulating dysfunction during the cortical visual processing of striate and extrastriate on visual cortex in-between attacks. |
| 2005 | First Report of Tomato chlorosis virus and Tomato infectious chlorosis virus in Tomato Crops in France. | Plant Dis | Since 2002, yellowing symptoms associated with high levels of white-fly populations have been observed in plants of protected tomato crops in France. Symptomatic plants exhibited interveinal yellowing areas in older leaves, followed by generalized yellowing. Symptoms were not observed in young plants or fruits. Trialeurodes vaporariorum populations were generally abundant in spring, and Bemisia tabaci (established in France for approximately 10 years) became predominant in summer and fall. To check for the presence of Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV), two whitefly-transmitted criniviruses known to induce yellowing symptoms, 696 samples were collected in the major tomato-growing areas; 573 samples from southern France and 123 samples from northern France. Total RNA was extracted from each sample and analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Primers specific to ToCV (2) and TICV (1,3) were used to amplify either part of the heat-shock-like protein gene HSP70h (both viruses) or part of the diverged coat protein gene (CPd), (TICV only). A 439-bp DNA fragment was obtained with ToCV primers in 178 samples from southern France collected mainly from mid-spring to early fall from 2002 to 2004. Three RT-PCR products amplified from samples collected from diverse growing areas were sequenced and showed 99 to 100% sequence identity with published ToCV sequences from Spain (GenBank Accession Nos. AF215818, AF233435, and AF215817), Portugal (GenBank Accession No. AF234029), Sicily (GenBank Accession No. AY048854), and the United States (GenBank Accession No. AF024630). Considering the high frequency of ToCV-infected samples (41 positive samples of 112 samples collected in 2002, 71 of 295 collected in 2003, and 66 of 166 collected in 2004), this virus appears to be well established in southern France but remains absent in the northern regions. The presence of TICV was tested in 485 samples using the CPd-specific primers or the HSP70h-specific primers. The virus was detected in only two samples from Nice (southeastern France) in 2003 with both primer pairs. The CPd DNA fragment (700 bp) from one of these samples was sequenced, showing 98.9% sequence identity with a TICV Japanese isolate (AB085603). Results of these assays suggest that in contrast to ToCV, TICV is not yet broadly established in France. This difference could be associated with the specificity of the vectors, since ToCV is transmitted by B. tabaci and T. vaporariorum, while TICV is transmitted only by T. vaporariorum (4). References: (1) R. H. Li et al. Plant Dis. 82:84, 1998. (2) D. Louro et al. Eur. J. Plant Pathol. 1065:589, 2000. (3) A. M. Vaira et al. Phytoparasitica 30:290, 2002. (4) G. C. Wisler et al. Plant Dis. 82:271, 1998. |
| 2003 | Nucleotide sequence and genome organization of Cucumber yellows virus, a member of the genus Crinivirus. | J Gen Virol | The genome of Cucumber yellows virus (CuYV), isolated in Japan from cucumber (Cucumis sativus L.), was completely sequenced and shown to be bipartite. CuYV RNA1 consisted of 7889 nucleotides and encompassed seven open reading frames (ORFs), which is typical of the Closteroviridae, including a heat-shock protein 70 homologue, a coat protein and a diverged coat protein (CPd). CuYV RNA2 consisted of 7607 nucleotides and included two ORFs: ORF1a potentially encoded a polyprotein containing putative papain-like protease, methyltransferase and helicase domains, and ORF 1b potentially encoded an RNA-dependent RNA polymerase, which is probably expressed via a +1 ribosomal frameshift. The size and organization of the CuYV genome are similar to those of Lettuce infectious yellows virus (LIYV), the type member of the genus Crinivirus in the family Closteroviridae, indicating that CuYV is a member of that genus, although CuYV differed in several points from LIYV. |
| 2002 | Major pulmonary embolism: review of a pathophysiologic approach to the golden hour of hemodynamically significant pulmonary embolism. | Chest | Major pulmonary embolism (PE) results whenever the combination of embolism size and underlying cardiopulmonary status interact to produce hemodynamic instability. Physical findings and standard data crudely estimate the severity of the embolic event in patients without prior cardiopulmonary disease (CPD) but are unreliable indicators in patients with prior CPD. In either case, the presence of shock defines a threefold to sevenfold increase in mortality, with a majority of deaths occurring within 1 h of presentation. A rapid integration of historical information and physical findings with readily available laboratory data and a structured physiologic approach to diagnosis and resuscitation are necessary for optimal therapeutics in this "golden hour." Echocardiography is ideal because it is transportable, and is capable of differentiating shock states and recognizing the characteristic features of PE. Spiral CT scanning is evolving to replace angiography as a confirmatory study in this population. Thrombolytic therapy is acknowledged as the treatment of choice, with embolectomy reserved for those in whom thrombolysis is contraindicated. |
| 2001 | The effect of the transfusion of stored RBCs on intestinal microvascular oxygenation in the rat. | Transfusion | Although it is known that the transfusion of stored RBCs does not always improve tissue O(2) consumption under conditions of limited tissue oxygenation, the efficiency of O(2) delivery to the microcirculation by stored RBCs has never been determined.In a rat hemorrhagic shock model, the effects of resuscitation with fresh or 28-day-old RBCs stored in CPD plasma, saline-adenine-glucose-mannitol, and CPDA-1 plasma were investigated. Systemic hemodynamic and intestinal oxygenation measures were monitored. Intestinal microvascular PO(2) was determined with the O(2)-dependent quenching of palladium-porphyrin phosphorescence, and the RBC deformability was measured with a Laser-assisted optic rotational cell analyzer.Hemodynamic and oxygenation measures were significantly decreased during hemorrhagic shock. Intestinal oxygen consumption and mesenteric venous pO(2) were restored with the transfusion of both fresh and stored RBCs, except for CPD-stored RBCs. The intestinal microvascular pO(2) improved only with the transfusion of fresh RBCs. Deformability of the stored RBCs was significantly decreased.In contrast to that of fresh RBCs, the transfusion of stored RBCs did not restore the microcirculatory oxygenation, possibly because of impaired O(2) unloading, but, except for CPD-stored RBCs, the storage-induced changes were not enough to impair intestinal VO(2) and mesenteric venous pO(2). |
| 1999 | Comparison of Adsol and CPDA-1 blood preservatives during simulated massive resuscitation after hemorrhage in swine. | Transfusion | In recent years, there has been a change from the use of blood stored in CPDA-1 to the use of red cells (RBCs) stored in electrolyte mixtures, such as Adsol (AS-1 RBCs). However, because Adsol contains mannitol, as well as increased amounts of glucose relative to CPD and CPDA-1, concerns have been expressed as to possible harmful effects (recipient hyperglycemia, inappropriate osmotic diuresis) that it might induce under conditions of massive RBC transfusion.A hemorrhagic shock animal model was used to evaluate the effects of large-volume infusion of CPDA-1 or Adsol on glucose homeostasis and on urinary output under conditions that were devoid of extensive surgical manipulation. Hemorrhage was induced in 10 female Pitman-Moore mini-pigs to maintain mean arterial blood pressure at 55 mmHg for 90 minutes. After the return of autologous RBCs plus 1 L of 0.9-percent sodium chloride, the animals were given solution equivalent to the solute load in either 20 units of CPDA-1 whole blood (63 mL x 20 = 1260 mL) or 20 units of AS-1 RBCs (100 mL x 20 = 2000 mL) over a period of 90 minutes. Animals were monitored to determine physiologic and blood chemical responses to infusion of the solutions and to determine if there was hyperglycemia or inappropriate diuresis in the Adsol-treated group.Animals that received CPDA-1 developed significant hypocalcemia, arterial hypotension, and elevated blood glucose concentrations; two of five animals died of circulatory collapse. In contrast, glucose metabolism in the Adsol recipients was well-regulated, serum ionized calcium concentration was not significantly altered, and all animals survived. No evidence of inappropriate diuresis was observed.Administration of large amounts of Adsol was not associated with hyperglycemia or inappropriate osmotic duiresis in hemorrhaged and resuscitated minipigs. These data suggest that fewer physiologic changes may be associated with the massive transfusion of AS-1 RBCs than with that of CPDA-1 whole blood. |
| 1998 | Nucleotide sequence of the 3'-terminal two-thirds of the grapevine leafroll-associated virus-3 genome reveals a typical monopartite closterovirus. | J Gen Virol | The RNA genome of grapevine leafroll-associated closterovirus-3 (GLRaV-3) was cloned as a cDNA generated from GLRaV-3-specific dsRNA, and a partial genome sequence of 13154 nucleotides (nt) including the 3' terminus was determined. The sequenced portion contained 13 open reading frames (ORFs) potentially encoding, in the 5'-3' direction, proteins of > 77 kDa (ORF1a; helicase, HEL), 61 kDa (ORF1b; RNA-dependent RNA polymerase, RdRp), 6 kDa (ORF2), 5 kDa (ORF3, small transmembrane protein), 59 kDa (ORF4; heat shock protein 70, HSP70), 55 kDa (ORF5), 35 kDa (ORF6; coat protein, CP), 53 kDa (ORF7; diverged coat protein, CPd), 21 kDa (ORF8), 20 kDa (ORF9), 20 kDa (ORF10), 4 kDa (ORF11), 7 kDa (ORF12), and an untranslated region of 277 nt. ORF1b is probably expressed via a +1 ribosomal frameshift mechanism, most similar to that of lettuce infectious yellows virus (LIYV). Phylogenetic analysis using various gene sequences (HEL, RdRp, HSP70 and CP) clearly demonstrated that GLRaV-3, a mealybug-transmissible closterovirus, is positioned independently from aphid-transmissible monopartite closteroviruses (beet yellows, citrus tristeza and beet yellows stunt) and whitefly-transmissible bipartite closterovirus (lettuce infectious yellows, LIYV). However, another alleged mealybug-transmissible closterovirus, little cherry virus, was shown to be more closely related to the whitefly-transmissible LIYV than to GLRaV-3. |
| 1998 | A multi-laboratory evaluation of in vitro platelet assays: the tests for extent of shape change and response to hypotonic shock. Biomedical Excellence for Safer Transfusion Working Party of the International Society of Blood Transfusion. | Transfusion | There is no consensus regarding the use of specific in vitro tests for the assessment of the quality of platelet components. A literature review found that the platelet discoid shape as measured photometrically by the extent of shape change (ESC) and hypotonic shock response (HSR) correlated well with in vivo viability. The purpose of this study was to determine whether multiple research laboratories can perform the ESC and HSR assays in an accurate, reproducible manner, with acceptable sensitivity and comparable results.Eleven laboratories conducted five identical experiments, each with a different unit of platelet-rich plasma (PRP). For each experiment, 2 half-units of PRP were prepared and stored overnight: 1 half-unit at 20 to 24 degrees C in CPD (CPD-PRP) and the other at 1 to 6 degrees C with 2 mg per mL of EDTA (cold EDTA-PRP) added to produce spherical platelets with reduced HSR. Platelet suspensions having different proportions of the two PRPs were prepared and evaluated in duplicate by ESC and HSR assays, and morphologically scored by microscopy. One-way ANOVA and Duncan multiple-range tests were performed to determine significant differences in assay results for suspensions having different proportions of CPD-PRP.Comparable ESC (mean range: 20-28% for CPD-PRP and 1-6% for cold EDTA-PRP) and HSR (mean range: 58-81% for CPD-PRP and 12-31% for cold EDTA-PRP) measurements were obtained by nine laboratories. Duplicate testing showed high reproducibility of ESR and HSR results /in all laboratories. A 25-percent difference in the proportion of CPD-PRP (indicative of a difference of approximately 25% in the proportions of discoid and spherical platelets) was detected with a sample size of five (p<0.05) for both the ESC and HSR assays. A high correlation was found for the ESC assay and morphology score (r = 0.93, n = 345).Multiple laboratories were able to obtain comparable results with the ESC and HSR tests. They were able to show that the tests can be performed in an accurate, reproducible manner and with acceptable sensitivity. |
| 1990 | Comparison of platelet concentrates stored in plasma and in three plasmafree media. | Platelets | Platelet concentrates (PC) were stored for 7 days at 22°C in either CPDA-1 plasma (control) or one of three plasma-free media, namely a Tyrodes solution (BTST) fortified with citrate and with additional phosphate for buffering effect; Plasma-Lyte A solution (PCD) fortified with dextrose and citrate; and Ringers/CPD-50 (10:1) solution (RCPD) buffered with phosphate, all with a pH of 7.4. Parameters monitored included platelet count, pH, hypotonic shock response, aggregation response to single and paired agonists, and release by platelets of endogenous LDH, ATP and 5HT. BTST was shown to be a satisfactory plasma substitute over the entire storage period. PCD gave similar results over 4 days, although the decrease in pH was more marked. In all test systems RCPD was shown to be a poor plasma substitute. |
| 1989 | Properties of platelet concentrates prepared after extended whole blood holding time. | Transfusion | Extension of the maximum holding time for whole blood collected into a CPD-ADSOL system from 6 to 8 hours at ambient temperature under conditions that cause the temperature of the blood to decrease to 20 to 24 degrees C would facilitate the preparation of platelet concentrates (PCs). In this study, the properties of CPD-PCs prepared and stored for 5 days in PL-732 containers after various initial holding periods were assessed in two laboratories, designated Laboratory A and Laboratory B. Laboratory A found higher platelet-rich plasma (PRP) volumes (276 +/- 25 vs. 249 +/- 19 mL) and platelet yields (76 +/- 18 vs. 66 +/- 18 x 10(9) platelets) with 8-hour holds (n = 10) than with 1- to 2-hour holds (n = 10), although only the difference in PRP volumes was significant (p less than 0.05). No significant difference was observed in autologous in vivo recovery (54 +/- 11 vs. 47 +/- 9%) or survival (167 +/- 37 vs. 170 +/- 25 hrs), as calculated by gamma function using 111In as radiolabel. Laboratory B also found higher PRP volumes (304 +/- 31 vs. 279 +/- 37 mL) and platelet yields with 8-hour holds (n = 12) than with a 6-hour holds (n = 10) (88 +/- 26 vs. 77 +/- 27 x 10(9) platelets). No significant differences were found in morphology score, the extent of release of beta-thromboglobulin, the discharge of lactate dehydrogenase, or hypotonic shock response.(ABSTRACT TRUNCATED AT 250 WORDS) |
| 1989 | In vitro and in vivo evaluation of platelet concentrates after cotton wool filtration. | Vox Sang | Removal of leukocytes from platelet concentrates (PC) may decrease the incidence of alloimmunization to HLA antigens on white cells and prevent or delay refractoriness. Cotton wool filtration, which effectively removes leukocytes from PC with minimal platelet loss, may cause platelet or complement activation. In this study, the effect of cotton wool filtration (Imugard IG-500) on platelet activation, in vitro function, posttransfusion survival, and complement activation was investigated. Five paired autologous in vivo percent recovery and survival studies were performed with fresh (6-8h) PC or with 5-day-stored PC prepared from CPD-anticoagulated blood and stored in PL-732(TM) containers using standard methods. In the paired design, PC from the same donor were filtered on one occasion, and not filtered on another, prior to labeling with indium-III-oxine and reinfusion. Percent recovery and survival were then determined by the standardized method. Filtration had no statistically significant effect on percent recovery on PC stored for 6-8h or for 5 days. There was, however, a slight decrease in survival hours of the filtered PC which was statistically significant at 6-8h (203 +/- 14 vs. 179 +/- 18h; p less than 0.05) but not at 5 days (166 +/- 28 vs. 132 +/- 27h; p greater than 0.05). Samples taken before and after filtration of single units (fresh and 5-day) and pooled units (3-day) for measurement of release of granular content (beta-thromboglobulin), lysis (LDH), and in vitro viability - ATP, extent of shape change with ADP, and hypotonic shock response (3-day only)-demonstrated no effect of filtration on these parameters.(ABSTRACT TRUNCATED AT 250 WORDS) |
| 1987 | Platelet storage lesion in second-generation containers: correlation with platelet ATP levels. | Vox Sang | The nature of platelet lesion occurring with storage of platelet concentrates (PC) in second-generation containers was investigated using various storage media and storage periods up to 14 days. In CPD-plasma (control medium), the changes which occurred progressively during storage were loss of discoid shape, microscopic platelet aggregate formation, fragmentation and the appearance of disintegrated, 'balloon' forms. By day 14 less than 10% of the platelets were discoid in shape, the platelet count had decreased by 23%, and there was a 5-fold increase in the amount of lactate dehydrogenase in plasma. Associated with this was a decrease in the platelet oxygen consumption rate, D(O2), loss of cellular ATP and extent of ADP-induced shape change, and a decrease in the hypotonic shock response. These parameters decreased at a similar rate, with a 50% decrease (t1/2) at days 7-9. They correlated highly with each other during storage and also with a fall in pH. At day 14 of storage, mean pH was 6.1 +/- 0.3. To evaluate the effect of pH stabilization during storage, 4 mEq sodium bicarbonate was added to PC in CPD-plasma. Although pH maintenance was much improved, 7.2-6.6 during 14 days of storage, the same in vitro lesions developed, although more slowly. The t1/2 of the same parameters was prolonged for approximately two days. When PC were stored in a plasma-free physiologic salt solution whose salt composition was similar to CPD-plasma, the t1/2 of the parameters increased to 11-15 days of storage, although the platelets eventually developed the same in vitro lesions.(ABSTRACT TRUNCATED AT 250 WORDS) |
| 1986 | In vitro effect on stored red blood cells and platelets after a 15-hour delayed refrigeration of whole blood prior to component preparation in CPD-AD. | Vox Sang | We extended the time of keeping whole blood at 20-24 degrees C to 15 h (overnight) after phlebotomy for preparing platelet concentrates. We have evaluated the in vitro characteristics of platelets and blood cells prepared from whole blood drawn into CPD-AD, an anticoagulant containing 0.4 mM adenine and 1.5 times more dextrose than CPD. We studied in vitro red cell and platelet function of blood cooled either within 4 h after collection or after a 15-hour delay. In vitro platelet function measured as hypotonic shock reaction, aggregation response to ADP and collagen and 14C-serotonin uptake were not significantly different after preparation and after a 5-day storage period. Units held at room temperature for 15 h after blood collection exhibited a level of 2,3-DPG that was 45% of that exhibited by red cells held for 15 h at 1-6 degrees C. All other in vitro parameters of red cell concentrates measured during 35 days of storage were not significantly different. Based on these in vitro data blood drawn into CPD-AD might be kept up to 15 h at room temperature prior to refrigeration in order to prepare platelet concentrates. |
| 1976 | Cryobiology overview of red cell preservation: achievements and prospective. | Prog Clin Biol Res | Preserved red cells are transfused to improve the delivery of oxygen to tissue. The preserved red cells must circulate to increase the red cell volume and improve the oxygen carrying capacity. The correlation between oxygen transport function and the red cell 2,3 DPG level has been known since 1967. but it was only recently that the importance of the oxygen delivering capacity of transfused red cells during the immediate posttransfusion period became apparent. Red cells with low 2,3 DPG levels and increased affinity for oxygen will increase cardiac output and/or decrease venous pO2 for at least 4 hours after transfusion. Oxygen transport function is maintained longer in red cells stored in CPD than in ACD-stored red cells. Supplementation of CPD red cells with inosine, ascorbate; dihydroxyacetone and other substances helps to maintain the oxygen transport function during storage at 4 C. Liquid-stored red clls can be biochemically modified before freeze-preservation to increase the 2,3 DPG levels to to 1-1/2 to 2 times normal; these red cells have decreased oxygen affinity. The red cells have acceptable posttransfusion survival and improved oxygen releasing capacity for at least 72 hours after transfusion. The well-being of certain patients may be placed in jeopardy of they are given preserved red cells with increased affinity for oxygen. The patient may not be able to meet the accompanying demand for increased blood flow, and the venous-capillary oxygen tension may fall to a critical level. Clearly, patients in hemorrhagic and septic shock, those subjected to extracorporeal circulation during cardiac surgery, and anemic patients with myocardial or cerebrovascular insufficiency are best treated with red cells that have 150% normal 2,3 DPG levels. |
| Blood components in the treatment of acute blood loss: use of freeze-preserved red cells, platelets, and plasma proteins. | Anesth Analg | To avoid untoward reactions from blood transfusions and to make best use of the limited supply of blood, anesthesiologists and surgeons have many newly developed means at their disposal. Blood components should be separated from whole blood at the time of collection and prepared for either liquid or freeze-preservation. Citrate-phosphate-dextrose (CPD) blood should be separated into its components at room temperature within 4 hours of collection for greatest service from each collected unit. Red cell concentrates with hematocrits of 70 volumes percent can be prepared from the whole blood at the time of collection and frozen either shortly thereafter or after storage at 4 degrees C. for up to 3 weeks. Red-cell levels of 2, 3-diphosphoglycerate (2, 3-DPG) and adenosine triphosphate (ATP) can be increased by a rejuvenation process prior to freeze-preservation with either 40 percent W/V glycerol and storage at minus 80 degrees C. or with 20 percent W/V glycerol and storage at minus 150 degrees C. While hemorrhagic shock can best be managed with fresh whole blood, such blood is often not available; liquid- and freeze-preserved products serve as best substitutes. When previously-frozen washed red cells are used, crystalloid, colloid, coagulation factors, and platelets may also be required. Platelet concentrates stored at 4 degrees C. provide platelets that are hemostatically effective immediately upon infusion but have poor circulation. Platelet concentrates stored at 22 degrees C. provide platelets that have good circulation but upon transfusion have impaired hemostatic effectiveness. The coagulation factors and oncotic properties of plasma protein necessary for proper treatment of patients in hemorrhagic shock can be met by an adequate supply of fresh-frozen plasma and albumin. When liquid-stored red-cell concentrates or whole blood is given, filters must be used to remove the accumulated amorphous material, although the actual effects of the infused microaggregates are not yet known. |
| 1974 | The left shifted oxyhemoglobin curve in sepsis: a preventable defect. | Ann Surg | Nine patients with severe sepsis were studied to determine causes for any alterations in oxygen dissociation. Seven of the patients had oxyhemoglobin curves shifted to the left of expected and diminished DPG levels. These deficiences were not corrected in one case. The other eight patients survived or expired with normal to elevated P(50T) and DPG levels. In this study, three factors occurring either individually, in concordance, or in sequence were present when P(50T) was decreased. Correction of these deficiencies lead to normalization and, in one case, exceedingly high P(50T) and DPG levels. Where hypophosphatemia, acidosis, and transfusion of DPG deficient blood were avoided, no such change occurred. Hypophosphatemia is a common occurrence in the seriously ill patient whether or not hyperalimentation is used and may occur in spite of phosphate supplementation. Blood transfusions with CPD as the preservative are effective in reducing the severity of this disorder by the addition of an inorganic phosphate load. Septic shock itself had no untoward effect on oxygen dissociation. This held true even in the terminal stages of the disease process. |