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Establishment and characterization of clonal cell lines derived from a fibrosarcoma of the H2-K/V-JUN transgenic mouse. A model of H2-K/V-JUN mediated tumorigenesis.

We used fibrosarcoma from an H2-K/V-JUN transgenic mouse to derive a series of three immortal cell lines (JUN-1, -2, and -3). The cell lines exhibit strikingly different behavior regarding phenotype transformation. Features examined include contact inhibition and density limitation of growth, proliferation, invasiveness, motility, and organization of the microfilament system. Overall, JUN-2 and JUN-3 represent extreme phenotypes, with JUN-2 having a phenotype indicative of low-level cellular transformation and JUN-3 meeting all the criteria of the transformed phenotype. JUN-1 cells can also be regarded as transformed, but to a lesser extent than JUN-3. Their phenotype is in the majority of characteristics intermediary between JUN-2 and JUN-3. The transformation status is inversely related to the expression of the V-JUN transgene, which is the highest in JUN-2, lower in JUN-1 and very low in JUN-3. This might be related to the MHC class I promoter driving its expression and to the general observation of repression of MHC class I genes coupled with cellular transformation. Based on this premise, we present a model of H2-K/V-JUN-mediated tumorigenesis, in which v-jun-conditioned transformation represents merely an initial phase of tumorigenesis. Later during tumor progression, additional oncogenes are activated and/or tumor suppressor genes inactivated, leading on the one hand to further exacerbation of the transformed phenotype, and on the other hand to the repression of the H2-K/V-JUN transgene (fixed in JUN-3). We believe that the system of JUN cell lines can be valuable for further molecular analysis of transformation-related traits.

Actins↗

Development of a congeneic line of the GR mouse strain without early mammary tumours.

Based on the previous observation that Mtv-2 controls the appearance of pregnancy-dependent mammary tumours and the early expression of mammary tumour virus (MTV)-antigens in the milk, an attempt was made to develop a congeneic GR line, without this locus, by introducing genetic material of the C57BL/10 strain. The cross-intercross system was used for this purpose. The mice of the even-numbered generations were selected for the absence of the Mtv-2 locus with the early maammary tumour (EMT) test and during the later cycles with the immunodiffusion (ID) test for MTV-antigens in the milk. After six cycles of cross-intercross, brother-sister mating was started with mice selected for the absence of Mtv-2. After two brother-sister matings the milk-transmitted MTV was eliminated by foster-nursing. Of the foster-nursed subline 233 mice (16 of the first, 118 of the second and 99 of the third generation) were tested for the presence of Mtv-2 with one or two of the following three methods: (1) ID-test, (2) EMT-test and (3) by examination of the development of spontaneous mammary tumours in breeding females. The tests were negative in 226 mice. This indicates that Mtv-2 was absent in nearly all mice of the congeneic subline. The positive reactions in the seven other tested mice were weaker than that observed in the GR strain and in GR mice foster-nursed by BALB/c. The presence of the Mtv-2 gene in the seven positive mice of the congeneic line is doubtful. With the ID test, viral antigens were detectable in 50% of the milk samples from the first lactation period of mice of the segregating backcross I population [GR congeneic X (GR congeneic X GR)], indicating a one-gene difference between the congeneic GR line and the progenitor GR strain.

Animals↗

[The rolandic line: a simple baseline for the identification of the central region. An MRI study and functional validation].

INTRODUCTION: The identification of the central region--i.e. the central sulcus, the pre- and post-central gyri, the paracentral lobule--on MRI and angiographic images may be necessary prior to stereotactic procedures such as biopsies or resection of centrally located tumors, depth electrode recordings for presurgical evaluation of drug-resistant epilepsies, or radiosurgery of arteriovenous malformations. Stereotactic methods, such as the Talairach's proportional grid based on the bicommissural system, demonstrated the statistical position of the central sulcus according to the Ac-Pc, Vac and Vpc baselines. However, the course and the spatial position of this sulcus have remarkable individual differences that sometimes make the sulcus difficult to identify on serial sagittal MRI or lateral angiographic images. In order to facilitate this identification, the authors propose a new oblique baseline, the rolandic (R) line. MATERIAL AND METHODS: The stereotactic MRI and angiography of 22 patients were reviewed for this study. Eleven of these patients had stereotactic biopsies for a low-grade tumor located in the central region, while eleven others had multiple intracerebral electrodes implantation and depth EEG recording (SEEG: stereoelectroencephalography) in the presurgical evaluation of drug-resistant partial epilepsy, prior to epileptogenic cortex resection. The Ac-Pc, Vac, Vpc baselines and segments of the central sulcus were drawn from the mid-sagittal and lateral T1-weighted MRI images and reported on an individual graph. Surface and deep margins as well as axis of the central sulcus were also reported along with corpus callosum baselines as defined by Olivier et al.: horizontal plane, anterior and posterior callosal planes. The rolandic line was then traced from the graph:it joined the intersection point between the anterior callosal plane and an orthogonal line passing through the floor of the temporal fossa, and the intersection point between posterior callosal plane and an orthogonal line passing through the top of the hemisphere. The rolandic line was then superimposed on any sagittal MRI image or lateral stereotactic angiographic film. Finally, the spatial position of electrode contracts through which electrical stimulations elicited motor and/or sensory responses, either from central electrode implanted for motor fibers identification prior to stereotactic biopsies or from SEEG electrodes implanted for epileptogenic zone identification and cortical mapping, was reported on the individual graph. Angular and linear measurements were taken from the graph, between the rolandic line, the central sulcus axis, the Ac-Pc and callosal baselines, and the central sulcus limits (top, bottom, anterior and posterior margins). RESULTS: Graph measurements indicated that the rolandic line was significantly closer to the inferior part of the central sulcus than to its superior part (average distance between the line and the inferior point of the sulcus: 1.86 +/- 1.87 mm; average distance between the line and the superior point of the sulcus: 4.5 +/- 2.3 mm; p < 0.001-t test); similarly, the rolandic line was closer to the deep margin of the sulcus rather than to its superficial border (average distance between the line and the most anterior point of the sulcus: 11.43 +/- 3.16 mm; average distance between the line and the most posterior point of the sulcus: 7.95 +/- 4.14 mm; p < 0.01-t test). In 90% of the cases, the rolandic line followed the deep or middle part of the sulcus, with an average angle of 4.18 degrees +/- 2.53 degrees between the line and the sulcus axis. The spatial position of the electrode contacts that elicited motor/or sensory responses to stimulations correlated topographically well in all cases with the position of the motor and sensory fibers defined according to the central sulcus, baselines and reference to stereotactic atlases. (ABSTRACT TRUNCATED)

Adolescent↗

Characterisation of a series of human immunodeficiency virus isolates derived sequentially from a single patient.

Five HIV-1 isolates were obtained sequentially from a single seropositive individual during the later stages of AIDS. Four of these isolates were adapted to grow in a continuous human T-lymphocytic cell line. Comparative biological and biochemical studies of the virus isolates were made using persistently infected cultures or virus derived from these systems respectively. The data obtained clearly shows that viruses with different biological properties can be isolated from the same individual at different times during the course of clinical AIDS.

Acquired Immunodeficiency Syndrome↗

The bilingual brain: cerebral representation of languages.

The present article deals with theoretical and experimental aspects of language representation in the multilingual brain. Two general approaches were adopted in the study of the bilingual brain. The study of bilingual aphasics allows us to describe dissociations and double dissociations between the different subcomponents of the various languages. Furthermore, symptoms peculiar to bilingual aphasia were reported (pathological mixing and switching and translations disorders) which allowed the correlation of some abilities specific to bilinguals with particular neurofunctional systems. Another approach to the study of the bilingual brain is of the experimental type, such as electrophysiological investigations (electrocorticostimulation during brain surgery and event-related potentials) and functional neuroanatomy studies (positron emission tomography and functional magnetic resonance imaging). Functional neuroanatomy studies investigated the brain representation of languages when processing lexical and syntactic stimuli and short stories. Neurophysiologic and neuroimaging studies evidenced a similar cerebral representation of L1 and L2 lexicons both in early and late bilinguals. The representation of grammatical aspects of languages seems to be different between the two languages if L2 is acquired after the age of 7, with automatic processes and correctness being lower than those of the native language. These results are in line with a greater representation of the two lexicons in the declarative memory systems, whereas morphosyntactic aspects may be organized in different systems according to the acquisition vs learning modality.

Brain↗

Changes in the protein metabolism of the CNS of a teleost following stimulation of the lateral-line organ.

Unilateral stimulation of the lateral-line organ (LLO) of the teleost Scardinius erythrophthalmus by current water caused an increase in the afferent fibre activity in the stimulated organ of about 3.5 times as compared with the non-stimulated LLO. There was an increase of [3H] histidine incorporation as compared to controls following stimulation applied either at the beginning (1 h) or at the end (1.5-6 h) of various incorporation times. Following a 1 h stimulation period, the different areas of the LLO-system (lateral nerve, medulla oblongata, cerebellum and valvulae cerebelli), as well as the optic tectum, subtectum and spinal cord showed a significant increase of protein labelling; whereas after 12 h post-incorporation times only the lateral nerve showed highly significant differences as compared to controls. In animals stimulated at the end of a 12 h pre-incorporation period (1.5-6 h) there was a significant increase of protein labelling in all investigated structures of the brain as compared to controls.

Action Potentials↗

Evaluation of the longitudinal arch of the foot with hallux valgus using a newly developed two-dimensional coordinate system.

The purposes of the present study were to establish a method for the evaluation of the longitudinal arch, using a two-dimensional coordinate system, and to clarify the main point of flattening of the foot in patients with hallux valgus. Measurements on a weight-bearing lateral radiograph of the foot were performed in 186 feet of 110 women with symptomatic hallux valgus and in 93 normal feet of 72 women. A two-dimensional coordinate system was used to perform the measurements. In this coordinate system, the line connecting the lowest point of the first metatarsal head and the lowest point (O) of the calcaneus is fixed as the x-axis, and the line that is perpendicular to the x-axis and intersects the x-axis at point O is fixed as the y-axis. The mean values of the y-coordinates at the points of the medial longitudinal arch of the foot in the patients with hallux valgus were significantly lower than those in the normal subjects (P < 0.001). The values were noticeably lower at points on the navicular. However, no change was observed in the posterior talus body. The new coordinate system indicated that the main points of flattening of the foot in patients with hallux valgus were the plantar flexion of the talus and the sinking of the navicular.

Adolescent↗

The distribution of excitatory amino acid binding sites in the brain of an electric fish, Apteronotus leptorhynchus.

The distribution of three types of exitatory amino acid receptors was examined in the brain of a high frequency weakly electric fish, Apteronotus leptorhynchus, by localizing the binding sites of ligands selective for mammalian kainic acid (KA), quisqualate (AMPA) and N-methyl-D-aspartate (NMDA) receptors. All three binding sites were densest within the forebrain and in certain hypothalamic nuclei (nucleus tuberis anterior, inferior lobe). The core of the dorsal forebrain (dorsal centralis) had a very high density of NMDA binding sites and only moderate levels of AMPA and KA binding sites, while this was reversed for the dorsolateral forebrain. The AMPA and NMDA binding sites were found throughout the brain while KA binding sites were relatively restricted and were absent from most of the brainstem. The cerebellar molecular layer contained a very high density of KA and AMPA binding sites but almost no NMDA binding sites; the granular layer had a low density of AMPA and NMDA binding sites but was lacking in KA binding sites. All three types of binding sites were found within the electromotor system (nucleus electrosensorius and prepacemaker nucleus) at sites where the iontophoresis of glutamate causes species-specific behaviours. KA binding sites were found at only two sites along the electrosensory afferent pathways: (1) in the molecular layer of the electrosensory lateral line lobe, associated with a feedback pathway emanating from granule cells of the overlying cerebellum, and (2) in the lateral nucleus praeminentialis dorsalis, associated with a descending pathway emanating from the torus semicircularis. NMDA and AMPA binding sites are found throughout the electrosensory pathways. Within the electrosensory lateral line lobe the NMDA binding sites were predominantly associated with the feedback pathways terminating in its molecular layer and not with the deep neuropil layer containing primary electroreceptor afferents.

Animals↗

The reconstitution and activity of the small multidrug transporter EmrE is modulated by non-bilayer lipid composition.

The ability of multidrug transport proteins within biological membranes to recognise a diverse array of substrates is a fundamental aspect of antibiotic resistance. Detailed information on the mechanisms of recognition and transport can be provided only by in vitro studies in reconstituted bilayer systems. We describe the controlled, efficient reconstitution of the small multidrug transporter EmrE in a simple model membrane and investigate the effect of non-bilayer lipids on this process. Transport activity is impaired, in line with an increase in the lateral pressure within the bilayer. We demonstrate the potential of this lateral pressure modulation method as a general approach to the folding and assembly of membrane proteins in vitro, by recovering functional transporter from a partly denatured state. Our results highlight the importance of optimising reconstitution procedures and bilayer lipid composition in studies of membrane transporters. This is particularly pertinent for multidrug proteins, and we show that the use of a sub-optimal lipid bilayer environment or reconstitution method could lead to incorrect information on protein activity.

Amino Acid Sequence↗

Lateral flagella of Aeromonas species are essential for epithelial cell adherence and biofilm formation.

Mesophilic Aeromonas strains express a single polar flagellum in all culture conditions and produce lateral flagella on solid media. Such hyperflagellated cells demonstrate increased adherence. Nine lateral flagella genes, lafA-U for Aeromonas hydrophila, and four Aeromonas caviae genes, lafA1, lafA2, lafB and fliU, were isolated. Mutant characterization, nucleotide and N-terminal sequencing demonstrated that the A. hydrophila and A. caviae lateral flagellins were almost identical, but were distinct from their polar flagellum counterparts. The aeromonad lateral flagellins exhibited higher molecular masses on SDS-PAGE, and this aberrant migration was thought to result from post-translational modification through glycosylation. Mutation of the Aeromonas lafB, lafS or both A. caviae lateral flagellins caused the loss of lateral flagella and a reduction in adherence and biofilm formation. Mutations in lafA1, lafA2, fliU or lafT resulted in strains that expressed lateral flagella, but had reduced adherence levels. Mutation of the lateral flagella loci did not affect polar flagellum synthesis, but the polarity of the transposon insertions on the A. hydrophila lafTlU genes resulted in non-motility. However, mutations that abolished polar flagellum production also inhibited lateral flagella expression. We conclude that Aeromonas lateral flagella: (i) play a role in adherence and biofilm formation; (ii) are distinct from the polar flagellum; (iii) synthesis is dependent upon the presence of a polar flagellum filament; and (iv) that the motor proteins of the polar and lateral flagella systems appear to be shared.

Aeromonas↗

Soft tissue profile changes after maxillary advancement with distraction osteogenesis by use of a rigid external distraction device: a 1-year follow-up.

PURPOSE: This study was conducted to evaluate the soft tissue profile changes after maxillary advancement with distraction osteogenesis (DO). PATIENTS AND METHODS: Sixteen subjects underwent maxillary advancement with rigid external distraction after a high Le Fort I osteotomy. There were 11 male and 5 female patients, ages 5.2 to 25.7 years. The subjects included 9 with unilateral cleft lip and palate (UCLP), 4 with bilateral CLP, 2 with facial clefts and bilateral CLP, and 1 with cleft palate. Pretreatment and posttreatment lateral cephalograms were compared to evaluate the changes in soft tissue profile. A line 7 degrees below the SN plane was used as the horizontal coordinate, and a perpendicular line through Sella was used as the vertical coordinate in an XY coordinate system. RESULTS: The preoperative facial concavity (N'SnPg') was reduced by 15.59 degrees, and the nasal tip moved 3.75 mm forward and 2.05 mm upward. These changes were positively correlated with the change of ANS position. The soft-tissue-to-hard-tissue ratio was 0.53:1 for nasal tip and ANS. The ratio was negatively correlated with the age of the patient. The ratio of soft tissue A point to skeletal A point was 0.96:1 and for the incisal edge to vermilion border of the upper lip it was 0.8:1. The soft tissue B point and Pg did not change significantly with maxillary distraction. However, the nasolabial angle increased by 4.96 degrees, the upper lip curvature flattened by 0.65 mm, and the lower lip curvature was accentuated by 0.89 mm after distraction. The amount of upper incisal exposure increased from 1.1 to 5.01 mm in the rest position. CONCLUSION: Maxillary DO improved the soft tissue profile by increasing nasal projection, normalizing the nasolabial angle, and making the upper lip more prominent. More upper anterior tooth show in the rest position was obtained, but the upper lip length did not change. The concave facial profile became convex, with improved facial balance and aesthetics.

Adolescent↗

A mathematical model of line bisection behaviour in neglect.

Subjects with left hemispatial neglect frequently demonstrate an array of abnormal behaviours on line bisection tasks. They misbisect long horizontal lines to the right of true midline. They bisect short lines to the left of true midline. They exaggerate the left-sided length of lines when placing the endpoints for 'invisible' lines, and they underestimate the length of the left side of long lines that are shown to them bisected accurately. No current theory of neglect explains all these features of line bisection behaviour. A mathematical model of line bisection behaviour in neglect is presented that proposes that subjects bisect lines at the point where they perceive the 'salience' of the two line segments created by their bisection mark to be equal. Salience is determined by the brain's attentional systems which map salience amplitude to spatial position following a bell shaped distribution. Right hemisphere strokes simulated by decreasing the 'height' and "breadth' of the right hemisphere salience to position function produced all of the above features of clinical neglect subjects' line bisection behaviour. Neglect may be conceived of as damage to brain systems performing mappings between stimulus characteristics (such as spatial location) and salience.

Attention↗

Anticonvulsive effects of kappa-opioid receptor modulation in an animal model of ethanol withdrawal.

Although the neurochemical mechanisms contributing to alcohol withdrawal seizures are poorly understood, withdrawal seizures probably reflect neuronal hyperexcitability resulting from adaptation to chronic alcohol. Altered kappa-Opioid receptor (KOP-R) signaling has been observed in multiple seizure types; however, a role for this system in ethanol withdrawal seizures has not been systematically characterized. We hypothesized that pharmacological manipulations of the KOP-R would alter withdrawal in mice selectively bred for differences in ethanol withdrawal severity. Withdrawal Seizure-Prone (WSP) and Withdrawal Seizure-Resistant (WSR) mice were made physically dependent using chronic ethanol vapor inhalation, and the effects of the KOP-R antagonist nor-binaltorphimine or agonist U-50,488H on withdrawal severity were examined. Pretreatment with nor-binaltorphimine significantly increased handling-induced convulsion (HIC) severity in withdrawing WSR mice, with no observable effects in withdrawing WSP mice. In contrast, U-50,488H significantly decreased HIC severity in WSP mice, with no effects in WSR mice. During extended withdrawal (i.e. hours 12+), a rebound hyperexcitability was observed in WSP mice given agonist. Thus, administration of a KOP-R antagonist increased withdrawal severity in mice normally resistant to withdrawal seizures, while a KOP-R agonist reduced convulsion severity in animals susceptible to withdrawal seizures. These observations are consistent with differences in the KOP-R system observed in these lines at the molecular level, and suggest the KOP-R system may be a promising therapeutic target for management of ethanol withdrawal seizures. Finally, these findings underscore the importance of determining the potential for rebound increases in withdrawal severity during later withdrawal episodes.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

The orphan nuclear receptors NURR1 and NGFI-B modulate aromatase gene expression in ovarian granulosa cells: a possible mechanism for repression of aromatase expression upon luteinizing hormone surge.

Ovarian granulosa cells play pivotal roles in many aspects of ovary functions including folliculogenesis and steroidogenesis. In response to FSH and LH, the elevation of intracellular cAMP level in granulosa cells leads to activation of multiple ovarian genes. Here, we report findings from a genome-wide study of the cAMP-responsive gene expression profiles in a human granulosa-like tumor cell line, KGN. The study identified 140 genes that are either activated or repressed by 2-fold or greater after stimulation by the adenylyl cyclase activator forskolin. The induction patterns of some cAMP-responsive genes were further analyzed by quantitative real-time PCR. Consistent with previous observations, the LH-responsive genes, such as the nuclear receptor 4A subfamily (NURR1, NGFI-B, and NOR-1), were rapidly but transiently induced, whereas the FSH-responsive gene CYP19 encoding aromatase was induced in a delayed fashion. Interestingly, ectopic expression of NURR1 or NGFI-B severely attenuated the cAMP-responsive activation of the ovary-specific aromatase promoter. Reduction of the endogenous NURR1 or NGFI-B by small interfering RNA significantly elevated aromatase gene expression. The cis-elements responsible for NURR1/NGFI-B-mediated repression were mapped to the minimal aromatase promoter sequence that confers camp responsiveness. Furthermore, the DNA-binding domain of NURR1 was required for the repression. Taken together, these results strongly suggest a causal relationship between the rapid decline of aromatase mRNA and induction of nuclear receptor subfamily 4A expression, which concomitantly occur upon LH surge at the later stages of ovarian follicular development.

Aromatase↗

Neuronal sensitivity to microsecond time disparities in the electrosensory system of Gymnarchus niloticus.

To perform the jamming avoidance response (JAR), the weakly electric fish Gymnarchus detects time disparities on the order of microseconds between electrosensory signals received by electroreceptors in different parts of the body surface. This paper describes time-disparity thresholds of output neurons of the electrosensory lateral line lobe (ELL), where the representation of timing information is converted from a time code to a firing-rate code. We recorded extracellular single-unit responses from pyramidal cells in the ELL to sinusoidally modulated time disparity with various depths (0-200 micros). Threshold sensitivity to time disparities measured in 123 units ranged from 0.5 to 100 micros and was < or =5 micros in 60% of the units. The units from pyramidal cells in the inner and outer cell layers of the ELL responded equally well to small time disparities. The neuronal thresholds to time disparities found in the ELL are comparable with those demonstrated in behavioral performance of the JAR. The sensitivity of ELL units to small time disparities was unaffected when the center of the cyclic time-disparity modulation was shifted over a wide range (up to 250 micros), indicating an adaptation mechanism for steady-state time disparities that preserves the sensitivity to small dynamic changes in time disparities. Phase-locked input neurons, which provide time information to the ELL by phase-locked firing of action potentials, did not adapt to steady-state time shifts of sensory signals. This suggests that the adaptation emerges within the ELL.

Action Potentials↗

Postsplenectomy recurrence of thrombocytopenia with an accessory spleen.

Autoimmune thrombocytopenic purpura (AITP) is an autoimmune disorder that results from antiplatelet autoantibodies; these autoantibodies cause platelet destruction in the reticluoendothelial system. Oral corticosteroid therapy is the first line treatment. Splenectomy is the major treatment modality after the failure of more conservative medical therapy. Approximately 15% of the patients will relapse either soon after splenectomy or, as is less common, many years later. The presence of an accessory spleen should be sought. We experienced a patient with a known diagnosis of autoimmune thrombocytopenic purpura who had a worsening thrombocytopenia 11 years after splenectomy. This patient was diagnosed with an accessory spleen. Accessory splenectomy was performed with only a transient elevation of the platelets. We report here on this case with a review of the literature.

Adult↗

[Analysis of the changes of movement function and viability in human spermatozoa induced by reactive oxygen species].

OBJECTIVES: To analyze the changes of movement function and viability in human spermatozoa induced by reactive oxygen species(ROS), and to prove whether ROS were one of the causes of the movement dysfunction of spermatozoa. METHODS: Spermatozoa with normal physiological functions selected from semen samples by Percoll gradient centrifugation technique were regarded as normal sperm models. ROS were generated by hypoxanthine-xanthine oxidase system and then incubated with normal sperm models under aerobic environment. After model spermatozoa were incubated with ROS, movement parameters of spermatozoa were analyzed by computer-assisted semen analysis (CASA) system. RESULTS: Compared with the control group, after model spermatozoa were incubated with ROS for 30 minutes, motility, curvilinear velocity(VCL), straight line velocity (VSL) and average path velocity (VAP) of the spermatozoa were significantly decreased (P < 0.001), but amplitude of lateral head displacement (ALH) was insignificant (P > 0.05). When incubated with ROS for 60 minutes, spermatozoa almost lost movement function and all movement parameters of spermatozoa inclined to zero. CONCLUSIONS: When normal spermatozoa were incubated with ROS, movement functions of sperm were decreased. It was demonstrated that ROS were one of the causes of the movement dysfunction of spermatozoa.

Adult↗

[Lumbosacral dermal sinus as entry point of purulent meningitis. Report of 3 cases].

Defects of neural tube closure, although minimal, can provide access for infections of the central nervous system. All skin alterations in rear middle line of the body, however, minimal, must be carefully investigated as they could give access to bacterial meningitis. We present three new cases of dermal lumbosacral sinus which went unnoticed in the neonatal period later becoming the access point for bacterial meningitis.

Bacterial Infections↗