Knee lymphocutaneous fistula secondary to knee arthroplasty.

Lower limb lymphorrhea secondary to a surgical procedure is a rare but difficult-to-solve complication. In lower limb, this entity is frequently associated with vascular procedures around the inguinal area. We report on a case of a knee lymphocutaneous fistula secondary to a knee revision arthroplasty. To our knowledge, no previous reports regarding this complication have been published.

Sialyloligosaccharides in human and bovine milk and in infant formulas: variations with the progression of lactation.

Several lines of research support a role for human milk oligosaccharides in the defense of breast-fed infants against pathogens. Some ofthese oligosaccharides contain at least one moiety of sialic acid and are, thus, termed sialyloligosaccharides. These constitute a significant component (>1 g/L) of human milk. It is well established that milk composition varies among species, and previous reports have indicated that one ofthe differences between human and bovine milk is precisely their contents of sialyloligosaccharides. Because most infant formulas are manufactured with bovine milk components, it follows that formula-fed and breast-fed infants ingest dissimilar quantities of these carbohydrate structures. To ascertain these differences and their impact along lactation, the contents of oligosaccharide-bound sialic acids and major sialyloligosaccharides in samples of human and bovine milk (obtained at different lactation stages) were determined. In addition, infant formulas were assayed for their sialyloligosaccharide contents. Seven sialyloligosaccharides were identified in human milk; namely, 3'-sialyl-3-fucosyllactose and sialyllacto-N-tetraoses (a and b+c), the predominant structures at all lactation stages. Five sialyloligosaccharides were identified in bovine milk, of which 6'-sialyllactosamine and 3'-sialyllactose were the most abundant. In addition, sialyloligosaccharides in human and bovine milk decreased along lactation, and infant formulas did not contain significant amounts of sialyloligosaccharides. The results point to the general conclusion that regarding both qualitative and quantitative aspects, milk from humans and cows and infant formulas have different oligosaccharide contents. In this sense, bottle-fed infants are subject to reduced sialyloligosaccharide intake as compared to breast-fed infants.

Distribution of bovine milk sialoglycoconjugates during lactation.

The sialoglycoconjugate content of human milk has been extensively studied. However, little attention has been paid to the changes occurring in these compounds in bovine milk during lactation. Since sialoglycoconjugates are very abundant in milk from the early stages of lactation, they have been suggested to be important for the nutrition of the newborn during the first months of life. The distribution of sialoglycoconjugates (expressed as glycoconjugate-bound sialic acid) from four different stages of lactation (colostrum, transitional, mature, and late-lactation milks) was investigated in four Spanish-Brown cows. All the fractions studied (total sialic acids, glycoproteins, oligosaccharides, casein, and gangliosides) showed a similar trend. We found the highest values in the colostrum, these decreasing in transitional and mature milks and increasing again in late-lactation milk. We also found a selective change in the relative contents of glycoprotein- and oligosaccharide-bound sialic acids. In mature milk, the latter increased up to 80% (59% in colostrum) and the former decreased to 3.9% (35.3% in colostrum). It would appear that the decrease in oligosaccharide-bound sialic acid is compensated by the increase in glycoprotein-bound sialic acid. From these results, it is deduced that newborn infants or calves fed with infant formulas or milk replacers, respectively, should be supplemented with sialoglycoconjugates to approximate the composition of human and cow milk as far as is practicable.

Different patterns of structural luteolysis in the human corpus luteum of menstruation.

Structural luteolysis is a complex process responsible for the elimination of the corpus luteum (CL). The aim of this study was to analyse the luteolytic process of the CL of menstruation. For this, we have morphologically studied 654 ovaries from 340 cycling women. Apoptotic cells were observed almost exclusively during the perimenstrual period and were extremely scarce at advanced stages of involution. Steroidogenic luteal cells surviving to the perimenstrual apoptotic wave underwent characteristic degenerative changes, consisting of intense cytoplasmic vacuolation, expression of macrophage markers and accumulation of lipofuscin pigment, and they persisted for long periods of time. Accumulation of corpora albicantia (CA) was observed in only 25% of a subset of 168 women, whereas 28% showed involuting CL without hyalinization, consisting of clusters of pigment-filled cells, and 46.4% showed ovaries with a mixture of CA and involuting CL without hyalinization or involuting CL with intermediate features. Evolution of the CL towards CA seemed to be related to the presence of a large, blood-filled cavity. The data from this study suggested that different patterns of structural luteolysis exist during CL involution, and that the final fate of the involuting CL is dependent on the presence of a large, central, blood-filled cavity.

Immunolocalization of glutaredoxin in the human corpus luteum.

Glutaredoxin (Grx) is a small protein with oxidoreductase activity which is involved in the cellular defence against oxidative stress. Corpus luteum (CL) regression has been related to the generation of reactive oxygen species (ROS). We have studied the presence of glutaredoxin in the human ovary during the ovulatory cycle using polyclonal antibodies developed against recombinant human Grx. Immunostaining was only detected between days 15 and 23 of the cycle and was localized exclusively in the corpus luteum. Grx-positive cells corresponded to granulosa-derived luteal cells (GLC) whereas the remaining luteal cell types were not immunostained. In general, Grx immunoreactivity was parallel to the functional activity of the CL. Most GLC were immunostained on days 15-16 of the cycle, whereas on days 17-19 immunoreaction was found mainly at the inner and outer aspects of the granulosa lutein layer (GLL). After this stage only isolated GLC showed Grx immunoreactivity and no reaction was found from day 23 of the cycle onward. In two CL of pregnancy that were also studied, isolated GLC showed Grx immunoreactivity. Loss of Grx immunoreactivity was coincident with the appearance of morphological signs of structural luteolysis, such as shrinkage of the GLL and the presence of apoptotic cells. These data suggest that Grx, as a cellular antioxidant, plays an important role in the mechanisms of human CL development.

A quantitative study of changes in the human corpus luteum microvasculature during the menstrual cycle.

Endothelial cells are the most abundant cell type in the corpus luteum (CL), and changes in blood vessels have been proposed to play a pivotal role in CL regression. We have studied quantitatively the changes in the human granulosa-luteal microvasculature in CL of various ages: young (Days 17-19 of the cycle), mature (Days 20-24), old (Days 25-27), early regressing (follicular phase of the following cycle), and late regressing (luteal phase of the following cycle). Blood vessels were identified by immunohistochemical staining for the endothelial cell marker CD34. Because of the anisotropy of blood vessels, both vertical and transverse sections of the granulosa-lutein layer (GLL) were used to estimate relative (volume, surface, and length densities) and absolute (mean cross-sectional area) vascular variables. Full luteinization from young to mature CL was accompanied by a 61% increase in the mean cross-sectional area of vascular profiles and a 52% increase in the mean volume of granulosa-lutein cells, as an estimator of changes in the volume of the GLL. In old and early regressing CL, there was a progressive increase in relative structural vascular variables, due to the shrinkage of the GLL, whereas the mean cross-sectional area of capillaries showed a 53% decrease from mature to old CL. Finally, in late regressing CL, there was a decrease in most relative structural variables, in spite of the increasingly shrunken GLL. The decrease in the capillary diameter found at the late luteal phase most likely leads to a decreased blood flow, and early changes in blood vessels could initiate and/or accelerate CL regression.

Macrophages, cell proliferation, and cell death in the human menstrual corpus luteum.

We studied the presence and numbers of macrophages in the different compartments of the human menstrual corpus luteum (CL) in relation to the proliferative activity and apoptosis in luteal cells. Macrophages were recognized by immunohistochemical demonstration of the lysosome-associated glycoprotein CD68, and proliferating cells by immunohistochemical detection of the cell cycle-related protein Ki67 and by counting mitotic cells. In general, changes in the number of macrophages were parallel to the functional activity of the CL. Macrophage numbers increased up to the end of the early luteal phase, remained relatively unchanged during the midluteal phase, and decreased at the late luteal phase. Furthermore, macrophages showed prominent morphological changes during the cycle. They showed round or elongated cytoplasm during the early and late luteal phases, and dendritic features in the midluteal phase. Proliferating cells were very abundant on Days 15-16 and showed a significant decrease thereafter. Most proliferating cells corresponded to stromal (mainly vascular) cells. However, about 5% of granulosa-lutein cells and about 15% of theca-lutein cells were proliferating during the early and midluteal phases. Regression of the CL at the late luteal phase was associated with both a decrease in the number of proliferating cells and an increase in the number of apoptotic cells, which were highly increased on Days 25-27 of the cycle. The number of macrophages was not related to cell proliferation nor to cell death during the luteal phase. The observed changes in both macrophage number and morphology suggest the existence of a bidirectional communication between macrophages and steroidogenic cells in the human CL, or regulation of both cell populations by similar mechanisms.

Developmental changes in UDP-N-acetylglucosamine 2-epimerase activity of rat and guinea-pig liver.

The activity of UDP-N-acetylglucosamine 2-epimerase was determined in the liver of rats and guinea-pigs of different ages. The activity of this enzyme in rats was low at birth, increased to a maximum value on day 15, and fell gradually until day 30. Thereafter, it increased up to the 60th day. The activity profile of the enzyme from guinea-pig liver was very similar. However, guinea-pig activity was 2-5 times lower than in rats. Both rats and guinea-pigs displayed similar liver sialic acid contents which increased from birth to 2 months of age. Rats also showed a N-glycolylneuraminic acid content that decreased from birth to 2 months. From these results we can inferred that postnatal UDP-N-acetylglucosamine 2-epimerase activity seems to be correlated with age and the developmental states of rats and guinea-pigs.

Changes in ganglioside and sialic acid contents of goat milk during lactation.

The ganglioside content of goat milk has been determined from d 1 after parturition to d 60 of lactation. Marked changes occurred in milk over the course of lactation; the highest ganglioside content occurred in d-1 colostrum and then decreased to the end of the period studied. At least seven different ganglioside species were detected; three gangliosides containing sialyllactosylceramide accounted for 66 to 92% of the total lipid-bound sialic acid; this result reflected a very simple core structure of goat milk gangliosides. The most abundant ganglioside, II3(N-acetylneuraminic acid)2-lactosylceramide, was about 35 to 56%. The sialic acid content exhibited a trend similar to that of gangliosides; during early lactation sialic acid content was higher than in mature milk. Fat, protein, and total solids were high at initiation of lactation and decreased thereafter. However, lactose content remained almost unchanged during the period studied.

Alterations induced by fascioliasis and cirrhosis on the biliary excretion of cefmetazole in Wistar rats.

1. Alterations induced by fascioliasis and cirrhosis on the biliary excretion of cefmetazole have been studied in Wistar rats. 2. Both infestation with Fasciola hepatica and experimental cirrhosis originated a significant decrease in the biliary excretion and in bile flow increase induced by the drug. 3. Administration of the beta-lactam antibiotic induced a lower degree of uncoupling of biliary lipid secretion in the cirrhotic and fasciolotic animals, but the effect was evident in all experimental groups.

Gangliosides in bovine milk. Changes in content and distribution of individual ganglioside levels during lactation.

Bovine milk undergoes changes in its ganglioside contents during the different stages of lactation. These contents are higher in colostrum (7.5 mg of lipid-bound NeuAc/kg) than in transitional (2.3 mg) or mature (1.4 mg) milk. The sialic acid content of milk follows a similar profile to that of gangliosides with the highest content during the first few days post partum followed by a gradual decrease towards the end of the period studied. When the individual distribution of gangliosides was examined throughout the course of lactation, several changes were also found. GD3 is the major ganglioside (about 60-70%) found; its content decreases from the first to the fifth day, increasing towards the end of the period considered. GM3, GD3 and GT3, sialyllactosylceramide-containing gangliosides account for 80-90% of the total lipid-bound NeuAc content. The most striking change in the ganglioside pattern was the gradual increase in G3.

Hepatic transport of bilirubin in rats with streptozotocin-induced diabetes.

This study was undertaken to evaluate the effects of streptozotocin-induced diabetes on the hepatic transport of bilirubin in male Wistar rats. Rats were pretreated with streptozotocin (60 mg/kg i.p.) to induce uncontrolled diabetes. Six days later endogenous biliary excretion and plasma bilirubin concentration were significantly enhanced compared to control animals (+36% and +46%, respectively), while the blood levels of free hemoglobin remained unchanged. Following a bilirubin load, the maximal biliary excretion of the pigment (Tm) in diabetic animals was significantly enhanced compared to control animals (+49%). Liver and plasma bilirubin concentrations at the end of bilirubin administration were significantly reduced (-28% and -30%, respectively). Bilirubin UDP-glucuronosyltransferase activity and UDP-glucose concentration in liver were significantly enhanced (+31% and +81%, respectively), as was the biliary excretion of unconjugated bilirubin (+37%) and bilirubin mono- (+38%) and diconjugates (+53%). When streptozotocin-diabetic rats were treated with insulin, the parameters of bilirubin transport and metabolism were significantly reduced compared to diabetic animals receiving no hormone replacement. In summary, our data indicate that in short-term streptozotocin-diabetic rats there is increased bilirubin production as well as enhanced hepatic conjugation and subsequent biliary excretion of the pigment. These effects appear to be a direct consequence of diabetes.

Changes in hepatic cytosolic glutathione S-transferase enzymes induced by clotrimazole treatment in rats.

1. The influence of the antifungal agent clotrimazole on cytosolic glutathione S-transferase activities was studied in male Wistar rats. 2. Animals received clotrimazole by gastric lavage for 3 days (75 mg/kg per day). Hepatic glutathione S-transferase activity was determined with five different substrates: 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), p-nitro-benzyl chloride (PNBC), ethacrynic acid (EA) and trans-4-phenyl-3-buten-2-one (TPBO). 3. The largest increases in glutathione S-transferase activity were found with CDNB, DCNB and PNBC (+61%, +50% and +50%, respectively, when expressed per mg of cytosolic protein). Enzyme activity toward EA was induced to a lower extent (+33%). Changes in the formation of the conjugate of TPBO were relatively small (+22%). 4. These data indicate a differential induction of glutathione S-transferase isoenzymes and suggest that clotrimazole is a phenobarbital-type inducer of enzyme activity.

Sex-related differences in the hepatobiliary transport of phenolsulfonphthalein in the rat.

Sex-related differences in the hepatobiliary transport of phenolsulfonphthalein (PSP) were investigated in male and female Wistar rats. Maximal biliary excretion of unconjugated PSP was significantly higher in females while the excretion of the conjugated dye and liver UDP-glucuronosyltransferase activity toward PSP were higher in male animals. Orchidectomy decreased enzyme activity and excretion of the conjugate, whereas ovariectomy produced the opposite effect. Both in gonadectomized males and females maximal biliary excretion of the unconjugated dye was significantly reduced. Testosterone treatment increased the excretion of both conjugated and unconjugated PSP and transferase activity in orchidectomized males. Combined treatment of gonadectomized females with estradiol plus progesterone led to excretions of both conjugated and unconjugated PSP and UDP-glucoronosyltransferase activities similar to those found in control rats. These data indicate the existence of sex-related differences in the conjugation and biliary excretion of PSP in the rat and its modulation by sex hormones.

Postnatal development of the hepatobiliary transport of phenolsulfonphthalein in rats.

1. The postnatal development of the biliary excretion of phenolsulfonphthalein (PSP) was studied in male Wistar rats. 2. Following i.v. injection of PSP at 200 mumol/kg body wt, a maximal biliary excretion of 175 +/- 10 nmol/min/100 g body wt and 32 +/- 5 nmol/min/100 g body wt was reached for unconjugated and conjugated PSP, respectively, in the adult group. 3. The maximal biliary excretion of conjugated PSP was significantly lower in the 20-, 30- and 40-day-old groups as compared to the adults. The excretion of unconjugated dye was also significantly lower in 20- and 30-day-old rats. 4. The postnatal development of PSP excretion was unrelated to changes in the activity of UDP-glucuronosyltransferase. The importance of other factors is also discussed.