Metabolism of brain glycolipid fatty acids

The metabolism of the fatty acid moieties of brain cerebrosides, sulfatides, and gangliosides is reviewed and discussed. The methodology involved in the isolation of the fatty acids is described briefly. It seems clear now that most of these acids are made by chain elongation of intermediate length fatty acids by addition of acetate residues. The unsaturated acids are made by desaturation of the intermediate length acids (palmitic, heptadecanoic, stearic) followed by chain elongation. The hydroxy acids are made directly from the corresponding nonhydroxy acids, saturated, unsaturated, and odd‐numbered. All the hydroxy acids undergo oxidative decarboxylation to yield fatty acids containing one less carbon atom. The odd‐numbered acids are also made from propionate, which is elongated to intermediate length acids and then to longer acids. The major intermediate length “primer” acid seems to be palmitate, but there is evidence that the stearate used for cerebroside synthesis is also madede novo from acetate. The ganglioside fatty acids were found to turn over somewhat faster than the other fatty acids. Two metabolic pools for the cerebroside acids were found, one with a very high turnover rate, the other with a very low turnover rate.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141473/1/lipd0047.pd

Adenoviral targeting of malignant melanoma for fluorescence-guided surgery prevents recurrence in orthotopic nude-mouse models.

Malignant melanoma requires precise resection in order to avoid metastatic recurrence. We report here that the telomerase-dependent, green fluorescent protein (GFP)-containing adenovirus OBP-401 could label malignant melanoma with GFP in situ in orthotopic mouse models. OBP-401-based fluorescence-guided surgery (FGS) resulted in the complete resection of malignant melanoma in the orthotopic models, where conventional bright-light surgery (BLS) could not. High-dose administration of OBP-401 enabled FGS without residual cancer cells or recurrence, due to its dual effect of cancer-cell labeling with GFP and killing

Precise navigation surgery of tumours in the lung in mouse models enabled by in situ fluorescence labelling with a killer-reporter adenovirus.

BackgroundCurrent methods of image-guided surgery of tumours of the lung mostly rely on CT. A sensitive procedure of selective tumour fluorescence labelling would allow simple and high-resolution visualisation of the tumour for precise surgical navigation.MethodsHuman lung cancer cell lines H460 and A549 were genetically transformed to express red fluorescent protein (RFP). Tumours were grown subcutaneously for each cell line and harvested and minced for surgical orthotopic implantation on the left lung of nude mice. Tumour growth was measured by fluorescence imaging. After the tumours reached 5 mm in diameter, they were injected under fluorescence guidance with the telomerase-dependent green fluorescent protein (GFP)-containing adenovirus, OBP-401. Viral labelling of the lung tumours with GFP precisely colocalised with tumour RFP expression. Three days after administration of OBP-401, fluorescence-guided surgery (FGS) was performed.ResultsFGS of tumours in the lung was enabled by labelling with a telomerase-dependent adenovirus containing the GFP gene. Tumours in the lung were selectively and brightly labelled. FGS enabled complete lung tumour resection with no residual fluorescent tumour.ConclusionsFGS of tumours in the lung is feasible and more effective than bright-light surgery

Tumor-targeting adenovirus OBP-401 inhibits primary and metastatic tumor growth of triple-negative breast cancer in orthotopic nude-mouse models.

Our laboratory previously developed a highly-invasive, triple-negative breast cancer (TNBC) variant using serial orthotopic implantation of the human MDA-MB-231 cell line in nude mice. The isolated variant was highly-invasive in the mammary gland and lymphatic channels and metastasized to lymph nodes in 10 of 12 mice compared to 2 of 12 of the parental cell line. In the present study, the tumor-selective telomerase dependent OBP-401 adenovirus was injected intratumorally (i.t.) (1 × 108 PFU) when the high-metastatic MDA-MB-231 primary tumor expressing red fluorescent protein (MDA-MB-231-RFP) reached approximately 500 mm3 (diameter; 10 mm). The mock-infected orthotopic primary tumor grew rapidly. After i.t. OBP-401 injection, the growth of the orthotopic tumors was arrested. Six weeks after implantation, the fluorescent area and fluorescence intensity showed no increase from the beginning of treatment. OBP-401 was then injected into high-metastatic MDA-MB-231-RFP primary orthotopic tumor growing in mice which already had developed metastasis within lymphatic ducts. All 7 of 7 control mice subsequently developed lymph node metastasis. In contrast, none of 7 mice which received OBP-401 had lymph node metastasis. Seven of 7 control mice also had gross lung metastasis. In contrast, none of the 7 mice which received OBP-401 had gross lung metastasis. Confocal laser microscopy imaging demonstrated that all control mice had diffuse lung metastases. In contrast, all 7 mice which received OBP-401 only had a few metastatic cells in the lung. OBP-401 treatment significantly extended survival of the treated mice

Fluorescence-guided surgery of a highly-metastatic variant of human triple-negative breast cancer targeted with a cancer-specific GFP adenovirus prevents recurrence.

We have previously developed a genetically-engineered GFP-expressing telomerase-dependent adenovirus, OBP-401, which can selectively illuminate cancer cells. In the present report, we demonstrate that targeting a triple-negative high-invasive human breast cancer, orthotopically-growing in nude mice, with OBP-401 enables curative fluorescence-guided surgery (FGS). OBP-401 enabled complete resection and prevented local recurrence and greatly inhibited lymph-node metastasis due to the ability of the virus to selectively label and subsequently kill cancer cells. In contrast, residual breast cancer cells become more aggressive after bright (white)-light surgery (BLS). OBP-401-based FGS also improved the overall survival compared with conventional BLS. Thus, metastasis from a highly-aggressive triple-negative breast cancer can be prevented by FGS in a clinically-relevant mouse model

Catecholamines in amniotic fluid as indicators of intrapartum fetal stress.

Catecholamines were measured in the amniotic fluid and in the first voided newborn urine obtained from appropriate-for-date infants of term deliveries. Catecholamine values in the amniotic fluid and urine were nearly equal when expressed in terms of creatinine. Significant positive correlations were observed between the amniotic fluid and urine of norepinephrine and epinephrine. In normal cases (n = 32) that underwent uneventful vaginal delivery, the 95% confidence limits for norepinephrine and epinephrine in the amniotic fluid were 1.53 to 2.33 ng/ml and 0.16 to 0.30 ng/ml, respectively. In cases of moderate stress (n = 12), only norepinephrine showed significantly higher values than the normal cases, while in cases of severe stress (n = 12), norepinephrine became more significantly high, and epinephrine was found to be elevated significantly. A significant difference was noted in the incidence of fetal stress between the infants with more than and those with less than 2.30 ng/ml of norepinephrine, the upper limits of the normal 95% confidence limits. However, for epinephrine such a significant difference was not noted. It was concluded that amniotic fluid catecholamines are of fetal origin and reflect fetal sympathoadrenal activity directly, even during labor, and that their level may be a good indicator of fetal condition and stress.</p

Effects of L-dopa or dopamine on human decidual prostaglandin synthesis.

L-Dopa and three catecholamines in the amniotic fluid before and after labor were measured to confirm the amniotic fluid catecholamine levels at the end of gestation. L-Dopa values were higher than those of three catecholamines, and dopamine which was the predominant catecholamine, rose significantly after the onset of labor. Then, to evaluate the effects of L-dopa or dopamine on prostaglandin synthesis, strips of human decidua vera obtained from fetal membranes at the time of elective cesarean sections before the onset of labor were incubated in Krebs-Ringer buffer in the presence of L-dopa or dopamine. When L-dopa was added, the net production of prostaglandin(PG)F was significantly greater than that of the control at each incubation time. On the other hand, the significant rise was observed only after 10 min of incubation for PGE2 production. Dopamine had a stimulatory effect on PGF synthesis only after 15 and 30 min of incubation, and it also stimulated the release of PGE2 at each incubation time. These results suggest that dopamine and L-dopa in amniotic fluid stimulate the production of prostaglandin by the decidua in humans.</p

Improved Resection and Outcome of Colon-Cancer Liver Metastasis with Fluorescence-Guided Surgery Using In Situ GFP Labeling with a Telomerase-Dependent Adenovirus in an Orthotopic Mouse Model.

Fluorescence-guided surgery (FGS) of cancer is an area of intense development. In the present report, we demonstrate that the telomerase-dependent green fluorescent protein (GFP)-containing adenovirus OBP-401 could label colon-cancer liver metastasis in situ in an orthotopic mouse model enabling successful FGS. OBP-401-GFP-labeled liver metastasis resulted in complete resection with FGS, in contrast, conventional bright-light surgery (BLS) did not result in complete resection of the metastasis. OBP-401-FGS reduced the recurrence rate and prolonged over-all survival compared with BLS. In conclusion, adenovirus OBP-401 is a powerful tool to label liver metastasis in situ with GFP which enables its complete resection, not possible with conventional BLS

Fetal and neonatal excretion of free and conjugated ritodrine.

The ability of the human fetus and neonate to conjugate and excrete ritodrine, a beta 2-sympathomimetic drug, was investigated. Free and conjugated ritodrine concentrations in the plasma, amniotic fluid and urine were measured in 11 mother-infant pairs, to whom intravenous ritodrine had been administered before elective cesarean section at term. Ritodrine was determined by HPLC with electrochemical detection. At delivery, conjugated ritodrine values were significantly higher than those for the free form in maternal and fetal plasma. There were significant positive correlations between the concentrations in the maternal and umbilical vein plasma for both free and conjugated ritodrine. In the amniotic fluid, the total ritodrine concentrations were much higher than those in the fetal plasma, the conjugated form accounting for 90.2% of the total. Furthermore, the percentages of conjugated ritodrine in the amniotic fluid and neonatal urine were significantly higher than the percentage in the maternal urine on the day of birth. In the neonatal urine, the concentrations of free and conjugated ritodrine decreased rapidly after birth as did those in the maternal urine, on day 3 postpartum being less than 2% of the values on the day of parturition. These results indicate that the fetus at term is capable of forming conjugated metabolites of ritodrine and of excreting free and conjugated ritodrine in its urine.</p