Non-song vocalizations of pygmy blue whales in Geographe Bay, Western Australia.

Non-song vocalizations of migrating pygmy blue whales (Balaenoptera musculus brevicauda) in Western Australia are described. Simultaneous land-based visual observations and underwater acoustic recordings detected 27 groups in Geographe Bay, WA over 2011 to 2012. Six different vocalizations were recorded that were not repeated in a pattern or in association with song, and thus were identified as non-song vocalizations. Five of these were not previously described for this population. Their acoustic characteristics and context are presented. Given that 56% of groups vocalized, 86% of which produced non-song vocalizations and 14% song units, the inclusion of non-song vocalizations in passive-acoustic monitoring is proposed.

Studies on the flavins in rat liver mitochondrial outer membranes.

The incorporation of radioactivity derived from [2-14C] riboflavin into the flavins of rat liver mitochondrial outer membranes was studied. These membranes were found to contain about 0.6 nmol of non-covalently bound flavins per mg protein; the majority is in the form of FAD (73%) and FMN (24%). The membranes also contain about 1.5 nmol per mg of covalently bound flavins. After labeling, radioactive flavins appeared in the non-covalently bound flavins for about 4 h. Most of this radioactivity was in FAC (77%). Neither the rate nor extent of this labeling was affected by cycloheximide (1 mg/kg) administered 30 min prior to the radioactive riboflavin. With the covalently bound flavins, radioactivity was incorporated into the coenzymes for at least 18 h, but the rate of incorporation was much slower. After cycloheximide, radioactive flavins continued to appear in covalently bound flavins for about 2 h, but then stopped. Labeling of both types of flavins after [14C] riboflavin was considerably slower than the incorporation of [3H] leucine into outer membrane proteins. These results suggest that with flavoproteins from the mitochondrial outer membranes, the incorporation of flavins occurs after synthesis of the various apoenzymes is complete.

Monoamine oxidase A and monoamine oxidase B activities are catalyzed by different proteins.

Monoamine oxidases A and B (amino: oxygen oxidoreductase (deaminating) (flavin-containing), EC 1.4.3.4) have been identified in the outer membranes of rat liver mitochondria by their covalent reaction with the inhibitor, [3H]pargyline. On analysis by polyacrylamide gel electrophoresis under denaturing conditions. Monoamine oxidase A was found to migrate more slowly that monoamine oxidase B. Proteins which correspond to monoamine oxidases A and B (as identified by the electrophoretic distribution of covalently bound [3H]pargyline) were excised from the gels. Subsequent analysis showed that both monoamine oxidase A and monoamine B had been highly purified by this procedure. Electrophoretic analysis of the peptides produced by limited proteolysis with bovine trypsin, alpha-chymotrypsin, Staphylococcus aureus V8 proteinase and cyanogen bromide indicate that monoamine oxidases A and B have different amino acid sequences.

The effect of choline deficiency on the outer membranes of rat liver mitochondria.

The outer membranes of mitochondria prepared from the liver of rats kept 12 days on a choline-deficient diet were analyzed for changes in phospholipid and protein content. The total amount of phospholipid in the outer membranes was not affected by the deficiency. There was, however, a significant decrease in the amount of phosphatidylcholine and an increase in phosphatidylethanolamine. The alterations in the membrane phospholipids were reflected in a reduction in the fluorescence of the membrane probe, 8-anilino-1-naphthalene sulfonate. Choline deficiency also affected the protein composition of the outer membranes as judged by electrophoretic analysis; however, the activity of several enzymes which serve as markers for the outer membrane was not affected by the deficiency.

Glutamine-enriched parenteral nutrition regulates the activity and expression of intestinal glutaminase.

The aim of this study was to examine the effect of glutamine-enriched parenteral nutrition on the activity, expression and distribution of glutaminase mRNA within the small intestine of rats. Central venous lines were inserted into 30 male Wistar rats before they were fed for 6 days with either: (a) conventional parenteral nutrition, (b) 2.5% glutamine-enriched parenteral nutrition, or (c) rat food ad libitum. Jejunal glutaminase activity per milligram of dry matter was greatest in the animals fed rat food (0.94+/-0.29), intermediate in the glutamine supplemented rats (0.69+/-0.19) and least in the rats nourished with conventional parenteral nutrition (0.55+/-0.24) (P<0.05). The data for glutaminase expression exhibited a similar trend (P<0.05). In situ hybridisation analysis confirmed that glutaminase is expressed in the mucosa along the whole length of the small intestine. It was concluded that provision of glutamine alters the activity and expression of glutaminase in intestinal enterocytes. The results suggest that glutamine increases glutaminase activity by promoting the accumulation of intestinal glutaminase mRNA.

Phase I pharmacokinetic trial of perillyl alcohol (NSC 641066) in patients with refractory solid malignancies.

Perillyl alcohol (POH) is a monoterpene with anticarcinogenic and antitumor activity in murine tumor models. Putative mechanisms of action include activation of the transforming growth factor beta pathway and/or inhibition of p21ras signaling, leading to differentiation or apoptosis. In this Phase I trial, 17 patients took POH p.o. three times daily for 14 days of each 28-day cycle. The starting dose of POH was 1600 mg/m2/dose, with escalations to 2100 and 2800 mg/m2/dose in subsequent cohorts. Chronic nausea and fatigue were dose-limiting toxic effects at 2800 mg/m2. Grade 1-2 hypokalemia was common at 2100 and 2800 mg/m2. Although POH could not be detected in plasma, two of its metabolites, dihydroperillic acid (DHPA) and perillic acid (PA), were measured in plasma and urine on days 1 and 15 after the first and last doses of POH, respectively. Both area under the concentration versus time curve and peak plasma concentration (Cmax) values increased with dose and exhibited high intersubject variability. Day 15 DHPA Cmax values ranged from a mean +/- SD of 22.6+/-12 microM at 1600 mg/m2/dose to 42.4+/-15.24 microM at 2800 mg/m2/dose. Corresponding mean +/- SD Cmax values for PA were 433.2+/-245.8 and 774.1+/-439.6 microM. One patient treated at the 2800 mg/m2/dose had markedly prolonged plasma levels of both PA and DHPA and developed grade 3 mucositis. POH treatment did not consistently alter the expression of p21ras, rap1, or rhoA in peripheral blood mononuclear cells obtained from patients treated at the highest dose level. The metabolites PA and DHPA did not change expression or isoprenylation of p21ras in MCF-7 breast or DU145 prostate carcinoma cells at concentrations that exceeded those achieved in patient plasma after POH treatment. We conclude that POH at 1600-2100 mg/m2 p.o. three times daily is well tolerated on a 14-day on/14-day off dosing schedule. Inhibition of p21ras function in humans is not likely to occur after POH administration at safe doses of the present oral formulation.

The role of glutaminase in the small intestine.

Glutaminase is the enzyme which hydrolyses glutamine, the main respiratory fuel of the intestine, to yield glutamate and ammonia. Glutaminase has a central role in intestinal metabolism: the products of the reaction catalyzed by glutaminase can be transaminated, catabolized to yield energy or used for the biosynthesis of pyrimidine nucleotides. Experimental treatments which deprive the intestine of glutamine induce intestinal atrophy. In this review, attention is paid to the role of glutaminase in intestinal metabolism. Background information on the structure, kinetics and distribution of glutaminase precede a discussion of the metabolism of glutamine within the intestine. In closing, we review the factors known to regulate glutaminase activity and emphasise that the regulation of glutaminase within the intestine is poorly understood.

The in vitro insertion of monoamine oxidase B into mitochondrial outer membranes.

Bovine monoamine oxidase (MAO) B has been synthesized in vitro using a reticulocyte lysate translation system directed by bovine liver poly(A)+ RNA. The newly synthesized enzyme apparently lacks a cleavable N-terminal extension, but MAO B is readily incorporated into mitochondria or isolated mitochondrial outer membranes prepared from rat liver. ATP is not required for the binding of the newly synthesized enzyme to the outer membranes, but is necessary for the insertion of MAO B into these membrane vesicles. The ATP is not required to generate a mitochondrial membrane potential as assembly occurs under conditions that preclude either the formation or the maintenance of the potential. MAO B will bind to but not become incorporated into outer membrane vesicles which have been treated with trypsin, suggesting that the insertion of MAO B also depends on protein factors present on the outer membranes.

Persistent localized pulmonary interstitial emphysema and lymphangiectasia: a causal relationship?

The mechanism responsible for persistent regional pulmonary interstitial emphysema under tension, which occasionally complicates assisted ventilation, has not been explained. It is difficult to comprehend how interstitial air may cause lobar hyperinflation and not escape to the mediastinum or pleura. Histologic examination of resected emphysematous lung in two neonates with this type of interstitial emphysema revealed air dissecting from the interstitial connective tissue into markedly dilated lymphatics. It is proposed that intralymphatic air dissection causing emphysematous lymphangiectasia may be responsible for fixation of the lung in the emphysematous state.

Gomez-Lopez-Hernandez syndrome: expansion of the phenotype.

Gomez-Lopez-Hernandez syndrome (cerebello-trigeminal-dermal dysplasia) is a condition that includes abnormalities of the cerebellum (rhombencephalosynapsis), cranial nerves (trigeminal anesthesia), and scalp (alopecia). Seven patients with this condition have been documented since 1979. We now report a male with Gomez-Lopez-Hernandez syndrome who, at the age of 19 years, is the oldest patient identified to date. He has been followed since birth, allowing us to report on the progression of his physical findings and psychiatric problems including hyperactivity, depression, self-injurious behavior and bipolar disorder. In addition, he has short stature and growth hormone deficiency.

Urinary basic fibroblast growth factor: a noninvasive marker of progressive cystic renal disease in a child.

Autosomal recessive polycystic kidney disease (ARPKD) is a hereditary condition with an estimated incidence of 1 in 20,000 live births. Various growth factors have been implicated in the causation of this disease. We describe a child with ARPKD whose levels of urinary basic fibroblast growth factor (bFGF) were markedly elevated. The concentrations of bFGF increased further following right nephrectomy, in response to the compensatory growth of the remaining kidney. We hypothesize that measurement of urinary bFGF may be useful as a noninvasive marker to assess progression of cystic renal development.

Gene expression profiles of giant hairy naevi.

BACKGROUND: Congenital neomelanocytic naevi appear in nearly 1% of newborns. Giant hairy naevi (GHN) are uncommon lesions covering large areas of the body. They are of concern because they have the potential to transform into malignant melanomas. AIMS: To describe gene expression profiles of GHN and nearby normal skin from patients with GHN and normal control skin (from patients with cleft lip/palate). METHODS: Tissues from three patients with GHN and two normal controls were studied for differences in gene expression profiles. Total RNA was isolated from normal skin near the hairy naevus, GHN, and skin from normal controls. The RNA samples were subjected to probe labelling, hybridisation to chips, and image acquisition according to the standard Affymetrix protocol. RESULTS: There were 227 genes affected across all samples, as determined by DNA microarray analysis. There was increased expression of 22 genes in GHN compared with nearby normal skin. Decreased expression was noted in 73 genes. In addition, there was increased expression of 36 genes in normal skin near GHN compared with normal control skin, and decreased expression of five genes. Categories of genes affected were those encoding structural proteins, proteins related to developmental processes, cell death associated proteins, transcription factors, growth factors, stress response modulators, and collagen associated proteins. Changes in mRNA expression were checked by reverse transcription polymerase chain reaction. CONCLUSIONS: Genetic profiles of GHN may provide insight into their pathogenesis, including their potential for malignant transformation. Such information may be useful in improving the understanding and management of these lesions.

Surgical aspects of systemic necrotizing vasculitis.

Thirty patients with systemic necrotizing vasculitis (SNV) of the polyarteritis nodosa type were reviewed. Eleven patients (36%) had only gastrointestinal manifestations of the disease. Four patients (13%) had only pulmonary involvement. Seven patients (23%) had both gastrointestinal and pulmonary manifestations of SNV. With use of an approved protocol from the National Institute of Allergy and Infectious Diseases of cyclophosphamide and prednisone, remission was attained in 29 of 30 patients (96%). However, 18 patients (60%) underwent major surgical procedures for either diagnosis or complications of the disease. Six patients underwent eight thoracotomies; six operations were required for diagnosis and two for drainage of right-sided empyemas. Ten patients underwent 14 exploratory laparotomies. Six abdominal explorations were performed for diagnostic purposes, and eight were required for treatment of complications. Two patients required digital amputations because of ischemic necrosis. Two patients (11%) developed five postoperative complications, one of whom subsequently died. Arteriographic studies were performed in 16 patients (53%). Vascular abnormalities consistent with those described for SNV were demonstrated in six cases (38%). There was no correlation of clinical manifestations, subsequent therapy, and arteriographic findings with regard to the development of intra-abdominal complications. Pulmonary complications occurred only in patients who had pulmonary infiltrates secondary to SNV.

Incidence, management, and outcome of childhood empyema: a prospective study of children in Cambodian refugee camps.

To determine the incidence, outcome, and optimal management of empyema, all children less than 15 years of age admitted to Khao-I-Dang Hospital with a diagnosis of empyema during a 23-month period were prospectively studied. Khao-I-Dang Hospital provides care to 137,000 Cambodian children residing in eight refugee camps along the Thai-Cambodian border. Ninety-eight children with empyema were identified, for an annual incidence of 0.37 cases per 1,000 children. All patients had chest tubes inserted on admission, and all were treated with parenteral antibiotics, which included chloramphenicol in 92% of the patients and cloxacillin in 72%. Patients were hospitalized a mean of 30 days, and chest tubes were in place for a mean of 12 days. Surgery was performed on four patients who had bronchopleural fistulas that persisted for more than 14 days. Only one (1%) of the 70 patients treated with cloxacillin required thoracotomy, compared with three (11%) of the 28 patients who did not receive cloxacillin (P = 0.07). In a multiple regression analysis, the presence of pneumatoceles or mediastinal shift on admission chest radiograph, a history of tuberculosis in the family, and an age of more than five years were predictive of a longer duration of chest tube drainage. No patient died in the hospital, and only one patient died in the six months following discharge from the hospital. Chest radiographs that were obtained six months after discharge in 25 patients were all essentially normal, despite marked abnormalities on chest radiographs obtained at discharge. In summary, conservative medical management with the use of chest tubes for these 98 children with empyema resulted in a mortality rate of 1.0%, and should be considered as an effective alternative to the surgical management of patients presenting with this complication.

Prothrombin plasma clearance is not mediated by hepatic asialoglycoprotein receptors.

The hepatic asialoglycoprotein receptor (AGPR) system can efficiently internalize and degrade circulating glycoproteins which lack terminal sialic acids on their carbohydrate chains. Since pro-thrombin is a glycosylated plasma protein, possible involvement of AGPR system in its clearance from circulation was evaluated. The half lives of bovine 125I-prothrombin and 125I-asialoprothrombin, injected intravenously into rats, were 192 and 1.8 minutes, respectively. Asialoprothrombin appeared to be cleared by the hepatic AGPRs since 33% of it accumulated in the liver at 30 minutes and its clearance was competitively blocked by simultaneous administration of increasing amounts of asialofetuin. Only 5% of prothrombin accumulated in the liver at 3 hours and injections asialofetuin in amounts capable of saturating the AGPR for the duration of four asialoprothrombin half lives had no effect on the disappearance of prothrombin. Our observations indicate that, although asialoprothrombin is readily cleared from plasma by the AGPR system, prothrombin is not. Thus these receptors do not appear to be involved in physiological processes that control prothrombin half life.

Starvation alters the activity and mRNA level of glutaminase and glutamine synthetase in the rat intestine.

The metabolism of glutamine, the main respiratory fuel of enterocytes, is governed by the activity of glutaminase and glutamine synthetase. Because starvation induces intestinal atrophy, it might alter the rate of intestinal glutamine utilization. This study examined the effect of starvation on the activity, level of mRNA, and distribution of mRNA of glutaminase and glutamine synthetase in the rat intestine. Rats were randomized into groups and were either: (1) fed for 2 days with rat food ad libitum or (2) starved for 2 days. Standardized segments of jejunum and ileum were removed for the estimation of enzyme activity, level of mRNA, and in situ hybridization analysis. The jejunum of the fed rats had a greater activity of both enzymes per centimeter of intestine (P < 0.01), a greater glutaminase specific activity (1.97 +/- 0.45 vs. 1.09 +/- 0.34 micromol/hr/mg protein, P < 0.01), and a lower level of glutaminase and glutamine synthetase mRNA. The ileum of the fed rats had a greater activity of glutamine synthetase per centimeter of intestine (162.9 +/- 50.6 vs. 91.0 +/- 23.1 nmol/hr/cm bowel, P < 0.01), a lower level of glutaminase mRNA, and a greater level of glutamine synthetase mRNA. In situ hybridization analysis showed that starvation does not alter the distribution of glutaminase and glutamine synthetase mRNA in the intestinal mucosa. This study confirms that starvation decreases the total intestinal activity per centimeter of both glutaminase and glutamine synthetase. More importantly, the results indicate that the intestine adapts to starvation by accumulating glutaminase mRNA. This process prepares the intestine for a restoration of intake.

Identification of marine organism extracts active at the EGF binding site of human A431 cells.

Using a high throughput radioligand binding assay, we assessed aqueous ethanol extracts from 2885 marine organisms representing 17 phyla from the Indo-Pacific for their capacity to influence [125I]epidermal growth factor binding to human A431 cells in culture. Initial screening employed extracts pooled from five unrelated organisms to cells incubated at 37 degrees C for 20 min. Positive leads from the low stringency screening were pursued using extracts from individual organisms. Extracts from 57 organisms significantly inhibited radioligand binding, five organisms caused the cells to detach from the substrate, while extracts from two organisms brought about an increase in bound radiolabel. To discriminate between the mechanisms of action of the extracts, active organisms were also tested for their capacity to affect radioligand binding in the cells when incubated at 4 degrees C. Those organisms acting only at 37 degrees C were considered to have a cellular site of action, while those also active at the low temperature were considered to exert their effects more directly on the receptor binding event. The acute biochemical activity elicited by the positive organisms was distributed widely between taxa and between geographic regions. The approach provides a sensitive, high volume assay for detecting bioactive substances within marine organisms.

The effect of branched-chain amino acid-enriched parenteral nutrition on gut permeability.

In situations of catabolic stress, the gut becomes atrophic and has a diminished barrier function as evidenced by an increased permeability to a variety of molecules. It is known that the parenteral administration of branched-chain amino acids (BCAA) reduce gut atrophy. The aim of this study was to examine the effect of BCAA-enriched solutions of parenteral nutrients on gut permeability. A secondary aim was to observe the association between gut permeability and variables that have been used to assess jejunal atrophy. Central venous lines were inserted into 30 rats before randomization to receive nutritional support with: (1) a conventional parenteral solution (CPN), (2) A 2.0% BCAA-enriched solution (BCAA), or (3) rat food ad lib (Rat Food). The rats were assessed after 7 d for nutritional status, gut morphology, and gut permeability ratio (ratio of the permeability to 14C raffinose and 3H mannitol). We found that rats in the Rat Food Group lost the least amount of weight, had the least amount of jejunal atrophy, and had better preservation of barrier function as determined by gut permeability. When compared with the CPN Group, the BCAA Group had better preservation of jejunal morphology and protein content (p < 0.05), but a similar gut permeability. A cross-correlation matrix demonstrated a significant negative correlation between permeability to mannitol and mucosal weight, mucosal protein content and mucosal DNA content. Branched-chain amino acid-enriched parenteral nutrition reduced gut atrophy but not the gut permeability associated with parenteral nutrition. In the parenterally nourished rat model, atrophy of the jejunum is associated with increased permeability to small molecules.

Platelet monoamine oxidase activity in panic disorder.

Platelet monoamine oxidase (MAO) activity was studied in 21 patients meeting Research Diagnostic Criteria for panic disorder and in 12 healthy controls. Platelet MAO activity in females in both patient and control groups tended to be higher than that in males, but the results did not reach statistical significance. Platelet MAO activity was significantly decreased in panic disorder patients compared to controls.

Effects of glutamine infusion on colonic anastomotic strength in the rat.

Glutamine is one of the primary respiratory fuels of the colon. However, it is not included in commercial preparations of parenteral nutrients because of its short shelf life. It has been suggested that colonic atrophy induced by conventional parenteral nutrition can be reversed by the intravenous infusion of fresh solutions of glutamine. This study evaluated the hypothesis that glutamine-enriched parenteral nutrition would enhance the strength of a standard colonic anastomosis in undernourished rats. After surgery, the rats were randomized to receive 6 days of postoperative support with either rat chow, conventional parenteral nutrition, or parenteral nutrition containing 1.2% glutamine. Measurement of colonic bursting tension failed to demonstrate any significant differences between the groups under study. In conclusion, the administration of 1.2% glutamine-enriched parenteral nutrition failed to influence the healing of colonic anastomoses in undernourished rats.