Quality and safety of hemp meal as a protein supplement for nonlactating dairy cows.

Hemp seed meal may be a suitable protein supplement for dairy cows, but its quality and safety as a dairy cow feed has not yet been fully investigated. As a result, dry matter intake (DMI), rumen fermentation, blood metabolites, total-tract digestibility, and concentrations of cannabinoids in blood plasma, urine, muscle, and adipose tissues were compared among nonlactating Holstein dairy cows receiving a basal partial mixed ration that was supplemented with either 10.2% dry matter (DM) hemp meal (HM treatment), 13.5% DM canola meal (CM treatment), or 6.25% DM hemp meal and 6.16% DM canola meal (HC treatment). Diets were formulated to be isoenergetic and isonitrogenous. Six nonlactating, nonpregnant Holstein cows were used in a repeated 3 × 3 Latin square design trial with three 3-wk experimental periods. The first 2 weeks of each served as adaptation. Sample and data collection occurred during the third week of each period. Neither the partial mixed ration nor canola meal contain cannabidiol (CBD), cannabidiolic acid (CBDA), d9-tetrahydrocannabinol (THC), or tetrahydrocannabinolic acid A (THCA). However, the hemp meal contained 3.0, 4.4, 0, and 0.1 µg/g DM of CBD, CBDA, THC, and THCA, respectively. Treatment did not affect DMI, pH, concentrations of volatile fatty acids or ammonia in the rumen, total-tract digestibilities of DM and crude protein, or blood plasma concentrations of glucose, urea, ß-hydroxybutyrate, and nonesterified fatty acids. Hence, based on these metabolites, treatment did not affect the nutritional status of the cows. However, the total-tract neutral detergent fiber digestibility of the CM treatment (43%) was higher than that of the HM treatment (38%). No cannabinoids were detected in blood plasma, rumen fluid, and urine. Cannabinoids were also not detected in kidney, liver, urine, muscle, or adipose tissues at the end of the experiment when cows had undergone all treatments. Feces from all treatments did not contain detectable concentrations of THC or THCA, but feces of cows on the HC treatment contained 0.42 and 0.40 µg/g DM of CBD and CBDA, respectively. Feces of cows on the HM treatment contained 0.68 and 0.67 µg/g DM of CBD and CBDA, respectively. This indicated that most ingested CBD and CBDA were not absorbed but instead were excreted in the feces. Our data show dietary inclusion rates of up to 10.2% of DM. We find that hemp meal is a high-quality and safe protein supplement for nonlactating dairy cows.

Comparing dehulled hemp meal and canola meal as a protein supplement for lactating dairy cows.

Effects of replacing canola meal with dehulled hemp meal in the diet of lactating dairy cows on the dry matter intake (DMI), milk production, milk fatty acid profile, blood metabolites, total-tract nutrient digestibility, and transfer of cannabinoids were determined in 12 lactating, nonpregnant Holstein cows. These cows were used in a 3 × 3 Latin square design with three 3-wk experimental periods consisting of 2 wk of adaptation and 1 wk of sampling. Cows received basal partial mixed rations supplemented with either 15% dry matter (DM) canola meal (CM15), 15% DM dehulled hemp meal (HM15), or 7.5% DM dehulled hemp meal and 7.5% DM canola meal (CM7.5HM7.5). Diets were formulated to be isoenergetic and isonitrogenous, but the HM15 and CM7.5HM7.5 diets contained, on average 1.2 percentage units more crude protein (CP) that the CM15 diet. The CP of the dehulled hemp meal contained less soluble protein than that of canola meal. Hence, the intake of soluble protein did not differ among diets. Canola meal contained less crude fat than hemp seed meal (3.46% vs. 8.25% DM). The lipid fraction of canola meal fat contained more oleic acid (C18:1 cis-9; 47.3 vs. 14.9 g/100 g of fatty acids, FA) and vaccenic acid (18:1 cis-11; 13.7 vs. 1.2 g/100 g of FA) and less linoleic acid (C18:2n-6; 21.9 vs. 55.7 g/100 g of FA) and α linolenic acid (C18:3n-3; 3.2 vs. 8.9 g/100 g of FA) than the lipid fraction of hemp seed meal. The hemp seed meal contained 4.9 µg/g cannabidiol, 5.1 µg/g cannabidiolic acid, and 0.1 µg/g tertahydroxycannabinolic acid A. Treatments did not differ in DMI, yields of milk, milk protein and milk fat, total-tract neutral detergent fiber digestibility, and blood plasma concentrations of ß-hydroxybutyrate and nonesterified FA. Apparent total-tract DM digestibility was lowest in the HM15 treatment, whereas the CP digestibility and the concentrations of urea in blood, urine, and milk were lowest in the CM15 treatment. Cannabinoids were not detected in urine, milk, and blood plasma. Replacing canola meal with hemp seed meal increased milk fat contents of polyunsaturated fatty acids (PUFA), which were 3.42, 3.90, and 4.25 g/100 g of FA for the CM15, CM7.5HM7.5, and HM15 treatments, respectively. Especially, the milk fat contents of 18:2n-6 (1.99 vs. 1.56 g/100 g FA) and 18:3n-3 (0.31 vs. 0.43 g/100 g FA) were increased by hemp meal feeding. Especially, the milk fat contents of 18:2n-6 (1.99 vs. 1.56 g/100 g FA) and 18:3n-3 (0.31 vs. 0.43 g/100 g FA) were increased by hemp meal feeding. Our data show that hemp seed meal is a suitable and safe replacement for canola meal as a feed for lactating dairy cows and that this replacement increases CP digestibility and urea in urine, milk, and blood plasma, as well the PUFA content of milk fat.

Leonotis Leonurus improves the crosstalk between peripheral tissues both in vivo and in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Unravelling the anti-diabetic mechanism of action of L. leonurus at adipose, liver, muscle and pancreatic level. AIMS: To investigate the mechanism of action of an organic extract of L. leonurus and marrubiin at the gene level in adipose, liver and muscle tissues of an obese rat model and in a co-culture model. MATERIALS AND METHODS: Obese Wistar rats were fed a cafeteria diet for eight weeks, treated with an extract of L. Leonurus, marrubiin, sulfonylurea and aspirin for two weeks and the level of gene expression of selected markers were investigated across different tissues. The effects mediated by the different treatments were investigated in co-culture cell models involving 3T3-L1 (fat), Chang (liver), C2C12 (muscle) and INS-1 (pancreatic) cells under both normal and hyperglycemic conditions. RESULTS: L. leonurus extract mediated a significant increase in PPAR gamma, glucokinase, FAS and UCP2 gene expression in adipose tissue, whilst the opposite was observed in the liver. At the muscle level, a significant increase in FAS gene expression was observed relative to the obese control rats. Furthermore, the extract as well as marrubiin, modulated improvements in the adipokine profile. The co-culture models showed that the effect mediated by the extract was dependent on, the tissue type as well as the glycemic conditions. CONCLUSIONS: L. Leonurus extract as well as marrubiin exhibit anti-diabetic properties where the mechanism of action is mainly at the adipose tissue level. The increase in expression of the genes of interest mentioned above potentially play a protective role towards the liver and possibly towards the muscle tissues as well.

Biological effects of THC and a lipophilic cannabis extract on normal and insulin resistant 3T3-L1 adipocytes.

Type 2 diabetes, a chronic disease, affects about 150 million people world wide. It is characterized by insulin resistance of peripheral tissues such as liver, skeletal muscle, and fat. Insulin resistance is associated with elevated levels of tumor necrosis factor alpha (TNF-alpha), which in turn inhibits insulin receptor tyrosine kinase autophosphorylation. It has been reported that cannabis is used in the treatment of diabetes. A few reports indicate that smoking cannabis can lower blood glucose in diabetics. Delta(9)-tetrahydrocannabinol (THC) is the primary psychoactive component of cannabis. This study aimed to determine the effect of a lipophilic cannabis extract on adipogenesis, using 3T3-L1 cells, and to measure its effect on insulin sensitivity in insulin resistant adipocytes. Cells were cultured in Dulbecco's modified eagle medium (DMEM) with 10% fetal bovine serum (FBS) and differentiated over a 3 day period for all studies. In the adipogenesis studies, differentiated cells were exposed to the extract in the presence and absence of insulin. Lipid content and glucose uptake was subsequently measured. Insulin-induced glucose uptake increased, while the rate of adipogenesis decreased with increasing THC concentration. Insulin-resistance was induced using TNF-alpha, exposed to the extract and insulin-induced glucose uptake measured. Insulin-induced glucose was increased in these cells after exposure to the extract. Semiquantitative real time polymerase chain reaction (RT-PCR) was performed after ribonucleic acid (RNA) extraction to evaluate the effects of the extract on glucose transporter isotype 4 (GLUT-4), insulin receptor substrate-1 (IRS-1) and IRS-2 gene expression.

Feasibility of thyroid ultrasonography in field studies in a developing country, Ghana.

This study was carried out to determine the feasibility of using ultrasonography for goitre estimation in the field situation. It is a cross sectional study that was conducted using community based cluster sample. Thyroid sonography was performed on 112 randomly selected school children aged 10 -15 years from two districts in the Greater Accra area of Ghana, using normative values for thyroid volume recommended by WHO/ICCIDD. The mean age of all subjects was 13.5 years+/-0.13 SEM. The male to female ratio was 1:1. The mean height and weight of the children were 1.5 metres+/-0.9 SEM and 38.1 Kg+/-0.7 SEM, respectively. The mean body surface area was 1.27 m2+/-0.2. There were no significant gender differences in their ages (13.6yrs+/-0.2 SEM, 13.4yrs+/-0.1 SEM, respectively) and height (1.50m+/-1.6 SEM, 1.46m+/-1.7 SEM, respectively). The girls (40.0kg+/-1.2 SEM) weighed more than the boys (35.6kg+/-1.1). The mean and median urinary iodine concentration were 82.4+/-8.5 SEM and 67.9 ug/l, respectively. All the children examined had normal thyroid sonogram. The thyroid volumes ranged from 3.6 ml to 15.3ml. The mean thyroid volume was 7.0ml+/-0.2 SEM and the thyroid volume was higher in the girls (7.5ml+/-0.3 SEM) than the boys (6.5ml+/-0.2 SEM). The criteria of thyroid volume per age and sex yielded a goitre prevalence of 1.8 %. In contrast, the criteria of thyroid volume by surface area yielded a goitre prevalence of 8 %. Our study has shown that it is feasible to employ ultrasonography for field studies to determine goitre prevalence in school children in a developing country such as Ghana. However, the best criteria for goitre in children in Ghana, requires to be confirmed in future studies.

A phase II study of sequential combination chemotherapy with cyclophosphamide, prednisone, and 2-chlorodeoxyadenosine in previously untreated patients with chronic lymphocytic leukemia.

In an earlier study of previously untreated patients with chronic lymphocytic leukemia (CLL), we used a concomitant combination of chlorambucil and 2-chlorodeoxyadenosine and reported overall (OR) and complete (CR) remission rates of 80% and 20%, respectively. After a median follow-up of 5 years, more than 80% of the responders have had a relapse. In the current phase II study of 27 previously untreated patients with CLL, we used a sequential combination of six cycles of intravenous cyclophosphamide (1 g/m2) plus oral prednisone (100 mg/m2 per day for 5 days) followed by two to six cycles of 2-chlorodeoxyadenosine (5 mg/m2 per day for 5 days). The OR and CR rates were 96% and 33%, respectively. After a median follow-up of 29 months, 35% of the responders have had a relapse. Progression-free survival was significantly better in CR patients than in those with partial remission. However, minimal residual disease was phenotypically detected in four of the nine CR patients. Despite the fact that the current OR and CR rates are superior to those seen in a historical cohort treated with a concomitant schedule, a longer follow-up period is needed to assess the durability of these remissions, and a controlled trial is necessary to estimate the impact on overall survival and toxicity.

Phase II evaluation of continuous-infusion 5-fluorouracil, leucovorin, mitomycin-C, and oral dipyridamole in advanced measurable pancreatic cancer: a North Central Cancer Treatment Group Trial.

At present there remains a need for more effective systemic therapy in advanced pancreatic cancer. Some studies have suggested that infusional chemotherapy schedules and biomodulation of 5-fluorouracil (5-FU) may improve the therapeutic outcome in advanced colon cancer. One such regimen that uses continuous infusion 5-FU, weekly leucovorin, daily dipyridamole, and intermittent mitomycin-C has activity in both colon and unresectable pancreatic carcinoma. The intent of this trial was to test the effectiveness of this four-drug regimen in advanced pancreatic cancer. Patients received 5-FU 200 mg/m2 daily by continuous infusion, leucovorin 30 mg/m2 IV weekly, mitomycin-C 10 mg/m2 day 1, and dipyridamole 75 mg orally four times daily for 5 weeks. After a 1-week break, treatment cycles were repeated every 6 weeks. Eligibility included biopsy-proven advanced measurable pancreatic cancer, Eastern Cooperative Oncology Group performance status 0 and 2, and no prior systemic chemotherapy. Of 46 evaluable patients, 9 partial responses and 1 complete tumor response were seen, for an overall response rate of 22% (95% confidence interval 11-36%). The median survival in the group of 50 patients registered to this trial was 4.6 months, with a range of 0.33 to 40.2 months. Toxicity was manageable, with the most common toxicities (> or =grade III National Cancer Institute Common Toxicity Criteria) being anorexia (13%), stomatitis (17%), and hand-foot syndrome (13%). Of note, little severe hematologic toxicity and no significant headaches were reported. Although some patients did respond, the therapeutic results are not encouraging enough to take this regimen to phase III testing.

Endemic goitre and urinary iodine levels in rural communities in the Bolgatanga and Builsa districts of the upper east region of Ghana.

OBJECTIVE: To ascertain the severity of IDD in some rural communities in upper east of Ghana and to urge the establishment of intervention and control measures for IDD. SUBJECTS AND STUDY DESIGN: A total of 1061 subjects, made up of about an equal number of children (8-14 years) and women of childbearing age (15-45 years) from seven Sekoti villages and five Builsa villages of the upper east of Ghana were examined for goitre by the palpation method. In addition, every tenth subject examined, provided urine for urinary iodine determination. RESULTS: 68.8% of the subjects had goitre; 9.9% had visible goitre. The goitre rates of the children from Builsa (77.2%) were significantly higher than those from Sekoti (59.1%) [z = 4.5; p < 0.001]. The overall prevalence of goitre and visible goire in women in the two areas were 70.8% and 15.4% respectively. The women of Sekoti had more goitres (76.6%) but less visible goitres (8.8%) than those of Builsa (63.5% and 21.9%) [p < 0.001). The median urinary iodine level for the two survey areas was 1.6 micrograms/dl. 72% of subjects and urinary iodine less 2 micrograms/dl/24% had urinary iodine levels in the range 2-5 micrograms/dl and the remainder had urine iodine in the range 5-10 micrograms/dl. CONCLUSIONS: These findings indicate severe IDD in Sekoti and Builsa areas requiring urgent action. Further studies are indicated to determine the cause(s) of the IDD endemia.

Effect of matrix lipid chain length on liposomes containing cholesterol and ganglioside GM1: implications in drug delivery.

Incorporation of ganglioside GM1 into liposomes composed of phosphatidylcholine (PC) and cholesterol significantly increases their blood circulation time. Dipalmitoyl PC (DPPC) and distearoyl PC (DSPC) are the most commonly used saturated phospholipids employed in the preparation of long-circulating liposome formulations. Physical studies were performed on different formulations of these liposomes in an attempt to understand the effects of the matrix lipid chain length and GM1 concentration on the use of these liposomes as long-circulating drug delivery systems. The GM1/PC mixtures existed in different physical states: a lamellar state with components exhibiting miscibility (which is the desired state for drug delivery), a mixed micellar phase, and also a smectic mesophase. GM1 is miscible with DSPC bilayers up to a concentration of 25 mol%, beyond which conversion into the metastable mesophase occurs. By 30 mol%, solubilization into the mixed micellar state occurs. With DPPC, the metastable mesophase occurs at 33-38 mol% of GM1, with mixed micelles being formed at higher concentrations. The addition of cholesterol led to an inhibition of micelle formation. This study also indicates that GM1 stabilizes DPPC bilayers while destabilizing DSPC bilayers. On the basis of the interactions of GM1 with DPPC and DSPC, a new hypothesis for stabilization of PC bilayers is proposed, which also explains the destabilization of PC with C18.0 and higher. The longest circulating GM1 liposome formulations are predicted from this study.

Effects of polyethyleneglycol chain length and phospholipid acyl chain composition on the interaction of polyethyleneglycol-phospholipid conjugates with phospholipid: implications in liposomal drug delivery.

PURPOSE: The purpose of this study was to investigate polyethyleneglycol(PEG)-phosphatidylethanolamine(PE) conjugate interaction with phospholipid bilayers, in an attempt to explain the dependence of liposome circulation time on formulation. METHODS: Differential scanning calorimetry, electron microscopy, dynamic light scattering and NMR were the major methods used in the study. RESULTS: Mixtures of PEG-phospholipid conjugates and phosphatidylcholine existed in three different physical states: a lamellar phase with components exhibiting some miscibility, a lamellar phase with components phase separated, and mixed micelles. Beyond 7 mol-percent of PEG(1,000-3,000)-dipalmitoyl phosphatidylethanolamine (DPPE), and 11 mol% PEG(5,000)-DPPE in dipalmitoyl phosphatidylcholine (DPPC), a strong tendency towards mixed micelle formation was observed. All concentrations of PEG(12,000)-DPPE and PEG(5,000)-DPPE beyond 8 mol% formed phase separated lamellae with phosphatidylcholine. Decreasing the acyl chain length from C(16:0) to C(14:0) caused a decrease in tendency towards micelle formation and phase separation. These tendencies increased upon increasing acyl chain length to C(18:0). Phase separation was at least partly due to PEG chain-chain interaction. This was supported by an increased fraction of PEG chains exhibiting a fast NMR transverse relaxation in DPPC/PEG(5,000)-DPPE mixtures as compared to that in distearoyl phosphatidylcholine (DSPC)/PEG(5,000)-dioleoyl-PE (DOPE). CONCLUSIONS: These phenomena are discussed in relation to both bilayer and steric stabilization of liposomes, and the lack of prolonged circulation with certain formulations is discussed.

Interaction of polyethyleneglycol-phospholipid conjugates with cholesterol-phosphatidylcholine mixtures: sterically stabilized liposome formulations.

PURPOSE: The purpose of this study was to investigate polyethyleneglycol (PEG)-phosphatidylethanolamine (PE) conjugate interaction with cholesterol-phospholipid mixtures in an attempt to explain the effect of cholesterol on liposome circulation time. METHODS: Differential scanning calorimetry, NMR, electron microscopy, dynamic light scattering and fluorescence spectroscopy were the major methods used. RESULTS: Studies performed in the absence of cholesterol indicated the formation of three distinct physical states depending on the chain length of PEG in PEG-PE. Mixed micelle formation was observed at concentrations of PEG(1,000)-DPPE above 7 mol-% of lipid. Phase separated lamellae were observed at all concentrations of PEG( 12,000)-DPPE (Bedu-Addo et al. Pharm. Res. 13:710-717 (1996)). Upon incorporation of high concentrations of cholesterol >30 mol% into the lipid bilayer, the formation of phase separated lamellae was completely inhibited and the formation of mixed micelles significantly reduced. At high concentrations of PEG(1,000)-PE, solubilization of the bilayer occurred with preferential solubilization of cholesterol over phosphatidylcholine. Maximum steric stabilization (surface protection) was observed with low concentrations of short chain PEG-PE and high concentrations of cholesterol. CONCLUSIONS: The study provides a physical mechanism for the following observations: the blood circulation time is significantly increased or decreased with liposomes highly enriched with cholesterol or PEG-PE respectively.

Pneumococcal sepsis and meningitis in adults with sickle cell disease.

Patients with sickle cell disease are predisposed to infection caused by Streptococcus pneumoniae. However, there has been only one published case of bacteremic pneumococcal meningitis in an adult with sickle cell anemia. We report here the cases of six adults with sickle cell disease, pneumococcal sepsis, and meningitis. Five patients were male and one was female. Their ages ranged from 18 to 34 years (mean, 25.7 years). Five patients had the SS and one had the SC hemoglobin phenotype. Only one patient had received pneumococcal vaccine (14 valent). This vaccine did not protect against the pneumococcal serotype causing his infection. All patients had high fever (mean, 39.8 degrees C [103.7 degrees F]) on admission; five had generalized weakness and four had neck stiffness. Leukocyte counts were greater than 30,000/mm3 in all patients. Streptococcus pneumoniae was isolated from the blood and the cerebrospinal fluid in all patients. The cerebrospinal fluid showed pleocytosis in six patients, an elevated protein level in five, and hypoglycorrhachia in two. Complications included renal failure in four patients, disseminated intravascular coagulation in one, and seizures in another. Two patients died. Pneumococcal sepsis and meningitis are uncommon in adults with sickle cell disease, but they carry a high morbidity and mortality. Wider use of the new 23-valent polysaccharide vaccine in these patients is recommended.