Effect of pyridoxine on tumor necrosis factor activitiesin vitro

Clinical trials with tumor necrosis factor (TNF) as an antitumor agent have so far given rather disappointing results. In this study we show that the naturally occuring vitamin B₆ compound, pyridoxine, enhances TNF-induced cytolysis of three subclones of a mouse fibrosarcoma cell line (WEHI 164). The degree of pyridoxine-induced enhancement of TNF cytotoxicity seems to be dependent on the cells sensitivity to TNF, as the enhancement was much more pronounced in the relatively TNF resistant subclone act-R(cl.12)-WEHI 164, than in the very TNF sensitive subclone WEHI 164 clone 13. Furthermore, our study shows that pyridoxine, in contrast to its enhancing effect on TNF-induced cytotoxicity, rather inhibits TNF-induced growth of human FS-4 fibroblasts. Pyridoxine also enhances lymphotoxin (LT)-induced tumor cell killing and inhibits LT-induced fibroblast growth. Pyridoxine is a relatively non-toxic agentin vivo. Our results suggest that a combination of TNF and pyridoxine may be more efficient than TNF alone, in the treatment of cancer patients.     

The effect of insecticides on the distribution of foracing parasitoids,Diaeretiella rapae [Hym.: Braconidae] on plants

Foraging aphid parasitoids,Diaeretiella rapae M'Intosh, were exposed to sublethal doses of the insecticides pirimicarb, permethrin and malathion on brusslls sprouts plants. Observations on wasp distribution over time revealed that wasps spent less time on sprayed plants, relative to controls and, while on these plants, tended to concentrate activity on unsprayed surfaces. For permethrin and malathion, pesticide residues reversed the stereotypic upward foraging pattern of the wasp. Negative consequences of sublethal pesticide doses for parasitoid foraging efficiency are discussed.      

Resonance Raman characterization of the di-heme protein cytochrome c4 from Pseudomonas stutzeri

 Di-heme Pseudomonas stutzeri cytochrome c ₄ has been characterized by electronic absorption and resonance Raman spectroscopies in the ferric and ferrous forms at pH 7.5 and at room temperature. The data indicate that the two hemes are inequivalent. It is proposed that the N-terminal contains a more relaxed heme as a consequence of the relative orientation of the methionine and histidine ligands with respect to the N-Fe-N directions of the heme plane. This causes a weakening of the Fe-S bond with concomitant partial dissociation of the methionine and the formation of an Fe-aquo bond. Heme group relaxation is further accompanied by less distortion of the heme group than that associated with cytochrome c, expansion of the "core" and a negative shift of the redox potential. Received: 17 December 1996 / Accepted: 6 March 1997     

Shift Work Disorder in a Random Population Sample – Prevalence and Comorbidities

Few studies have investigated the presence of shift work disorder (SWD) in the general community. We addressed many of the limitations in this literature and present new findings. SWD has been treated as an ‘all or none’ construct but we propose the need to consider the ‘severity’ of the disorder. Using random digit dialling, we randomly recruited 1163 participants. Participants completed an extensive battery of scales and questions concerning work, health and individual differences. Three questions based on the criteria from the International Classification for Sleep Disorders were used to categorise participants with SWD (n = 176). In addition, we asked participants whether SWD interfered with aspects of their life and high ratings were used to define severe shift work disorder (SSWD). The prevalence of SWD was 32.1% among night workers and 10.1% in day workers (p<.001). SSWD was present in 9.1% of night workers and 1.3% of day workers (p<.001). Adjusted logistic regression analyses found significant associations between SWD and night work (OR  = 3.35, CI 2.19-5.12), weekly work hours (OR  = 1.02, CI 1.00–1.04), short sleep (≤6 h; OR  = 2.93, CI 1.94–4.41), languidity (OR  = 1.11, CI 1.06–1.16) and resilience (OR  = 0.56, CI 0.43–0.81). Night work, short sleep, languidity, and hypertension were significantly associated with SSWD. Overall, participants with SSWD slept 0.80 h less than other participants (p<.001). Night work, short sleep and languidity were associated with both SWD and SSWD. Day workers with SWD symptoms reported significantly shorter sleep duration, higher levels of languidity and worked longer working hours compared to day workers without SWD.     

Pregnancy associated glycoproteins (PAG) in postpartum cows, ewes, goats and their offspring

Determination of plasma concentrations of pregnancy associated glycoproteins (PAG) has been used for early pregnancy diagnosis in cows. However, this is complicated by the presence of PAG in plasma for an extended period postpartum. The main objective of the present study was to investigate the postpartum elimination rates of pregnancy associated glycoproteins (PAG) in sheep, goats and cows in order to gain background information applicable to the use of PAG for pregnancy diagnosis in domestic ruminants. A second objective was to investigate whether PAG are transferred to the foetus and newborn, by measuring plasma PAG concentrations in calves, lambs and goat kids before and after colostrum feeding. PAG in the blood at parturition were eliminated by a first order process in the cows and ewes, while a two-step log-linear decline occurred in the goats. Estimated postpartum half-life of plasma PAG in the cows and ewes was 9 and 4.5 days, respectively. In the goats, half-lives were 3.6 and 7.5 days in the initial fast and terminal slow phase. Basal levels were reached 80-90 days postpartum in cows. Plasma PAG concentration can be used for pregnancy diagnosis from day 28 after AI, provided that the time interval from calving to AI is >60 days. Using a heterologous antibody RIA, we found 4 ng/mL to be the appropriate cut-off. Due to the presence of PAG residues from the previous gestation, the interval from AI to pregnancy diagnosis should increase by approximately 0.5 days beyond 28 days for each day of AI closer to calving than 60. Measurements in newborn ruminants suggested that PAG enter the foetal blood in utero and that colostral PAG are transferred to the newborn. Following the peak plasma concentration observed 1 day after birth in most of the animals, PAG were rapidly eliminated in a log-linear fashion.     

Biologically-active flavonoids from Gossypium arboreum

A new flavonol glycoside, gossypetin 8-O-rhamnoside, was isolated from flower petals of Gossypium arboreum along with quercetin 7-O-glucoside, quercetin 3-O-glucoside and quercetin 3′-O-glucoside. These compounds showed antibacterial activity against Pseudomonas maltophilia and Enterobacter cloacae.     

Detection of Small Numbers of Campylobacter jejuni and Campylobacter coli Cells in Environmental Water, Sewage, and Food Samples by a Seminested PCR Assay

A rapid and sensitive assay was developed for detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples. Water and sewage samples were filtered, and the filters were enriched overnight in a nonselective medium. The enrichment cultures were prepared for PCR by a rapid and simple procedure consisting of centrifugation, proteinase K treatment, and boiling. A seminested PCR based on specific amplification of the intergenic sequence between the two Campylobacter flagellin genes, flaA and flaB, was performed, and the PCR products were visualized by agarose gel electrophoresis. The assay allowed us to detect 3 to 15 CFU of C. jejuni per 100 ml in water samples containing a background flora consisting of up to 8,700 heterotrophic organisms per ml and 10,000 CFU of coliform bacteria per 100 ml. Dilution of the enriched cultures 1:10 with sterile broth prior to the PCR was sometimes necessary to obtain positive results. The assay was also conducted with food samples analyzed with or without overnight enrichment. As few as ≤3 CFU per g of food could be detected with samples subjected to overnight enrichment, while variable results were obtained for samples analyzed without prior enrichment. This rapid and sensitive nested PCR assay provides a useful tool for specific detection of C. jejuni or C. coli in drinking water, as well as environmental water, sewage, and food samples containing high levels of background organisms.     

PDL1 Expression on Plasma and Dendritic Cells in Myeloma Bone Marrow Suggests Benefit of Targeted anti PD1-PDL1 Therapy

In this study we set out to investigate whether anti PDL1 or PD–1 treatment targeting the immune system could be used against multiple myeloma. DCs are important in regulating T cell responses against tumors. We therefore determined PDL1 and PDL2 expression on DC populations in bone marrow of patients with plasma cell disorders using multicolour Flow Cytometry. We specifically looked at CD141⁺ and CD141^- myeloid and CD303⁺ plasmacytoid DC. The majority of plasma cells (PC) and DC subpopulations expressed PDL1, but the proportion of positive PDL1+ cells varied among patients. A correlation between the proportion of PDL1⁺ PC and CD141⁺ mDC was found, suggesting both cell types could down-regulate the anti-tumor T cell response.     

Cytokine regulation of interleukin 6 production by human endothelial cells

The influence of recombinant (r) human tumor necrosis factor alpha (rTNF-alpha), r human interleukin 1 beta (rIL-1 beta), and r human interferon gamma (rIFN-gamma) on the production of interleukin 6 (IL-6) by human endothelial cells (HEC) was investigated. The addition of 1-100 U/ml of either rTNF-alpha or rIL-1 beta to cultures of HEC monolayers caused a dose-related increase in IL-6 production as detected after 24 hr of incubation. In contrast to rIL-1 beta and rTNF-alpha, the use of up to 1000 U/ml of rIFN-gamma caused only a moderate increase in IL-6 production. However, significantly greater quantities of IL-6 were produced by HEC monolayers subjected to 1000 U/ml of rIFN-gamma in combination with 1-100 U/ml of rTNF-alpha. Furthermore, the addition of graded concentrations of human transforming growth factor beta (TGF-beta) to cultures resulted in a dose-related inhibition of rIL-1 beta- and rTNF-alpha-induced IL-6 production by HEC. The results demonstrate that rIL-1 beta and rTNF-alpha share the ability to stimulate HEC for production of IL-6 and indicate that TGF-beta may act as an immunosuppressive agent, at least partially, through its ability to inhibit the action of TNF-alpha and IL-1 on endothelial cells.