From Genome to Field-Observation of the Multimodal Nematicidal and Plant Growth-Promoting Effects of Bacillus firmus I-1582 on Tomatoes Using Hyperspectral Remote Sensing.

Root-knot nematodes are considered the most important group of plant-parasitic nematodes due to their wide range of plant hosts and subsequent role in yield losses in agricultural production systems. Chemical nematicides are the primary control method, but ecotoxicity issues with some compounds has led to their phasing-out and consequential development of new control strategies, including biological control. We evaluated the nematicidal activity of Bacillus firmus I-1582 in pot and microplot experiments against Meloidogyne luci. I-1582 reduced nematode counts by 51% and 53% compared to the untreated control in pot and microplot experiments, respectively. I-1582 presence in the rhizosphere had concurrent nematicidal and plant growth-promoting effects, measured using plant morphology, relative chlorophyll content, elemental composition and hyperspectral imaging. Hyperspectral imaging in the 400-2500 nm spectral range and supervised classification using partial least squares support vector machines successfully differentiated B. firmus-treated and untreated plants, with 97.4% and 96.3% accuracy in pot and microplot experiments, respectively. Visible and shortwave infrared spectral regions associated with chlorophyll, N-H and C-N stretches in proteins were most relevant for treatment discrimination. This study shows the ability of hyperspectral imaging to rapidly assess the success of biological measures for pest control.

Drought-induced expression of aquaporin genes in leaves of two common bean cultivars differing in tolerance to drought stress.

Aquaporin proteins are part of the complex response of common bean (Phaseolus vulgaris L.) to drought which affects the quality and quantity of yield of this important crop. To better understand the role of aquaporins in common bean, drought-induced gene expression of several aquaporins was determined in two cultivars, the more drought tolerant Tiber and the less tolerant Starozagorski cern. The two bean cultivars were selected among 16 European genotypes based on the tolerance to drought determined by time needed for plants to wilt after withholding irrigation and yield at harvest. The expression patterns of two plasma membrane intrinsic proteins, PvPIP1;2 and PvPIP2;7, and two tonoplast intrinsic proteins, PvTIP1;1 and PvTIP4;1 in leaves of 21 day old plants were determined by RT-qPCR in both cultivars under three degrees of drought stress, and under rehydration and control conditions. Gene expression of all four examined aquaporins was down-regulated in drought stressed plants. After rehydration it returned to the level of control plants or was even higher. The responses of PvPIP2;7 and PvTIP1;1 during drought and rehydration were particularly pronounced. The gene expression of PvPIP2;7 and PvTIP4;1 during drought was cultivar specific, with greater down-regulation of these two aquaporins in drought tolerant Tiber. Under drought stress the relative water content and water potential of leaves were higher in Tiber than in Starozagorski plants. The differences in these physiological parameters indicate greater prevention of water loss in Tiber during drought, which may be associated with rapid and adequate down-regulation of aquaporins. These results suggest that the ability of plants to conserve water during drought stress involves timely and sufficient down-regulation of gene expression of specific aquaporins.

Radiosensitizing effect of intratumoral interleukin-12 gene electrotransfer in murine sarcoma.

BACKGROUND: Interleukin-12 (IL-12) based radiosensitization is an effective way of tumor treatment. Local cytokine production, without systemic shedding, might provide clinical benefit in radiation treatment of sarcomas. Therefore, the aim was to stimulate intratumoral IL-12 production by gene electrotransfer of plasmid coding for mouse IL-12 (mIL-12) into the tumors, in order to explore its radiosensitizing effect after single or multiple intratumoral gene electrotransfer. METHODS: Solid SA-1 fibrosarcoma tumors, on the back of A/J mice, were treated intratumorally by mIL-12 gene electrotransfer and 24 h later irradiated with a single dose. Treatment effectiveness was measured by tumor growth delay and local tumor control assay (TCD(50) assay). With respect to therapeutic index, skin reaction in the radiation field was scored. The tumor and serum concentrations of cytokines mIL-12 and mouse interferon γ (mIFNγ) were measured. Besides single, also multiple intratumoral mIL-12 gene electrotransfer before and after tumor irradiation was evaluated. RESULTS: Single intratumoral mIL-12 gene electrotransfer resulted in increased intratumoral but not serum mIL-12 and mIFNγ concentrations, and had good antitumor (7.1% tumor cures) and radiosensitizing effect (21.4% tumor cures). Combined treatment resulted in the radiation dose-modifying factor of 2.16. Multiple mIL-12 gene electrotransfer had an even more pronounced antitumor (50% tumor cures) and radiosensitizing (86.7% tumor cures) effect. CONCLUSIONS: Single or multiple intratumoral mIL-12 gene electrotransfer resulted in increased intratumoral mIL-12 and mIFNγ cytokine level, and may provide an efficient treatment modality for soft tissue sarcoma as single or adjuvant therapy to tumor irradiation.

Large protruding bladder stone.

We present a unique case of a large urinary bladder stone protruding through the external urethral meatus in a 77-year-old woman, which was causing acute urinary retention, complicated by bilateral hydronephrosis, and was removed under topical anesthesia in the emergency department. Epidemiology, etiology, clinical presentation and management of urinary bladder stones are briefly discussed.

Potentiation of electrochemotherapy by intramuscular IL-12 gene electrotransfer in murine sarcoma and carcinoma with different immunogenicity.

BACKGROUND.: Electrochemotherapy provides good local tumor control but requires adjuvant treatment for increased local response and action on distant metastasis. In relation to this, intramuscular interleukin-12 (IL-12) gene electro-transfer, which provides systemic shedding of IL-12, was combined with local electrochemotherapy with cisplatin. Furthermore, the dependence on tumor immunogenicity and immunocompetence of the host on combined treatment response was evaluated. MATERIALS AND METHODS.: Sensitivity of SA-1 sarcoma and TS/A carcinoma cells to electrochemotherapy with cisplatin was tested in vitro. In vivo, intratumoral electrochemotherapy with cisplatin (day 1) was combined with a single (day 0) or multiple (days 0, 2, 4) intramuscular murine IL-12 (mIL-12) gene electrotransfer. The antitumor effectiveness of combined treatment was evaluated on immunogenic murine SA-1 sarcoma in A/J mice and moderately immunogenic murine TS/A carcinoma, in immunocompetent BALB/c and immunodeficient SCID mice. RESULTS.: Electrochemotherapy in vitro resulted in a similar IC(50) values for both sarcoma and carcinoma cell lines. However, in vivo electrochemotherapy was more effective in the treatment of sarcoma, the more immunogenic of the tumors, resulting in a higher log cell kill, longer specific tumor growth delay, and also 17% tumor cures compared to carcinoma where no tumor cures were observed. Adjuvant intramuscular mIL-12 gene electrotransfer increased the log cell kill in both tumor models, potentiating the specific tumor growth delay by a factor of 1.8-2 and increasing tumor cure rate by approximately 20%. In sarcoma tumors, the potentiation of the response by intramuscular mIL-12 gene electrotransfer was dose-dependent and also resulted in a faster onset of tumor cures. Comparison of the carcinoma response to the combined treatment modality in immunocompetent and immunodeficient mice demonstrated that the immune system is needed both for increased cell kill and for attaining tumor cures. CONCLUSIONS.: Based on the comparison of the antitumor effectiveness of electrochemotherapy to intratumoral cisplatin administration, we can conclude that the fraction of cells killed and the tumor cure rate are higher in immunogenic sarcoma tumor compared to moderately immunogenic carcinoma tumor. The tumor cell kill and cure rate depend on the immune response elicited by the destroyed tumor cells, which might depend on the tumor immunogenicity. The effect of adjuvant intramuscular mIL-12 gene electrotransfer is dependent on the amount of IL-12 in the system and the immune competence of the host, as demonstrated by the dose-dependent increase in the cure rate of SA-1 tumors after multiple intramuscular mIL-12 gene electrotransfer and in the differential cure rate of TS/A tumors growing in immunocompetent and immunodeficient mice.

Performance of nuclear matrix protein 22 urine marker and voided urine cytology in the detection of urinary bladder tumors.

BACKGROUND: Cystoscopy with urinary cytology is the gold standard for the diagnosis and follow-up of patients with tumors of the urinary bladder. The aim of the study was to evaluate the performance of the nuclear matrix protein 22 (NMP22) tumor marker test, BladderChek® point-of-care test and voided urinary cytology for the detection and follow-up of bladder tumors. METHODS: NMP22 was measured using an ELISA assay in stabilized voided urine and using the BladderChek® test. Voided urinary cytology was performed on urine samples. Results were compared to cystoscopic findings and histopathological examination results after transurethral resection of the bladder lesion. RESULTS: For the prediction of malignant histopathological result, sensitivity and specificity were 45.2% and 75.0%, respectively, for NMP22 at a cut-off of 7.5 kU/L, 17.7% and 100% for the BladderChek® test and 37.0% and 100% for voided urine cytology. For the prediction of suspicious or positive cystoscopic finding, sensitivity and specificity were 40.4% and 72.1%, respectively, for NMP22 at a cut-off of 7.5 kU/L, 14.8% and 93.8% for the BladderChek® test and 26.8% and 98.1% for voided urine cytology. CONCLUSIONS: The NMP22 quantitative test showed higher sensitivity and lower specificity compared with voided urine cytology, whereas the sensitivity of the BladderChek® test was low. We could not recommend any of the three non-invasive tests as a replacement for cystoscopy for the diagnosis or follow-up of urinary bladder tumors.

Correlations between the ICIQ-UI short form and urodynamic diagnosis.

AIMS: Urinary incontinence is a common symptom in many diseases as well as in general population. To measure the impact of urinary incontinence (UI) on individual's quality of life the English short form International Consultation on Incontinence questionnaire (ICIQ-UI Short Form) has been developed, which consists of three scored and one self-diagnostic question. Our aim was to translate the ICIQ-UI Short Form into Slovene, re-validate it, and test the diagnostic value of the questionnaire in determining the type of UI. METHODS: ICIQ-UI Short Form was translated into Slovene and re-validated. In the validation process we included urology and gynecology outpatient clinics attendants, elderly institutionalized patients, and a community sample. One group of patients underwent urodynamic testing and the results were compared to their responses to question six of the ICIQ-UI Short Form. Both sets of answers were compared using the Kappa (kappa) statistics. RESULTS: The Slovene version of the ICIQ-UI Short Form was found to be valid (excellent test-retest reliability, good internal consistency, good responsiveness to change, and reflected well underlying theories). The questionnaire correlated well with urodynamic findings. The perceived cause of leakage (Question 6) was a good indicator of objectively proven cause of incontinence (Kappa value 0.77). CONCLUSIONS: We found that the Slovene version of the ICIQ-UI Short Form is a valid and reliable questionnaire for assessment of UI. In addition to its ability to assess the impact of UI on individual's quality of life it also has a diagnostic value of the cause of incontinence.