Iron deficiency down-regulates the Akt/TSC1-TSC2/mammalian Target of Rapamycin signaling pathway in rats and in COS-1 cells

Iron deficiency (ID) is one of the most commonly known forms of nutritional deficiencies. Low body iron is thought to induce neurologic defects but may also play a protective role against cancer development by cell growth arrest. Thus, ID may affect cellular pathways controlling cell growth and proliferation, the mechanism of which is still not fully understood. The serine/threonine protein kinase Akt and its downstream target, the mammalian Target of Rapamycin (mTOR), is known to play a crucial role in the regulation of cell growth and survival. Therefore, we hypothesized that Akt/mTOR pathway could be influenced by ID. Three-week-old male Wistar-strain rats were divided into 3 groups and the 2 groups had free access to a control diet (C group) or an iron-deficient diet (D group). The third group (PF group) were pair-fed the control diet to the mean intake of the D group. After 4 weeks, rats were killed and their brains were sampled. In separate experiments, COS-1 cells were cultured with or without the iron chelator deferoxamine. Western blots of brain samples and COS-1 lysates were used to analyze the expression and phosphorylation state of Akt, TSC2, mTOR, and S6 kinase proteins implicated in the Akt/mTOR pathway. Using 2 different ID models, we show for the first time that iron deficiency depresses Akt activity in rats and in COS-1 cells, leading to a decrease in mTOR activity.     

Significance of prenatal diagnosis in a patient with a huge neck tumor.

Due to the great advancements in fetal ultrasonography, the number of prenatal diagnoses are increasing, greatly contributing to improved neonatal surgery. It is now relatively easy to detect huge fetal cystic masses, and we have experienced three cases with a huge neck tumor detected in utero, one case with teratoma and two cases with cystic hygroma. Each case was complicated by neonatal asphyxia, and the neonate needed resuscitation by means of endotracheal intubation. The infant with teratoma unfortunately died of respiratory distress due to compression of the trachea before a perinatal team could be organized. Although the remaining cases with cystic hygroma were treated by a perinatal team, one died 19 hours after birth and the other has survived with the aid of endotracheal intubation in the hospital for three years. In addition, all four cases of cystic hygroma detected antenatally in our institute, which were not delivered, also had fetal hydrops which suggested a general lymphatic derangement. Cystic hygroma detected in utero is considered to be different from that detected after birth, since the former is associated with genetic lymphatic derangement. Prenatal diagnosis enables such patients to survive the perinatal period, but may not improve the prognosis of fetal cystic hygroma so much.     

Tinea faciei caused by Microsporum canis in a newborn

A case of tinea faciei caused by Microsporum canis in a 14-day-old infant is reported. The incubation period was seen to be 1 week. This was a familial infection which also affected the infant's grandmother and their pet cat and dog. Topical treatment with clotrimazole controlled the infection in the baby. A review of 14 cases (including our own) of dermatophytosis in newborn infants reported in the Japanese literature showed that newborn infants might be infected by several of these agents.     

Evaluation of image-diagnosing methods of enlarged parathyroid glands in chronic renal failure

Three noninvasive image-diagnosing methods, computed tomography (CT), scintigraphy with²⁰¹T1C1 and^99mTcO_h4 ^–, and ultrasonography (US), were preoperatively performed on 50 patients with chronic renal failure and secondary hyperparathyroidism who underwent total parathyroidectomy and parathyroid autograft. The detection rates of the 3 methods on the 191 excised parathyroid glands were compared according to weight and location. CT detected 57.1% of all glands and 78.6% of 103 glands weighing over 500 mg. Scintigraphy detected 51.8% and 75.7%, and US detected 42.4% and 53.4%, respectively. The detection rate of upper glands was best with CT at 58.9% and 89.1%; that of lower glands was best with scintigraphy at 65.3% and 80.4%. Although the combination of the 3 methods diagnosed 69.6% and 89.5%, CT and scintigraphy, the best 2 combinations, visualized 67.5% and 88.3%.

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Resumen Tres métodos diagnósticos no invasivos, la tomografía computadorizada (TC), la centelleografía con²⁰¹T1C1 y^99mTcO₄ y la ultrasonografía (US) fueron realizados preoperatoriamente en 50 pacientes con falla renal crónica e hiperparatiroidismo secundario sometidos a paratiroidectomía y autotransplante paratiroideo. Las tasas de detección de los 3 métodos fueron comparados sobre las 191 glándulas paratiroideas resecadas en relación a los pesos y a los sitios de ubicación. La TC detectó el 57.1% del total de glándulas y el 78.6% de aquellas glándulas (103) con pesos superiores a 500 mg. La centelleografía detectó 51.8% y 75.7%, y la US 42.4% y 53.4% respectivamente. La tasa de detección para las glándulas superiores fue optima con TC, con 58.9% y 89.1%; la de las glándulas inferiores fue óptima con centelleografía, con 65.3% y 80.4%. Aunque la combinación de los 3 metodos diagnosticó el 69.6% y 89.5%, la TC y la centelleografía, la mejor de las combinaciones, visualizó el 67.5% y el 88.3% respectivamente.

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Résumé Trois méthodes d'imagerie non invasives, la tomodensitométrie, la scintigraphie (avec T1C1²¹⁰ et TcO₄ ^99m), et l'ultrasonographie ont été pratiquées avant l'intervention chez 50 malades qui présentaient une insuffisance rénale chronique compliquée d'hyperparathyroïdisme secondaire et qui furent traités par parathyroïdectomie totale et autogreffe parathyroïdienne. Les taux de détection de ces 3 méthodes concernant 191 glandes parathyroïdes réséquées ont été évalués en fonction du poids et du siège des lésions. La tomodensitométrie a permis de découvrir 57.1% de toutes les glandes et 78.6% des glandes dont le poids dépassait 500 mg; la scintigraphie 51.8% et 75.7%; l'ultrasonographie 42.4% et 53.4%. Le taux de détection des glandes supérieures fut plus élevé avec la tomodensitométrie: 58.9% et 89.1%; celui des glandes inférieures le fut avec la scintigraphie: 65.3% et 80.4%. Si la combinaison des 3 méthodes permet le diagnostic dans 69.6% et 89.5% des cas la tomodensitométrie associée seulement à la scintigraphie donne des résultats très voisins, les taux respectifs étant de 67.5% et de 88.3%.

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Presented at the International Association of Endocrine Surgeons in Paris, September 1985.     

Effects of simultaneous increases in dietary phosphorus and magnesium concentrations on nephrocalcinosis and kidney function in female rats.

The effects of simultaneous increases in dietary phosphorus (P) and magnesium (Mg) concentrations while maintaining a constant P:Mg ratio on nephrocalcinosis and kidney function in female rats was investigated. Female Wistar rats were fed a control diet (3.12 g P, 0.51 g Mg per kg diet) or a diet having either 3 times the control P and Mg concentrations (3-fold diet; 9.25 g P and 1.42 g Mg per kg diet) or 5 times the control concentrations (5-fold diet; 14.97 g P and 2.37 g Mg per kg diet) for 21 d. The three experimental diets all had same P:Mg molar ratios (control diet; 4.81, 3-fold diet; 5.11, 5-fold diet; 4.96). The 3-fold diet had no significant influence on kidney calcium (Ca), Mg or P concentrations. However, kidney Ca, Mg and P concentrations were significantly higher in rats fed the 5-fold diet than in rats fed the control or 3-fold diets. No significant differences in creatinine clearance were observed among the three groups. Urinary albumin and beta 2-microglobulin excretion were higher in rats fed the 5-fold diet than in rats fed the control or 3-fold diets, while the 3-fold diet had no significant influence on the urinary albumin and beta 2-microglobulin excretion. These results suggest that absolute concentrations of dietary P and Mg are important factors with regard to the development of nephrocalcinosis and diminished kidney function.     

Excitation of squid giant axons in hypotonic and hypertonic solutions

Excitability of intracellularly perfused squid giant axons was maintained in hypotonic solutions (down to 300 mOSM) and in hypertonic solutions (up to about 10 OSM), when osmolalities of internal and external solutions were adjusted to be equal with glycerol, glucose, or sucrose. Molar concentrations of ions were kept constant during one series of experiments. The resting potential and the amplitude of the action potential did not change in both hypotonic and hypertonic solutions. With reduction of osmolality, the duration of action potential decreased and the maximum rate of rise and conduction velocity increased. By raising osmolality, the duration was prolonged and the maximum rate of rise and the conduction velocity decreased. Effects of osmolality change were almost reversible. However, these effects were not directly related to the osmolality change but seemed to be related to the viscosity change of the solutions. When the osmolality of external solution was raised with NaCl (up to 2.6 M NaCl), the overshoot increased in porportion to the logarithm of the NaCl concentration. The slope of increase was about 50 mV/decade. However, the resting potential showed little change. With increase of the NaCl concentration, the duration of the action potential increased.     

Gastric juvenile polyposis associated with germline SMAD4 mutation

We treated a 39-year-old woman with hypoproteinemia and anemia who had profuse gastric polyposis. Radiographic and endoscopic examination showed numerous polyps restricted to the stomach. The patient had pulmonary arteriovenous malformations in the left lung. Histological examination of the resected stomach revealed the gastric polyposis to be composed of cystic dilatation of the glands with small areas of adenocarcinoma. These findings were compatible with gastric juvenile polyposis (GJP) accompanied by gastric cancer. Analysis of genomic DNA revealed that the patient had truncating mutation of SMAD4, a responsible gene for juvenile polyposis (JP). Our case suggests that SMAD4 is possibly a responsible gene for GJP.     

Osteoclast-like cells formed in long-term human bone marrow cultures express a similar surface phenotype as authentic osteoclasts

Long-term cultures of human bone marrow form multinucleated cells (MNC) with many functional characteristics of osteoclasts including: expression of tartrate-resistant acid phosphatase, appropriate responses to osteotropic hormones, calcitonin-induced contraction and formation of resorption lacunae on calcified matrices. However, it is unclear if these cells express similar surface antigens as expressed by authentic osteoclasts, since they form on plastic surfaces in the absence of bone. Bone may be required to complete the differentiation process for osteoclasts. Therefore, we have examined the surface phenotype of MNC and compared it with that of osteoclasts freshly isolated from bone, to determine if MNC express similar surface antigens, and if MNC express antigens which identify their cellular origin. Similar to bone-derived osteoclasts, MNC formed in long-term human bone marrow culture expressed osteoclast-specific antigens (detected by monoclonal antibodies 13c2 and 23c6) and did not express Fc receptors, T cell specific antigens, most myeloid antigens or mature macrophage antigens. In contrast to authentic osteoclasts, MNC reacted with a monoclonal antibody (Mol) which identifies an antigen present on myeloblasts, monocytes, granulocytes, and null cells from human peripheral blood and bone marrow. MNC also reacted with the monoclonal antibody My11, which is present on CFU-GM, the granulocyte-macrophage colony-forming cell, the probable precursor for MNC. These data demonstrate that MNC formed in long-term human marrow cultures express a similar surface phenotype to osteoclasts. This phenotype is different from that expressed by macrophage polykaryons. In addition, MNC also expressed monocyte-related antigens (My11, Mol), suggesting that are derived from or related to the monocytic lineage.     

Demonstration of amelogenin in the enamel-free cusps of rat molar tooth germs: immunofluorescent and immunoelectron microscopic studies.

The enamel-free cusps of 1-4 day-old rat mandibular first molars were investigated using the monoclonal antibody En3 against rat amelogenin at light and electron microscopic levels in order to clarify whether the enamel-free cusp is virtually devoid of enamel. At 1 day after birth, there were presecretory ameloblast-like cells (PALCs), which were short and were not polarized, at the cusp tips. They were close to the outer enamel epithelium. Hematoxylin positive enamel matrix was not distinctly observed in the enamel-free cusp by light microscopy, but almost continuous immunofluorescence for amelogenin was detected at the interface between PALCs and dentin. The penetration of immunopositive material toward the dental pulp was also observed in the enamel-free cusp. At 4 day after birth, both in the frontal section and in the horizontal section, almost continuous immunofluorescence was recognized at the interface between PALCs and dentin in the enamel-free cusp. The penetration of amelogenin toward the dental pulp was not seen in the enamel-free cusp. By immunoelectron microscopy, immunolabelling was recognized in the Golgi apparatus of PALCs, in a layer of amorphous material at the interface between PALCs and dentin, and in stippled material-like substance in the intercellular space between PALCs. Although no basement membrane was observed beneath PALCs, they did not have Tomes' processes. These investigations suggest that PALCs in the enamel-free cusp differentiate into the secretory cells and that they can synthesize and secrete the amorphous material containing amelogenin at the interface between PALCs and dentin. The penetration of amelogenin toward the dental pulp might play a role in the interaction between PALCs and odontoblasts in the enamel-free cusp and/or the initiation of mineralization of predentin.