Immunological investigations in inbred mice chronically infected with cytopathogenic agents isolated from human cases of Balkan endemic nephropathy

An experimental chronic infection was induced in inbred A2G and CBA mice by repeated administration of four cytopathogenic agents isolated from human cases of Balkan endemic nephropathy (BEN). The slight humoral and cell-mediated immune responses to the BEN agents recorded in A2G mice were associated with autoimmunity and hypersensitivity phenomena. In contrast, the normal, intense immune response observed in CBA mice was not accompanied by any immunopathological changes. In both mouse lines the chronic infection with human BEN agents led to a secondary immune deficiency against sheep red blood cells or tuberculin.     

Comparative h NMR metabolomic urinalysis of people diagnosed with balkan endemic nephropathy, and healthy subjects, in romania and bulgaria: a pilot study

(1)H NMR spectroscopy of urine has been applied to exploring metabolomic differences between people diagnosed with Balkan endemic nephropathy (BEN), and treated by haemodialysis, and those without overt renal disease in Romania and Bulgaria. Convenience sampling was made from patients receiving haemodialysis in hospital and healthy controls in their village. Principal component analysis clustered healthy controls from both countries together. Bulgarian BEN patients clustered separately from controls, though in the same space. However, Romanian BEN patients not only also clustered away from controls but also clustered separately from the BEN patients in Bulgaria. Notably, the urinary metabolomic data of two people sampled as Romanian controls clustered within the Romanian BEN group. One of these had been suspected of incipient symptoms of BEN at the time of selection as a 'healthy' control. This implies, at first sight, that metabolomic analysis can be predictive of impending morbidity before conventional criteria can diagnose BEN. Separate clustering of BEN patients from Romania and Bulgaria could indicate difference in aetiology of this particular silent renal atrophy in different geographic foci across the Balkans.     

Beta-endorphin: a common factor in the antihypertensive action of clonidine-type imidazolines in spontaneously hypertensive rats

The hypotensive action of two novel imidazolinic alpha-adrenergic agonists (ICI-106270 and UK-14304) with similar pharmacological properties to clonidine was shown in spontaneously hypertensive (SH) rats. The antihypertensive effect of the clonidine-type agents was prevented by either peripheral administration of the opiate antagonist naloxone or by intracerebroventricular (i.c.v.) injection with a specific antibody against human beta-endorphin (BEN). A dose-response relationship was found for the hypotensive effect of i.c.v. given BEN in SH rats, the low blood pressure being significantly reversed by further treatment with either naloxone or anti-beta h-endorphin. These data confirm and extend the notion of a BEN mediation in the antihypertensive action of clonidine-type alpha-adrenergic agonists in SH rats.     

Serum antibodies to papova viruses (BK and SV 40) in subjects from the area with Balkan endemic nephropathy

The presence of antibodies to BK virus and SV40 was investigated in 63 patients with Balkan endemic nephropathy (BEN) and in 83 apparently healthy subjects from the endemic area. Serum antibodies to BK virus were detected in 95.2% of the former and in 74.7% of the latter, high antibody levels being prevalent in the age groups 41-60 years. Antibodies to SV40 were absent in the BEN patients and their frequency in the healthy subjects (27.7%) was much lower than that previously recorded in healthy persons from other zones of Romania (40%). The results obtained plead for a prevalence of BK virus infection in the endemic area with BEN.     

Cytopathogenic protein in filtrates from cultures of Propionibacterium acnes isolated from patients with Kawasaki disease.

Propionibacterium acnes may have a role in Kawasaki disease. Filtrates from cultures of P acnes isolated from cervical lymph node biopsy specimens and blood samples from patients with Kawasaki disease were studied and compared with samples from control subjects. After inoculation of human embryo liver cells with filtrates from the patients a cytopathogenic effect and vacuolation were seen. A specific cytopathogenic substance was found in only the filtrates of cultures from patients with Kawasaki disease; it was a protein of about isoelectric point 7.0 with a molecular weight of about 100,000 daltons. The amount of IgG antibody to this cytopathogenic protein was measured by enzyme linked immunosorbent assay (ELISA) in serum samples taken from 63 patients in the acute phase of Kawasaki disease (mean 5.2 (SD 1.1) days after onset of illness), 45 in the subacute phase (mean 23.6 (3.3) days), and 51 in the convalescent phase (mean 18.5 (4.1) months) and from 102 control subjects matched for age. Titres of IgG antibody were significantly raised in patients with Kawasaki disease, particularly in the acute and subacute phases of the illness, compared with in the control subjects. Titres of IgG antibodies to cytopathogenic protein were found to be low in normal children below the age of 4 years but they increased with age thereafter. This may explain why outbreaks of Kawasaki disease, which is most common in children aged under 4, occur every three years.     

Ochratoxin A and β2-microglobulin in BEN patients and controls

Ochratoxin A (OTA) is a mycotoxin naturally occurring in different foods. OTA is arguably a risk factor for Balkan endemic nephropathy (BEN). The aims of this study are to (1) test the OTA-BEN association in BEN-groups and controls and (2) determine whether urine β2-microglobulin, a marker of impaired ability of the kidneys to re-absorb, is related to OTA. BEN patients had significantly higher OTA serum levels. Within the offspring, OTA was significantly related to higher β2-microglobulin excretion. OTA (2005/2006) was related to a higher incidence of BEN after 2008, providing further evidence that OTA is a risk factor for BEN.     

Laboratory investigations in Balkan endemic nephropathy

Serum samples from three patients with Balkan endemic nephropathy (BEN) were inoculated intraperitoneally to guinea pigs. After a very long incubation period (58-273 days) the animals developed an experimental disease characterized by apathy, tremor, convulsions and a fatal outcome. The disease could be serially propagated, the same clinical symptoms being recorded at each of the three passages performed. The morphological features of the experimental disease included glomerular and tubular lesions and the presence of interstitial fibrous and lymphoplasmocytic reactions. Different hypotheses on the etiology of BEN are discussed.     

Pathomorphology of Balkan endemic nephropathy.

Within the county of Slavonski Brod, Yugoslavia hyperendemic areas of Balkan endemic nephropathy (BEN) have been recognized for a long time. As the Department of Pathology and Forensic Medicine of the Medical Centre in Slavonski Brod is responsible for all diagnostic post-mortem examinations in the county, and at the same time is engaged in surgical pathological diagnosis, a considerable amount of material relating to BEN and concomitant urinary-tract tumours has been collected during a 16-year period. This material has been classified and used for pathological, anatomical and histological investigations. In this paper the most relevant findings are described and briefly discussed, without the intention to speculate on possible aetiology of BEN. As the origin of BEN and of the high frequency of urinary tumours among the people from the endemic areas are still unexplained, and uniform criteria for the pathomorphological diagnosis of BEN are badly needed, the proposal to set up an archive encompassing cases from all the centres investigating BEN is supported.     

The role of lecithin cholesterol acyltransferase and organic substances from coal in the etiology of Balkan endemic nephropathy: a new hypothesis.

Balkan endemic nephropathy (BEN) occurs in Serbia, Bulgaria, Romania, Bosnia and Herzegovina, and Croatia. BEN has been characterized as a chronic, slowly progressive renal disease of unknown etiology. In this study, we examined the influence of soluble organic compounds in drinking water leached from Pliocene lignite from BEN-endemic areas on plasma lecithin-cholesterol acyltransferase (LCAT) activity. We found that changes for all samples were the most prominent for the dilution category containing 90% plasma and 10% of diluting media. Water samples from BEN villages from Serbia and Romania showed higher LCAT inhibiting activity (p=0.02) and (p=0.003), respectively, compared to deionised water and non-endemic water. A secondary LCAT deficiency could result from this inhibitory effect of the organic compounds found in endemic water supplies and provide an ethiopathogenic basis for the development of BEN in the susceptible population.     

Nebulized drug admixtures: effect on aerosol characteristics and albuterol output.

Although not recommended, co-administration of drugs separately prescribed for nebulization is done in real life. The impact of this practice on drug output and aerosol characteristics is poorly understood. We studied the effect of drug admixtures (DA) on aerosol characteristics and drug output of nebulized albuterol delivered by a continuous output (CONT) and a breath enhanced nebulizer (BEN). Albuterol was nebulized alone (ALB) and combined with cromolyn sodium (A+CRO), ipratropium bromide (A+IB), tobramycin (A+TOB), flunisolide (A+FLU), and n-acetylcysteine (A+NAC). A BEN (PARI LC Plus) and a CONT (Hudson T UP-DRAFT II) were tested at 8 liters per minute (Lpm) for 2 and 5 min, respectively. Albuterol output and aerosol characteristics were determined by impaction and chemical analysis. Mass median aerodynamic diameter (MMAD; microm) A+CRO reduced MMAD from 2.57 (ALB) to 1.29 with CONT. A+FLU increased MMAD from 2.71 (ALB) to 3.40 with BEN. Geometric standard deviation (GSD) A+CRO increased GSD from 2.66 (ALB) to 3.36 with CONT. GSD was 2.33 for ALB and was not changed by DA with BEN. BEN generated a smaller and less heterodisperse aerosol than CONT. Respirable fraction (RF%) was 74% for ALB and was not changed by DA with CON. A+TOB and A+FLU decreased RF% from 75%, to 70% and 67% (respectively) with BEN. Respirable mass (RM; microg) for ALB was 935 and was not changed by DA with CONT. A+IB and A+FLU increased RM from 917 (ALB) to 1172 and 1240, respectively, with BEN. Co-nebulization of albuterol with other drugs can affect its output and aerosol characteristics. In vivo data is needed to asses the clinical implications of our findings.     

Relevance of a rat model of papillary necrosis and upper urothelial carcinoma in understanding the role of ochratoxin A in Balkan endemic nephropathy and its associated carcinoma.

Ochratoxin A is nephrotoxic and has been implicated in the genesis of Balkan endemic nephropathy (BEN), a condition that leads to end-stage renal disease and upper urothelial tumours. This compound induces renal parenchymal carcinoma in male mice only, and is not considered to be a potent carcinogen nor is there experimental evidence of its propensity to cause upper urothelial carcinoma. There is, however, evidence that exposure to more than one mycotoxin may be an important factor in the clinical spectrum of BEN. Analgesic nephropathy is clinically different, but is also associated with an upper urothelial carcinoma. The combination of urothelial initiation and an acute papillary necrosis in rats produces upper urothelial carcinoma. This two-stage experimental model offers the potential to assess the role of ochratoxin A in BEN-associated upper urothelial carcinoma under experimental conditions.     

Ochratoxin A and beta2-microglobulinuria in healthy individuals and in chronic interstitial nephropathy patients in the centre of Tunisia: a hot spot of Ochratoxin A exposure

Ochratoxin A (OTA) is a nephrotoxic mycotoxin considered to be the causal agent of the Balkan endemic nephropathy (BEN). In Tunisia, a chronic interstitial nephropathy (CIN) of unknown aetiology, resembling BEN, has been characterised wherein OTA seems to be implicated too. However, despite the considerable number of investigations conducted so far, the role of OTA in the outcome of this human nephropathy is still uncertain. In this study, an attempt is being made to consolidate the link between OTA and the Tunisian CIN of unknown aetiology. Blood OTA and beta(2)-microglobulinuria levels were measured in several groups of healthy individuals and patients having different renal diseases of known and unknown aetiologies (100 nephropathy patients and 40 healthy subjects). The high blood OTA and beta(2)-microglobulinuria levels seem to be strongly associated to the CIN of unknown aetiology. Our results support the involvement of this nephrotoxic agent in the outcome of this particular human nephropathy and underline furthermore the importance of beta(2)-microglobulinuria in the characterization of this disease.     

Aristolochic acid and 'Chinese herbs nephropathy': a review of the evidence to date.

Chinese herbs nephropathy (CHN) is a rapidly progressive interstitial nephropathy reported after the introduction of Chinese herbs in a slimming regimen followed by young Belgian women. It is characterised by early, severe anaemia, mild tubular proteinuria and initially normal arterial blood pressure in half of the patients. Renal histology shows unusual extensive, virtually hypocellular cortical interstitial fibrosis associated with tubular atrophy and global sclerosis of glomeruli decreasing from the outer to the inner cortex. Urothelial malignancy of the upper urinary tract develops subsequently in almost half of the patients. Suspicion that the disease was due to the recent introduction of Chinese herbs in the slimming regimen was reinforced by identification in the slimming pills of the nephrotoxic and carcinogenic aristolochic acid (AA) extracted from species of Aristolochia. This hypothesis was substantiated by the identification of premutagenic AA-DNA adducts in the kidney and ureteric tissues of CHN patients. Finally, induction of the clinical features (interstitial fibrosis and upper urothelial malignancy) typical of CHN in rodents given AA alone removed any doubt on the causal role of this phytotoxin in CHN, now better called aristolochic acid nephropathy (AAN). AAN is not restricted to the Belgian cases. Similar cases have been observed throughout the world, but AA is sometimes incriminated on the basis of the known content of AA in the herbs. The possibility remains that in some individuals in whom AA has not been demonstrated, other phytotoxins might be implicated. Biological and morphological features of AAN are strikingly similar to those reported in another fibrosing interstitial nephropathy of still unknown aetiology, Balkan endemic nephropathy (BEN). Interestingly, AA was incriminated as the cause of BEN many years ago, a hypothesis yet to be fully explored. The intake of AA and the presence of tissular AA-DNA adducts in patients with an unequivocal diagnosis of BEN remains to be demonstrated. The tragic phenomenon of CHN, recognised only 10 years ago, has been at the root of significant research and progress both in nephrology and oncology. It has provided a fascinating opportunity to understand the link between a fibrosing interstitial nephropathy and urothelial carcinoma. It allows the categorisation of interstitial nephritis on the basis of histological findings, of initiating toxic substances and of associated clinical features. Finally, it has led to the withdrawal in several countries of a previously unsuspected carcinogenic and nephrotoxic substance.     

Ochratoxin A in blood and its pharmacokinetic properties.

Since there are pathomorphological similarities between porcine mycotoxic nephropathy caused by ochratoxin A and Balkan endemic nephropathy (BEN), it has been suggested that the same aetiological agent has a role in BEN. Based on the results from several field and experimental studies carried out on pigs, an appropriate analytical method of monitoring possible human exposure to ochratoxin A was developed. The toxicokinetic properties of the toxin were species specific, although in all the animal species studied (with the exception of fish), as well as in humans, two binding proteins were found in the plasma. The monkey had the longest elimination half-life of the toxin, 510 hr, in contrast to the fish whose elimination half-life was only 0.68 hr. The fish kidney displayed a specific pattern of distribution. In the laying quail the most prominent observation was the accumulation of labelled ochratoxin A in egg yolk. Generally, [14C]ochratoxin A was eliminated rapidly from the quail body, but had a long retention time in the circulating blood in the mouse. Although the elimination of ochratoxin A from the body depending on its binding to plasma constituents, the existence of enterohepatic circulation might have been partially responsible for its prolonged retention and elimination from the body of mammals. The toxicokinetic profile of ochratoxin A did not contradict the mycotoxic hypothesis in the aetiology of BEN.     

Deciphering the canonical blockade of activated Hageman factor (FXIIa) by benzamidine in the coagulation cascade: A thorough dynamical perspective

The experimental inhibitory potency of benzamidine (BEN) paved way for further design and development of inhibitors that target β‐FXIIa. Structural dynamics of the loops and catalytic residues that encompass the binding pocket of β‐FXIIa and all serine proteases are crucial to their overall activity. Employing molecular dynamics and post‐MD analysis, this study sorts to unravel the structural and molecular events that accompany the inhibitory activity of BEN on human β‐FXIIa upon selective non‐covalent binding. Analysis of conformational dynamics of crucial loops revealed prominent alterations of the original conformational posture of FXIIa, evidenced by increased flexibility, decreased compactness, and an increased exposure to solvent upon binding of BEN, which could have in turn interfered with the essential roles of these loops in enhancing their procoagulation interactions with biological substrates and cofactors, altogether resulting in the consequential inactivation of FXIIa. A sustained interaction of the catalytic triad residues and key residues of the autolysis loop impeded their roles in catalysis which equally enhanced the inhibitory potency of BEN toward β‐FXIIa evidenced by a favorable binding. Findings provide essential structural and molecular insights that could facilitate the structure‐based design of novel antithrombotic compounds with enhanced inhibitory activities and low therapeutic risk.     

Effect of pine seed shell extract on microbial and viral infection.

Pretreatment of mice with ammonia extract of seed shell of Pinus Koraicenis, via the intraperitoneal or intravenous route, effectively protected them from lethal infection of Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus. The pine seed shell extract also moderately inhibited syncytium formation and cytopathogenic effect induced by human immunodeficiency virus (HIV) infection in cultured human lymphotropic virus type I (HTLV-1) positive MT-4 cells. These data suggest a medicinal potential of pine seed shell extract against opportunistic infection in HIV patients.     

葛根素对高血压患者血浆内皮素和一氧化氮的影响

目的：检测正常人与高血压病（ＥＨ）患者血浆内皮素、一气体氮的变化及葛根素对其的干预效应。方法：ＥＨ对照组口服苯磺酸氨氯地平５ｍｇ／ｄ,或合和盐酸苯那普利１０ｍｇ／ｄ,每日一次,１５ｄ为一疗程。ＥＨ伍用治疗组同时合用５％ＧＮＳ２５０ｍｌ＋葛根素注射液４００ｍｇ静滴,两组治疗前后检测血浆ＥＴ、ＮＯ水平变化。结果：ＥＨ各组血浆ＥＴ较正常人组均显著增高（Ｐ〈０．０１）,血浆ＮＯ水平及ＮＯ／ＥＴ比值除轻度Ｅ     

Geographically-based evaluation of multidrug resistance trends among Streptococcus pneumoniae in the USA: findings of the FAST surveillance initiative (2003-2004)

Surveillance initiatives to track Streptococcus pneumoniae resistance trends are important for understanding the current in vitro effectiveness of available antimicrobial agents. The antimicrobial susceptibility profiles of S. pneumoniae (n = 1479 isolates) collected from 17 geographical areas across the USA (2003–2004) were analysed; 36.8% of isolates were resistant to one or more agents (24.4% were multidrug-resistant, i.e. resistant to two or more antimicrobial classes). Multidrug resistance involved resistance to β-lactams, macrolides, tetracycline and trimethoprim/sulphamethoxazole, but rarely fluoroquinolones (>96% of multidrug-resistant isolates were fluoroquinolone-susceptible). Multidrug resistance rates were prominent regardless of the geographical region surveyed. As this trend continues, the empirical therapeutic options for S. pneumoniae infections will diminish and there will be an ongoing need to evaluate the effectiveness of potent fluoroquinolones such as gemifloxacin.     

派瑞松乳膏中3种药物透皮特性的研究

目的:评价派瑞松乳膏中曲安奈德(TACA)、苯甲酸(BEN)、硝酸益康唑(ECN)3种药物的透皮特性.方法:采用Franz扩散池法,考察药物经完整皮肤和去角质层皮肤的体外透皮能力,并采用了胶带剥离、皮肤萃取法分别获取了皮肤角质层、去角质层皮肤样本,用HPLC法测定了样本中的药物含量.结果:经过24 h透皮吸收,TACA和BEN的去角质层皮肤渗透量分别为完整皮肤的1.5和1.3倍,ECN的渗透量基本为零.8h透皮实验,TACA高、中、低3个浓度角质层中药物含量基本相同,真皮层则存在浓度依赖现象;ECN在皮肤各层和接受室均检测不到;BEN在角质层和真皮层中的分布与TACA相似,但透过量比TACA大.结论:角质层是皮肤渗透的重要屏障,派瑞松乳膏应用于皮肤溃疡、受损或者婴幼儿皮肤仍需谨慎.