Role of Light and Medium Composition on Growth and Valepotriate Contents in Valeriana glechomifolia Whole Plant Liquid Cultures

A system for growing in liquid medium whole plants of Valeriana glechomifolia, endemic to southern Brazil and capable of accumulating bioactive valepotriates, is described. Murashige and Skoog (MS) and Gamborg B5 (B5) media (1.0×, 0.3× and 0.1× strength) without phytohormones were evaluated after four weeks of culture in relation to growth and valepotriate yield. Plants grown in 1.0× MS displayed greatest growth and valepotriate yields and the study of the light condition showed that plants grown under light and dark had similar weight increase and maximum valepotriate yield, 27.2 mg/g DW and 25.0 mg/g DW, respectively. Valtrate was the most abundant valepotriate, followed by acevaltrate and didrovaltrate.     

The interplay among psychopathology,personal resources,context‐related factors and real‐life functioning in schizophrenia: stability in relationships after 4 years and differences in network structure between recovered and non‐recovered patients

Improving real‐life functioning is the main goal of the most advanced integrated treatment programs in people with schizophrenia. The Italian Network for Research on Psychoses previously explored, by using network analysis, the interplay among illness‐related variables, personal resources, context‐related factors and real‐life functioning in a large sample of patients with schizophrenia. The same research network has now completed a 4‐year follow‐up of the original sample. In the present study, we used network analysis to test whether the pattern of relationships among all variables investigated at baseline was similar at follow‐up. In addition, we compared the network structure of patients who were classified as recovered at follow‐up versus those who did not recover. Six hundred eighteen subjects recruited at baseline could be assessed in the follow‐up study. The network structure did not change significantly from baseline to follow‐up, and the overall strength of the connections among variables increased slightly, but not significantly. Functional capacity and everyday life skills had a high betweenness and closeness in the network at follow‐up, as they had at baseline, while psychopathological variables remained more peripheral. The network structure and connectivity of non‐recovered patients were similar to those observed in the whole sample, but very different from those in recovered subjects, in which we found few connections only. These data strongly suggest that tightly coupled symptoms/dysfunctions tend to maintain each other's activation, contributing to poor outcome in schizophrenia. Early and integrated treatment plans, targeting variables with high centrality, might prevent the emergence of self‐reinforcing networks of symptoms and dysfunctions in people with schizophrenia.     

Tissue culture propagation of Mentha spp. by the use of axillary buds

A method is described for rapid multiplication from axillary buds of six Mentha species. Nodal segments from one-year old plants were grown on Murashige and Skoog medium (BMS), supplemented with BAP (1.0; 2.0 mg/l) and KIN (1.0 mg/l) and kept at 28 ± 1°C under fluorescent light for 30 days. After this period, several shoots (15–20 shoots per explant) with roots were produced which were placed in soil for further growth.     

Adjuvant adoptive immunotherapy with tumour-infiltrating lymphocytes and modulated doses of interleukin-2 in 22 patients with melanoma, colorectal and renal cancer, after radical metastasectomy, and in 12 advanced patients

 Adoptive tumour infiltrating lymphocytes (TIL) in combination with a modulated dosage of interleukin-2 (IL-2) can be used with acceptable toxicity in the treatment of immunogenic tumours. Following an experience of reinfusion in advanced melanoma, colorectal and renal cancer patients, treatment was given to disease-free patients after metastasectomy. The high risk of relapse and favourable ratio between reinfused TIL and possible microscopic residual disease determined this choice of adjuvant treatment. A group of 12 patients with advanced disease (7 melanoma, 4 colorectal carcinoma, 1 kidney carcinoma) were treated with TIL (median 5.8×10¹⁰ cells) and IL-2 (West’s schedule) modulated towards a lower dosage (from 12 to 6 MIU/day) in order to maintain an acceptable level of toxicity. As treatment was well tolerated, it was offered to another 22 patients in an adjuvant setting after metastasectomy (11 melanoma, 10 colorectal carcinoma, 1 renal cancer), the median dose of TIL reinfused being 4.95×10¹⁰ cells. No objective response was observed in advanced patients: all patients progressed after a median of 1.5 months (0–8 months) and median survival was 8 months (3–22+ months). Thirteen patients from the second group are still disease-free after a median of 23+ months (9+–47+ months). The remaining 9 patients relapsed after a median of 5 months (3–18 months). Toxicity was moderate as clinical and hepatic/renal function parameters were used to assess the need for dose reductions. Consequently, there was great diversity in IL-2 dosages administered. In particular, there seemed to be a difference in IL-2 doses administered between disease-free cases and those who progressed (17.5 MIU/day versus 7 MIU/day in melanoma patients; 11.2 MIU/day versus 7.1 MIU/day in colorectal cancer patients). By contrast, no differences were observed between number of TIL reinfused and clinical response. Phenotypical characteristics of reinfused TIL were similar to those reported in the literature: 97% were CD3 and 92% were CD8. Aspecific cytolytic activity was evaluated on 12 cases whereas, in 2 melanoma cases, autologous tumour tissue was available for the specific cytotoxicity test. Perforin levels in TIL measured at the end of culture were generally high or very high. Cytokine levels were measured on the supernatant at the end of culture, with an estreme variability in results. Finally, ζ chain and p56^lck were histologically assessed on the resected tissue from which TIL were cultivated. There were virtually none of the former and a complete absence of the latter, which concurs with data reported in the literature. The same immunocytochemical analysis was carried out on TIL at the end of culture. This time an almost complete restoration of both functions was seen, especially in melanoma patients, who are still free from disease. The study is on-going and it has been decided to focus on disease-free patients after metastasectomy in order to increase the number and possibility of clinical and histological correlations.     

Microsatellite instability and mutations of p53 and TGF-‚ RII genes in gastric cancer

To investigate the molecular mechanism of gastric carcinogenesis, we analyzed genetic instability and p53 gene mutations in 40 primary gastric carcinomas. Tumor samples were from untreated patients with no family history suggestive of genetic predisposition to cancer. We screened six microsatellite loci by the polymerase chain reaction (PCR) method, and exons 5–8 of the p53 gene by the PCR-based denaturing gradient gel electrophoresis and sequencing techniques. Microsatellite instability was detected in 32.5% (13/40), and gene mutations in 40% (16/40), of the tumors analyzed. No statistically significant associations were found between genetic alterations and clinico-pathological variables (with the exception of diffusion of lymph node metastases, which was inversely associated with the presence of microsatellite alterations; P < 0.01). Interestingly, a negative association was found between genetic instability and p53 gene mutations: 11 out of 13 tumors showing instability proved to carry a nonmutated p53 gene versus 2/13 carrying a mutated gene (P = 0.03). These observations suggest that genetic instability and p53 gene mutations play a crucial role in the gastric carcinogenic process, but likely act through distinct pathways during cancer development. However, genetic instability is not in and of itself neoplastic. Therefore, we investigated whether insertion/deletion mutations of the polyadenine tract within the transforming growth factor-β type II receptor gene (TGF-βRII) were frequently present in gastric tumors with an RER+ (replication error) phenotype. We found RII mutations in 8/40 (20%) samples: mutations were present in 7/13 (54%) RER+ tumors versus 1/27 (4%) RER– cases (P < 0.001). Received: 14 May 1996 / Revised: 13 June 1996     

The use of molecular assays to identify plant pathogenic organisms vectored by biological control agents

The planthopper Prokelisia marginata VanDuzee (Homoptera: Delphacidae) has beenconsidered for the biological control of theweed Spartina alterniflora Loisel(Poaceae) in Willapa Bay, Washington, U.S.A. Prokelisia marginata is a stenophagousphloem-feeding insect with the potential totransmit bacterial plant diseases that could bemoved by less-specific vectors to other plantspecies. Initial assays with PCR primers thatare putatively specific for phytoplasmas gavepositive results in Spartina. However,subsequent analyses did not indicate thetransmission of the pathogen by theplanthopper. We sequenced the 16S ribosomalRNA (rRNA) gene of the bacterial species thatgave positive results in PCR. Comparisons withsequences available in GenBank suggested thatthe positive results using the putativelyspecific PCR primers were due to the presenceof such bacteria as Pseudomonas, Holomonas, Vibrio, and Acinetobacter. We did not find phytoplasmasin either Spartina or the planthopperP. marginata.