Pulmonary cell response to bacterial challenge in mutant mice with some hereditary alterations resembling cystic fibrosis

Animal models make it possible to perform studies that normally can not be done in human beings. In the present work, the cellular response pattern of the lungs both to the environment and to a challenger strain of bacteria were analyzed in two mouse mutations-cribriform degeneration ($\text{cri}$) and motheaten (me)-. The mice were submitted to an infective aerosol containing $\text{Staphylococcus aureus}$ and were killed either immediately or 4 h after exposure; pulmonary washings were performed in these animals as well as in non-infected controls. The results showed neither quantitative differences in the total cell count nor in cell viability between the different groups. However, there were qualitative changes in the differential cell content characterized by a neutrophilic response in $\text{cri}$/$\text{cri}$ mice and a giant cell response in $\text{me}$/$\text{me}$ mice. Since most of the cystic fibrosis patients suffer from chronic lung infection, with a persistently high proportion of neutrophils, these findings point to the $\text{cri}$/$\text{cri}$ mouse as a promising animal model for cystic fibrosis studies.     

Kallikrein and amylase contents in tissues from a mutant mouse model for human cystic fibrosis

Kallikrein and amylase activities are decreased in the pancreas and salivary glands from $\text{cri/cri}$ homozygote mutant mice. Kallikrein is decreased in the $\text{cri/cri}$ kidney too. With reference to nucleic acid concentrations there is no difference between control and mutant mice. The previously described electrolyte abnormalities of the cribriform degeneration $\text{(cri)}$ mutant mouse, could be due to the abnormal activity of the kallikrein-kinin system on the transport mechanism of tubular cells in the organs mentioned. These findings represent a new step on our efforts to develop a useful animal model for human cystic fibrosis research.     

Administration of heparin causes in vitro release of non-esterified fatty acids in human plasma

The objective of this study was to investigate the extent to which $\text{in vitro}$ hydrolysis of endogenous triglycerides contributes to the elevated concentrations of non-esterified fatty acids (NEFAs) which have been reported after heparin administration. Heparin is known to induce the release of lipases which hydrolyze endogenous substrate both $\text{in vivo}$ and $\text{in vitro}$. Four patients undergoing diagnostic cardiac catheterization, who routinely receive heparin, were studied. Blood samples were obtained before and at 5 and 30 minutes after an intravenous bolus of heparin (46 U/kg) was administered. Determinations of NEFAs in plasma were carried out immediately and again various times after the samples had incubated at 24°C and at 0°C. In addition, an aliquot of each sample was frozen quickly, stored for 5–7 days, thawed, and incubated at 24°C for 180 minutes. As expected, there were no significant increases after incubation in the concentrations of NEFAs in the samples obtained before heparin administration. In contrast, in the samples obtained after heparin administration, incubation at 24°C produced significant increases in the concentrations of NEFAs. For example, in the plasma samples obtained 5 minutes after administration of heparin, concentrations of NEFAs increased 50, 160 and 300% after 5, 60, and 180 minutes of incubation compared to pre-heparin concentrations. When assayed immediately, the concentrations of NEFAs increased only 15% over pre-heparin concentrations. Incubating the samples at 0°C slowed lipase activity. Freezing the samples stopped the lipase activity; however, when the thawed samples were incubated at 24°C, concentrations of NEFAs continued to rise. This study suggests that much of the reported increases in the $\text{in vivo}$ concentrations of NEFAs after administration of heparin may be due to $\text{in vitro}$ formation from continued lipase activity on endogenous substrate. Moreover, studies relating increases in the concentrations of NEFAs after administration of heparin to changes in drug binding to plasma proteins should be re-examined for possible $\text{in vitro}$ artifacts.     

Fat absorption in cystic fibrosis mice is impeded by defective lipolysis and post-lipolytic events

Cystic fibrosis (CF) is frequently associated with progressive loss of exocrine pancreas function, leading to incomplete digestion and absorption of dietary fat. Supplementing patients with pancreatic lipase reduces fat excretion, but it does not completely correct fat malabsorption, indicating that additional pathological processes affect lipolysis and/or uptake of lipolytic products. To delineate the role of such (post) lipolytic processes in CF-related fat malabsorption, we assessed fat absorption, lipolysis, and fatty acid uptake in two murine CF models by measuring fecal fat excretion and uptake of oleate- and triolein-derived lipid. Pancreatic and biliary function was investigated by determining lipase secretion and biliary bile salt (BS) secretion, respectively. A marked increase in fecal fat excretion was observed in cftr null mice but not in homozygous DeltaF508 mice. Fecal BS loss was enhanced in both CF models, but biliary BS secretion rates were similar. Uptake of free fatty acid was delayed in both CF models, but only in null mice was a specific reduction in lipolytic activity apparent, characterized by strongly reduced triglyceride absorption. Impaired lipolysis was not due to reduced pancreatic lipase secretion. Suppression of gastric acid secretion partially restored lipolytic activity and lipid uptake, indicating that incomplete neutralization of gastric acid impedes fat absorption. We conclude that fat malabsorption in cftr null mice is caused by impairment of lipolysis, which may result from aberrant duodenal pH regulation.     

On the metabolic function of heparin-releasable liver lipase

Intravenous administration of specific antibody against heparin-releasable liver lipase (liver lipase) induced a 75% inhibition of the enzyme activity $\text{in situ}$. Administration of the antibody resulted in an increase of high density lipoprotein (density range 1.050–1.13 g/ml; HDL₂) phospholipid levels (20% after 1 h; 54% after 4 h). Short-term (1 h) treatment with antibody had no significant effect on any of the other lipoprotein components. After long-term (4 h) treatment the free cholesterol level of HDL₂ and all components in the very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL) fraction were elevated (1.5–2.0 fold). In the low density lipoprotein (LDL) fraction only the phospholipid level was affected (increased by 72%). All lipid components in the HDL₃ fraction were decreased by the antibody treatment, but this decrease was only statistically significant for the cholesterolesters. The rate of removal of iodine-labeled high density lipoprotein (HDL) and LDL from serum was not affected by the antibody treatment.These results suggest that liver lipase may promote phospholipid removal $\text{in vivo}$ and show that a lowering of liver lipase $\text{in situ}$ has profound consequences for serum lipoprotein metabolism.     

On the dual localization of lipoprotein lipase in rat heart. Studies with a modified perfusion technique

Rat hearts were perfused $\text{in vitro}$ using a modified $\text{Langendorff}$ technique, allowing the separate collection of coronary- and interstitial effluents. When heparin was added to the perfusion medium lipoprotein lipase was found in the coronary, as well as in the interstitial effluents. The relative amounts of lipase activity in both effluents varied with the feeding conditions of the animals, being high in the coronary effluent during fasting and high in the interstitial effluent during feeding. When glucagon (2.10^?7 M) was included in the perfusion medium, no differences between fasted and fed animals were obtained. The apparent K_m of the interstitial lipase was lower than that of the lipase found in the coronary effluent. The results are discussed in the light of the localization of lipoprotein lipase in rat hearts $\text{in situ}$.     

Effect of adding albumin to solutions of cholecystokinin on pancreatic protein output and pancreatic polypeptide release

In four conscious dogs with chronic gastric and pancreatic Thomas fistulas we studied the effect of 99% pure cholecystokinin-33 (CCK-33) solutions on pancreatic secretion and PP release. CCK-33 was dissolved in 0.154 M NaCl alone or in the same solution containing 1 g per 100 ml dog albumin. The response of pancreatic protein output to increasing doses of CCK-33 (0.5, 1, 2, 4 IDU/kg per h) was significantly $\text{(P < 0.05)}$ higher when CCK was dissolved in NaCl with albumin than in NaCl alone. These results were confirmed by measuring CCK immunoreactivity in samples from tips of infusion lines by a gastrin radioimmunoassay. Release of pancreatic polypeptide (PP) following increasing doses of CCK-33 was also significantly $\text{(P < 0.05)}$ elevated when CCK was dissolved in an albumin-containing solution. There was a significant $\text{(P < 0.02)}$ correlation between plasma concentrations of PP and pancreatic protein output.This study suggests that albumin should be added to CCK-33 solutions to preserve biological activity. The biological effect of CCK-33 may be substantially underestimated if albumin is omitted.     

Absence of Diabetes and Pancreatic Exocrine Dysfunction in a Transgenic Model of Carboxyl-Ester Lipase-MODY (Maturity-Onset Diabetes of the Young)

Background

CEL-MODY is a monogenic form of diabetes with exocrine pancreatic insufficiency caused by mutations in CARBOXYL-ESTER LIPASE (CEL). The pathogenic processes underlying CEL-MODY are poorly understood, and the global knockout mouse model of the CEL gene (CELKO) did not recapitulate the disease. We therefore aimed to create and phenotype a mouse model specifically over-expressing mutated CEL in the pancreas.

Methods

We established a monotransgenic floxed (flanking LOX sequences) mouse line carrying the human CEL mutation c.1686delT and crossed it with an elastase-Cre mouse to derive a bitransgenic mouse line with pancreas-specific over-expression of CEL carrying this disease-associated mutation (TgCEL). Following confirmation of murine pancreatic expression of the human transgene by real-time quantitative PCR, we phenotyped the mouse model fed a normal chow and compared it with mice fed a 60% high fat diet (HFD) as well as the effects of short-term and long-term cerulein exposure.

Results

Pancreatic exocrine function was normal in TgCEL mice on normal chow as assessed by serum lipid and lipid-soluble vitamin levels, fecal elastase and fecal fat absorption, and the normoglycemic mice exhibited normal pancreatic morphology. On 60% HFD, the mice gained weight to the same extent as controls, had normal pancreatic exocrine function and comparable glucose tolerance even after resuming normal diet and follow up up to 22 months of age. The cerulein-exposed TgCEL mice gained weight and remained glucose tolerant, and there were no detectable mutation-specific differences in serum amylase, islet hormones or the extent of pancreatic tissue inflammation.

Conclusions

In this murine model of human CEL-MODY diabetes, we did not detect mutation-specific endocrine or exocrine pancreatic phenotypes, in response to altered diets or exposure to cerulein.     

The in vitro effect of growth hormone on adipose tissue lipoprotein lipase in rats

The $\text{in vitro}$ effect of growth hormone on adipose tissue lipoprotein lipase was studied in rats. Epididymal adipose tissue was incubated with human growth hormone in the presence of heparin. Growth hormone at a concentration of 0.1 μg per ml decreased by approximately 20% (p<0.005) the heparin-releasable lipoprotein lipase in rat adipose tissue. Discussion was focussed on the reciprocal changes caused by growth hormone of the activities of lipoprotein lipase and hormone-sensitive lipase in the rat adipose tissue.     

Studies on the activity of brown adipose tissue in suckling,pre-obese, obob mice

The properties and activity of brown adipose tissue have been investigated in suckling, pre-obese, $\text{ob}\text{ob}$ mice in order to determine whether decreased thermogenesis in the tissue precedes the development of obesity in this mutant. At 14 days of age there was no difference between the $\text{ob}\text{ob}$ and normal animals in the total amount of interscapular brown adipose tissue, and the DNA content, protein content, and cytochrome oxidase activity of the tissue were similar in the two groups of mice. Respiration rates of brown adipose tissue mitochondria in the presence of albumin were, however, greater in the normal than the $\text{ob}\text{ob}$ animals, although after the addition of GDP to recouple the mitochondria there was no difference between the two groups. The mitochondrial membrane potential, measured with [³H]methyltriphenylphosphonium, was less affected by exogenous GDP in $\text{ob}\text{ob}$ mice than in normal animals. GDP binding to brown adipose tissue mitochondria, an index of the proton conductance pathway, was much greater in normal than in $\text{ob}\text{ob}$ mice at both 10 and 14 days of age; the decreased GDP binding in the mutant animals was found to result from a reduction in the number of binding sites. It is concluded that brown adipose tissue mitochondria of pre-obese $\text{ob}\text{ob}$ mice are more tightly coupled than those of normal siblings, and that the activity of the ‘thermogenic’ proton conductance pathway is lower in the mutant animals. A decrease in thermogenesis in brown adipose tissue is therefore an early event in the development of the $\text{ob}\text{ob}$ mouse and precedes the appearance of obesity.     

Rapid isolation of pancreatic islets from collagenase digested pancreas by sedimentation through Percol at unit gravity.

A Percoll solution with a density of 1.045 g/ml was used to separate pancreatic islets and exocrine tissue from collagenase-digested human and $\text{ob/ob}$ mouse pancreases by sedimentation at unit gravity. Most exocrine tissue from the mouse was found to range in density from 1.015 to 1.045 g/ml whereas the denser islets lay in a narrower range of 1.065–1.070 g/ml. Up to 400 islets were obtained from each mouse pancreas and 140 islets from 4 g of human pancreas; the isolated islets being essentially free from contamination with exocrine tissue. Glucose-stimulated insulin release was the same whether the mouse islets were isolated with or without Percoll. The simplicity of the method makes it suitable for large-scale islet isolation, a feature of potential importance for the treatment of diabetes by islet transplantation.     

Direct fluorometric analysis of benzo(a)pyrene metabolite formation by mouse liver microsomes

We have examined the metabolites produced by in vitro incubation of benzo(a)pyrene with 3-methylcholanthrene-induced mice liver microsomes. Our objective was to observe directly a possible difference in microsomal enzyme systems of animal models having different susceptibility to chemical carcinogens. The metabolites produced by the two animal models,$\text{C57BL}\text{6J}$ and $\text{DBA}\text{2}$ mice, were analyzed by a highly sensitive, “three-dimensional” fluorescence plotting technique. The fluorescence spectra of the total ethyl acetate-soluble metabolites clearly indicate that the metabolites produced by $\text{DBA}\text{2}$ enzymes were predominantly monohydroxylated benzo(a)pyrene while those produced by the liver microsomes of $\text{C57BL}\text{6J}$ were highly enriched with the 7,8-dihydrodihydroxybenzo(a)pyrene type.     

Evidence for dosage compensation of an X-linked gene in the 6-day embryo of the mouse.

The time at which dosage compensation of an X-linked gene in the mouse is established has been estimated by measuring the activity levels of two glycolytic enzymes, phosphoglycerate kinase (EC 2.7.2.3) and triosephosphate isomerase (EC 5.3.1.1), during early development of embryos from $\text{X}\text{X}$ and $\text{X}\text{O}$ mice. During preimplantation development the level of phosphoglycerate kinase in embryos from $\text{X}\text{X}$ mice was constant for the first 48 hr of development (2.55–2.71 nmoles/hr/embryo) and then dropped to one-half the earlier level (1.44 nmoles/hr/embryo) by 72 hr of development. The general developmental profile of phosphoglycerate kinase was similar in embryos from $\text{X}\text{O}$ mice; however, the absolute level of enzyme activity was reduced to approximately 1.4 nmoles/hr/embryo during the first 48 hr of development and to 0.9 nmoles/hr/embryo by 72 hr of development. These differences in phosphoglycerate kinase levels between embryos from $\text{X}\text{X}$ and $\text{X}\text{O}$ mice disappeared between the blastocyst and egg cylinder stages (150 hr postplug) during which time the activity levels had increased to 183 and 193 nmoles/hr/egg cylinder for embryos from $\text{X}\text{O}$ and $\text{X}\text{X}$ mice, respectively.The activity level for triosephosphate isomerase in embryos from $\text{X}\text{X}$ and $\text{X}\text{O}$ mothers did not differ at any stage of development; this suggests that the gene coding for triosephosphate isomerase is autosomal. In addition the level of activity remained constant during preimplantation development (approximately 3 nmoles/hr/embryo) and then, like phosphoglycerate kinase, increased 100-fold between the blastocyst and egg cylinder stages.The frequency distribution of the activity ratio (triosephosphate isomerase to phosphoglycerate kinase) in the egg cylinder of individual embryos from both $\text{X}\text{X}$ and $\text{X}\text{O}$ mothers was determined in order to detect differences among $\text{X}\text{X}\text{,}\text{X}\text{O}$ and $\text{X}\text{Y}$ embryos. These frequency distributions were unimodal, and hence these results coupled with the gene dosage differences observed during preimplantation development indicate that dosage compensation for an X-linked gene has been established in the 6-day mouse embryo.     

The phosphorylation of troponin B by phosphorylase b kinase in skeletal muscle of mice carrying the phosphorylase b kinase deficiency gene

In skeletal muscle of animals with the phosphorylase $\text{b}$ kinase deficiency gene there is < 1% of the normal activity to convert phosphorylase $\text{b}$ to $\text{a}$ in the presence of Ca⁺⁺, Mg⁺⁺, and ATP (1). Correspondingly, there is < 1% of the normal activity to phosphorylate phosphorylase $\text{b}$. Nevertheless, under the same conditions, these extracts catalyze the phosphorylation of troponin at a rate 57% of normal. Phosphorylase $\text{b}$ converting activity can be sedimented from skeletal muscle of control mice by centrifugation. This fraction isolated from I strain skeletal muscle extracts phosphorylates troponin at a rate 29–39% of the control. EGTA¹ (15 mM) inhibits troponin phosphorylation by 50–60% in this fraction from both strains. The EGTA inhibition is reversed by 15 mM Ca⁺⁺. Thus the phosphorylase $\text{b}$ kinase in skeletal muscle of animals with the phosphorylase $\text{b}$ kinase deficiency gene can phosphorylate troponin B, although it shows little or no activity with phosphorylase as a substrate. This observation is consistent with the normal muscle contractility of I strain animals.     

Pancreatic polypeptide response to ethanol in humans and dogs

We studied the effect of a drink of various concentrations of pure ethanol and several commonly ingested alcoholic beverages on plasma levels of immunoreactive pancreatic polypeptide in six healthy human volunteers and compared the results to a protein-rich meal. A drink of distilled water (250 ml) and of pure ethanol (250 ml or 125 ml in the case of 40% v/v ethanol) in concentrations (4, 10, 20, and 40%, v/v) normally present in beer, wine, liquor and whisky did not stimulate plasma pancreatic polypeptide levels above basal. Neither beer, red and white wine (250 ml each) nor whisky (125 ml) caused an increase in basal plasma pancreatic polypeptide levels. The 90-min integrated plasma pancreatic polypeptide response to the protein-rich meal was significantly reduced by an additional drink of 250 ml of white wine ($\text{5987 ± 1315}$ versus $\text{4126 ± 809}\text{pmol · min}{}^{\mathrm{?1}}\text{· 1}{}^{\mathrm{?1}}$). An intravenous infusion of ethanol (300 mg · kg^?1 over 30 min) did not increase plasma pancreatic polypeptide levels above basal.In six dogs with gastric and duodenal fistulas the infusion of pure ethanol into a peripheral vein, into the stomach or into the duodenum did not alter plasma pancreatic polypeptide levels. When ethanol (200 ml of either 1.8, 10 or 40%, v/v) was given as an intragastric bolus injection, only 40% ethanol caused an increase in the mean 90-min integrated plasma pancreatic polypeptide response which was only one-twelfth of the pancreatic polypeptide response to an oral mixed meat meal (35 g · kg^?1). We conclude that in man neither an intravenous infusion nor a drink of ethanol in concentrations normally present in beer, wine and whisky, release pancreatic polypeptide. Also, beer, red and white wine and whisky have no effect on plasma pancreatic polypeptide concentrations. In dogs, a large amount of intragastric ethanol was needed to produce a very small rise in plasma pancreatic polypeptide levels. These results do not favour the hypothesis that, in man and dog, pancreatic polypeptide is the hormonal mediator of the ethanol induced inhibition of exocrine pancreatic secretion.     

Abnormal intracellular lipid processing contributes to fat malabsorption in cystic fibrosis patients

A common feature of cystic fibrosis (CF) is the functional derangement of the exocrine pancreas, which affects output of pancreatic lipase. This condition results in severe dietary malabsorption due to the poor hydrolysis of triacylglycerol (TG) in the lumen of the small intestine. Despite the benefits of pancreatic enzyme supplements, patients with CF present with persistent intestinal fat malabsorption. The aim of the present investigation was to determine whether defects in the intracellular phase of lipid transport occur in this pathophysiology in addition to the known disturbed digestive processes. Our hypothesis was tested by incubating intestinal biopsies from six CF and six healthy subjects with radiolabeled lipid and protein precursors. Lipid esterification and secretion were markedly decreased by 22-31% and 38-42%, respectively, in CF samples, as noted by the low incorporation of [(14)C]palmitic acid into TGs, phospholipids, and cholesteryl esters in patients' duodenal explants and culture media compared with controls (100%). Accordingly, the output of TG-rich lipoproteins was substantially reduced (P < 0.05), and a similar trend was observed for high-density lipoproteins. Because intestinal lipoprotein assembly/secretion shows an absolute requirement for apolipoprotein (apo) B-48, radioactive labeling experiments were performed; these experiments demonstrated a significantly (P < 0.05) diminished synthesis of apoB-48 (40%) and apoA-I (30%). Given the critical role of microsomal triglyceride transfer protein in the formation of apoB-containing lipoproteins, its activity was determined and not found to be altered in CF intestinal tissue. Together, these results suggest that CF malabsorption may also be caused by defects in mucosal mechanisms leading to abnormal lipoprotein delivery into the blood circulation.     

LSD and phenithylamine hallucinogens: New structural analogy and implications for receptor geometry

The hallucinogen analog $\text{trans}$-2-(2,5-dimethoxy-4-methylphenyl)-cyclopropylamine (DMCPA) was resolved into its two optical isomers. Examination of selected behavioral profiles in mice and cats clearly showed that the levorotatory isomer of DMCPA possesses stereoselective activity when compared with the dextro isomer. The results parallel those obtained using the isomers of the known hallucinogen, DOM (STP) in the same animal models. Comparison of the optical rotatory dispersion (ORD) curves for the N-(5-bromosalicylidene) derivatives of DMCPA and $\text{trans}$-2-phenylcyclopropylamine (tranylcypromine) of known absolute configuration established the configuration of DMCPA to be (-)-1R,2S. This stereoselective activity and proof of absolute configuration lend strong support to a new model of the hallucinogen receptor. The proposed model suggests possible structural similarities between LSD and phenethylamine hallucinogens.     

Changes in the exocrine pancreas secondary to altered small intestinal function in the CF mouse

The exocrine pancreas of the cystic fibrosis (CF) mouse (cftr(m1UNC)) is only mildly affected compared with the human disease, providing a useful model to study alterations in exocrine function. The CF mouse pancreas has approximately 50% of normal amylase levels and approximately 200% normal Muclin levels, the major sulfated glycoprotein of the pancreas. Protein biosynthetic rates and mRNA levels for amylase were not altered in CF compared with normal mice, and increases in Muclin biosynthesis and mRNA paralleled the increased protein content. Stimulated pancreatic amylase secretion in vitro and in vivo tended to be increased in CF mice but was not statistically significant compared with normal mice. We show for the first time that the CF mouse duodenum is abnormally acidic (normal intestinal pH = 6.47 +/- 0.05; CF intestinal pH = 6.15 +/- 0.07) and hypothesize that this may result in increased signaling to the exocrine pancreas. There were significant increases in CF intestinal mRNA levels for secretin (310% of normal, P < 0.001) and vasoactive intestinal peptide (148% of normal, P < 0.05). Furthermore, CF pancreatic cAMP levels were 147% of normal (P < 0.01). These data suggest that the CF pancreas may be chronically stimulated by cAMP-mediated signals, which in turn may exacerbate protein plugging in the acinar/ductal lumen, believed to be the primary cause of destruction of the pancreas in CF.     

Hymenolepis nana: Immunogenicity of a lumen phase of the direct cycle and failure of autoinfection in BALBc mice

When $\text{BALB}\text{c}$ mice were given $\text{BALB}\text{c}$ mouse-derived cysticercoids (cysts) of Hymenolepis nana, only $\text{1}\text{43}$ mice became autoinfected, whereas most ($\text{31}\text{38}$) of dd mice given the same infection became massively autoinfected with mature worms. When $\text{BALB}\text{c}$ mice initially given cysts were challenged with eggs on Day 7, just before the patency of the primary infection, there was normal development into cysts, but almost none of them developed into adult worms. Thus, the failure of autoinfection of H. nana in $\text{BALB}\text{c}$ mice was not a result of failure of eggs to differentiate into cysts in the intestinal tissue, but a result of failure of these cysts to develop into adult worms in the lumen. The reasons why autoinfection does occur in dd and other strains of mice and not in the $\text{BALB}\text{c}$ strain are discussed in terms of the difference in onset of the late response in these strains of mice, ie., the response that is acquired after egg inoculation, and is directed against the lumen phase of cyst challenges. It is strongly suggested that (1) the lumen phase which follows cyst inoculation is highly immunogenic, but clearly differs from tissue phase which follows egg inoculation, (2) the autoinfection which occurs in some strains of mice is therefore not a result of no or poor immunogenicity of the lumen phase but is due to a delay of onset of the late response with the result that a secondary generation may mature, and (3) in other strains of mice, including $\text{BALB}\text{c}$, which acquire the late response within 15 days of initial egg inoculation, autoinfection normally does not occur after cyst infections.     

Intestinal calcium and phosphate transport in genetic hypophosphatemic mice

Serum phosphate, serum calcium, intestinal phosphate and intestinal calcium transport were measured in normal ($\text{C57BL}\text{6J}$ +/Y) and genetic (X-linked) hypophosphatemic mice ($\text{C57BL}\text{6J}\text{Hyp}\text{Y}$). The hypophosphatemic mice had low serum phosphorus levels and dramatically decreased intestinal phosphate transport compared with normal controls. On the other hand, normal and hypophosphatemic mice had equivalent levels of intestinal calcium transport. The hypophosphatemic mice did illustrate a slightly decreased serum calcium concentration, however. Administration of 1,25-dihydroxy-vitamin D₃, the principal active metabolite of vitamin D, stimulated intestinal calcium transport but not intestinal phosphate transport in the genetic hypophosphatemic mice. The results obtained are consistent with the hypothesis that the primary metabolic disturbance in familial hypophosphatemia involves a defect in phosphate transport mechanisms.