Variations in WGA-binding proteins in tumours growing in different implantation sites

The protein/glycoprotein composition of Triton X100 extracts of four metastatic tumour lines have been compared by gel electrophoresis after the implantation into different sites of the body. No consistent differences in the patterns were noted after staining with Coomassie Blue or treating with concanavalin A. In contrast, after treating with wheat germ agglutinin (WGA), the patterns varied with site. This was most marked with the ML and M5 tumours, where the overall binding was much lower for some sites. Although WGA binding with the LL and BL6 tumours was generally higher than with the ML and M5, reduced binding was still evident for some sites. Reduced WGA binding was also observed for extracts of secondary tumour growing in the liver or lungs.     

Hyperkalaemia in patients in hospital.

Significant hyperkalaemia occurred in 406 out of 29 063 patients admitted to a major Scottish teaching hospital in one year (1.4%). Mortality was higher in these patients than in control patients and was strongly correlated with the severity of the hyperkalaemia. Overall seven deaths were directly due to hyperkalaemia (out of 58 deaths among patients with hyperkalaemia). Factors contributing to a poor prognosis were severity and speed of onset of hyperkalaemia and the presence of appreciable renal impairment. Patients with hyperkalaemia were older and more likely to be male; this trend was present in all diagnostic subcategories. Genitourinary disease, gastrointestinal disease, and cancer were significantly more common among the patients with hyperkalaemia than the controls. Hyperkalaemia due to drug treatment was invariably mild and non-fatal, whereas genitourinary disease was often associated with moderate to severe hyperkalaemia, which in two cases proved fatal. Use of electrocardiographic monitoring was rare, and although the treatment of hyperkalaemia was effective, it was often used when not required. Hyperkalaemia is a potential hazard in diabetic ketoacidosis, and use of potassium supplements should be carefully monitored during correction of the acidosis.     

国内双歧杆菌制剂预防小儿继发性腹泻临床疗效的Meta分析

目的系统评价国内双歧杆菌制剂临床预防小儿继发性腹泻的效果。方法按照系统评价的要求检索CBMd isc、VIP、CNK I以及万方数据库等,获得18篇符合纳入标准的文献,共计患儿4050例,对其进行M eta分析,并评价M eta分析结果的稳定性和发表偏倚。结果异质性检验χ^2=34.60,P=0.007〈0.05,采用随机效应模型进行M eta分析,合并RR=0.41,95%C I为0.35～0.49,总体效应检验,Z=10.39,P〈0.00001,差异具有非常显著性,固定效应模型RR值和95%C I与随机效应模型完全一致,剔除小样本报道后的合并RR=0.42,95%C I为0.35～0.50,与剔除前的结果基本一致,且本研究的发表偏倚得到了很好地控制。结论从现有的临床证据来看,双歧杆菌制剂能降低小儿继发性腹泻的发生率,对预防小儿继发性腹泻起到了满意的效果。     

Micronuclei in human lymphocytes irradiated in vitro or in vivo

Venous blood from healthy donors or from patients with various lympho- and myeloproliferative diseases was incubated in vitro in the presence of cytochalasin B for the induction of binucleated lymphocytes. The time at which cytochalasin B was added depended on the proliferation rate of the lymphocytes. Proliferation was monitored using a semiautomatic microscope photometer/computer system. The background level of micronuclei in binucleated lymphocytes of the patients before radiotherapy was statistically indistinguishable from that of healthy persons. Blood from both groups was irradiated in vitro for the study of the dose-response relationship. The dose-response curves were very similar up to 3.75 Gy, and a somewhat lower micronucleus frequency was found in lymphocytes of patients after a 5-Gy exposure. These in vitro results were compared with in vivo exposure after total-body irradiation of leukemic patients. Due to heavy medication that accompanied radiation therapy, only two doses (1.25 and 2.5 Gy) could be checked after in vivo exposure. There was no statistically significant difference between in vitro and in vivo results after 1.25 Gy, but a slightly lower number of micronuclei was observed after in vivo exposure to 2.5 Gy.     

Estrogen metabolism in hyperthyroidism and in cirrhosis of the liver.

Estrogen metabolism was studied in spontaneous hyperthyroidism (Graves disease) and in alcoholic cirrhosis of the liver. The plasma concentration of estradiol-17beta (PCE2) was increased in men with hyperthyroidism. Although the metabolic clearance rate of estradiol-17beta (MCRE2) was reduced, the production rate (PR) of the steroid was increased above normal. The MCRE2 was also decreased in women with hyperthyroidism but the PCE2 and PRE2 was unchanged from normal. The conversion ratio of estradiol-17beta (CRE2E1) was increased in both hyperthyroid men and women. The PCE2 was significantly increased in men with cirrhosis of the liver. The MCRE2 was normal and this resulted in an increase in the PRE2 in this disorder. The CRE2E1 was significantly higher than normal. The plasma concentration of estrone (E1) was elevated in men with both disorders.     

Iminoglycinuria in a child in Czechoslovakia

Summary The child with iminoglycinuria is in our observation. Hyperprolinuria was seen at 5 months by screening program. The child was repeatedly examined in the hospital and was seen last time at 16 months. IQ was 67, in the urine were excessive amounts of glycine, proline a hydroxyproline. In the blood aminoacids were in normal levels. In the child was noticed the same increase of proline in the blood as in the control child of the same age following loading test with L-proline, indicating normal intestinal absoption. Both parents and father's sister's 2-year-old mentally retarded child exhibit excessive glycinuria. The father, his sister, father's father, and grandfather are partialy, congenitaly deaf.     

EPID in vivo dosimetry in RapidArc technique

Aim

The aim of the study was to estimate the dose at the reference point applying an aSi-EPID device in the course of patient treatment.

Materials and methods

The method assumes direct proportionality between EPID signal and dose delivered to the patient reference point during the treatment session. The procedure consists of treatment plan calculation for the actual patient in the arc technique. The plan was realized with an elliptic water-equivalent phantom. An ionization chamber inside the phantom measured the dose delivered to the reference point. Simultaneously, the EPID matrix measured the CU distribution. EPID signal was also registered during patient irradiation with the same treatment plan. The formula for in vivo dose calculation was based on the CU(g) function, EPID signal registered during therapy and the relation between the dose and EPID signal level measured for the phantom. In vivo dose was compared with dose planned with the treatment planning system.Irradiation was performed with a Clinac accelerator by Varian Medical Systems in the RapidArc technique. The Clinac was equipped with an EPID matrix (electronic portal image device) of aSi-1000. Treatment plans were calculated with the Eclipse/Helios system. The phantom was a Scanditronix/Wellhöfer Slab phantom, and the ionization chamber was a 0.6 ccm PTW chamber.

Results

In vivo dose calculations were performed for five patients. Planned dose at the reference point was 2 Gy for each treatment plan. Mean in vivo dose was in the range of 1.96–2.09.

Conclusions

Our method was shown to be appropriate for in vivo dose evaluation in the RapidArc technique.     

Gene organization in the region containing a new gene involved in chromosome partition in Escherichia coli.

A new mutation, parC, causing abnormal chromosome segregation was identified in two thermosensitive mutants of Escherichia coli. The thermosensitive growth of the mutants was corrected by pLC4-14 in the Clarke-Carbon collection. This plasmid carries a putative gene which can suppress the cell division defect due to ftsI (pbpB) and has hence been termed sufI (sui). The nearness of parC to metC was confirmed, and cotransduction frequency of parC was 59% with metC and 20% with glc. The parC-sufI region was analyzed by subcloning the chromosome region of pLC4-14. The parC and the sufI gene products were electrophoretically identified as proteins of 75 and 55 kilodaltons (kDa), respectively. The allelism of parC+ on pLC4-14 to parC1215 was confirmed by cloning parC1215. The sufI gene appeared to be dispensable for cell viability, and overproduction of its product caused suppression of ftsI. An essential gene coding for a 25-kDa protein was found between the parC and the sufI gene. These three genes were transcribed in the same direction and may be organized into an operon, with parC to the proximal side and with internal promoters at least for the distal genes. The localization of the gene products was examined in maxicells. The sufI protein was synthesized as a precursor which could be chased into a mature form. The major part of the mature form was found in the soluble fraction. The 25-kDa protein was found almost exclusively in the membrane fraction. The parC protein was associated with the membrane fraction in the presence of Mg2+ but found in the soluble fraction when Mg2+ was sequestered with EDTA.     

Parasympathetic involvement in PAF-induced contraction in canine trachealis in vivo

We studied the contractile response elicited by platelet-activating factor (PAF) administered intra-arterially into the tracheal circulation of 34 dogs in vivo. A method that avoided tachyphylaxis encountered in prior investigations was developed for isometric measurement of multiple dose-response effects. PAF was a very potent contractile agent; active tension was elicited with 10(-11) mol ia PAF. To determine the mechanism by which contraction was induced, dose-response curves were generated in groups of five animals each treated with either 0.5 mg/kg (approximately 1.5 X 10(-5) mol) iv + 10(-3) mg/kg (3 X 10(-8) mol) ia atropine, 5 mg/kg iv indomethacin (INDO), or 7.5 mg/kg iv hexamethonium (HEX). After pretreatment with atropine, contraction still was elicited with 10(-11) mol ia PAF. However, maximal contraction was only 16.2 +/- 2.74 g/cm (vs. 35.7 +/- 5.74 g/cm for untreated controls; P less than 0.02). The dose at which maximal contraction was elicited after atropine was 10(-7) mol ia (vs. 1.9 X 10(-9) mol for controls; P less than 0.001). Pretreatment with INDO caused minimal attenuation, and HEX had no effect on the response elicited by ia PAF. We demonstrate a method for assessing the effects of PAF in central airways that avoids tachyphylaxis and permits dose-response studies in the same animal. We also demonstrate that PAF is an extremely potent mediator that elicits tracheal smooth muscle contraction at least in part by postganglionic activation of parasympathetic nerves. A direct contractile effect of PAF which is not related to secretion of products of the cyclooxygenase pathway is also suggested.     

Absorption of hexose and pentose sugars in vivo in perfused intestinal segments in the fowl.

1. Rates of absorption of two hexose (D-glucose and D-galactose) and two pentose (D-xylose and D-arabinose) sugars were measured by in vivo perfusion, in jejunum, ileum and (distal) caecum, in immature hens conditioned to either a standard (ST) or "high fibre" (ST + 20% grass) diet. 2. Each bird was tested in one intestinal segment with all four (U-14C-labelled, 10 mM) sugars, with either the hexoses preceding the pentoses or vice versa. 3. With all treatments, absorption rates of the hexoses were alike, as were those of the pentoses. Hexose absorption was twice as fast as pentose absorption in jejunum and ileum with both dietary pretreatments, whereas in caecum hexose and pentose rates were similarly high, except when pentose (and its associated fluid transfer) was apparently inhibited by prior hexose absorption with the ST diet. 4. With the ST diet, hexose absorption (per unit length and dry weight) was faster in caecum than in jejunum and ileum, and pentose absorption was also fastest in caecum when all pentose data from testing after hexose were excluded. 5. With the ST/grass diet, hexose absorption was faster in jejunum than in ileum and caecum when expressed per unit length, and pentose absorption was fastest in caecum on a dry weight basis. 6. Hexose absorption was faster in jejunum and slower in caecum with the ST/grass pretreatment than with ST. However, the dietary comparison was not conclusive because it involved birds form (two) different hatches (of similar age and weight) tested at different times.     

Seasonal distribution of Vibrio parahaemolyticus in freshwater environs and in association with freshwater fishes in Calcutta

The seasonal distribution of Vibrio parahaemolyticus in freshwater environs and in association with freshwater fishes was studied in 1982 and 1983. The occurrence of this organism in water and sediments at the three sites studied was very infrequent and was restricted to the summer months, although it was not always isolated during these months. The association of V. parahaemolyticus with plankton was chiefly confined to the summer months and progressively declined with the onset of monsoons, remaining below detectable levels during the postmonsoon and winter months. The incidence and counts of V. parahaemolyticus were consistently higher in association with plankton than with water and sediment samples. V. parahaemolyticus could be recovered throughout the period of investigation from freshly caught and market samples of freshwater fishes. The highest recovery rate of this halophile from fishes was invariably from fecal samples. Most of the strains isolated in this study were untypable, and those which could be typed were predominantly serotypes encountered in the environment. All the isolates were Kanagawa negative. From this study, it could be concluded that the survival of V. parahaemolyticus in freshwater ecosystems is transient and dependent on a biological host.     

Orthobunyaviruses in the Caribbean: Melao and Oropouche virus infections in school children in Haiti in 2014

We report the identification of two orthobunyaviruses, Melao virus (MELV) and Oropouche virus (OROV), in plasma specimens from Haitian children with acute febrile illness who presented during outbreaks caused by alpha- and flaviviruses in 2014. Heretofore not described as a human pathogen, MELV was isolated in cell culture from the plasma of five case patients. OROV RNA was detected in the plasma of an additional child, using an unbiased sequencing approach, with phylogenetic inference suggesting a close relationship with strains from Brazil. Abdominal pain was reported by four case patients with MELV infections, with lymphadenopathy noted in two cases. Our findings document the occurrence of these orthobunyaviruses within the Caribbean region and highlight the critical importance of surveillance with viral genome sequence analyses to identify outbreaks caused by these and other emerging viruses.     

Apoptosis in experimental myocardial infarction in situ and in the perfused heart in vitro

We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions.     

The role of carbohydrates in sperm-egg interaction in rats

The first step in fertilization is the interaction of the capacitated sperm with the zona pellucida. It has been proposed that the initial interaction, as in other types of cell adherence, is due to complementary interacting sites on the opposing surfaces of the gametes. This work intended to investigate the role of carbohydrates in sperm-egg binding in rats. Ejaculated sperm was collected from uterine horns of mated females. The sperm was suspended in Rat Fertilization Medium at final concentrations of 3-7 times 10(5) sperm/ml. After 5 1/2 h of sperm incubation, eggs were added to the sperm suspensions concomitantly with various carbohydrates to achieve a final concentration of up to 50 mM. The eggs were separated after 30 min, and the number of sperm bound to the zona pellucida was counted. Among a variety of monosaccharides tested at 50 mM concentration, it was found that alpha-methyl-mannoside was the most potent inhibitor (producing 80% inhibition); less potent was D-mannose and even less, L-fucose. A combination of alpha-methyl-mannoside and L-fucose showed a synergistic effect. Mannan was not more effective as an inhibitor than the monosugar mannose, while fucoidin was extremely potent, causing over 90% inhibition of binding at 0.1%. We assume the presence of macromolecules containing sugars on the zona pellucida because inhibition of sperm binding to this layer was observed: a) after preincubation of mannan or fucoidin with sperm, but not with the eggs; and b) after pretreatment of the egg with specific enzymes. The results obtained in this study in the rat are consistent with the hypothesis that carbohydrates are critical for the sperm-egg interaction.     

Toxoplasmosis in Sheep

The presence of Toxoplasma gondii in the diaphragm was correlated with the dye test titres in 174 sheep. In 94 of these the presence of the parasite was also correlated with the haemoglobin (Hb) type. T. gondii was recovered from 3 % of the sheep with titres < 1/16, compared with 30 % of those with titres 1/16 and 70 % of those with titres ≥ 1/32. The results indicate that the distinction between serologically positive and negative individuals at a final serum dilution of 1/16 is justified. Some evidence was found that the parasite is easier to recover from dye test positive mature sheep than from dye test positive lambs. Of the 174 sheep, 143 were examined at random among 186 sheep culled or cast for age during a 4-year period from 1 flock in which the prevalence of Toxoplasma antibodies was representative for flocks in the southern Norway, and T. gondii was recovered from 53 (37 %) of these. It was concluded that 10—15 % of the lamb carcases and 25—37 % of the carcases from mature sheep in this country have T. gondii in their muscles detectable by the peptic digestion technique. A possible genetical influence on the infection was indicated by the higher frequency of recoveries of T. gondii from sheep with Hb type B than from sheep with the Hb types A or AB, but the number of individuals with Hb type B was too small to demonstrate statistically significant differences. The epidemiological importance of infected sheep carcases is discussed.     

Inhalation anesthesia in adult cattle

An inhalation technique was used for anesthesia during ileal cannulation in five adult cows. Following sedation with intravenous acepromazine, anesthesia was induced intravenously with thiopental sodium in 5% glyceryl guaiacolate solution. Endotracheal intubation was performed and anesthesia maintained with halothane in oxygen via a circle system with a precision vaporizer. In all cases, induction was smooth and no difficulties were experienced during the maintenance of anesthesia. Total anesthesia time was 1.5 to 2.5 hours. Following completion of the surgical procedure, which was performed with the animal in left lateral recumbency, each cow was rolled to a sternal position and supported, if necessary. The endotracheal tube was left in place, with oxygen administration continued, until the animal was able to swallow. Recoveries were rapid and all animals were ambulatory within 30 minutes after completion of the surgery. The only post-operative complication due to anesthesia was transient mouth soreness in two cases, attributed to the use of a mouth speculum during intubation.     

Poikilocytosis in Cancer Patients

The incidence of poikilocytosis in 100 patients with cancer of various organ systems was found to be 12 per cent. Poikilocytosis was more frequently seen in patients with adenocarcinoma of the gastrointestinal tract and with inoperable metastatic carcinoma than in patients with other malignant lesions. Anemia was associated in three cases, but in nine instances the patient was not significantly anemic at admission. Two of these patients may have been dehydrated. Uremia was not a factor in any of the cases.Fifty per cent of the patients exhibiting poikilocytosis died or were in a terminal condition within one month after the observation was made, as compared with 21 per cent of the patients who did not have evidence of poikilocytosis. These findings show that poikilocytosis may occur in cancer patients without anemia or uremia; and they indicate that its presence is an unfavorable prognosis sign.     

Importance of guanine nitration and hydroxylation in DNA in vitro and in vivo

Guanine (Gua) modification by nitrating and hydroxylating systems was investigated in DNA. In isolated calf thymus DNA, 8-NO(2)-Gua and 8-oxo-Gua were dose-dependently formed with peroxynitrite, and 8-NO(2)-Gua was released in substantial amounts. Myeloperoxidase (MPO) with H(2)O(2) and NO(2)(-) reacted with calf thymus DNA to form 8-NO(2)-Gua dose dependently without release of 8-NO(2)-Gua. The frequency of strand breaks was higher than the sum of 8-NO(2)-Gua and 8-oxo-Gua, particularly in the MPO-treated DNA, indicating the importance of other types of damage. The activation of human neutrophils and lymphocytes with phorbol ester did not induce 8-NO(2)-Gua and 8-oxo-Gua in their nuclear DNA. However, 8-NO(2)-Gua was found in calf thymus DNA co-incubated with activated neutrophils in the presence of NO(2)(-). No significant formation of 8-NO(2)-Gua was found in liver DNA from mice treated with Escherichia coli lipopolysaccharide. The incubation of peroxynitrite or MPO-H(2)O(2)-NO(2)(-)-treated DNA with formamidopyrimidine glycosylase (Fpg) released 8-oxo-Gua, but not 8-NO(2)-Gua, indicating that 8-NO(2)-Gua is not a substrate for Fpg. Although 8-NO(2)-Gua was generated in isolated DNA by different nitrating systems, other types of damage were formed in abundance, and the lesion could not be found reliably in nuclear DNA, suggesting that the biological importance is limited.     

Sister chromatid exchange in lymphocytes in myelodysplastic syndrome

Sister chromatid exchange (SCE), percentage of first, second, third mitoses, blastic transformation index and mitotic index in patients with myelodysplastic syndrome (MDS) (3 with refractory anaemia, 2 with refractory anaemia with sideroblasts, 1 with refractory anaemia with excess of blasts, 4 with refractory anaemia with excess of blasts in transformation) and in 15 healthy volunteers were estimated. Three types of lymphocytes cultures were set up: first with phytohemaglutinin (PHA), second with PHA and bromodeoxyuridine (BRdU), third with BRdU. In healthy persons the SCE frequency was negatively correlated to proliferating rate index, but in MDS such correlation was not found. The lymphocytes cell cycle duration based on percentage of mitoses was longer in MDS patients than in controls. The results of our studies show the disturbances of lymphocytes during cell cycle division resulting in higher SCE frequency and lower proliferating rate compared to controls.     

Mutagenicity study of fried sausages in Salmonella, Drosophila and mammalian cells in vitro and in vivo

The basic extract of pan-fried sausages was studied for mutagenic potential in seven test systems. Mutagenic activity was high in the standard Ames assay in the Salmonella typhimurium strains TA1538 and TA98 in presence of S9 mix. In vivo, in the intrasanguine host-mediated assay with strain TA98 on Aroclor-pretreated mice, the mutagenic activity of the extract was low. A borderline activity was seen in the SCE assay in vitro with V79 Chinese hamster cells in presence of S9 mix. No significant mutagenic action was found in the gene-mutation assay for thioguanine resistance with V79 cells, the Drosophila sex-linked recessive lethal test, the micronucleus test and the mammalian spot test.