Polyphenol oxidase can cross thylakoids by both the Tat and the Sec-dependent pathways: a putative role for two stromal processing sites

Polyphenol oxidase (PPO; EC 1.10.3.2 or EC 1.14.18.1), a thylakoid-lumen protein encoded by a nuclear gene, plays a role in the defense of plants against both herbivores and pathogens. Although previously reported to be a Tat ( t win- a rginine-dependent t ranslocation) protein, the import of PPO by isolated chloroplasts was inhibited by azide, a diagnostic inhibitor of the Sec-dependent pathway. Import of PPO inhibited thylakoid translocation of a Tat protein and did not affect translocation of Sec-dependent proteins. In contrast, a pre-accumulated iPPO competed with Sec-dependent but not with Tat proteins. A previously reported second processing step in the stroma removes a twin-Arg that is part of a 'Sec-avoidance' motif in the thylakoid targeting domain of PPO. When the second processing site was mutated, the import of the resulting precursor showed Sec-dependent characteristics. The PPO transit peptide could drive thylakoid translocation of a Tat protein in the dark. Azide inhibited the secretion of a PPO intermediate that lacks a twin-Arg to the periplasm of Escherichia coli , but had no effect on the export of the intermediate containing the twin-Arg. PPO is synthesized in plants in response to wound and pathogen-related signals and it is possible that when the Tat pathway is unable to translocate adequate amounts of newly synthesized PPO, translocation is diverted to the Sec-dependent pathway by processing the intermediate at the second site and removing the twin-Arg.     

Some Problems in the Chemotherapy of Solid Tumours

Chemotherapy of solid tumours is considered with respect to cell vulnerability, toxicity, tissue penetrability, tumour disruption and anatomical location. Chemotherapy to date has dealt primarily with the first two factors. Even when the cell is vulnerable to the agent used and host toxicity can be controlled, the other factors can lead to treatment failures. It is suggested that combinations of cytotoxic agents may be of more value than single agents. Timing and dosage are also considered. It is recommended that terminal cases and extremely toxic patients not be submitted to chemotherapy. In 78 patients with bronchogenic carcinoma, to whom nicotinic acid was given as a possible aid to tissue penetrability by nitrogen mustard, there was some indication that further similar studies might be of some value. Specific cases are summarized to illustrate the importance of various factors in the treatment of individual patients.     

Clinical aspects of atypical mycobacterial infection.

Summary: Atypical mycobacterial infections may give rise to various clinical difficulties. Case reports of six patients--three adult patients with pulmonary lesions, two children with cervical lymphadentis and one patient in whom the atypical mycobacterium appeared to be present as a commensal--illustrate these difficulties. Determination of the significance of the organism and differentiation of condition from tuberculosis and others requires consideration of the clinical picture, the results of skin testing, histologic features and cultural characteristics. Three patients, including the two children, were treated with a combination of surgery and chemotherapy, with satisfactory results. An elderly patient with chronic bronchitis and a pulmonary infection due to M. kansasii was treated successfully with antiuberculosis agents alone. Chemotherapy is being tried on a fifth patient with cavitary disease due to M. intracellulare, but is seems likely that an operation will also be required.     

Increased Functional Connectivity between Prefrontal Cortex and Reward System in Pathological Gambling

Pathological gambling (PG) shares clinical characteristics with substance-use disorders and is thus discussed as a behavioral addiction. Recent neuroimaging studies on PG report functional changes in prefrontal structures and the mesolimbic reward system. While an imbalance between these structures has been related to addictive behavior, whether their dysfunction in PG is reflected in the interaction between them remains unclear. We addressed this question using functional connectivity resting-state fMRI in male subjects with PG and controls. Seed-based functional connectivity was computed using two regions-of-interest, based on the results of a previous voxel-based morphometry study, located in the prefrontal cortex and the mesolimbic reward system (right middle frontal gyrus and right ventral striatum). PG patients demonstrated increased connectivity from the right middle frontal gyrus to the right striatum as compared to controls, which was also positively correlated with nonplanning aspect of impulsiveness, smoking and craving scores in the PG group. Moreover, PG patients demonstrated decreased connectivity from the right middle frontal gyrus to other prefrontal areas as compared to controls. The right ventral striatum demonstrated increased connectivity to the right superior and middle frontal gyrus and left cerebellum in PG patients as compared to controls. The increased connectivity to the cerebellum was positively correlated with smoking in the PG group. Our results provide further evidence for alterations in functional connectivity in PG with increased connectivity between prefrontal regions and the reward system, similar to connectivity changes reported in substance use disorder.     

Reduction in inter-hemispheric connectivity in disorders of consciousness

Clinical diagnosis of disorders of consciousness (DOC) caused by brain injury poses great challenges since patients are often behaviorally unresponsive. A promising new approach towards objective DOC diagnosis may be offered by the analysis of ultra-slow (<0.1 Hz) spontaneous brain activity fluctuations measured with functional magnetic resonance imaging (fMRI) during the resting-state. Previous work has shown reduced functional connectivity within the "default network", a subset of regions known to be deactivated during engaging tasks, which correlated with the degree of consciousness impairment. However, it remains unclear whether the breakdown of connectivity is restricted to the "default network", and to what degree changes in functional connectivity can be observed at the single subject level. Here, we analyzed resting-state inter-hemispheric connectivity in three homotopic regions of interest, which could reliably be identified based on distinct anatomical landmarks, and were part of the "Extrinsic" (externally oriented, task positive) network (pre- and postcentral gyrus, and intraparietal sulcus). Resting-state fMRI data were acquired for a group of 11 healthy subjects and 8 DOC patients. At the group level, our results indicate decreased inter-hemispheric functional connectivity in subjects with impaired awareness as compared to subjects with intact awareness. Individual connectivity scores significantly correlated with the degree of consciousness. Furthermore, a single-case statistic indicated a significant deviation from the healthy sample in 5/8 patients. Importantly, of the three patients whose connectivity indices were comparable to the healthy sample, one was diagnosed as locked-in. Taken together, our results further highlight the clinical potential of resting-state connectivity analysis and might guide the way towards a connectivity measure complementing existing DOC diagnosis.