Fatal respiratory infection due to ST308 VIM-1-producing Pseudomonas aeruginosa in a lung transplant recipient: case report and review of the literature.

BACKGROUND: Data regarding the prevalence of metallo-ß-lactamases (MBLs) among Pseudomonas aeruginosa isolates in cystic fibrosis patients are scarce. Furthermore, there is limited knowledge on the effect of MBL production on patient outcomes. Here we describe a fatal respiratory infection due to P. aeruginosa producing VIM-type MBLs in a lung transplant recipient and the results of the subsequent epidemiological investigation. CASE PRESENTATION: P. aeruginosa isolates collected in the index patient and among patients temporally or spatially linked with the index patient were analyzed in terms of antibiotic susceptibility profile and MBL production. Whole-genome sequencing and phylogenetic reconstruction were also performed for all P. aeruginosa isolates producing VIM-type MBLs. A VIM-producing P. aeruginosa strain was identified in a lung biopsy of a lung transplant recipient with cystic fibrosis. The strain was VIM-1-producer and belonged to the ST308. Despite aggressive treatment, the transplant patient succumbed to the pulmonary infection due to the ST308 strain. A VIM-producing P. aeruginosa strain was also collected from the respiratory samples of a different cystic fibrosis patient attending the same cystic fibrosis center. This isolate harbored the blaVIM-2 gene and belonged to the clone ST175. This patient did not experience an adverse outcome. CONCLUSIONS: This is the first description of a fatal infection due to P. aeruginosa producing VIM-type MBLs in a lung transplant recipient. The circulation of P. aeruginosa isolates harboring MBLs pose a substantial risk to the cystic fibrosis population due to the limited therapeutic options available and their spreading potential.

Bone involvement in young adults with cystic fibrosis awaiting lung transplantation for end-stage respiratory failure.

Patients with cystic fibrosis awaiting lung transplantation for end-stage respiratory failure have high prevalence of reduced bone mineral density and fragility fracture. Suboptimal 25-hydroxyvitamin D levels could significantly contribute to the development of cystic fibrosis-related bone disease. INTRODUCTION: The assessment of the prevalence of cystic fibrosis-related bone disease (CFBD) and its associated risk factors in young adults with cystic fibrosis (CF) awaiting lung transplantation for end-stage respiratory failure. METHODS: Clinical characteristics, bone mineral density (BMD), the parameters of calcium metabolism, including vitamin D (25OHVitD) levels, and the presence of fragility fractures were evaluated in 42 CF patients (24 females, age 34.0 ± 8.4 years) consecutively referred as lung transplant candidates. RESULTS: Mean 25OHVitD levels (54.9 ± 26.2 nmol/L) were below the reference range and hypovitaminosis D (25OHVitD < 75 nmol/L) was found in 34 patients (81%) and daily calcium intakes (median 550 mg/day) were lower than recommended. A BMD below the expected range for age (Z-score of - 2.0 or lower) and at least one prevalent fragility fracture were found in 22 patients (52.4%) and 18 patients (45.2%), respectively. The coexistence of low BMD and the presence of fracture was observed in 13 patients (31.0%). In these patients, the prevalence of nephrolithiasis was higher than in the remaining ones (p = 0.046). The presence of kidney stones was associated with a worse bone status and with severe vitamin D deficiency. In the whole sample, femoral BMD Z-scores were directly correlated with albumin-adjusted calcium (p < 0.05) and 25OHVitD levels (p < 0.01). CONCLUSIONS: Despite the improvement of CF care, CFBD is still highly prevalent in young adults awaiting lung transplantation for end-stage CF. Suboptimal 25OHVitD levels could significantly contribute to the development of CFBD. The presence of nephrolithiasis could be an additional warning about the need for a careful evaluation of bone health in CF patients.

Extracorporeal lung perfusion and ventilation to improve donor lung function and increase the number of organs available for transplantation.

INTRODUCTION: Ex vivo lung perfusion (EVLP) has been validated as a valuable technique to increase the pool of organs available for lung transplantation. MATERIAL AND METHODS: After a preclinical experience, we obtained permission from the Ethics Committee of our institution to transplant lungs after EVLP reconditioning. ABO compatibility, size match, and donor arterial oxygen pressure (PaO(2))/fraction of inspired oxygen (FiO(2)) ≤ 300 mm Hg were considered to be inclusion criteria, whereas the presence of chest trauma and lung contusion, evidence of gastric content aspiration, pneumonia, sepsis, or systemic disease were exclusion criteria. We only considered subjects on an extra corporeal membrane oxygenation (ECMO) bridge to transplantation with rapid functional deterioration. Using Steen solution with packed red blood cells oxygenated with 21% O(2), 5% to 7% CO(2) was delivered, targeted with a blood flow of approximately 40% predicted cardiac output. Once normothermic, the lungs were ventilated with a tidal volume of 7 mL/kg a PEEP of 5 cmH(2)O and a respiratory rate of 7 bpm. Lungs were considered to be suitable for transplantation if well oxygenated [P(v-a) O(2) > 350 mm Hg on FiO(2) 100%], in the absence of deterioration of pulmonary vascular resistance and lung mechanics over the perfusion time. RESULTS: From March to September 2011, six lung transplantations were performed, including two with EVLP. The functional outcomes were similar between groups: at T72 posttransplantation, the median PaO(2)/FiO(2) were 306 mm Hg (range, 282 to 331 mm Hg) and 323 mm Hg (range, 270 to 396 mm Hg) (P = 1, EVLP versus conventional). Intensive care unit ICU and hospital length of stay were similar (P = .533 and P = .663, respectively) with no mortality at 60 days in both groups. EVLP donors were older (49 ± 6 y versus 21 ± 7 y, P < .05), less well oxygenated (184 ± 6 mm Hg versus 570 ± 30, P < .05), displaying higher Oto scores (9.5 ± 0.7 versus 1.7 ± 1.5, P < .05). CONCLUSIONS: The first 6 months of the EVLP program allowed us to increase the number of organs available for transplantation with short-term outcomes comparable to conventional transplantations.

Lung transplantation and posterior reversible encephalopathy syndrome: a case series.

Posterior reversible encephalopathy syndrome is a neurological problem characterized by headache, altered mental status, focal neurological deficits, visual disorders, and seizures. The disorder is related to a number of diseases including calcineurin inhibitor therapy in solid organ transplantation. The incidence of posterior reversible encephalopathy syndrome in lung transplantation patients is unclear; probably the majority of the cases are unreported. The authors have described a case series constituted of four patients presenting posterior reversible encephalopathy syndrome after bilateral lung transplantation. The cases had in common complicated surgery and a posttransplant course characterized by hypertension, hypomagnesemia and acidosis. Invasive mechanical ventilation, calcineurin inhibitor discontinuation, aggressive antihypertensive therapy, and electrolyte regulation led to near complete recovery of symptoms.

Non-invasive continuous positive airway pressure in monolateral lung transplant patient with pneumonia and IPF.

Patients who undergo lung transplantation are prone to develop lower respiratory tract infections, leading to severe acute respiratory failure (ARF). Endotracheal intubation may not be indicated in these patients in light of a higher rate of mortality due to infections. The application of non-invasive ventilation could play a role in bridging these patients through the episode of ARF waiting for medical treatment to have effect. We report the evidence of morphological and physiological effects of the application of non-invasive continuous positive airway pressure during ARF sustained by pneumonia in a patient who underwent left lung transplantation because of idiopathic pulmonary fibrosis (IPF). We studied the effects of the application of positive end-expiratory pressure on both the right native lung affected by IPF and the transplanted lung affected by pneumonia.

Practices and attitudes towards quality assurance, inspection and accreditation in blood collection establishments in the European community.

In the framework of a European Community (EC)-supported project, a survey of practices and attitudes towards quality assurance, inspection and accreditation in Blood Collection Establishments (BCEs) in the EC member states was carried out. Analysis of 352 responses to a structured questionnaire revealed a preference for national standards over international, and an introspective and reactive view to quality management. Four broad categories of operational performance in relation to safety were formed: initial, repeatable, managed and optimising, with the majority of responses (209) being characterized as initial. Although a direct relationship between the size of the BCE and the range and level of quality management practices is apparent in the data, further analysis shows that small BCEs have much higher ratio of personnel per blood unit collected/processed than large BCEs and thus seem to have an inherent potential for improvement. Overall, a clear preference for inspection and accreditation by professional peers at the national level was indicated.

Relationship of the time of storage and transfusion reactions to platelet concentrates from buffy coats.

BACKGROUND: Transfusion reactions to platelet concentrates prepared from buffy coats (BC-PCs) were reviewed to determine the effect of some variables of BC-PC preparation and storage: time of BC storage before BC-PC preparation (1-2 days); time of BC-PC storage before transfusion (1-5 days); no white cell reduction versus laboratory and bedside BC-PC white cell reduction. STUDY DESIGN AND METHODS: A multiple linear logistic regression model was used by which the relative effect of one variable is expressed as the relative risk of transfusion reaction against a baseline level (1-day storage, no white cell reduction). RESULTS: During the 14 months of study, a total of 2707 BC-PC transfusions were given to 192 patients; 37 reactions (1.4%) were reported in 25 patients (13%). The transfusion reactions were febrile, nonhemolytic in 23 cases; allergic in 5; febrile and allergic in 2; and other in 7. The relative risk of transfusion reaction to BC-PCs prepared from BCs stored for 2 days was 1.98 times that to BC-PCs prepared from BCs stored for 1 day (p = 0.07). The relative risk of transfusion reaction of 5-day-old BC-PCs was 10.7 times that of 1-day-old BC-PCs (p = 0.001). The relative risk of transfusion reactions of BC-PCs white cell-reduced in the laboratory and at the bedside were 0.65 (p = 0.3) and 1.87 (p = 0.1) times, respectively, that of non-white cell-reduced BC-PCs. CONCLUSION: Time of storage seems to be an important variable associated with BC-PC transfusion reaction.

Sexual transmission of hepatitis C virus to a repeat blood donor.

BACKGROUND: Sexual transmission of hepatitis C virus (HCV) can occur, albeit inefficiently, and this represents a possible cause of community-acquired infections. This study describes a case of asymptomatic HCV infection acquired by a repeat blood donor from her sexual partner. CASE REPORT: A female repeat blood donor showed anti-HCV seroconversion and a slight elevation in alanine aminotransferase. She had a normal physical examination and no clinical symptoms. She admitted a sexual partnership with a man with chronic HCV infection. Genotyping showed subtype 3a infection in both. Nucleotide sequence analysis of the hypervariable region of the viral envelope was performed on five clones obtained from the donor and the partner. Five blood donors with subtype 3a infection were analyzed as controls. The mean homology among clones was 99.3 percent (95% CI, 98.9-99.7) in the donor and 96.8 percent (95% CI, 94.4-99.2) in the partner, which suggests a more recent infection in the woman. The mean homology between donor and partner was 93.4 percent (95% CI, 93.1-93.8), which is different from that between donor and controls (76.2%; 95% CI, 73.3-79.1; difference between means, 17.2%; 95% CI, 16.0-18.4). This suggests that the infection was transmitted to the donor from her sexual partner. Sexual intercourse is the most probable route of transmission, because parenteral risk factors were absent. CONCLUSION: Heterosexual transmission of HCV can occur in the absence of a long-lasting contact, and the infection can be asymptomatic. It remains to be determined whether the sexual partners of HCV-infected subjects should be deferred from blood donation.

The incidence and risk factors of community-acquired hepatitis C in a cohort of Italian blood donors.

To assess the incidence and source of community-acquired hepatitis C virus (HCV) infection among subjects at low risk for blood-borne diseases, we prospectively studied a cohort of 16,515 repeat blood donors over a mean follow-up time of 36 months. Second- and third-generation methods were used for hepatitis C virus antibody (anti-HCV) testing. HCV RNA was determined in the serum of anti-HCV-positive donors by reverse-transcription polymerase chain reaction. Liver biopsy was performed in the viremic subjects. Risk factors for HCV infection were identified by a psychosocial questionnaire in the whole cohort. During follow-up, 5 donors became infected with HCV. The incidence was 1 per 10,000 person-years (95% confidence interval, 0.3-2.4 per 10,000). During the 6 months before seroconversion, four subjects (80%) underwent medical or surgical percutaneous procedures, compared with 26.5% in the entire donor cohort (difference between frequencies, 53.5%; CI: 18.9-89.1). One seroconverting donor had sexual intercourse with an infected subject. Only 1 infected donor developed clinically evident acute hepatitis. HCV RNA remained detectable in 4 of 5 subjects for 8 to 36 months after seroconversion, and liver biopsy showed chronic hepatitis in all cases. Thus, new cases of hepatitis C occur among individuals without a history of known risk factors, some of which may be caused by nosocomial exposure.

Use of SERVQUAL to assess clinicians' satisfaction with the blood transfusion service.

BACKGROUND AND OBJECTIVES: Limited information is available on the level of satisfaction of clinicians with services delivered by blood banks. The purpose of this study was to evaluate the satisfaction of clinicians with our blood transfusion service. MATERIALS AND METHODS: We prepared a questionnaire based on SERVQUAL, a method used to measure customers' appreciation of quality of service, by assessing the gap between perceived and expected quality. The questionnaire consisted of 14 items grouped according to five dimensions of quality of service: assurance, empathy, responsiveness, reliability, tangibles. Clinicians were asked to give two scores on a scale from 1 to 7 for each item, score (e) representing what they expected from an 'excellent' service, score (r) how they graded the service received. We considered wide differences in scores of service expectation and receipt for a question to be indicative of either service above expected levels (r > e) or service below expectation (r < e); similar scores for both expected and received service (within 1 point on the grading scale) were taken to indicate that the service received was that which was expected. RESULTS: A total of 184 questionnaires (49%) were returned. For the 14 items considered, the proportion of clinicians expressing levels of satisfaction similar to or above expectation ranged from 67 to 96%. Three critical areas, which clinicians considered important (expectation scores 6-7) were associated with satisfaction below expectation in more than 20% of responders. They were: clarity of procedures, clarity of blood request forms, and convenience of blood request and issuing times, which were rated as important by 77, 80 and 72% of clinicians, respectively. CONCLUSION: SERVQUAL was useful to gather information on the level of clinicians' satisfaction with our transfusion service.

Thyroid vascularization by color doppler ultrasonography in Graves' disease. Changes related to different phases and to the long-term outcome of the disease.

To investigate possible correlations between thyroid vascularization and activity of Graves' disease, we measured blood flow (TBF) at the inferior thyroid artery and intraparenchymal vascularization (number of vessels per square centimeter) by color Doppler ultrasonography (CDU) on Graves' patients at different phases of the disease. We studied 88 patients cross sectionally: 22 untreated; 17 euthyroid after 6 months of methimazole; 49 euthyroid at drug withdrawal after 12 to 24 months of treatment. The patients of the latter group were followed up for 29.1 +/- 6.3 months after discontinuation of treatment. On the day of CDU examination, free triiodothyronine (FT3), free thyroxine (FT4), thyrotropin (TSH), antiperoxidase and anti-TSH receptor (TRAb) antibodies were measured. Vascularization indices were significantly higher in the Graves' patients than in controls. In the patients euthyroid under treatment, vascularization was not significantly lower than in the untreated group, but TBF and vessel number both appeared clearly reduced in the patients tested at drug withdrawal. The vascularization indices at drug withdrawal were significantly higher in the patients who relapsed than in those in stable remission: TBF (mL/min) 50.6 +/- 36.8 vs. 23.8 +/- 17.5, p = 0.001; vessel number/cm2 1.8 +/- 0.8 vs. 0.8 +/- 0.5, p = 0.002. A multivariate analysis, evaluating the predictive value of vascularization, hormonal and immunological parameters for relapse, demonstrated a significant predictive value for TRAb (RR 8.2; p = 0.001) and a weak predictive value for TBF (RR 1.1; p = 0.02). In conclusion, CDU examination confirms that thyroid hypervascularization in Graves' disease is not related to thyroid hormone circulating levels. The association of increased TBF and high levels of TRAb in the relapsing forms of disease suggests that thyroid hypervascularization is probably related to the activity of the underlying autoimmune processes.

A micro colorimetric assay using cryopreserved monocytes to evaluate antibody-mediated red cell-monocyte interaction.

BACKGROUND AND OBJECTIVE: A number of in vitro assays based on the interaction of red cells with monocytes have been used to determine the clinical significance of red cell antibodies. When used in our laboratory, one of these assays (the monocyte-macrophage phagocytosis assay-MMPA), was very time consuming and showed great variability. METHODS: We set up a monocyte phagocytosis colorimetric assay (MPCA), using standard microtiter plate wells coated at 37 degrees C for 1 hour with monocytes from healthy donors. After washing the wells to remove non-adherent monocytes, test red cells are added to the wells. Sensitized red cells bind to the monocytes, which are lysed after incubation to measure red cell phagocytosis. This is done by hemoglobin detection in the lysate through reaction with o-phenylenediamine and absorbance evaluation with a colorimeter. The results are expressed as the phagocytosis index (PI), which is calculated with the following formula: PI = [1-(A450 unsensitized red cells/A450 sensitized red cells)] x 100. In this study we determined: the source of MPCA variability; the precision of MPCA results; the correlation between MMPA and MPCA results; the MPCA reference values and the MPCA and MMPA execution times. RESULTS: MPCA variability depended largely on the monocyte source. The smallest variation coefficient of the results of replicate assays (19-21%) was found using pooled, cryopreserved monocytes. When performed with a pool of cryopreserved monocytes from 10 subjects, the SD of PI values obtained in replicate assays showed little variation (11-13) over the range of anti-D concentrations tested (from 18.75 to 300 ng/mL). A linear correlation coefficient r of 0.96 was obtained when MPCA and MMPA were performed in parallel, and the 95th centile of PI reference values determined with red cells of 40 non-transfused surgical patients free of irregular red cell antibodies was 7. MPCA execution time was 56% of that needed to perform MMPA. INTERPRETATION AND CONCLUSIONS: These studies show that MPCA is an easy and reproducible assay which allows objective and automated evaluation of red cell phagocytosis.

Influence of different hepatitis C virus genotypes on the course of asymptomatic hepatitis C virus infection.

BACKGROUND & AIMS: The association of liver disease with hepatitis C virus (HCV) genotypes mainly refers to patients with serious liver damage; little information is available on symptomless carriers. The aim of this study was to investigate the correlation of genotypes with clinical course, risk factors for infection, and antibody to HCV reactivity in asymptomatic subjects. METHODS: One hundred nine viremic blood donors with at least 1 year of follow-up were studied; 41 underwent liver biopsy. Genotypes were determined by line-probe assay. RESULTS: Genotype 1 was found in 47 (43.1%), genotype 2 in 48 (44%), genotype 3 in 8 (7.3%), genotype 4 in 2 (1.8%), and coinfections in 4 (3.7%). The relative risk (RR) for a raised pattern of alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyltranspeptidase was 2.1 (confidence interval [CI], 1.4-3.2), 1.7 (CI, 1.2-2.4), and 2.8 (CI, 1.6-4.9) in subjects with genotype 1 vs. 0.4 (CI, 0.2-0.7), 0.4 (CI, 0.3-0.7), and 0.4 (CI, 0.2-0.8) in subjects with genotype 2. Chronic hepatitis was found in 68%; the RR of chronic hepatitis was similar for genotypes 1 and 2 (RR, 1.1 [CI, 0.8-1.7] vs. RR, 1.0 [CI, 0.7-1.6]). Reactivity to NS4-derived antigens was infrequent in type 2-infected subjects. CONCLUSIONS: Genotype 2 was as frequent as genotype 1 but associated with less liver function impairment. The high prevalence of chronic hepatitis should be considered in counseling viremic asymptomatic donors.

Hepatitis C virus RNA and liver histology in blood donors reactive to a single antigen by second-generation recombinant immunoblot assay.

The clinical significance of single band reactivity (indeterminate pattern) at anti-hepatitis C virus (HCV) second-generation recombinant immunoblot assay (RIBA-2) was investigated in symptomless subjects with normal liver function tests to obtain data for their counseling and clinical management. Serum and hepatic HCV RNA were determined by the nested polymerase chain reaction, and liver histology was evaluated in 40 symptomless blood donors with stable indeterminate RIBA-2 pattern, including 38 reactive to c22-3. All but one had normal alanine aminotransferase (ALT) levels. Two new immunoblot tests, RIBA-3 and INNO-LIA HCV Ab III (LIA-III), which incorporate additional HCV antigens, were also done to assess whether they could identify the viremic subjects. Ten cases (25%, confidence interval 12 to 38) were HCV RNA positive. Three of the HCV RNA-positive and none of the HCV RNA-negative subjects had chronic hepatitis. RIBA-2 strong intensity of reaction (score > 2+) was observed in all the HCV RNA-positive and in 12 HCV RNA-negative subjects. RIBA-3 and LIA-III gave positive results in 9 of 10 and 10 of 10 HCV RNA-positive, but also in 8 of 30 and 24 of 30 HCV RNA-negative subjects. A c-22-3 reactivity score of 4+ by RIBA-3 and E2/NS1 reactivity by LIA-III were both strongly associated with HCV RNA (P < .001). Based on relatively high prevalence of chronic hepatitis in our series (30%), apparently healthy subjects with stable indeterminate RIBA-2 pattern and HCV RNA positivity should be considered for liver biopsy independently of ALT profile.(ABSTRACT TRUNCATED AT 250 WORDS)

Human lymphocyte antigens in Graves' disease: correlation with persistent course of disease.

Graves' disease is a thyroid autoimmune disorder associated with specific human lymphocyte antigen (HLA) alleles, characterized by an unpredictable long-term course. To investigate possible relations between HLA phenotype and outcome of the disease, the authors typed for HLA antigens in 105 patients with Graves' disease with different course of disease. All patients were treated with antithyroid drugs for at least 12 months; 29 patients had stable remission 24 or more months after withdrawal of treatment; 76 patients had persistent disease--66 unremitting/relapsing hyperthyroidism, 10 stable hypothyroidism--36 or more months after onset of disease. The following findings emerged from this study: 1) HLA B8 and DR3 were increased significantly in Graves' patients versus 6,682 control subjects from the same geographic area (23.80% vs 12.01%, odds ratio [OR] 1.98, and 31.43% vs 18.00%, OR 1.75, respectively); the antigen combinations B8-DR3, B8-Cw7-DR3, and A1-B8-Cw7-DR3 were significantly more frequent in Graves' patients vs control subjects; in addition, these combinations were present exclusively in patients with persistent disease (B8-DR3 28.95%, OR 7.14, B8-Cw7-DR3 27.63%, OR 11.24, and A1-B8-Cw7-DR3 18.42%, OR 11.29). These data provide evidence that not only susceptibility to Graves' disease, but also persistent activity of the autoimmune process, producing either hyperthyroidism or stable hypothyroidism, is associated with specific HLA antigen phenotypes.

HLA-DRB1 compatibility in cadaver kidney transplantation: correlation with graft survival and function.

The introduction of genomic HLA-DR typing has stimulated a re-evaluation of the role of HLA-DR compatibility on cadaver kidney transplantation. We retrospectively studied the influence of HLA-DRB1 matching on the survival of 416 patients using univariate and Cox regression analysis as well as its influence on the occurrence of rejection episodes and on creatinine level at the 3rd month in the 198 recipients for whom these data were available. The following parameters were also considered: HLA-A,B compatibility, donor and recipient age, graft number, pre-transplant blood transfusions and panel reactive antibodies (PRA). Twenty-four month graft survival was 100% for transplants with zero mismatches (n = 47), 87.9% for those with one mismatch (n = 191) and 81.3% for those with two mismatches (n = 178). In the Cox model, HLA-DRB1 matching was the most significant variable influencing graft survival (47% of chi 2 P = 0.001), followed by HLA-A,B matching (23%, P = 0.02) and donor age (19%, P = 0.04). Ninety-two percent of the patients with zero mismatches experienced no rejection episodes in the first 3 posttransplant months compared with 62% and 41% of patients with one and two mismatches, respectively. Mean creatinine level (mg/dl) was 1.2, 1.4, and 1.5 in patients with zero, one, and two mismatches, respectively. Should these results be confirmed by prospective studies, HLA-DRB1 compatibility will have to be considered as an organ allocation criterion.