Purity and stability of online-prepared hemodiafiltration fluid after storage.

BACKGROUND/AIMS: Previous studies have suggested that online hemodiafiltration (OL-HDF) fluid can be used as dialysate for continuous renal replacement therapies, and thus HDF costs can be reduced. The aims of this study were to determine the purity of OL-HDF fluid and to verify the stability of the electrolyte composition and acid-base balance during its storage. METHODS: OL-HDF fluid was collected in 70 individual bags and stored for up to 7 days. The following tests were performed daily in 10 bags: natural visible precipitation (macrocrystallization), sample collection for chemical analysis and fluid culture, limulus amebocyte lysate endotoxin test, standard culture of NALGENE® filters after passing of the fluid, and molecular analysis of bacterial DNA. RESULTS: The values of pH and pCO(2) showed a significant change starting at 24 h (p < 0.001); after 72 h, their values were beyond the measurable range. Coefficient of variation for pCO(2) was as high as 25.7%. Electrolyte composition (Na(+), K(+), Cl(-), Ca(2+) and glucose) showed a statistically significant difference over time (p < 0.05); however, their coefficients of variation were low (1.7, 1.4, 0.6, 2.3 and 0.9%, respectively), which might not be considered clinically significant. Negative results were obtained at all points by fluid and filter cultures, endotoxin test and molecular analysis. No macrocrystallization was observed at any time point. CONCLUSIONS: We demonstrate the microbiological purity of OL-HDF fluid stored for up to 7 days. The electrolyte composition was stable, except for a relevant change in pCO(2) and consequently in pH (first noted at 24 h), emphasizing the need to reassess the acid-base balance in multilayer plastic bags in future studies.

Stone composition: where do we stand?

Kidney stones are a common disorder of the urinary tract. Nephrolithiasis is a morbid and expensive disease. The prevalence and incidence are estimated at 5-10% and 100-300/100,000/year, respectively. Relapses occur in 50-70% of all cases. For these reasons, prevention of stone formation is of great importance. Knowing the composition of the calculus is thus fundamental for a more complete evaluation of the metabolic study. The nature of the calculus in fact helps the physician to find a convenient metaphylaxis consisting of both sanitary and therapeutic measures. Study of the composition of urinary stones remains one of the most interesting aspects of the lithiasic pathology today. Presently crystallographic examination constitutes one of the most precise and less expensive methodologies to identify the nature of the concretion. This method also allows the urologist to catalogue the typology of the lithiasis during endoscopy.

Transurethral resection of prostate: technical progress by bipolar Gyrus plasma-kinetic tissue management system.

AIM: We report our experience about bipolar plasma-kinetic resection of the prostate for the treatment of bladder outlet obstruction due to benign prostatic hyperplasia (BPH), considering intraoperative variables and short-term METHODS: Three hundred and eighty-nine patients affected by symptomatic BPH, underwent transurethral resection of prostate using the bipolar plasma-kinetic technique from Decem-ber 2001 to July 2004 in a prospective fashion. Bipolar resections of the prostate were performed using Plasma-Kinetic Tissue Management System (Gyrus Medical Ltd., UK). Preoperative, digital rectal examination, haemoglobin level, total PSA, transrectal ultrasound, International-Prostate Symptom Score (I-PSS) and uroflowmetry parameters were recorded. Patients were assessed for safety and efficacy, evaluating intraoperative and postoperative complications and measuring both the IPSS and the maximum flow rates (Qmax) after 12 months. RESULTS: The mean operative time was 89 min (range 48-121 min); the mean prostatic resected weight (measured by an electronic scales) was 49.6 g (3267 g). The mean decrease of haemoglobin level was 1.1 g/dL (range 0.5-1.9), with a mean catheterization time of 1.3 days (range 1-5). Qmax increase ranged from 120 to 230% (mean 190%) 12 months after surgery. Correspondingly, IPSS decrease ranged from 48% to 86% (mean 79%). Postoperative acute urinary retention, urethral strictures, bladder neck sclerosis and urinary incontinence were recorded in 1.57%, 2.57%, 1.28% and 0.77%, respective. CONCLUSION: The transurethral resection of prostate using a bipolar plasma-kinetic device represent a safe and effective option for the treatment of symptomatic BPH.

First Report of Leaf Spot and Blight of Strelitzia reginae Caused by Burkholderia gladioli in Italy.

During the summer of 2005, a new disease of bird of paradise (Strelitzia reginae Aiton) was observed on young seedlings (2 to 3 months old) in a nursery located in Giarre (Catania) in eastern Sicily. Symptoms included brown, water-soaked leaf spots that first appeared after seedling emergence and then gradually enlarged and became necrotic. Occasionally, during wet conditions, seedlings were completely blighted, resulting in total loss. The disease was observed on 10% of the 3,000 plants present in one nursery. A single bacterial colony was consistently isolated on King's medium B (KB) supplemented with 0.01% cycloeximide from surface-sterilized, brownish lesions and water-soaked leaf tissues. The isolates were purified on nutrient agar (NA). Three bacterial strains isolated from three different symptomatic plants were used for pathogenicity and identification tests on S. reginae plants. Five plants were inoculated per bacterial strain by spraying the leaves with a buffer phosphate suspension (0.1 M) at 106 CFU/ml prepared from KB plates incubated for 24 h at 28°C and wounding the leaves (four wounds per leaf) with a sterile needle. The same number of noninoculated plants was used as control. All plants were covered with plastic bags and maintained in a greenhouse at 25 ± 1°C with 95 to 100% relative humidity until symptoms occurred 3 to 4 days later. All three bacterial strains tested were virulent and caused symptoms identical to those observed in the nursery. No symptoms were observed in control plants. Koch's postulates were fulfilled by the reisolation of the three strains from inoculated plants. The strains were gram-negative, aerobic rods, grew aerobically, were white and nonmucoid on yeast dextrose calcium carbonate agar, nonfluorescent on KB, produced diffusible nonfluorescent pigment on KB, and were oxidase and urease negative. All strains utilized glucose, arabinose, mannose, mannitol, N-acetylglucosamine, gluconate, caprate, malate, citrate, and phenyl acetate and none of the strains produced indole or acidified glucose. Using the API 20NE test strips (bioMérieux, Marcy l'Etoile, France) incubated at 28°C for 24 to 48 h, all strains were initially identified as Burkholderia cepacia. On the basis of the nutrient profiles revealed by the BIOLOG system (Microlog System Release 4.2, Hayward, CA), the strains were identified as B. gladioli (Severini 1913) Yabuuchi et al. 1993. The index of probability was 100% and the index of similarity was 0.75%. For molecular identification of strains, 16S rDNA was amplified by using species-specific primers Eub-16-1 and Gl-16-2457, obtaining a polymerase chain reaction (PCR) product of 463 bp (1). PCR analysis indicated that the strains belong to B. gladioli. Other bacteria have been previously reported in Italy as pathogens of Strelitzia spp. (2,3). To our knowledge, this is the first outbreak of leaf spot and blight caused by B. gladioli on S. reginae. References: (1) A. Bauernfeind et al. J. Clin. Microbiol. 37:1335, 1999. (2) P. Bella et al. J. Plant Pathol. 82:159, 2000; (3) G. Polizzi et al. Plant Dis. 89:1010, 2005.

Our experience with nifedipine and prednisolone as expulsive therapy for ureteral stones.

OBJECTIVES: We evaluated the effect of nifedipine associated with prednisolone in ureteral stone passage. MATERIAL AND METHODS: In our department we enrolled 50 patients with radiopaque ureteral stones. Stone size was 15 mm or less. The patients were divided into two groups: group I included 25 patients who received 30 mg oral treatment of slow-release nifedipine (for a maximum of 20 days) and 25 mg of prednisolone (for a maximum of 10 days) daily. Group II was made up of 25 patients who received 25 mg of prednisolone daily. On request, both groups could use non-steroidal anti-inflammatory drugs. RESULTS: The mean expulsion time was 6 days in group I and 10 days in group II. The average stone size was 12 mm in group I and 12.8 mm in group II. Six patients suspended therapy in group I (5 erythema, 1 stomachache), and seven in group II (3 because of intolerable pain, 4 stomachache). The expulsion success rates were 68% in group I and 81% in group II. CONCLUSIONS: For ureteral stones that do not cause an emergency situation, such as obstructive uropathy, infection or intolerable pain, we suggest expulsive medical treatment with nifedipine and prednisolone, if there are no contraindications to drug use.

ESWL: Comparing two analgesic techniques. Our experience.

OBJECTIVE: We compared the effects of intramuscular analgesia versus intramuscular analgesia associated with topical application of Luan (gel containing lidocaine 1%) on the efficacy of extracorporeal shockwave lithotripsy (ESWL). MATERIAL AND METHODS: From January 2001 to November 2002, 300 patients (mean age 46) were treated for kidney or upper ureteral radiopaque stones with ESWL, using a Dornier Compact Delta magneto lithotripter. The patients were divided into two groups: group A, 180 patients, received intramuscular analgesia with Ketorolac 30 mg, Tramadolo 100 mg and, during treatment, intravenous betametasone 4 mg. Group B, 120 patients, received the same pharmacological treatment associated with topical application of Luan. RESULTS: At 3 months, the stone-free rate in group A was 61% compared with 79% in group B. The energy of treatment was less than 13 kV in group A and less than 15 kV in group B. CONCLUSIONS: For ESWL treatment of kidney stones of 20 mm or less and ureteral stones of 15 mm or less, we suggest local analgesia with topical Luan and intramuscular analgesia. This approach increases the success rate of ESWL and reduces the discomfort associated with treatment.

Acute interstitial pneumonia: report of a series.

Four cases of acute interstitial pneumonia (AIP) are described with special emphasis on clinical background, lung imaging and bronchoalveolar lavage findings. A retrospective chart review of four patients with histologically-proven AIP, diagnosed between 1998 and 2000, was carried out. Clinical data, bronchoalveolar lavage (BAL) findings, high-resolution computed tomography (HRCT) and histological features were analysed. Three patients died and only one is in follow-up. HRCT showed areas of ground glass attenuation and alveolar consolidation in all patients. Histology, documented by open lung biopsy or autopsy specimens, was consistent with the organising form of diffuse alveolar damage pattern. BAL findings were characteristic, with a huge neutrophilia associated with scattered atypical type II pneumocytes collected in clusters with extracellular amorphous material (fragments of hyaline membranes) observed in two out of three cases. In this paper, four cases of acute interstitial pneumonia are reported in detail. The poor prognosis associated with this entity has been confirmed and the possible diagnostic role of the bronchoalveolar lavage is emphasised.

Transcription pattern of human herpesvirus 8 open reading frame K3 in primary effusion lymphoma and Kaposi's sarcoma.

Human herpesvirus 8 (HHV-8) is found in immunoblastic B cells of patients with multicentric Castleman's disease (MCD) and, predominantly in a latent form, in primary effusion lymphoma (PEL) cells and Kaposi's sarcoma (KS) spindle cells. Recent studies have shown that upon reactivation, HHV-8 expresses factors that downregulate major histocompatibility class I proteins and coactivation molecules and that may enable productively infected cells to escape cytotoxic T lymphocytes and natural killer cell responses. One of these viral factors is encoded by open reading frame (ORF) K3. Here we show that in PEL cells, ORF K3 is expressed through viral transcripts that are induced very early upon virus reactivation, including bicistronic RNA molecules containing coding sequences from viral ORFs K3 and 70. Specifically, we found that a bicistronic transcript was expressed in the absence of de novo protein synthesis, thereby identifying a novel HHV-8 immediate-early gene product. Several features of the RNA molecules encoding the K3 product, including multiple transcriptional start sites, multiple donor splicing sites, and potential alternative ATG usage, suggest that there exists a finely tuned modulation of ORF K3 expression. By contrast, ORF K3 transcripts are not detected in the majority of cells present in KS lesions that are latently infected by the virus, suggesting that there are other, as-yet-unknown mechanisms of immune evasion for infected KS spindle cells. Nevertheless, because HHV-8 viremia precedes the development of KS lesions and is associated with the recrudescence of MCD symptoms, the prompt expression of ORF K3 in productively infected circulating cells may be important for virus pathogenesis. Thus, molecules targeting host or viral factors that activate ORF K3 expression or inactivate the biological functions of the K3 product should be exploited for the prevention or treatment of HHV-8-associated diseases in at-risk individuals.

DNA ploidy and TLI in association with other prognostic parameters in breast cancer.

DNA ploidy and thymidine-labeling index (TLI) have been introduced as prognostic indicators to characterize the biological behavior of breast cancer for the selection of patients eligible for adjuvant therapy. The aim of this study was to evaluate the prognostic significance and correlation between ploidy status, TLI and other common histopathologic parameters such as histotype, stage, grading, lymph node metastasis, hormone receptors and recurrence. DNA ploidy and TLI were assessed for 68 breast cancer patients, 27 to 85 years of age. DNA histograms, analyzed with static cytometry, were: diploid in 21(31%) and non diploid in 47 (70%). High TLI values were observed in 16 cases (24%), medium values in 46 cases (68%) and low values in 6 cases (8.5%). DNA ploidy and TLI showed a statistically significant correlation, as independent parameters, with recurrence (respectively, p = 0.0267 and p < 0.0001). Therefore, DNA ploidy and TLI may be considered additional prognostic parameters to be utilized besides all the other clinical-pathologic data for the assessment of these lesions, and to plan therapeutic strategies.

Alpha interferon inhibits human herpesvirus 8 (HHV-8) reactivation in primary effusion lymphoma cells and reduces HHV-8 load in cultured peripheral blood mononuclear cells.

Infection by human herpesvirus 8 (HHV-8) is associated with the development of Kaposi's sarcoma (KS). Since regression of KS can be achieved by treatment of the patients with alpha interferon (IFN-alpha), we analyzed the effects of IFN-alpha or anti-IFN-alpha antibodies (Ab) on HHV-8 latently infected primary effusion lymphoma-derived cell lines (BCBL-1 and BC-1) and on peripheral blood mononuclear cells (PBMC) from patients with all forms of KS and from at-risk subjects. IFN-alpha inhibited in a dose-dependent manner the amplification of HHV-8 DNA in BCBL-1 cells induced to lytic infection with tetradecanoyl phorbol acetate (TPA). This effect was associated with the inhibition of the expression of HHV-8 nut-1 and kaposin genes that are induced early and several hours, respectively, after TPA treatment. In addition, IFN-alpha inhibited virus production and/or release from BCBL-1 cells. Inhibition of nut-1 and kaposin genes by IFN-alpha was also observed in BC-1 cells induced with n-butyrate. Conversely, the addition of anti-IFN-alpha Ab to TPA-induced BCBL-1 cells resulted in a larger number of mature enveloped particles and in a more extensive cytopathic effect due to the neutralization of the endogenous IFN produced by these cells. IFN was also produced by cultured PBMC from HHV-8-infected individuals, and this was associated with a loss of viral DNA during culture. However, the addition of anti-IFN-alpha Ab or anti-type I IFN receptor Ab promoted the maintenance of HHV-8 DNA in these cells that was associated with the detection of the latency-associated kaposin RNA. Finally, the addition of IFN-alpha reduced the HHV-8 load in PBMC. Thus, IFN-alpha appears to have inhibitory effects on HHV-8 persistent infection of PBMC. These results suggest that, in addition to inhibiting the expression of angiogenic factors that are key to KS development, IFN-alpha may induce KS regression by reducing the HHV-8 load and/or inhibiting virus reactivation.

Genetic disorders in normally androgenized infertile men and the use of intracytoplasmic sperm injection as a way of treatment.

OBJECTIVE: To determine the incidence of chromosomal abnormalities in normally androgenized infertile men with no other recognized causes of infertility or who had ever been submitted to other unsuccessful methods of treatment. DESIGN: Collaborative retrospective study of clinical experience collected by an endocrinologist and a geneticist over a 5-year period. SETTING: Outpatients at an endocrinology clinic. PATIENT(S): Infertile male patients with azoospermia (n = 23), oligozoospermia (n = 66), and normozoospermia (n = 14) presenting normal (n = 85) or subnormal (n = 18) testicular volume. INTERVENTION(S): All patients were submitted to cytogenetic analysis. MAIN OUTCOME MEASURE(S): Two patients were referred to intracytoplasmic sperm injection (ICSI), and in one case, a successful gemellar pregnancy ended up uneventful. Children's genetic testing were not performed according to parents' request. RESULT(S): Abnormal karyotypes were found in 11 (10.6%) patients. Chromosomal abnormalities were found in 17.3% of the 23 azoospermic patients, in 10.6% of the 66 oligozoospermic patients, and in none of the 14 normozoospermic patients. These disorders were found only in patients with normal testicular volumes and no more than 10 x 10(6) spermatozoa per mL. CONCLUSION(S): A high incidence of chromosomal abnormalities was found in a selected group of normally androgenized infertile male patients. The elevated rate of fertilization achieved in one patient indicates that ICSI is, at the moment, the only choice for treatment of male infertility because of chromosomal abnormalities.

Interlaboratory reproducibility in reporting inadequate cervical smears--a multicentre multinational study.

A set of 300 vaginal smears was interpreted by 13 cytologists from six European laboratories, who were requested to report inadequate and suboptimal smears. The set had been appropriately seeded to reach approximately 10% inadequate and 20% suboptimal smear frequency. According to the majority report, 230 smears were classified as adequate (76.7%), 43 as inadequate (14.3%), and 27 as suboptimal (9.0%). Agreement with the majority report ranged from 52% to 91% (average 78%). Kappa statistics for reporting inadequate smears showed a high level of agreement for five cytologists, and fair to good agreement for eight. In contrast, kappa statistics for reporting suboptimal smears showed fair to good agreement with the majority report only in five instances, whereas agreement was poor for eight cytologists. 'Inadequate smear' rates may be used to compare the quality of smears received in different laboratories, as there is a high level of agreement among cytologists as to what constitutes an inadequate smear. However, this is not true for "suboptimal smear" rates, and more precise reporting criteria must be defined and tested if an intermediate category is to be retained to report poor quality smears: more precise reporting criteria must be defined and tested if an intermediate category is to be retained to report poor quality smears.

Male infertility due to congenital adrenal hyperplasia: testicular biopsy findings, hormonal evaluation, and therapeutic results in three patients.

Testicular biopsy in three infertile male patients with congenital adrenal hyperplasia showed different grades of germinal epithelium maturation, and no Leydig cells in the interstitium. In two patients with severe germinal cell hypoplasia, very low serum gonadotropins showed no response to luteinizing hormone-releasing hormone stimulation and responded to long-term clomiphene citrate administration. It is hypothesized that an increase of hypothalamic estrogen production because of local androstenedione aromatization might be responsible for the luteinizing hormone-releasing hormone block observed in these patients. All patients were submitted to adrenal suppressive therapy, and in two cases a return of the fertility status was obtained.

The influence of soil particle adsorption on 2,3,7,8-tetrachlorodibenzo-p-dioxin biological uptake in the rabbit.

A comparative study on the biological uptake in the rabbit of 2,3,7,8-tetrachloridibenzo-p-dioxin (TCDD) in different formulations, including accident-contaminated Seveso soil, was attempted. On the whole, these results indicated that soil-borne TCDD had a bioavailability lower than that of free (solvent-borne) TCDD.