Analysis of cytokine immune response profile in response to inflammatory stimuli in mice with genetic defects in fetal and adult hemoglobin chain expression.

Injections of a crude fetal sheep liver extract (FSLE) containing fetal hemoglobin, MPLA, and glutathione (GSSH) reversed cytokine changes in aged mice. To investigate the role of fetal hemoglobin we derived mice with homzygous deletions for either of the two major ßchains, HgbßmaKO or HgbßmiKO. Hgbßmi is the most prominent fetal Hgbß chain, with Hgbßma more prominent in adult mice. Mice lacking another fetal Hgb chain, HgbεKO, died in utero. CHO cells transfected with cloned Hgb chains were used to produce proteins for preparation of rabbit heteroantibodes. Splenocytes from HgbßmaKO mice stimulated in vitro with Conconavalin A showed a higher IL-2:IL-4 ratio than cells from HgbßmiKO mice. Following immunization in vivo with ovalbumin in alum, HgbßmaKO mice produced less IgE than HgbßmiKO mice, suggesting that in the absence of HgbßmiKO mice had a predeliction to heightened allergic-type responses. Using CHO cells transfected with cloned Hgb chains, we found that only the fetal Hgb chain, Hgbε, was secreted at high levels. Secretion of Hgbßma or Hgbßmi chains was seen only after genetic mutation to introduce the two N-linked glycosylation sites present in Hgbε, but absent in the Hgbß chains. We speculated that a previously unanticipated biological function of a naturally secreted fetal Hgb chain may be partly responsible for the effects reported following injection of animals with fetal, not adult, Hgb. Mice receiving injections of rabbit anti-Hgbε but not either anti-Hgbßma or anti-Hgbßmi from day 14 gestation also showed a bias towards the higher IL-2:IL-4 ratios seen in HgbßmiKO mice.

Efficacy and safety of human placental extract for alcoholic and nonalcoholic steatohepatitis: an open-label, randomized, comparative study.

Human placental extract (HPE) is a traditional medicine that has been used for the symptomatic treatment of liver disease without any verifying clinical evidence. This study aimed to evaluate the efficacy and safety of HPE in patients with alcoholic or nonalcoholic steatohepatitis (ASH or NASH). We designed this clinical trial as a multicenter, open-label, randomized, comparative noninferiority study to improve the reliability of analyses. The enrollment criteria were limited to ASH or NASH patients with serum alanine aminotransferase (ALT) 1.5-fold higher than the normal level. Patients in the control group were treated with a commercially available mixture of liver extract and flavin adenine dinucleotide (LE­FAD). Intention-to-treat (ITT) analysis was applied to 194 patients, and per-protocol (PP) analysis was available for 154 patients. The rate of primary goal achievement of treatment efficacy was arbitrarily defined as 20% or greater improvement in ALT level compared with the pretreatment level and did not differ significantly between the HPE and control groups [62.9% (44/70) vs. 48.8% (41/84); p=0.0772]. ITT and modified ITT analysis showed results similar to those of PP analysis. Adverse drug reactions (ADRs) of minimal to moderate degree occurred in 3.1% of patients. The ADR and treatment compliance rates were similar in both groups. In conclusion, the clinical value of HPE in the treatment of ASH and NASH is equivalent to that of LE­FAD.

Liver hydrolysate assists in the recovery from physical fatigue in a mouse model.

It is reported that liver hydrolysate (LH) enhances liver function. However, the effects of LH on physical fatigue are unknown. The aim of this study was to investigate the effect of LH on alterations in locomotor activity and energy metabolism such as 5'-AMP-activated protein kinase (AMPK), glycogen content, and blood lactic acid, after forced walking. Adult male ddY mice were used. Locomotor activity, AMPK phosphorylation, and glycogen content in the liver and soleus muscle, as well as blood lactic acid were determined following LH treatment before and/or after forced walking. The locomotor activity significantly decreased after forced walking for 3 h. Two administrations of LH (30 or 100 mg/kg) significantly increased the locomotor activity, while a single administration either before or after forced walking did not show any specific effect. Administering LH twice activated AMPK in the liver and soleus muscle. Glycogen levels significantly decreased in both the liver and soleus muscle after forced walking, whereas the blood lactate level significantly increased. In contrast, administering LH twice increased muscle glycogen and decreased blood lactic acid. These findings indicate that LH produced an anti-fatigue effect and that this effect appears to involve the efficient glycogen utilization through activation of AMPK.

A comparative study of isolutrol versus benzoyl peroxide in the treatment of acne.

Isolutrol is the active principle isolated from aqueous tissue extracts of deep sea shark liver and gall-bladder. A previous study has demonstrated the ability of isolutrol to reduce hyperseborrhoea, which provides a rationale for its use in the treatment of acne. We have performed a double-blind clinical trial on 70 patients to evaluate the efficacy and skin tolerance of isolutrol 0.15 g/100 mL (Ketsugo) in the treatment of mild to moderate acne when compared with 5% benzoyl peroxide lotion. The results from this study showed that both isolutrol and benzoyl peroxide significantly improved patients' acne by reducing the number of inflamed lesions. Isolutrol did not significantly reduce the numbers of non-inflamed lesions whereas benzoyl peroxide did. Fewer side effects were experienced by patients treated with isolutrol when compared with benzoyl peroxide. These results indicate that isolutrol may be a useful adjunct in the treatment of acne, particularly in patients with inflamed lesions.

[Analysis of the clinical effect of Adelavin infusion during enflurane anesthesia on post-operative hepatic function].

The protective effect of Adelavin, which is made of liver essence, for maintaining postoperative hepatic function was analysed in 85 patients who had enflurane anesthesia. The patients who showed hepatic dysfunction preoperatively were excluded from this trial. Adelavin was infused at a speed of 0.1 mg.kg-1.hr-1 during anesthesia. The group who had upper abdominal surgery in the Adelavin group showed significantly lower incidence of postoperative hepatic dysfunction. The group who had only enflurane anesthesia in the Adelavin group showed significantly lower incidence of postoperative hepatic dysfunction. These results suggest that the Adelavin infusion has a significant protective effect on hepatic function after enflurane anesthesia.

Liver extract-folic acid-cyanocobalamin vs placebo for chronic fatigue syndrome.

Chronic fatigue syndrome is a recently defined entity for which clinical criteria were proposed by the Centers for Disease Control, Atlanta, Ga. A frequently advocated treatment in Southern California is an injectable solution of bovine liver extract containing folic acid and cyanocobalamin (LEFAC). We conducted a double-blind, placebo-controlled, crossover trial of intramuscular LEFAC in 15 patients who met the Centers for Disease Control criteria for chronic fatigue syndrome. Although patients responded to placebo and LEFAC by several criteria of functional status, no significant difference was apparent between response to placebo and that to LEFAC. The placebo response appeared to be strong.

Antigen-specific modulation of graft-versus-host reactions by two distinct placental factors.

We have previously shown that two distinct mouse placental fractions (PF) are potent immunomodulators in vivo. A 40 kDa PF induces a marked decrease of plaque forming cell (PFC) responses, while a 60 kDa PF increases them. Both effects are specific for the priming antigen. In the present study, these two PF are assayed on a cell-mediated response to allogeneic cells, i.e. in a local graft-versus-host reaction (LGVHR). Mice were primed with allogeneic cells in the presence of various amounts of the 40 kDa or 60 kDa PF, or liver extract (LE) as control. Six days later, their spleen cells were injected into the footpads of F1 recipients. Precise dose-response curves were established and the kinetics of the GVH response were carefully followed. Parallel with the modulation of PFC responses, the 40 kDa PF caused a potent inhibition of the LGVHR, while the 60 kDa PF greatly enhanced it. Both effects were specific for the alloantigens injected with the PF. Furthermore, we showed that these modulations were observed whatever the intensity of the GVH reaction, which varied according to the number of primed spleen cells transferred. This report also demonstrates that these PF can be greatly enriched by passage over affinity columns made of insolubilized lectins. The 40 kDa PF is retained on and can be eluted from columns of insolubilized concanavalin A (Con A) or wheat germ agglutinin (WGA), which indicates that it is a glycoprotein. Conversely, the 60 kDa PF does not bind to any of the above lectins and is probably not a glycoprotein. This biochemical purification step is also a good procedure for obtaining an even cleaner separation of the two fractions from each other. Thus, this paper demonstrates that both PF have important regulatory properties on specific cellular immune responses.

Enterogastrone-like effect of peptide YY is vagally mediated in the dog.

Intraluminal fat inhibits gastric secretion through as yet undetermined mechanisms which involve release of one or more hormonal enterogastrones. As intraluminal fat releases Peptide YY (PYY) in amounts sufficient to inhibit meal-stimulated acid secretion, this ileo-colonic peptide exhibits the characteristics required of an enterogastrone. The present study seeks to determine the mechanism by which PYY inhibits acid secretion by examining the effects of PYY on gastric acid stimulated by pentagastrin, histamine, and bethanechol. In addition, effects of PYY on the acid response to sham feeding and distention of a denervated gastric pouch were examined. A dose of PYY (400 pmol X kg-1 X h-1) was employed that reproduced blood levels observed after intestinal perfusion with oleic acid and inhibited the acid secretory response to an intragastric meal by 35 +/- 6%. This same dose of PYY maximally inhibited histamine- and pentagastrin-stimulated acid secretion by 28 +/- 7% (P less than 0.05), and 17 +/- 4% (P less than 0.05), respectively. Although PYY had no effect on bethanechol-stimulated secretion it markedly inhibited the secretory response to sham feeding, maximally reducing secretion by 90 +/- 4% (P less than 0.01). We speculate that PYY acts by inhibiting acetylcholine release from vagal nerve fibers rather than by inhibiting acetylcholine's action on the parietal cell. The demonstration that PYY virtually abolishes cephalic phase acid secretion while having little if any effect on the response to exogenous secretogogues gives PYY unique characteristics among the known hormonal inhibitors of gastric secretion.

Mechanisms of protein activation of the intestinal phase of gastric secretion.

To determine if the secretory response to protein in the gut is due to direct effects of absorbed amino acids on the parietal cells, to hormonal effects, or to a combination of both, Heidenhain pouch secretion and plasma amino nitrogen levels were measured during enteral infusions of casein or liver extract, as well as during intravenous infusions of amino acids, in three dogs with and three dogs without portacaval transposition. In the dogs without portacaval transposition, gastric secretory responses were slight and did not differ significantly; secretory responses were seen only during infusions in which elevations of plasma amino nitrogen values occurred. In dogs with portacaval transposition, secretory responses to both casein and liver extract were significantly higher (P less than 0.02) than was the response to intravenous amino acids, while plasma amino nitrogen levels rose to virtually identical levels during all three kinds of infusion. These data support the hypothesis that absorbed amino acids contribute to the intestinal phase of gastric secretion. The finding that casein and liver extract in the gut evoked greater gastric secretion responses than did intravenous amino acids, even though increases in plasma amino nitrogen levels were similar, suggests that the intestinal phase results form the combined effects of absorbed amino acids and hormonal influences rather than from the effects of amino acids alone.

Role of antral mucosa in intestinal phase of gastric secretion in dogs.

This study was performed to investigate the mechanisms and significance of the intestinal phase of gastric secretion. With infusion of 10% liver extract into the duodenum of dogs, serum gastrin levels of the right gastroepiploic vein increased significantly (p less than 0.01). With infusion of the pH 7.0 liver extract, acid and pepsin outputs in non- antrectomized dogs increased significantly (p less than 0.01), as did serum gastrin levels (p less than 0.01). Post-infusion acid output in these dogs was 48 +/- 16% of tetragastrin-stimulated output. With the same infusion, antrectomized dogs showed no significant increase of acid and pepsin outputs and serum gastrin levels. Acid output and serum gastrin levels in non- antrectomized dogs that were infused with pH 2.0 liver extract did not increase significantly. The values of almost all serum amino acids in both types of dogs increased slightly, but insignificantly, with pH 7.0 liver extract. We obtained the following conclusions. The intestinal phase of gastric secretion operates by the mediation of the antral mucosa, from which gastrin is released. Although this phase has some effect, the acid-secretory ability of this phase is masked by inhibitory systems under certain physical conditions.

Prolongation of skin allograft survival in ATS-treated mice by post-transplant injections of species antigenic extracts.

B10.A male mice were grafted with H-2-incompatible murine B10.A(2R) skin allografts and treated with antithymocyte serum on days 2, 4, and 7 after transplantation. Repeated injections of cell-free tissue extracts from livers or spleens of B10.A(2R) mice were given in the standard doses, starting on the day of transplantation or on day 14 or day 28 after transplantation. The standard doses were the equivalents of material extracted from 40 mg or 80 mg of wet weight of liver or spleen tissue. Almost all of the regimens used in which antigen injections were begun on day 14 or day 28 after transplantation were successful and led to a marked prolongation in skin allograft survival. In some experimental groups most of the grafts survived 100 days after grafting and 8--33% grafts showed long-term survival in individual groups. The mechanism of this tolerance is mediated by suppressor cells which were characterized by means of anti-Thy 1.2 antibodies as T lymphocytes. the in vitro experiments have shown that cytotoxic cell precursors may be present in long-term tolerant mice and that they may be reactive to the tolerated antigens after sensitization.

Role of H2-receptors in gastrogenic hyperglycemia and hyperinsulinemia in dogs.

Recent studies have demonstrated that an acidified liver meal stimulates insulin release and raises plasma glucose levels. To examine the mechanism of these changes, a liver extract test meal at either pH 7 or at pH 2 was instilled into the stomach of dogs with a bisected pylorus and a gastric fistula during the infusion of either cimetidine, a specific H2-receptor antagonist, or a saline control. In response to the meal at pH 2 insulin, glucagon and glucose levels rose significantly and were not significantly changed by the infusion of cimetidine. In response to the liver meal at pH 7 a late rise in plasma glucagon levels was observed while plasma insulin and glucose did not change significantly; however, during the infusion of cimetidine a significant rise in plasma insulin and glucose levels occurred. The present data suggest that H2-receptors participate in an inhibitory mechanism with respect to the insulin and glucose response during the gastric phase of a neutral meal, but they do not seem to be involved in the rise in insulin and glucose observed in response to an acidified gastric meal.

Stimulation of hepatic regeneration after partial hepatectomy by infusion of a cytosol extract from regenerating dog liver.

A cytosol liver extract was prepared from adult dog livers and from liver remnants that had been regenerating for one, two and three days after 72 per cent partial hepatectomy. Given intraportally, the most active of these cytosols did not stimulate proliferation in the livers of normal dogs. However, infused during a six hour period into the portal vein of test group dogs, the cytosol from 48 and, especially, 72 hour regenerating livers augmented the regeneration response ordinarily produced by 44 per cent partial depatectomy. The effect was delayed. It became identifiable 48 hours after infusion and rached a peak at 72 hours. Neither augmentation nor significant inhibition of the normal regeneration response was produced by cytosol from normal liver and 24 hour regenerating liver or by six hour infusion of insulin. The amplification effect of active cytosol was equivocal when the infusions were given intraperitoneally and was not demonstrable at all by the intravenous route. In these investigations, it is confirmed that there are growth control factors in regenerating liver but the nature or physiologic significance of the factor or factors has not been clarified.

Sympathectomy and prostaglandin deficiency do not prevent gastrogenic hyperglycaemia and hyperinsulinaemia.

To examine the mechanism of the recently reported effect of an acidified intragastric test meal on insulin release and glucose homeostasis, a liver extract test meal at either pH 2 or pH 7 was instilled into the stomach of normal dogs and dogs with a chemical sympathectomy or indomethacin-induced prostaglandin deficiency, all of which had a bisected pylorus and gastric fistula. In the normal dogs the instillation of the liver meal at pH 2 elicited a significant rise in plasma glucose, glucagon and insulin levels, while in response to the meal at pH 7 only glucagon rose significantly. This was not altered in chemically sympathectomized dogs, nor during the infusion of indomethacin. In all experiments gastrin or gastric glucagon release in response to the meal at pH 2 was either lower than or similar to the response to the meal at pH 7. These data suggest that the influence of the stomach upon islet cell function and glucose homeostasis does not depend on either adrenergic innervation or the presence of prostaglandings, but rather is mediated by a yet undetermined mechanism.

Growth-stimulating factor in regenerating canine liver.

Extracts from dog livers which had been regenerating for 24, 48, and 72 h after hepatectomy were infused for 6 h into the left portal vein of animals which had fresh portacaval shunts (Eck fistula) and which were killed 2 and 3 days later. The brief exposure to the 48-h and especially the 72-h regenerating liver extracts induced a delayed proliferative response predominantly in the left liver lobes, with a slight spillover effect to the right liver lobes but none to the kidney. The response reached its peak 3 days later. In the left but not the right liver lobes, both the 48-h and the 72-h regenerating liver extract reversed the atrophy ordinarily caused by Eck fistula in 3 days and partly prevented the ultrastructural hepatocyte deterioration characteristic of Eck fistula. The active liver extracts apparently contained a growth-control factor or factors which is (are) not insulin or glucagon.

[Allogeneic hepatic cell suspension in the treatment of liver insufficiency]. / Allogennaia kletochnaia vzves' pecheni pri lechenii pechenochnoi nedostatochnosti.

A study was made of a possibility of using isolated hepatocytes for the treatment of diseases of the liver in animals. The optimal dose of the cell suspension in intravascular, intraperitoneal, intrapleural and subcutaneous administration was determined; a reaction of the experimental animals to this biological substrate was studied. The efficacy of the isolated hepatocytes in the treatment of hepatic insufficiency is demonstrated; also a comparative assessment of the mentioned methods of the cell suspension administration is given.

Properties of hepatic tissue extracts: 2. An approach to the artificial liver.

In the previous paper [3], the lethal effect of normal rabbit liver extract intravenously infused into rabbits was described. By comparing to the course of destruction of the liver tissue in fulminant hepatitis, the toxicity of the liver extract was recently studied. Attempts were also made to adsorbing the liver extract by activated clay and detoxifying the liver extract with a formaldehyde solution, and the results suggested that the findings would be applicable as an approach to the artificial liver.