Microbiological safety assessment of silkworm farms: a case study.

Silkworms have been farmed for their silk since ancient times. After silk reeling, their chrysalides are consumed as food in several Asian countries. Despite the long rearing tradition of this insect, few studies have investigated the silkworm's microbiological safety all along the life cycle, focusing on detecting silkworm pathogens or on the safety of the dried chrysalis for food consumption. However, the in-farm rearing process, which takes around forty days, may affect the microbial load of the silkworm and of the rearing environment, as well as the quality of fresh cocoon and other performance parameters. No data is available on how microbial contamination changes during the rearing period and between different farmers. Furthermore, in light of the possible use of the chrysalis as food, it is crucial to understand how its microbial load varies according to the water content. To address these specific questions, we conducted an investigation involving the analysis of specific microbial indicators commonly used in the food chain. We collected environmental and silkworm samples from several farms. The examination covered the entire life cycle of silkworms, beginning with the first instar larvae and concluding with the scrutiny of both freshly harvested and dried pupae. Silkworm farms in Northeast Italy proved to be an appropriate model system for carrying out the experimentation. Additionally, an evaluation of rearing performance was conducted, with a focus on the quality of fresh cocoons and the survival rate of the insects.

TM9SF4 is a novel V-ATPase-interacting protein that modulates tumor pH alterations associated with drug resistance and invasiveness of colon cancer cells.

An inverted pH gradient across the cell membranes is a typical feature of malignant cancer cells that are characterized by extracellular acidosis and cytosol alkalization. These dysregulations are able to create a unique milieu that favors tumor progression, metastasis and chemo/immune-resistance traits of solid tumors. A key event mediating tumor cell pH alterations is an aberrant activation of ion channels and proton pumps such as (H+)-vacuolar-ATPase (V-ATPase). TM9SF4 is a poorly characterized transmembrane protein that we have recently shown to be related to cannibal behavior of metastatic melanoma cells. Here, we demonstrate that TM9SF4 represents a novel V-ATPase-associated protein involved in V-ATPase activation. We have observed in HCT116 and SW480 colon cancer cell lines that TM9SF4 interacts with the ATP6V1H subunit of the V-ATPase V1 sector. Suppression of TM9SF4 with small interfering RNAs strongly reduces assembly of V-ATPase V0/V1 sectors, thus reversing tumor pH gradient with a decrease of cytosolic pH, alkalization of intracellular vesicles and a reduction of extracellular acidity. Such effects are associated with a significant inhibition of the invasive behavior of colon cancer cells and with an increased sensitivity to the cytotoxic effects of 5-fluorouracil. Our study shows for the first time the important role of TM9SF4 in the aberrant constitutive activation of the V-ATPase, and the development of a malignant phenotype, supporting the potential use of TM9SF4 as a target for future anticancer therapies.

Low-dose oral methotrexate treatment in chronic progressive multiple sclerosis.

We aimed to further assess the safety and efficacy of low-dose oral methotrexate (LDOM) treatment for chronic progressive MS (CPMS). We studied 20 CPMS patients, including 16 with secondary progressive MS who had shown disease progression in the previous year. The mean follow-up was 23 months. The mean EDSS score was 6.3+/-1.1 before treatment and 6.4+/-1.1 after one year of treatment. At one year, 15 of 20 patients were still being treated, and 10 were stable. Twelve patients have completed 18 months of treatment, and eight are stable. Two patients stopped treatment because of side effects, two more because they did not perceive benefit, and one was lost to follow-up. Six patients had mild, transient increases in liver enzymes not requiring treatment interruption, and two had localized herpes zoster. Magnetic resonance imaging (MRI) performed before treatment and at one year remained unchanged in responders. We confirm that LDOM is safe in carefully selected and monitored CPMS patients. MTX is inexpensive and, given its anti-inflammatory and immunomodulatory properties, may be used as add-on therapy in non-responders to interferon beta, although hepatic toxicity may be a problem in long-term treatment.

Prednisone and plasma exchange improve suppressor cell function in chronic inflammatory demyelinating polyneuropathy.

We have recently demonstrated that ConA-induced suppressor cell function is defective in chronic inflammatory demyelinating polyneuropathy (CIDP). To assess whether this defect plays a role in disease activity and its reversal is important in recovery, we studied modifications of ConA-induced suppressor cell function induced by prednisone and plasma exchange in 20 patients with CIDP. We found a significant increase towards normal of ConA-induced suppressor cell function after treatment in concurrence with clinical improvement. Induction of suppression, presumably through favorable modifications of the cytokine network or other humoral mediators, might be one, among others, of the mechanisms through which prednisone and plasma exchange are effective in CIDP.