Pre-Kidney Transplant Lower Extremity Impairment and Post-Kidney Transplant Mortality.

Prediction models for post-kidney transplantation mortality have had limited success (C-statistics ≤0.70). Adding objective measures of potentially modifiable factors may improve prediction and, consequently, kidney transplant (KT) survival through intervention. The Short Physical Performance Battery (SPPB) is an easily administered objective test of lower extremity function consisting of three parts (balance, walking speed, chair stands), each with scores of 0-4, for a composite score of 0-12, with higher scores indicating better function. SPPB performance and frailty (Fried frailty phenotype) were assessed at admission for KT in a prospective cohort of 719 KT recipients at Johns Hopkins Hospital (8/2009 to 6/2016) and University of Michigan (2/2013 to 12/2016). The independent associations between SPPB impairment (SPPB composite score ≤10) and composite score with post-KT mortality were tested using adjusted competing risks models treating graft failure as a competing risk. The 5-year posttransplantation mortality for impaired recipients was 20.6% compared to 4.5% for unimpaired recipients (p < 0.001). Impaired recipients had a 2.30-fold (adjusted hazard ratio [aHR] 2.30, 95% confidence interval [CI] 1.12-4.74, p = 0.02) increased risk of postkidney transplantation mortality compared to unimpaired recipients. Each one-point decrease in SPPB score was independently associated with a 1.19-fold (95% CI 1.09-1.30, p < 0.001) higher risk of post-KT mortality. SPPB-derived lower extremity function is a potentially highly useful and modifiable objective measure for pre-KT risk prediction.

Intra-Abdominal Cooling System Limits Ischemia-Reperfusion Injury During Robot-Assisted Renal Transplantation.

Robot-assisted kidney transplantation is feasible; however, concerns have been raised about possible increases in warm ischemia times. We describe a novel intra-abdominal cooling system to continuously cool the kidney during the procedure. Porcine kidneys were procured by standard open technique. Groups were as follows: Robotic renal transplantation with (n = 11) and without (n = 6) continuous intra-abdominal cooling and conventional open technique with intermittent 4°C saline cooling (n = 6). Renal cortex temperature, magnetic resonance imaging, and histology were analyzed. Robotic renal transplantation required a longer anastomosis time, either with or without the cooling system, compared to the open approach (70.4 ± 17.7 min and 74.0 ± 21.5 min vs. 48.7 ± 11.2 min, p-values < 0.05). The temperature was lower in the robotic group with cooling system compared to the open approach group (6.5 ± 3.1°C vs. 22.5 ± 6.5°C; p = 0.001) or compared to the robotic group without the cooling system (28.7 ± 3.3°C; p < 0.001). Magnetic resonance imaging parenchymal heterogeneities and histologic ischemia-reperfusion lesions were more severe in the robotic group without cooling than in the cooled (open and robotic) groups. Robot-assisted kidney transplantation prolongs the warm ischemia time of the donor kidney. We developed a novel intra-abdominal cooling system that suppresses the noncontrolled rewarming of donor kidneys during the transplant procedure and prevents ischemia-reperfusion injuries.

[Robotic-assisted organ transplantation]. / Transplantation d'organes avec assistance robotique.

Advanced surgical procedures have traditionally been a domain of open surgery. However, minimally invasive approaches are evolving with the development of robotic technology which appears capable to overcome technical limitations of conventional laparoscopy. While traditionally perceived as impossible indications for minimally invasive surgery, reports on robotic organ transplantations have surfaced with promising results.

Dietary phosphate restriction in dialysis patients: a new approach for the treatment of hyperphosphataemia.

BACKGROUND AND AIM: Elevated serum phosphate and calcium-phosphate levels play an important role in the pathogenesis of vascular calcifications in uraemic patients and appear to be associated with increased cardiovascular mortality. We aimed to evaluate the effects of a partial replacement of food protein with a low-phosphorus and low-potassium whey protein concentrate on phosphate levels of dialysis patients with hyperphosphataemia. METHODS AND RESULTS: Twenty-seven patients undergoing chronic haemodialysis were studied for a 3-month period. In the intervention group (n = 15), food protein were replaced by 30 or 40 g of low-phosphorus and low-potassium protein concentrate aimed at limiting the phosphate intake. In the control group (n = 12) no changes were made to their usual diet. Anthropometric measurements, biochemical markers and dietary interviews were registered at baseline and during the follow-up period. From baseline to the end of the study, in the intervention group, serum phosphate and circulating intact parathyroid hormone levels lessened significantly (8.3 ± 1.2 mg/dL vs 5.7 ± 1.4 mg/dL and 488 ± 205 pg/ml vs 177 ± 100 pg/ml respectively; p < 0.05) with decreasing of phosphate and potassium intake. No significant differences were found in the control group. No significant changes were observed in serum albumin, calcium, potassium, Kt/V, body weight and body composition in both the intervention and control groups. CONCLUSION: Dietary intake of phosphate mainly comes from protein sources, so dietary phosphorus restriction may lead to a protein/energy malnutrition in a dialysis patient. A phosphorus-controlled diet plan including a nutritional substitute resulted in serum phosphate and intact parathyroid hormone decrease without nutritional status modifications in dialysis patients.

A food borne outbreak of Salmonella enterica serotype Brandenburg as a hint to compare human, animal and food isolates identified in the years 2005-2009 in Italy.

INTRODUCTION: There are only a few reported cases of Salmonella enterica serotype Brandenburg foodborne outbreaks in the literature. In Italy Brandenburg is consistently present among the top ten serotypes from human source, but at low prevalences. MATERIALS AND METHODS: Fifty-five S. Brandenburg isolates from human, animal, environmental and food sources, including twelve isolates from a foodborne outbreak, were genotyped by Pulsed-Field Gel Electrophoresis (PFGE). RESULTS AND DISCUSSION: Eight pulsogroups and 19 pulsotypes were detected, with a unique pulsotype being attributed to the outbreak strains. Molecular subtyping can reliably complement the epidemiological investigations. Moreover, mapping molecular types of Salmonella isolates from human and non-human source may greatly contribute to risk assessment, by tracking possible animal sources, so improving cost-effectiveness of the prevention and control strategies.

[The viability of kidneys tested by gadolinium-perfusion MRI during ex vivo perfusion]. / La viabilité des reins marginaux testée par perfusion de gadolinium sous IRM pendant leur réanimation.

INTRODUCTION: Marginal kidneys must be reanimated before their transplantation. Reanimation is conducted with hypothermic pulsatile perfusion. The tests used generally to demonstrate the viability is the vascular resistance which is not convenient for everybody. We have developed a magnetic resonance compatible perfusional technology allowing us to test the organs during the perfusion by Gd-perfusion MRI. METHODS AND RESULTS: We have used pigs' kidneys with no warm ischemic time to establish the basis in a normal kidney. After an eight-hour hypothermic pulsatile perfusion, kidneys are submitted to a Gd perfusion. First, we measure the anatomy of the vessels, then the distribution of Gd in the kidney. We obtain simultaneously a dynamic study of the organs where T0 represents the Gd bolus arrival in the cortex and TP the maximum saturation time of Gd. CONCLUSION: We have observed that a normal T0 is inferior to 30s and TP is inferior to one minute. We have compared these values with ATP resynthesis in these organs and found that they correlate. We hope for the future through that predictive use of Gd-MRI to avoid the clinical use of "too" marginal kidneys or the discard of good kidneys but not corresponding with the vascular resistance theory.

Differential effects of deuterium oxide on the fluorescence lifetimes and intensities of dyes with different modes of binding to DNA.

Deuterium oxide (D2O) increases both the fluorescence lifetime and the fluorescence intensity of the intercalating dyes propidium iodide (PI) and ethidium bromide (EB) when bound to nucleic acid structures. We have used spectroscopic analysis coupled with conventional and phase-sensitive flow cytometry to compare the alterations in intensity and lifetime of various DNA-binding fluorochromes bound to DNA and Chinese hamster ovary (CHO) cells in the presence of D2O vs phosphate-buffered saline (PBS). Spectroscopic and flow cytometric studies showed a differential enhancement of intensity and lifetime based on the mode of fluorochrome-DNA interaction. The fluorescence properties of intercalating probes, such as 7-aminoactinomycin D (7.AAD) and ethidium homodimer II (EthD II) were enhanced to the greatest degree, followed by the probes TOTO and YOYO, and the non-intercalating probes Hoechst 33342 (HO) and 4,6-diamidino-2-phenylindole (DAPI). The non-intercalating probe mithramycin (MI) gave unexpected results, showing a great enhancement of fluorescence intensity and lifetime in D2O, indicating that when staining is performed in PBS, much of the MI fluorescence is quenched by the solvent environment. Apoptotic subpopulations of HL-60 cells had a shorter lifetime compared to non-apoptotic subpopulations when stained with EthD II. These results indicate that accessibility of the dye molecules to the solvent environment once bound to DNA, leads to the differential enhancement effects of D2O on fluorescence intensity and lifetime of these probes.

Epidemiological evaluation by PCR ribotyping of sporadic and outbreak-associated strains of Salmonella enterica serotype Typhimurium.

Salmonella enterica serotype Typhimurium has a very large diffusion worldwide within human and non-human hosts. The simultaneous circulation in the same geographical areas of many bacterial clones requires the use of reliable, reproducible and highly discriminatory typing techniques for epidemiological studies. Molecular biological methods, such as plasmid profile analysis, restriction endonuclease digestion of plasmid and chromosomal DNA and hybridization-based procedures have proven to be useful tools for strain differentiation. More recently, detection of polymorphisms in the intergenic spacer regions of rRNA genes by polymerase chain reaction (PCR ribotyping) has been successfully applied to characterize bacterial strains. In this study, PCR ribotyping was performed on 45 epidemiologically related and unrelated strains of S. enterica serotype Typhimurium isolated in northern and southern Italy during 1992. Isolates were simultaneously characterized by traditional ribotyping. Results suggest that PCR ribotyping is a rapid, easy-to-perform and reproducible typing method able to determine relatedness among isolates of this serotype.

Epidemiology of Salmonella enterica serotype Enteritidis infections in southern Italy during the years 1980-1994.

Increased frequency of identification of Salmonella enterica serotype Enteritidis as a causative agent of sporadic and epidemic cases of infection in humans, along with isolation in many parts of the world of strains belonging in a large proportion to a few phage types, has made phage typing alone inadequate for epidemiological investigations. In southern Italy the epidemic increase in isolation of S. enterica serotype Enteritidis that has been observed since 1990 has been associated in approximately 80% of isolates with phage type 4 (PT-4), in agreement with the epidemiological observations from other European countries. We have applied polymerase chain reaction (PCR) ribotyping in association with phage typing to a sample of non-outbreak strains and to all the outbreak strains sent for identification and typing to the Southern Italy Centre for Enterobacterial Pathogens between 1980 and 1994 from hospital and public health laboratories. This technique identified 15 distinct profiles among the 405 strains examined. Whereas a single profile (PCR ribotype a1) appeared to be closely related to PT-8, and to characterize a high percentage of the strains circulating during the early non-epidemic years (1980-1985), 11 patterns were recognizable within PT-4, and 5 within PT-1. Some of these apparently emerged after 1990. This subdivision enabled attribution of the epidemic circulation of PT-4 to multiple clones of S. enterica serotype Enteritidis.

Molecular analysis of strains of Shigella boydii isolated in northern and southern Italy.

In the years 1981-1988, Shigella boydii played a very limited role in the aetiology of diarrhoeal disease in Italy. However, between September and November, 1985, 19 isolates of serotype 2 were recovered in northern Italy from a dysentery outbreak which occurred in a geriatrics hospital in Abbiategrasso (Milan, Lombardy) and seven were identified in southern Italy during the period January-July, 1986 from apparently unrelated infection cases occurring in Brindisi (Apulia). These isolates were compared by molecular methods to seven strains of S. boydii of serotype 2 isolated since 1981 from the same geographic areas. Plasmid DNA analysis showed a large variety of patterns, whereas hybridization of chromosomal DNA with E. coli rRNA identified only two different profiles, one of which was exclusively found in all isolates from the hospital outbreak. No differences were detected among rDNA patterns of the remaining strains of S. boydii, irrespective of their geographic origin. These findings are consistent with the hypothesis that the infrequent cases of infection from S. boydii of serotype 2 which occurred during the years under study could probably be attributed to two different bacterial clones. Hybridization procedure and detection of hybrids were simplified by replacement of radioactive labelling of rRNA by the use of photobiotin.

Alterations in the progression of cells through the cell cycle after exposure to alpha particles or gamma rays.

A G1-phase delay after exposure to alpha particles has not been report ed previously, perhaps because immortalized cell lines or cell lines from tumor cells were used in past studies. Therefore, we compared the effects of alpha particles (0.19 or 0.57 Gy) and approximately equitoxic doses of gamma rays (2 or 4 Gy) on progression of cells through the cell cycle in normal human skin fibroblasts. Cell cycle analyses were performed using flow cytometry by measuring incorporation of bromodeoxyuridine (BrdUrd) in each phase of the cell cycle up to 44 h after irradiation. We observed an alpha-particle-induced G1-phase delay in human skin fibroblasts even at the lowest dose, 0.19 Gy. At equitoxic doses, more pronounced and persistent G1-phase delays and arrests were observed in gamma-irradiated cultures in that increased fractions of the G1-phase cells remained BrdUrd- over the course of the study after gamma-ray exposure compared to cells exposed to alpha particles. In addition, G1-phase cells that became BrdUrd+ after gamma irradiation re-arrested in G1 phase, whereas BrdUrd+ G1-phase cells in alpha-particle-irradiated cultures continued cycling. In contrast, comparable percentages of cells were delayed in G2 phase after either alpha-particle or gamma irradiation. Both gamma and alpha-particle irradiation caused increases in cellular p53 and p2lCip1 shortly after the exposures, which suggests that the G1-phase delay that occurs in response to alpha-particle irradiation is dependent on p53 like the initial G1-phase delay induced by gamma rays.

Epidemiological analysis of strains of Salmonella enterica serotype Enteritidis from foodborne outbreaks occurring in Italy, 1980-1994.

Polymerase chain reaction (PCR-) ribotyping was performed on 243 strains of Salmonella enterica serotype Enteritidis isolated during the years 1980-1994 from 58 foodborne outbreaks occurring in different regions of Italy. The majority (37) of the outbreaks were attributed to phage type (PT) 4, followed by PT1 (seven outbreaks); the latter was identified in 1993 in Italy in epidemic strains of Enteritidis. In eight cases more than one phage type was recognised from a single event. Nine PCR-ribotypes (PCR-RTs) were detected, with a strong prevalence of PCR-RTs f7 and e5 (23 and 21 outbreaks, respectively). In two instances two distinct PCR-RTs were identified within strains from a single outbreak. All but one of the PT1 outbreaks were caused by PCR-RT f7, whereas PT4 outbreaks could be subdivided into six subsets. Clustering of isolates was consistent with data obtained from epidemiological investigations. PCR-ribotyping proved to be an effective and reliable tool for subtyping isolates of Enteritidis belonging to the most frequent phage types. Nevertheless, in terms of laboratory expertise and lack of inter-laboratory standardisation, this typing technique is best suited for reference laboratories.

Nutritional intervention in a hemodialysis pregnant woman: a case report.

Pregnancy in dialysis patients is a rare occurrence. When pregnancy does occur, the risk of spontaneous abortion, stillbirth and neonatal complications, such as prematurity and growth retardation, are fairly high. The authors describe their experience in the follow-up of a patient with chronic renal failure who became pregnant during regular dialysis treatment and followed nutritional care. The outcomes were successful and she gave birth to a healthy baby. It is emphasized that special dedication to the nutritional control enabled a good outcome of the pregnancy. The importance of the nutritionist intervention in the follow-up of dialysis patients with the integration of a multidisciplinary staff is stressed.

Can dietary intake influence plasma levels of amino acids in liver cirrhosis?

BACKGROUND: Modifications in plasma amino acid patterns in cirrhotics are attributed to impaired liver function, being more evident in alcoholic than in viral cirrhosis. AIM: To evaluate whether diet influences plasma amino acid concentrations in different aetiological groups of cirrhotics. PATIENTS: Study population comprised 40 patients with cirrhosis (25 virus- and 15 alcohol-related], all Child A, and 30 healthy subjects (controls). METHOD: A food frequency and quality questionnaire was utilized to determine dietary history and alcohol intake. Nutritional status was evaluated by anthropometric method. Amino acids were determined, on venous blood samples, using a specific analyzer while cysteine was evaluated by fluorescent high power liquid chromatography RESULTS: The total daily intake of calories, proteins, lipids, and carbohydrates was similar in all individuals. Food quality distinguished the cirrhotics from the controls, but not the different aetiological groups of cirrhotics. Plasma cysteine levels were significantly lower, while aromatic amino acids and methionine were significantly higher, in all cirrhotics (p<0.001 and p<0.01, respectively, versus controls). The decrease in cysteine and the increase in other amino acids were more marked in alcoholics (p<0.01). CONCLUSIONS: Ethanol intake, but not diet, further enhances the changes in plasma aromatic amino acids, methionine and cysteine induced by impaired liver function in patients with cirrhosis, suggesting a direct interference of alcohol in their metabolism.

Cell cycle-dependent protein expression of mammalian homologs of yeast DNA double-strand break repair genes Rad51 and Rad52.

Recently, human and rodent homologs of yeast repair genes Rad51 and Rad52 have been identified and proposed to play roles in DNA double-strand break (DSB) repair. In this study, cell cycle-dependent expression of human and rodent RAD51 and RAD52 proteins was monitored using two approaches. First, flow cytometric measurements of DNA content and immunofluorescence were used to determine the phase-specific levels of RAD51 and RAD52 protein expression in irradiated and control populations. The expression of both proteins was lowest in G0/G1, increased in S and reached a maximum in G2/M. No difference was found in the whole-cell level of RAD51 or RAD52 protein expression between gamma-irradiated and control cell populations. Second, cell cycle-dependent protein expression was confirmed by Western analysis of populations synchronized in G0, G1 and G2 phases. Analysis of V3, a hamster equivalent of SCID, indicates that the protein level increases of RAD51 and RAD52 from G0 to G1/S/G2 do not require DNA-PK.

Rehabilitation of fractures in the elderly.

Rehabilitation is the most important aspect of care after a fracture in an older person. Epidemiology, contributing factors, general principles of management are discussed in this article. Proper management requires knowledge of various mechanisms of injury, different forms of orthopedic treatments, interpretation of radiographs, and familiarity with available therapeutic modalities.

Outbreak of Salmonella enteritis bongori 48:z35:- in Sicily.

Salmonella bongori 48:z 35 :- was first isolated from a lizard in Chad in 1966 and was classified as a biochemically atypical strain of the subgenus I of Kauffmann. Successively, some additional strains with different antigenic formulas but similar bioche

Endemic presence of Salmonella enterica serotype Cerro in southern Italy.

Molecular typing of salmonella strains isolated between 1997 and 1999 in southern Italy and carried out by the Southern Italy Centre for Enteric Pathogens, has shown a high frequency of Salmonella enterica serotype Cerro. This serotype is extremely rare i

Clonal circulation of Salmonella enterica serotype Heidelberg in Italy?

Phenotypic and genetic characteristics of 21 strains of Salmonella serotype Heidelberg isolated in the years 1999-2003 from different sources in Italy were studied. Susceptibility patterns, plasmid analysis, and PFGE were used as epidemiological markers. Although non-homogeneous drug resistance patterns and plasmid profiles had been detected, PFGE patterns suggest the hypothesis of a nationwide clonal spread of this serotype associated with poultry.

Detection of Salmonella spp. in food by a rapid PCR-hybridization procedure.

A rapid and sensitive PCR-hybridization procedure for detection of Salmonella serovars in food samples was developed. This method is based on three subsequent steps: (1) extraction of nucleic acids from a 2 ml aliquot of the pre-enrichment medium used for the conventional culture method after 6 h of incubation at 37 degrees C; (2) amplification with primers selected from the sequences of invE and invA genes; (3) Southern blot and hybridization with a biotin labeled oligonucleotide probe. The entire procedure requires 30 h. The PCR-hybridization assay was able to detect as little as 50 fg of purified chromosomal DNA of S. typhimurium and 0.2 cfu g-1 of an artificially contaminated food sample. Of 245 food samples analyzed by culture and PCR-hybridization, 20 were positive by both methods and 16 were positive by PCR-hybridization only. None of the 209 PCR-negative samples tested positive by culture. The sensitivity, specificity, alpha and beta error values of the results of the PCR-hybridization procedure, compared with those of culture, were 100, 92.9, 0 and 7.1%, respectively. These results indicate that a short pre-enrichment and PCR-hybridization could be used as a screening test for the detection of Salmonella in food samples.