First Report of Pseudomonas syringae pv. coriandricola Causing Bacterial Leaf Spot on Carrot, Parsley, and Parsnip in Serbia.

During the spring of 2014, a severe leaf spot disease was observed on carrot (Daucus carota), parsley (Petroselinum crispum), and parsnip (Pastinaca sativa) on a 0.5-ha vegetable farm in Vojvodina Province, Serbia. The disease appeared under wet and cool conditions with 5 to 25% of plants infected for each of the three crops. Symptoms were characterized as brown angular leaf spots, ~2 mm in diameter, often limited by veins. Collected symptomatic leaves were rinsed and dried at room temperature, and leaf sections taken from the margin of necrotic tissue were macerated in sterile phosphate buffer and streaked onto nutrient agar with 5% (w/v) sucrose (NAS). After isolation, whitish, circular, dome-shaped, Levan-positive colonies consistently formed. Five strains from each host (carrot, parsley, and parsnip) were used for further study. Strains were gram-negative, aerobic, and positive for catalase and tobacco hypersensitive reaction but negative for oxidase, rot of potato slices, and arginine dihydrolase. These reactions corresponded to LOPAT group Ia, which includes Pseudomonas syringae pathovars (3). Repetitive extragenic palindromic sequence (Rep)-PCR fingerprint profiles using the REP, ERIC, and BOX primers (4) were identical for all strains. Sequence typing of the housekeeping genes gyrB and rpoD (1) was performed for three representative strains (one from each host). Sequences were deposited in the NCBI GenBank database as accessions KM979434 to KM979436 (strains from carrot, parsnip, and parsley, respectively) for the gyrB gene and KM979437 to KM979439 (strains from parsnip, parsley and carrot, respectively) for the rpoD gene. Sequences were compared with pathotype strain Pseudomonas syringae pv. coriandricola ICMP12471 deposited in the Plant Associated and Environmental Microbes Database ( http://genome.ppws.vt.edu/cgi-bin/MLST/home.pl ). BLAST analysis revealed 100% homology for gyrB and 99% homology for rpoD. Pathogenicity was tested with five representative strains from each host on four-week-old plants of carrot (cv. Nantes), parsley (cv. NS Molski), and parsnip (cv. Dugi beli glatki) using two methods: spraying the bacterial suspension (108 CFU ml-1) on the leaves until runoff (5) and injecting the bacterial suspension into leaves with a hypodermic syringe (2). Four plants were used per strain and method. Sterile distilled water was applied as a negative control treatment for each plant species. All plants were kept in a mist room with 100% humidity for 4 h, then transferred to a greenhouse at 25°C and 80% relative humidity and examined for symptom development over a period of three weeks. For all strains, inoculated leaves first developed water-soaked lesions on the leaves 5 to 7 days after inoculation (DAI); 14 DAI lesions became dark brown, often surrounded by haloes. No symptoms were observed on control plants inoculated with sterile distilled water. For fulfillment of Koch's postulates, re-isolations were done onto NAS. Re-isolated bacteria were obtained from each inoculated host and confirmed to be identical to the original isolates using the LOPAT tests and Rep-PCR fingerprinting profiles. Based on the pathogenicity test accompanied by completion of Koch's postulates, sequence analysis, and bacteriological tests, the strains were identified as P. s. pv. coriandricola. To our knowledge, this is the first report of bacterial leaf spot of carrot, parsley, and parsnip in Serbia. It may present a threat to production due to quality requirements for fresh market. References: (1) P. Ferrente and M. Scortichini. Plant Pathol. 59:954, 2010. (2) M. Gupta et al. Plant Dis. 97:418, 2013. (3) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (4) F. J. Louws et al. Appl. Environ. Microb. 60:2286, 1994. (5) X. Xu and S. A. Miller. Plant Dis. 97:988, 2013.

Lupus nephritis in Croatian children: clinicopathologic findings and outcome.

We report clinical and histopathological features, treatment and outcome of 37 Croatian children with biopsy-proven lupus nephritis seen over a 30-year period. The mean age at lupus nephritis presentation was 12.11 ± 2.59 years (range 4.66-17.0). The most frequent histopathological finding was class IV (37.8%), followed by class III (35.1%), class V (16.2 %) and class II (10.8 %) lupus nephritis. Compared with other classes there were more boys among patients with class IV lupus nephritis, and hypertension, nephrotic syndrome and decreased estimated glomerular filtration rate at presentation were more common. The median histopathological activity and total scores were highest in class IV lupus nephritis patients. The mean follow-up was 7.14 ± 4.71 years, ranging from 1.1 years to 21.0 years. Kaplan-Meier estimate S: of patient and kidney (without renal failure) survival rate S: were 90.5% and 87 % at five years. The renal survival rate of class IV lupus nephritis patients was found significantly lower compared with other histological classes combined. Decreased estimated glomerular filtration rate at the time of diagnosis, class IV lupus nephritis versus other lupus nephritis classes, and high total histological score were the parameters significantly associated with adverse outcome. The therapy with cyclophosphamide showed as superior to the therapy with azathioprine.

Redifferentiation and induction of tumor suppressors miR-122 and miR-375 by the PAX8/PPARγ fusion protein inhibits anaplastic thyroid cancer: a novel therapeutic strategy.

Anaplastic thyroid cancer (ATC) is an aggressive, fatal disease unresponsive to traditional therapies, generating a need to develop effective therapies. The PAX8/PPARγ fusion protein (PPFP) has been shown to favorably modulate tumor growth in follicular thyroid cancer, prompting our evaluation of its efficacy to inhibit ATC cell and tumor growth in vitro and in vivo. PPFP was constitutively expressed in five ATC cell lines: BHT-101, FRO, C-643, KTC-2 and KTC-3, and inhibited cell growth in four of five cell lines and xenograft tumor growth in four of four cell lines. PPFP-mediated growth inhibition involved multiple mechanisms, including upregulation of miR-122 and miR-375, associated with decreased angiogenesis and AKT pathway inactivation, respectively. Also, PPFP expression resulted in marked increase of thyroid-specific marker transcripts, including PAX8, thyroid peroxidase (TPO), sodium iodide symporter (NIS) and thyroglobulin, to varying degrees by activating their respective promoters, suggesting that PPFP induced cellular redifferentiation. Functional studies demonstrate that increased NIS messenger RNA is not associated with increased 125I uptake. However, ectopic expression of wild-type NIS-induced perchlorate-sensitive iodine uptake, suggesting that endogenous NIS in ATC cell lines is defective. As current treatment for ATC is only palliative, overexpression of PPFP may offer a novel therapeutic strategy for the treatment of ATC.

First Report of Cucumber mosaic virus Infecting Peperomia tuisana in Serbia.

Peperomia tuisana C.DC. ex Pittier (family Piperaceae) is an attractive succulent grown as an ornamental. Despite its tropical origins, it can be successfully grown indoors in any climate. In March 2012, three samples of P. tuisana showing virus-like symptoms were collected from a commercial greenhouse in Zemun (District of Belgrade, Serbia) in which estimated disease incidence was 80%. Infected plants showed symptoms including necrotic ringspots and line patterns that enlarged and caused necrosis of leaves. A serious leaf drop led to growth reduction and even death of the plant. Leaves from three symptomatic P. tuisana plants were sampled and analyzed by double-antibody sandwich (DAS)-ELISA using commercial diagnostic kits (Bioreba AG, Reinach, Switzerland) against the most common viral pathogens of ornamentals: Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV), and Impatiens necrotic spot virus (INSV) (1,2). Commercial positive and negative controls were included in each ELISA. Serological analyses showed that all plants were positive for CMV and negative for TSWV and INSV. The ELISA-positive sample (isolate 1-12) was mechanically inoculated onto five plants each of three test species as well as of healthy young P. tuisana using 0.01 M phosphate buffer (pH 7). Chlorotic local lesions on Chenopodium quinoa and severe mosaic and leaf malformations were observed on all inoculated Nicotiana tabacum 'Samsun' and N. glutinosa. Also, the virus was successfully mechanically transmitted to P. tuisana that reacted with symptoms identical to those observed on the original host plants. All mechanically inoculated plants were positive for CMV in DAS-ELISA. For further confirmation of CMV infection, reverse transcription (RT)-PCR was performed on extracts made from symptomatic P. tuisana, N. tabacum 'Samsun,' and N. glutinosa leaf materials. Total RNAs were extracted with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was carried out using One-Step RT-PCR Kit (Qiagen). A CMV-specific primer pair, CMVCPfwd and CMVCPrev (3), which amplifies an 871-bp fragment of the entire coat protein (CP) gene and part of 3'- and 5'-UTRs, were used for both amplification and sequencing. Total RNAs obtained from the Serbian CMV isolate (HM065510) and healthy P. tuisana were used as positive and negative controls, respectively. A product of the correct predicted size was obtained in all naturally and mechanically infected plants, as well as positive control. No amplicon was recorded in the healthy control. The amplified product derived from isolate 1-12 was purified (QIAquick PCR Purification Kit, Qiagen), directly sequenced in both directions, deposited in GenBank (KC505441), and analyzed by MEGA5 software (4). Sequence comparison of the complete CP gene (657 nt) revealed that the Serbian isolate 1-12 shared the highest nucleotide identity of 99.1% (99.5% amino acid identity) with the Japanese isolate (AB006813). To our knowledge, this is the first report on the occurrence of CMV in P. tuisana in Serbia. This is also an important discovery since P. tuisana is commonly grown together with other ornamental hosts of CMV, and thus could represent a serious threat for future expansion of CMV in the greenhouse floriculture industry in Serbia. References: (1) M. L. Daughtrey et al. Plant Dis. 81:1220, 1997. (2) S. Flasinski et al. Plant Dis. 79:843, 1995. (3) K. Milojevic et al. Plant Dis. 96:1706, 2012. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

Acute geriatric units.

Together with increase of population of elderly people, there is an increase of the number of hospital admission for emergencies, especially for elderly people. Serbia shares this need for Acute Geriatric Units (AGU). The National Programme of Health Care of Elderly People Improvement is planning to open the Geriatric Ward in every regional general hospital in Serbia. But in cities with several hospitals, there is a waste need for geriatric ward capable of taking acute care. So, there is a need for geriatric wards in Serbia nowadays. The main targerts for AGU should be: the comprehensive geriatric assessment, prevention of development of worsening of delirium, decubital wounds, incontinence, improving mobility and early planning of discharge. The multidisciplinary team, which includes physicians, nurses, physiatrist and social worker, is the best solution for getting this target day by day. Thus, one of the distinctive features of Acute Geriatric Units should be comprehensive geriatric assessment, the prevention of geriatric syndrome and early planning of discharging the elderly patient.

The principles of chemotherapy of colorectal cancer in elderly.

The prevalence of colorectal cancer (CRC) increases significantly with age, with 40% of patients in Europe being older than 74 years of age at the time of initial diagnosis. The individualized management of the older-aged patient with cancer is based on the answers to the following questions: 1) will the patient die of cancer or with cancer; 2) will the patient suffer cancer-related morbidity; and 3) is the patient able to handle the toxicity of treatment? More than chronological age, the following parameters are important when elderly patients are to be treated with antineoplastic agents: general condition, liver function, kidney function and bone marrow status. Frail elderly with malignant disease should not be treated with cytostatic therapy. In the case of fit elderly, the standard chemotherapy (i.e. FOLFOX) regimen could be administered. In elderly ineligible for combination chemotherapy, the capecitabine used orally, as a single-agent therapy, is an important therapeutic option for colorectal cancer.

Accomplishments and perspectives of immunological interventions in lymphoproliferative disorders.

The purpose of this review is to consider evidences accumulated in the last few years which might lead to a new therapeutic strategy for lymphoproliferative disorders. Tumor-targeted and immunologically designed therapy has already started. Clinical effectiveness also includes the primary and secondary prevention of these malignancies. The fortifying of body's defense through vaccination as primary prevention with tumor-specific cell surface antigen, has been seen over the past few years, and will lead to health patient's improvement. Data collected from clinical trials and in vitro analysis indicated that the immune system of patients could recognize and eliminate neoplastic cells while sparing normal cells. In B-cell lymphomas and myelomas, the tumor-idiotype (Id) produced by a single B-cell clone, has been used for vaccination. Therapeutic Id vaccination used two types of antigen-presenting cells as natural adjuvants for the induction of antigen-specific T cell response. Dendritic cells (DCs)--based vaccines are under active investigation and are entering clinical evaluation. Current research focuses on optimization of DCs source, choice and loading of antigen, mode of injection, as well as immune monitoring. Some preliminary results were obtained and no significant side effects of dendritic cells vaccination of lymphoma and myeloma patients have so far been reported. The over-estimation of clinical effectiveness, at this moment, is still limited by the small number of patients included in this kind of treatment. The challenge for the future will be to extend these early results to reproducible vaccination strategy according to current standards of good clinical practice (GCP).

Effects of X-ray irradiation on the overexpression of HER-2/Erb-B2 on breast cancer cell lines.

Irradiation is the conventional treatment modality for cancer patients. However, besides its cytotoxic effects on malignant cells it might also affect the biology of surviving cells. Since overexpression of HER-2 receptors on malignant cells is a prerequisite for the therapeutic efficacy of Herceptin, it seems important to know whether previous irradiation changes their overexpression. The experiments performed in this work were aimed to determine whether X-ray irradiation of MDA-MB-361 and MDA-MB-453 breast carcinoma cell lines, besides its cytotoxic action, affects the overexpression of HER-2 protein. Determination of the cytotoxic effect of X-ray irradiation was done using trypan blue test. The breast carcinoma cell responsiveness to herceptin treatment in the presence of 10% fresh human serum (from healthy volunteer's) in the presence or absence of 25 microg/ml of herceptin, in vitro before and after cell-irradiation, was evaluated by MTT test. The degree of HER-2 overexpression was determined by immunocytochemistry, using DAKO HercepTest. Preliminary results obtained in this work showed that X-ray irradiation, besides its cytotoxic effect on malignant cells, could lead to overexpression of HER-2 receptors on (initially by immunocytochemistry, HER-2 negative) tumor cells, indicating change in biology of treated tumor cells. Further investigation in this direction will probably be helpful to elucidate this task in order to improve the selection of irradiated patients for Herceptin therapy.

The changes of serum immunoglobulins in patients treated by a combination of anti-CD20 monoclonal antibody rituximab plus chlorambucil for low-grade non- Hodgkin's lymphoma.

PURPOSE: To assess whether the therapeutic application of monoclonal anti-CD20 antibody (rituximab) may affect normal B lymphocyte function, since CD20 is involved in their activation, proliferation and differentiation. PATIENTS AND METHODS: Serum immunoglobulins (Igs) concentrations and their possible relation to pretreatment levels of Igs and B-cells ratio as well as the T/B cell ratio were investigated in 9 patients with low-grade non Hodgkin's lymphoma (NHL) during the administration of rituximab (Mabthera(R)) and chlorambucil combination therapy. Serum Igs concentrations were determined by the radial immunodiffusion (RID) method and the number of B and T lymphocytes by flow cytometry analysis. RESULTS: The altered values of Igs concentration and B cell number were registered in each patient before therapy. Generally, the therapy did not normalize the pretreatment alterations of these parameters, though it depleted malignant clones from peripheral blood. Nevertheless, IgM and IgA concentrations have been considerably changed from baseline level (35-74%) in 5 patients. The concentration of IgA and IgM raised in 4, while the IgM declined from baseline values in 1 patient, irrespective of their therapeutic response. Four of these patients had a normal concentration of these Igs classes, and a profound B cell number alteration before therapy. CONCLUSION: The changes of IgM and IgA concentrations in relation to pretreatment B cell number, although found in a small number of patients, might deserve further investigation with an aim to study any interference of the anti-CD20-based therapy with lymphocytes function.

Use of a ruthenium(III), iron(II), and nickel(II) hexacyanometallate-modified graphite electrode with immobilized oxalate oxidase for the determination of urinary oxalate.

This paper describes the performance of a biosensor with an Ru(III), Ni(II), and Fe(II) hexacyanometallate-modified graphite electrode and immobilized oxalate oxidase for the determination of urinary oxalate. The addition of ruthenium enhances the electrochemical reversibility and chemical stability of the electrocrystallized layer and improves the sensitivity of the biosensor. Hydrogen peroxide, produced by the enzyme-catalyzed oxidation of oxalate, was measured at -50 mV vs an Hg Hg2CI2 3M KCl electrode in a solution of pH 3.6 succinic buffer, 0.1 M KCl, and 5.4mM ethylenediaminetetraacetic acid. The linear concentration range for the determination of oxalate was 0.18-280 microM. The recoveries of added oxalate (10-35 microM) from aqueous solution ranged from 99.5 to 101.7%, whereas from urine samples without oxalate (or with a concentration of oxalate below the detection limit) the recoveries of added oxalate ranged from 91.4 to 106.6%. The oxalate in 24 h urine samples, taken during their daily routine from 35 infants and children, was measured and found to range from 0.6 to 121.7 mg/L. There were no interferences from uric acid, acetylsalicylic acid, and urea in the concentration range investigated, but paracetamol and ascorbic acid did interfere. A good correlation (R2 = 0.9242) was found between values obtained for oxalate in real urine samples by 2 laboratories, with the proposed biosensor and ion chromatography, respectively.

The different level of immunological recovery after chemotherapy in leukemia and lymphoma patients.

Analysis of T lymphocyte subsets made a great progress in attempting their important role in host defence against malignant disease. As the major regulatory elements in the immune system, helper (CD4+) and suppressor (CD8+) T cells are required for recognizing a large scale of surface antigen on tumor cells and subsequently, for mediating the regression of cancers. Many data suggest that T cell immunity system is not intact in different malignant diseases. The malignant disorders of hemopoietic tissue are frequently associated with T cell functional impairment, the mechanism(s) of which is still unresolved. In recent years, defects in immunoregulatory T cell network have been repeatedly proposed as the cause of this alteration and might be considered as a parameter of immunodeficiency in those patients. The additional disturbance of T cell immunoregulatory system may be induced by different extrinsic factors. In this context the chemotherapeutic agents with their non-selective action, including an immunomodulatory activity, should be involved in the evaluation of lymphocyte subset changes during treatment. In this brief review we analyzed the data concerning the T cell subset changes, both numerical and functional, before and in course of the treatment of patients with different hematologic malignancies. According to this consideration, it might be suggested that the immune system of these patients have some capacity to recover from the suppressive effect of chemotherapy, although not sufficiently to be normalized. The long-term monitoring of a larger scale of immunological parameters may probably help us to resolve whether T lymphocyte subset changes are related only to the therapy or, in some instance, might be connected with compromised natural host defence mechanisms in oncohematologic patients.

[Otogenic herpes zoster--the Ramsay-Hunt syndrome]. / Otogeni herpes zoster--Ramsay-Huntov sindrom.

INTRODUCTION: Herpes zoster is a viral disease caused by a specific neurotropic virus-varicella zoster, similar to varicella virus, but not identical. Herpes zoster oticus was described by Letulle in 1882 and Körner in 1884, but particularly studied by Ramsay Hunt who reported it as a herpetic disease of ganglion geniculi in 1907. Herpes zoster oticus associated with facial nerve paralysis is most commonly called the Ramsay-Hunt syndrome. MATERIAL AND METHODS: In this work, cases of herpes zoster oticus associated with facial nerve paralysis are shown. At the ORL Clinic in Novi Sad, in the period from 1996-1997, 5 cases with Ramsay-Hunt syndrome were treated. The diagnostic procedure involved analysis of anamnestic data, clinical examination, complete cochleovestibular investigation with electronystagmography (ENG), topodiagnostic investigation of facial nerve (Schirmer's test, stapedial reflex, electrogustometry), electromyographic investigation (EMG), laboratory and virusologic investigations. According to many statistical data, paralysis of facial nerve due to herpes zoster is after Bell's paralysis the most common cause of the disease. The efflorescence of auricula, face and neck, which are typical manifestations of the disease, may precede facial nerve paralysis for about a week or more, and therefore may be disregarded and misdiagnosed with Bell's paralysis. The peripheral paralysis of this nerve in herpes zoster has an unfavorable course. More than 75% of patients have consequences of paralysis (paresis, hemispasm, synkinesia etc.). Regarding the unfavorable recovery period in herpes zoster, we managed our patients accordingly. CONCLUSION: Herpes zoster oticus is a common cause of peripheral facial nerve paralysis. The clinical course is not as favorable as in Bell's paralysis. It may be associated with sensorineural hearing disorder, vertigo and paralysis of other cranial nerves. The therapeutic procedures in Ramsay-Hunt syndrome include administration of conservative therapy and surgical intervention. We performed surgery in 2 and conservative therapy in 3 patients. Facial nerve decompression is indicated in persistent paralyses, or in cases without clear clinical signs of recovery after 6 weeks-2 months from the onset of the disease. The site of decompression is determined by topodiagnostic investigations.

[Mycotic disease of the mucous membranes of the head and neck]. / Mikoticno oboljenje sluzokoza organa glave i vrata.

INTRODUCTION: Candidiasis is usually a superficial infection of the moist areas of the body and is generally caused by Candida albicans. Visceral infections occur in diabetes, lymphomas and leukemias, malnutrition, avitaminosis and they are associated with antibiotic, corticosteroid and immunosuppressive therapy. Candida albicans was isolated from middle ear inflammation. The diagnosis is made on the basis of microscopic appearance of colonies and characteristic smell. Candidiasis is successfully treated with nystatin, imidazol derivatives (fluconazole, ketoconazole and intraconazole), amphotericin B, 5-fluorocystosine and 1% iodine solution. CASE DESCRIPTION: This is a case report of a 46-year-old patient with a persistent nasal, sinus and ear infection of unknown origin. The patient first received antibiotic and steroid therapy and trepanation of the right maxillary sinus was performed. As the patient's condition aggravated with increase of temperature and bad laboratory findings, he was hospitalized. Radiography revealed a pathological process in both maxillary sinuses and both mastoids, so mastoidectomy and left maxillary sinus trepanation were performed. Histopathological examination of the right mastoid revealed a mould infection. The immunologic status pointed to hypogammaglobulinemia IgG. The following diseases were excluded: systemic diseases, blood diseases, Reiter's syndrome, AIDS, Hepatitis B, other viral diseases, toxoplasmosis, trichinellosis, borreliosis, typhus, paratyphus and exanthematous typhus. The diagnosis of candidiasis caused by Candida crusei and Candida kefyr was made on the basis of macroscopic and microscopic findings and biochemical identification. Ketoconazole was introduced (400 mg/per day) as well as high doses of vitamins and povidone-iodine locally. After a period of remission the patient died due to myocarditis, sepsis, acute kidney failure associated with severe mucosal necrosis of the mouth, esophagus and throat. Differential diagnosis in fever of unknown origin must include the possibility of mycotic infection, whereas the therapy of mycotic diseases must include two antimycotics at the same time. DISCUSSION AND CONCLUSION: Candida albicans is often found in the oral cavity and skin as well as in intestines of 18% of healthy subjects. It is unknown why it causes clinical illness. Antibiotic therapy of bacterial infections enables candida colonization especially in immunosuppressed patients. In our patient two types were found: Candida krusei and Candida kefyr. It is of special importance to perform differential diagnosis in cases with fever of unknown origin in order to include the possibility of mycotic infections, whereas treatment of systemic fungal infections requires a team of physicians.

[Vasomotor skin tests in non-eosinophilic and eosinophilic long-term (perennial) nonallergic rhinitis]. / Vazomotorni kozni testovi u neeosinofilnom i eozinofilnom dugotrajnom ("perenijalnom") nealergijskom rinitisu.

INTRODUCTION: It has generally been assumed that "perennial" non-allergic rhinitis is a heterogeneous syndrome consisting of at least two groups: non-eosinopilic and eosinophilic. Opposite to non-eosinophilic group, eosinophilic group is characterized by nasal secretion eosinophilia, frequent evolution to nasal polyposis or complete ASA triad (nasal polyposis, intrinsic asthma and intolerance of non-steroidal anti-inflammatory drugs) and by a good response to treatment with anti-histamines and corticosteroids. These characteristics obviously separate eosinophilic from non-eosinophilic rhinitis and point out the importance of nasal secretion eosinophilia for the evolution and therapy of rhinitis. However, the distinction between eosinophilic and non-eosinophilic rhinitis can only be made by nasal cytology. Skin tests with vasomotor agents were carried out to characterize vasomotor skin reactivity in "perennial" non-allergic rhinitis and determine whether the patients with non-eosinophilic rhinitis differ from patients with eosinophilic rhinitis. METHODS: On the basis of the examination of nasal smears of eosinophils, 74 patients with "perennial" non-allergic rhinitis were divided into non-eosinophilic (n = 63) and eosinophilic group (n = 11). Nasal eosinophilic was considered significant when 20% and more of the cells in nasal smear were eosinophils. Skin reactivity to intracutaneous test with different concentrations of papaverine, metacholine, histamine and compound 48/48 was measured, as well as specific skin reactivity to control saline solution. Pathological skin reactivity to vasomotor agents was defined as hyporeactivity to papaverine (5 mg/mL), when wheal-and-flare skin reaction diameter was less than 15 mm; hyper-reactivity to metacholine (0.02, 0.2 and 2.0 mg/mL), when two of three wheal-and-flare skin reaction diameters were greater than 15, 25 and 31 mm, respectively; hyper-reactivity to histamine (0.01, 0.1, 1.0 and 10.0 micrograms/mL), when three of four wheal-and-flare skin reaction diameters were greater than 7, 13, 25 and 40 mm, respectively; and hyper-reactivity to compound 48/80 (0.01, 0.1, 1.0 and 10.0 micrograms/mL, when three of four wheal-and-flare skin reaction diameters were greater than 9, 16, 26 and 38 mm, respectively. RESULTS: Seventy four patients with "perennial" non-allergic rhinitis were included in the study. There were 51 females, age range from 18 to 57 yrs. (mean 37 yrs.) and 23 males, age range from 18 to 73 yrs. (mean 44 yrs.). The difference between the number of females and males was significant (p = 1.1 x 10(-3)), while no significant difference regarding the age between females and males was found (p = 0.122). Significant percentage of eosinophils was found in 15% of "perennial" non-allergic rhinitis patients, and they were classified into eosinophilic group (n = 11). In this group, the percentage of eosinophils varied from 20% to 80%, mean 35%. In non-eosinophilic group (n = 63), it ranged from 0% to 10%, mean 1%. No significant difference concerning sex and age between the two groups of the rhinitis patients was observed (Table 1). There was no significant intergroup difference for pathological skin reactivity to papaverine, metacholine, histamine, compound 48/80 and saline (Table 2). Total pathological skin reactivity to vasomotor agents, single and in combination, was found in 78% of non-eosinophilic and in 91% of eosiniophilic rhinitis patients (Table 3). In both, non-eosinophilic and eosinophilic groups, frequencies of total pathological skin reactivity to vasomotor agents was significantly greater than frequencies of total normal skin reactivity (p = 1.1 x 10(-5) and p = 0.007 respectively). However, the difference of total pathological skin reactivity to vasomotor agents between the two groups was not significant (p = 0.552). (ABSTRACT TRUNCATED)

Potential diagnostic, prognostic and therapeutic implications of some new regulatory proteins in hematologic malignancies.

Multiple cell surface receptors play an important role in the biology of hematological malignant cells. Like already clustered monoclonal antibodies, the use of new markers of differentiation provide some new information about their structure and function. Here we consider the role of few selected regulatory proteins that are most frequently involved in the processes of activation, cell differentiation and proliferation of human hematological malignant cells. These molecules, being either lineage-restricted or multi-lineage cell activation ones, are involved in cell-cell and cell-matrix communications. They are involved in various leukocyte functions such as mobility, interaction with endothelium, and homing. Since physiologic cell growth involves not only cell division, but also programmed-cell death, we considered some apoptosis-regulatory molecules implicated in the clinical heterogeneity of hematological malignancies. The presence or absence of staining for these molecules is not only an important discriminatory immunophenotypic feature, but it appears useful also for prognosis and particularly in an experimental therapeutical setting. The prognostic significance of cell-expression of these molecules has not yet been clearly established, but it might be helpful in the evaluation of stability and progression of the disease.

[Simultaneous extracranial and intracranial otogenic complications]. / Istovremena egzokranijalna i endokranijalna otogena komplikacija.

INTRODUCTION: Chronic otitis media is the most common cause of otologic complications. A simultaneous occurrence of extracranial and intracranial otologic complications is rare in clinical practice. In this paper, we are presenting a patient with exacerbation of chronic otitis media and associated otologic complications: peripheral facial nerve palsy and subdural cerebral abscess. CASE REPORT: A patient aged 56 who has suffered from chronic otitis media during the past 37 years, was admitted at the Clinic of Otorhinolaryngology due to worsening of the underlying disease. The clinical examination revealed chronic otitis media with granulation in the external auditory canal, pulsatile discharge and extracranial otologic complication--facial nerve paralysis. The preoperative diagnostic procedure included: cochleovestibular investigations (hearing disorder of mixed type on the left side with normal labyrinthine function), temporal bone radiography (sclerotic cell alteration of mastoid on the left), topodiagnosis of facial nerve). The laboratory finding confirmed increased number of leukocytes (21.7 x 10(9)/l), increased erythrocyte sedimentation rate (25/58 mm/h), increased fibrinogen (5.0 g/l) and presence of protein in urine. Chest and heart X-ray findings were normal. Staphylococcus aureus was isolated from ear discharge by microbiological investigation. Signs of meningitis were negative, the liquor was colorless, slightly stirred up with total number of cells 384 x 10(6)/l, sugar 2.7 mmol/l and total proteins from 0.82 g/l. Bacteriological liquor culture was negative. The ophthalmologic examination confirmed normal finding of the eye fundus and absence of increased cranial pressure. For further diagnostics CT (computerized tomography) and MRI of head (magnetic resonance imaging) were performed. The findings confirmed subdural abscess and suspected encephalitic foci of the left cerebral lobe. According to findings, surgery involving radical trepanation of the temporal bone, decompression of facial nerve, denudation of sigmoid sinus dural sinus and angle area and incision of dura mater of the cranial fossa posterior with drainage of subdural abscess (meningitis surgery) was performed. CONCLUSION: A simultaneous occurrence of extracranial and intracranial otologic complications accompanied by subdural abscess is rare in clinical practice. The mechanism of development and spreading of subdural abscess is very interesting. In this case, subdural abscess caused the reaction of dura mater, which has prevented spreading of the disease by cerebrospinal liquid at the onset. However, there is a possibility of its further spreading by blood vessels into the brain white substance, where encephalitic foci may develop and later brain tissue abscesses as well. The clinical course of subdural abscess may be atypical, without headache and increased cranial pressure. This kind of disease demands a complex diagnostic procedure, sufficient otosurgical and neurosurgical interventions, cooperation with infectologist and administration of antibiotic therapy.

Lyophilized whole human melanoma cells stimulate human PBMC proliferation and enhance suppressive action of PBMC toward survival of the same malignant cell line in vitro.

The goal of this work was to determine: a) do lyophilized human melanoma BG or Fem-X cells affect the proliferative capacity of normal human peripheral blood mononuclear cells (PBMC) and b) does the PBMC six-days preincubation in nutrient medium with FBS with, or without lyophilized human melanoma BG or Fem-x cells, affect their suppressive action on the survival of the same malignant cell line in vitro. In the aim to avoid any stimulating effect of FBS, other group of experiments were done in nutrient medium with human AB serum in order to determine: c) does the PBMC six-day-preincubation with lyophilized human melanoma BG or Fem-x cells affect their antiproliferative action on the corresponding malignant cell line in vitro and d) does the PBMC six-day preincubation with lyophilized normal PBMC, obtained from healthy volunteer (as a source of allogenous, but not of tumor antigens), affect their suppressive action on the survival of both melanoma BG and Fern-x cell lines in vitro. Results obtained in the presence of FBS in nutrient medium, showed that lyophilized BG cells induced a proliferation of the healthy PBMC, depending on the number of stimulating lyophilized cells. Lyophilized Fem-x cells induced healthy PBMC proliferation in lesser degree than lyophilized BG cells. This stimulation was almost constant, not dependent on the number of stimulating lyophilized Fem-x cells. Six-day stimulation in vitro by both lyophilized melanoma cells enhanced the suppressive action of PBMC on the survival of the corresponding malignant cell line. Experiments done in nutrient medium with normal human AB serum showed that six-day stimulation with lyophilized melanoma cells enhanced, again, the suppressive action of PBMC on the survival of the corresponding malignant cell line. Contrary, six day preincubation of normal PBMC with the lyophilized healthy PBMC (obtained from other healthy person) inhibited their suppressive action on the survival of both malignant cell lines in vitro.

The absence of correlation between immunoregulatory T cells and induced lymphoproliferative response in treated B-chronic lymphocytic leukemia patients.

BACKGROUND: Many data suggest T cell functional impairment in B-cell chronic lymphocytic leukemia (B-CLL). The mechanism responsible for this phenomenon is still unresolved. METHODS: In 88 B-CLL patients (RAI II-IV) the relationship between immunoregulatory T cells and PHA induced lymphoproliferative response (LPR) was analysed before and after the therapy. The number of peripheral blood CD3+, CD4+ and CD8+ T lymphocytes was determined by indirect immunofluorescence assay using monoclonal antibodies. LPR was estimated in whole blood culture method. RESULTS: The absolute number of CD3+, CD4+ and CD8+ cells in untreated CLL patients was much higher than in healthy controls (n = 26), but the percentages of these subpopulations, CD4/CD8 ratio and LPR to PHA were significantly (p < 0.00001) decreased. The chemotherapy induced a significant rise of CD3+ and CD4+ percentages (p < 0.006 < p < 0.022 respectively) in comparison to baseline levels, but their levels remained significantly (p < 0.00001) lower than the controls. The CD4/CD8 ratio was also elevated after the therapy (p < 0.048) but remained below the normal value as well. The absolute number of CD3+ and CD4+ T cells were normalized after treatment, while the CD8+ cells were still higher (p < 0.044) than controls. The increase of LPR has been registered after treatment, but it failed to reach the control values. We could not find any correlation between the number of immunoregulatory T cells and induced LPR (r = 0.07, for CD4+; r = 0.09 for CD8+ cells). CONCLUSIONS: These data indicate some profound lymphoid cell defect in CLL patients affecting CD8+ proliferation as well as LPR.