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PubMed1983

Immunocytochemical localization of cathepsin B in degenerating rat skeletal muscle induced by a local anesthetic, bupivacaine.

Experimental acute degenerative changes in skeletal muscle accompanied by a drastic increase in cathepsins B&L were induced in rats by intramuscular injection of a local anesthetic, bupivacaine. Cathepsins B&L have been implicated in the rapid disappearance of muscle fibers. Degenerating muscle showed a spotty fluorescence when stained with antibodies against cathepsin B, indicating that the increased cathepsin B did not originate from the muscle itself, but from invading phagocytes. We report here results showing that cathepsin B of nonmuscle cell origin is involved in the breakdown of myofibrillar proteins in acute bupivacaine-induced muscle degeneration.

Animals↗
PubMed1999

Helospectin-like peptides: immunochemical localization and effects on isolated cerebral arteries and on local cerebral blood flow in the cat.

Helospectin I and II and helodermin are nonamidated, vasoactive intestinal peptide (VIP)-like peptides, isolated from the salivary gland venom of the lizards Heloderma suspectum and Heloderma horridum. Helospectin I has 38 amino acid residues and differs from helospectin II in that it has an additional serine residue at the C-terminus. Numerous nerve fibers containing helospectin-like immunoreactivity (LI) and a few fibers containing helodermin-LI were present in the adventitia and at the adventitia-media border of cat cerebral arteries. In the sphenopalatine ganglion, numerous nerve cell bodies containing helospectin-LI were seen. Double immunostaining revealed that helospectin-LI nerve cell bodies coexisted with VIP-containing cell bodies. Radioimmunoassay showed high levels of helospectin-LI in extracts of cerebral vessels from the circle of Willis (27.4 pg/mg [wt/wt]). Helospectin I and II and helodermin (10(-10) to 10(-6) mol/L) produced concentration-dependent relaxations of feline middle cerebral arteries amounting to 50% to 80% of precontraction induced by U46619. The maximum effects and the potency were similar to that of VIP. Neither of these peptides elicited endothelium-dependent relaxations. Intracerebral microinjection of helospectin and helodermin produced a moderate concentration-dependent increase of the cerebral blood flow of alpha-chloralose anesthetized cats. The maximum increase (21 +/- 5%) was observed after the injection of 5 microg helodermin, whereas 16 +/- 7% was seen with helospectin I and 19 + 5% with helospectin II. The results suggest that helospectin/helodermin-like peptides co-localize with VIP in perivascular nerve fibers originating in the sphenopalatine ganglion. They seem to have strong and potent vasodilator effects.

Animals↗
PubMed2001

Amylin: localization, effects on cerebral arteries and on local cerebral blood flow in the cat.

Amylin and adrenomedullin are two peptides structurally related to calcitonin gene-related peptide (CGRP). We studied the occurrence of amylin in trigeminal ganglia and cerebral blood vessels of the cat with immunocytochemistry and evaluated the role of amylin and adrenomedullin in the cerebral circulation by in vitro and in vivo pharmacology. Immunocytochemistry revealed that numerous nerve cell bodies in the trigeminal ganglion contained CGRP immunoreactivity (-ir); some of these also expressed amylin-ir but none adrenomedullin-ir. There were numerous nerve fibres surrounding cerebral blood vessels that contained CGRP-ir. Occasional fibres contained amylin-ir while we observed no adrenomedullin-ir in the vessel walls. With RT-PCR and Real-Time-PCR we revealed the presence of mRNA for calcitonin receptor-like receptor (CLRL) and receptor-activity-modifying proteins (RAMPs) in cat cerebral arteries. In vitro studies revealed that amylin, adrenomedullin, and CGRP relaxed ring segments of the cat middle cerebral artery. CGRP and amylin caused concentration-dependent relaxations at low concentrations of PGF 2alpha-precontracted segment (with or without endothelium) whereas only at high concentration did adrenomedullin cause relaxation. CGRP8-37 blocked the CGRP and amylin induced relaxations in a parallel fashion. In vivo studies of amylin, adrenomedullin, and CGRP showed a brisk reproducible increase in local cerebral blood flow as examined using laser Doppler flowmetry applied to the cerebral cortex of the alpha-chloralose-anesthetized cat. The responses to amylin and CGRP were blocked by CGRP8-37. The studies suggest that there is a functional sub-set of amylin-containing trigeminal neurons which probably act via CGRP receptors.

Adrenomedullin↗
PubMed2006

Automated microaneurysm detection using local contrast normalization and local vessel detection.

Screening programs using retinal photography for the detection of diabetic eye disease are being introduced in the UK and elsewhere. Automatic grading of the images is being considered by health boards so that the human grading task is reduced. Microaneurysms (MAs) are the earliest sign of this disease and so are very important for classifying whether images show signs of retinopathy. This paper describes automatic methods for MA detection and shows how image contrast normalization can improve the ability to distinguish between MAs and other dots that occur on the retina. Various methods for contrast normalization are compared. Best results were obtained with a method that uses the watershed transform to derive a region that contains no vessels or other lesions. Dots within vessels are handled successfully using a local vessel detection technique. Results are presented for detection of individual MAs and for detection of images containing MAs. Images containing MAs are detected with sensitivity 85.4% and specificity 83.1%.

Algorithms↗
PubMed1995

Immunohistochemical localization of five members of the Kv1 channel subunits: contrasting subcellular locations and neuron-specific co-localizations in rat brain.

A large variety of potassium channels is involved in regulating integration and transmission of electrical signals in the nervous system. Different types of neurons, therefore, require specific patterns of potassium channel subunits expression and specific regulation of subunit coassembly into heteromultimeric channels, as well as subunit-specific sorting and segregation. This was investigated by studying in detail the expression of six different alpha-subunits of voltage-gated potassium channels in the rat hippocampus, cerebellum, olfactory bulb and spinal cord, combining in situ hybridization and immunocytochemistry. Specific polyclonal antibodies were prepared for five alpha-subunits (Kv1.1, Kv1.2, Kv1.3 Kv1.4, Kv1.6) of the Shaker-related subfamily of rat Kv channels, which encode delayed-rectifier type and rapidly inactivating A-type potassium channels. Their distribution was compared to that of an A-type potassium channel (Kv3.4), belonging to the Shaw-related subfamily of rat Kv channels. Our results show that these Kv channel alpha-subunits are differentially expressed in rat brain neurons. We did not observe in various neurons a stereotypical distribution of Kv channel alpha-subunits to dendritic and axonal compartments, but a complex differential subcellular subunit distribution. The different Kv channel subunits are targeted either to presynaptic or to postsynaptic domains, depending on neuronal cell type. Thus, distinct combinations of Kv1 alpha-subunits are co-localized in different neurons. The implications of these findings are that both differential expression and assembly as well as subcellular targeting of Kv channel alpha-subunits may contribute to Kv channel diversity and thereby to presynaptic and postsynaptic membrane excitability.

Animals↗
PubMed2005

Local radiotherapy as a complement to incomplete transcatheter arterial chemoembolization in locally advanced hepatocellular carcinoma.

PURPOSE: In order to determine the effect of additional radiotherapy (RT) after an incomplete transcatheter arterial chemoembolization (TACE) in an unresectable hepatocellular carcinoma (HCC), the treatment results of patients receiving TACE plus RT were analyzed and compared with those treated with TACE alone. MATERIALS AND METHODS: One hundred and five patients with an unresectable HCC were treated with TACE from January 1992 to December 2002. In 73 of these patients, the TACE was incomplete. Among them, TACE was repeatedly performed in 35 patients (TACE group), and the remaining 38 patients were also treated with local RT (TACERT group). The patients were either in stage III or IVa, Eastern Cooperative Oncology Group 2 or less, and Child-Pugh class A or B. The average frequency of TACE prior to RT was 2 and the RT was started within 7-10 days after the TACE. RESULTS: The 2-year survival rate was significantly higher in the TACERT than in the TACE group (36.8 % vs. 14.3%, P=0.001). According to the tumor size, the 2-year survival rates in the TACERT and TACE groups were 63% vs. 42% in 5-7 cm (P=0.22), 50% vs. 0% in 8-10 cm (P=0.03), and 17% vs. 0% in larger than 10 cm (P=0.0002) respectively. CONCLUSION: There was a significantly improved survival rate in the TACERT group of unresectable HCC patients than in the TACE group, particularly in case of tumors > or =8 cm in diameter. Therefore, RT in addition to TACE is strongly recommended for patients with an unresectable HCC.

Antibiotics, Antineoplastic↗
PubMed1987

Local retinal regions control local eye growth and myopia.

In chicks, visual deprivation leads to myopia and enlargement of the vitreous chamber of the eye. When chicks were raised with white translucent occluders over their eyes so that either the nasal half, the temporal half, or all of the retina was visually deprived, the resulting myopia (median = -15 diopters) was limited to the deprived part of the retina, regardless of which half of the retina was visually deprived; the nondeprived part remained nearly emmetropic. Correspondingly, the vitreous chamber was elongated only in the region of the visual deprivation, resulting in eyes with different asymmetric shapes depending on which retinal region was deprived. These results argue for a local regulation of ocular growth that is dependent on vision and suggest a hypothesis to explain the epidemiological association of myopia in humans with large amounts of reading. Because most nonfoveal retinal neurons have large receptive fields, they cannot resolve the individual letters on the printed page; this may lead to their activity being less during reading than during most other forms of visual stimulation. Thus, the impoverished stimulus situation of reading may lead to myopia, as do other types of visual form deprivation.

Animals↗
PubMed1995

Nonspiking local interneurons in insect leg motor control. II. Role of nonspiking local interneurons in the control of leg swing during walking.

1. Nonspiking local interneurons (NSIs) were recorded intracellularly in the mesothoracic ganglion of semi-intact locusts walking on a treadwheel. Interneurons were characterized by their connectivity to motoneurons. Their activity patterns in the step cycle and the effect current injection had on the leg movement were analyzed. We examined interneurons that provided excitatory or inhibitory synaptic drive to a subset of motoneurons active during the swing movement of walking. 2. Interneuron activity was observed to support or oppose the actual leg movement. Both supporting and opposing interneurons were active simultaneously, lending support to the idea that the actual motor output of walking is generated by the adjustment of parallel antagonistic pathways of signal processing. 3. The examined interneurons showed qualitatively the same patterns of activity during forward and backward walking. This indicates that swing movement in both situations may be generated by similar neuronal networks (although the mechanism of movement reversal remains unclear). 4. At least two functional types of NSIs could be distinguished. First, there were interneurons whose depolarization patterns showed distinct variability, often correlated with duration or amplitude of the swing movement. As a rule, current injection had minor, if any, effects on leg movement. Populations of these interneurons appear to be involved in the control of a coordinated swing movement by driving appropriate sets of muscle groups. The second type of NSIs showed more stereotyped activity patterns that varied relatively little with changes in the swing movement. Current injection had strong effects on the leg movement and could, for example, arrest the leg in the stance phase. These interneurons appear to be primarily involved in the trigger mechanism of leg swing.

Animals↗
PubMed1990

Relation between localization of coronary artery disease and local abnormalities in ventricular activation during exercise tests.

To examine whether or not the location of local abnormalities on body surface isochrone maps reflects the site of myocardial ischemia, 48 coronary artery disease patients without myocardial infarction were studied. Eighty-seven unipolar electrocardiograms distributed over the anterior chest and the back were recorded simultaneously before and after the submaximal treadmill exercise. For each lead, the duration from the QRS onset to the time of the most rapid decrease in QRS voltage was measured (index of ventricular activation [IVA]). Based o the data provided by these 87 leads, IVA isochrone maps (IVA map) in preexercise and in postexercise, as well as IVA maps showing the difference between preexercise and postexercise, were constructed. The IVA was defined as abnormal when it exceeded (mean + 2 SD) the normal range. We called the area with the abnormal IVA, the "+2SD area." In patients having a stenosis in the left anterior descending artery, the +2SD area in each map was located mainly on the left anterior chest, whereas in patients having a stenosis in the right coronary artery, the +2SD area in each map was located mainly on the right lower thoracic surface. Moreover, the +2SD area of patients with both left anterior descending and right coronary artery disease appeared on both the left anterior chest and the right lower thoracic surface. In patients with left circumflex artery disease, however, the location of the +2SD area did not suggest a stenotic site because of its small population. On the other hand, it was difficult to determine the ischemic site from the body surface distribution of ST segment depression.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Neutrophil sequestration and migration in localized pulmonary inflammation. Capillary localization and migration across the interalveolar septum.

Recent studies have raised the possibility that biophysical properties of the neutrophil, specifically cell size and deformability, along with the unique hemodynamic properties of the pulmonary microvasculature may be important determinants of neutrophil transit through the lung and lead to significant retention in capillaries. Additionally, accumulating evidence supports the concept that leukocytes may emigrate into inflammatory lesions through the pulmonary capillary wall, quite unlike the situation in the systemic circulation where the predominant site of margination and emigration is the postcapillary venule. To confirm and extent this suggestion, morphometric techniques were employed to determine the location and kinetics of neutrophil movement from the vascular compartment to the alveolar space in response to an inflammatory reaction initiated by local instillation of fragments of the activated form of the fifth component of complement (C5f). We found that the site of neutrophil sequestration and probable migration was almost exclusively the capillary. Neutrophil sequestration appeared first in the capillaries of the interalveolar septae, with negligible accumulation in arterioles and venules (less than that in interalveolar septae by a factor of 10(4) when corrected for the relevant surface areas). Accumulation in the airspaces lagged behind that in the interalveolar septa by about 60 min, placing an upper time-limit on the emigration process across the epithelial barriers. Interestingly, neutrophils samples by bronchoalveolar lavage represented only 1 to 2% of those neutrophils shown to be present in the airspaces of the lung by morphometric assessment, suggesting that bronchoalveolar lavage may sample only a subpopulation of the emigrated cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Spatial localization of transplanted hemopoietic stem cells: inferences for the localization of stem cell niches.

The spatial distribution of subpopulations of hemopoietic progenitor cells following syngeneic transplantation was investigated at the single-cell level. The location of infused hemopoietic progenitor cells within the femoral bone marrow of nonablated recipients was determined by 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester labeling of cells and in situ fixation by perfusion. Analysis performed over 15 hours after infusion demonstrated that the spatial distribution of transplanted marrow cells is not a random process. Although the majority of cells enter the bone marrow from the central marrow vessels, the subsequent localization within the bone marrow varied according to their phenotype. Candidate "stem cells" demonstrated selective redistribution and were significantly enriched within the endosteal region, whereas mature terminally differentiated and lineage-committed cells selectively redistributed away from the endosteal region and were predominantly in the central marrow region. Together, these data strongly support historical evidence of the presence of endosteal hemopoietic stem cell niches.

Animals↗
PubMed1993

Otitis externa: clinical comparison of local ciprofloxacin versus local oxytetracycline, polymyxin B, hydrocortisone combination treatment.

Thirty patients with otitis externa were included in a prospective, randomized, parallel group comparison study. Treatment regimens used were ciprofloxacin (2 mg/ml) as ear drops twice daily or drops containing oxytetracycline (5 mg/ml) polymyxin B (10,000 units/ml) and hydrocortisone (15 mg/ml) twice daily for 7 days. Evaluation was by clinical signs and symptoms and bacteriological findings from ear swabs prior to and 3 and 8 days after start of therapy, together with long-term follow-up if necessary. Sixteen patients were included in the ciprofloxacin and 14 in the combined antibiotic group. Six of 16 and 7 of 14 initial swabs, respectively, contained Pseudomonas aeruginosa. Mixed Gram-negative and Gram-positive pathogens were found in 3 patients in the ciprofloxacin group. All Gram-negative isolates were sensitive to ciprofloxacin. Microbiological evaluation showed rapid eradication of all bacterial pathogens in the ciprofloxacin group with 1 persistence of Candida albicans but eradication of Staphylococcus aureus in the same ear with marked clinical improvement. There was 1 case of supercolonization in the ciprofloxacin group with Candida parapsilosis. In the combined antibiotic group, there were 7 persistencies of which 5 were P. aeruginosa. No adverse effects were noted in either group. Treatment was assessed as a complete success in 14/16 and 5/14 cases, partial success in 2/16 and 4/14 cases and unsuccessful or indeterminate in 0/16 and 5/14 cases, respectively. Ciprofloxacin thus appears to be a useful and effective agent in the local therapy of otitis externa.

Administration, Topical↗
PubMed2005

Combined chemoradiotherapy regimens of paclitaxel and carboplatin for locally advanced non-small-cell lung cancer: a randomized phase II locally advanced multi-modality protocol.

PURPOSE: This phase II noncomparative randomized trial was conducted to determine the optimal sequencing and integration of paclitaxel/carboplatin with standard daily thoracic radiation therapy (TRT), in patients with locally advanced unresected stage III non-small-cell lung cancer (NSCLC). Survival data were compared with historical standard sequential chemoradiotherapy data from the Radiation Therapy Oncology Group. PATIENTS AND METHODS: Patients with unresected stages IIIA and IIIB NSCLC, with Karnofsky performance status > or = 70% and weight loss < or = 10%, received two cycles of induction paclitaxel (200 mg/m2)/carboplatin (area under the plasma concentration time curve [AUC] = 6) followed by TRT 63.0 Gy (arm 1, sequential) or two cycles of induction paclitaxel (200 mg/m2)/carboplatin (AUC = 6) followed by weekly paclitaxel (45 mg/m2)/carboplatin (AUC = 2) with concurrent TRT 63.0 Gy (arm 2, induction/concurrent), or weekly paclitaxel (45 mg/m2)/carboplatin (AUC = 2)/TRT (63.0 Gy) followed by two cycles of paclitaxel (200 mg/m2)/carboplatin (AUC = 6; arm 3, concurrent/consolidation). RESULTS: With a median follow-up time of 39.6 months, median overall survival was 13.0, 12.7, and 16.3 months for arms 1, 2, and 3, respectively. During induction chemotherapy, grade 3/4 granulocytopenia occurred in 32% and 38% of patients on study arms 1 and 2, respectively. The most common locoregional grade 3/4 toxicity during and after TRT was esophagitis, which was more pronounced with the administration of concurrent chemoradiotherapy on study arms 2 and 3 (19% and 28%, respectively). CONCLUSION: Concurrent weekly paclitaxel, carboplatin, and TRT followed by consolidation seems to be associated with the best outcome, although this schedule was associated with greater toxicity.

Aged↗
PubMed1996

Expression and localization of epidermal growth factor, transforming growth factor-alpha, and localization of their common receptor in fetal human lung development.

Transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), and their common EGF receptor have been shown to be involved in cell proliferation and lung maturation. The aim of the study was to determine the site of production of TGF-alpha and EGF mRNA and the cellular distribution of TGF-alpha/EGF proteins and EGF receptor, in fetal human lung. By using in situ hybridization with 35S-labeled cDNA probes in frozen sections from eight lungs from fetuses ranging from 12 to 33 wk of gestation, TGF-alpha and EGF mRNA transcripts appeared to be confined to the mesenchymal cells and mainly found in the dense connective tissue along the pleura, bronchi, and large vessels, but undetected in bronchial epithelial cells. The streptavidin-biotin immunoperoxidase method, applied to paraffin-embedded specimens from 39 fetuses ranging from 10 to 41 wk, showed that TGF-alpha, EGF, and EGF receptor exhibited a similar cellular distribution during the whole period of gestation. They were detected in the undifferentiated cells of the airway surface epithelium, mesothelial cells, smooth muscle, and a few mesenchymal cells, as early as 10 wk. After 12 wk, the immunoreactivity was strong in the ciliated, secretory, and basal cells, and in growing glands along the large airways, but proved lower in the distal airways. After 24 wk, the immunoreactivity remained in the airway epithelium, but was mainly localized in the apical domain of ciliated cells, in alveolar cells, and in the serous cells of the glands. The presence of TGF-alpha, EGF, and EGF receptor during the whole period of fetal lung development suggests that these factors are not only mitogenic, but can also be involved in epithelial maturation, through paracrine secretion, as most TGF-alpha and EGF mRNA transcripts are expressed in mesenchymal cells.

Antibody Specificity↗
PubMed2005

A novel isoform of human Golgi complex-localized glycoprotein-1 (also known as E-selectin ligand-1, MG-160 and cysteine-rich fibroblast growth factor receptor) targets differential subcellular localization.

The initial step in trafficking of leukocytes through the vascular endothelium is mediated by an adhesive interaction between molecules of the selectin family and their cognate receptors. Previously, a putative murine E-selectin ligand-1 (ESL-1) was identified and found to be identical to Golgi complex-localized glycoprotein-1 (GLG1), also known as MG-160, and to a previously identified basic fibroblast growth factor (bFGF)-binding protein known as cysteine-rich FGF receptor (CFR). We report here a novel variant of the human GLG1 gene product that we call GLG2, cloned from a human monocyte cDNA library. GLG2 encodes a polypeptide identical to GLG1 except with a unique 24-amino-acid extension at the C-terminus of its cytoplasmic domain. Transfection of chimeric constructs into human embryonic kidney epithelial 293 cells revealed that the cytoplasmic domains of GLG1 and GLG2 targeted the expression of each chimeric protein differentially, GLG1 to the cell surface and GLG2 to the Golgi. Genetic analysis suggests that GLG1 and GLG2 are the products of a single gene, the mRNA of which can be processed by alternative splicing to generate two different transcripts encoding either GLG1 or GLG2. Northern blot analysis showed that the relative amounts of the mRNAs for either isoform differ in a cell- and species-specific manner. These data suggest that alternative splicing of the GLG1 gene transcript might regulate the function of its product.

Alternative Splicing↗
PubMed1984

Localized effects of naloxone on local cerebral glucose utilization in rat cerebral nuclei with Met-enkephalinergic neurons.

The alterations of local glucose utilization in 101 cerebral nuclei following the subcutaneous administration of the specific opioid antagonist naloxone were measured using the quantitative autoradiographic 14C-deoxy-D-glucose technique in conscious rats. Met5-enkephalin-like immunoreactivity (MELI) and ME receptor binding (MERB) levels in 82 cerebral nuclei were assayed quantitatively by microdensitometry of fluorescence micrographs and autoradiographs of the brain slices. Naloxone administration significantly reduced glucose utilization rate in 18 lower brainstem nuclei including the n. tractus solitarii, n. dorsalis nervi vagi, substantia grisea centralis, n. parabrachialis dorsalis and ventralis, and n. interpeduncularis. These nuclei contained ME perikarya with high levels of MELI and MERB. However, naloxone did not alter glucose utilization of other lower brainstem nuclei containing ME neurons and all thalamic, epithalamic, hypothalamic and limbic nuclei with ME perikarya. The distribution and magnitude of the neuronal response of cerebral nuclei to naloxone were apparently not related to the distributions of ME neurons. The present study offers direct evidence for the selective action of naloxone per se in some lower brainstem nuclei with ME perikarya.

Animals↗
PubMed2004

Effect of lactobacilli administration in the vaginal tract of mice: evaluation of side effects and local immune response by local administration of selected strains.

Lactobacilli are the predominant microorganisms in the vaginal tract of human and some homeothermic animals. They can maintain the ecological equilibrium of the tract by protecting against pathogenic microorganisms. In the last few years, there has been an increased tendency to use probiotic microorganisms to restore the ecological equilibrium and to protect against infections. This principle has been widely applied to the gastrointestinal tract. More recently, some other studies have reported the application of probiotics in different tracts, for example, the urogenital or respiratory tract. One of the objectives of our group is to design probiotic products for the urogenital tract. With this purpose, lactobacilli were isolated from the human vagina, and later some of them were selected for their probiotic characteristics (production of antagonistic substances or adhesion capability). The application of probiotic products in the vaginal tract has been approached empirically; some pharmaceuticals containing these microorganisms are available in the United States or Europe or are protected under the patent process or intellectual property rights. There are not enough studies in humans or animals to determine whether their administration can produce some type of collateral or adverse effect. Using Balb/c mice as the experimental model, the object of the present work was to study (1) whether intravaginal administration of human lactobacilli can produce colonization of the tract; (2) whether such administration produces some type of adverse or collateral effect; and (3) whether probiotics are able to stimulate the local immune system. Keeping in mind that hormones can affect the colonization or persistence ability of microorganisms, and with the purpose of having all animals at the same point in the sexual cycle, animals were cycled with estradiol 48 h before inoculation with lactobacilli. They were then inoculated im with hormones 48 h before beginning microorganism inoculations. Later they were intravaginally inoculated with the appropriate dose of each Lactobacillus strains. The animals were sacrificed on different days after inoculation to perform the following studies: 1. Microbiological assays: To determine the number of lactobacilli in the tract (in vaginal washes or in organ homogenates), by plating the samples in selective media containing antibiotic (to differentiate the resident flora from those administered experimentally).

Animals↗