Carditis: a relevant marker of gastroesophageal reflux disease. Data from a prospective central European multicenter study on histological and endoscopic diagnosis of esophagitis (histoGERD trial).

The columnar-lined mucosa at the gastroesophageal junction may contain an inflammatory infiltrate, commonly referred to as carditis (or cardia gastritis). The etiology of carditis is not entirely clear since published data are conflicting. Some authors believe it to be secondary to gastroesophageal reflux disease (GERD) and others to Helicobacter pylori gastritis. This prospective study aims at clarifying the relationship between carditis and the histological, clinical, and endoscopic findings of GERD, in a large cohort of individuals negative for H. pylori infection. Eight hundred and seventy-three individuals (477 females and 396 males, median age 53 years) participated in this study. Biopsy material was systematically sampled from above and below the gastroesophageal junction. Reflux-associated changes of the esophageal squamous epithelium were assessed according to the Esohisto consensus guidelines. Grading of carditis was performed according to the Updated Sydney System, known from the histological evaluation of gastritis. In total, 590 individuals (67.5%) had chronic carditis. Of these, 468 (53.6%) had mild chronic inflammation, with 321 individuals (68.6%) showing no or minimal changes on endoscopic examination (Los Angeles Categories N and M). The presence of chronic carditis was associated with several GERD-related parameters of the esophageal squamous epithelium (P < 0.0001), and data retained statistical significance even when analysis was restricted to individuals with mild chronic carditis and/or endoscopically normal mucosa. Chronic carditis was also associated with the presence of intestinal metaplasia (P < 0.0001). In addition, chronic carditis had a statistically significant association with patients' symptoms of GERD (P = 0.0107). This observation remained valid for mild chronic carditis in all patients (P = 0.0038) and in those with mild chronic carditis and normal endoscopic mucosa (P = 0.0217). In conclusion, chronic carditis appears to be the immediate consequence of GERD, correlating with patients' symptoms and endoscopic diagnosis. These results are valid in individuals with nonerosive reflux disease, which indicates a higher sensitivity of histological diagnosis. Our findings may impact the routine assessment of reflux patients.

Neurexins: synaptic cell surface proteins related to the alpha-latrotoxin receptor and laminin.

A family of highly polymorphic neuronal cell surface proteins, the neurexins, has been identified. At least two genes for neurexins exist. Each gene uses alternative promoters and multiple variably spliced exons to potentially generate more than a 100 different neurexin transcripts. The neurexins were discovered by the identification of one member of the family as the receptor for alpha-latrotoxin. This toxin is a component of the venom from black widow spiders; it binds to presynaptic nerve terminals and triggers massive neurotransmitter release. Neurexins contain single transmembrane regions and extracellular domains with repeated sequences similar to sequences in laminin A, slit, and agrin, proteins that have been implicated in axon guidance and synaptogenesis. An antibody to neurexin I showed highly concentrated immunoreactivity at the synapse. The polymorphic structure of the neurexins, their neural localization, and their sequence similarity to proteins associated with neurogenesis suggest a function as cell recognition molecules in the nerve terminal.

SV2A and SV2B function as redundant Ca2+ regulators in neurotransmitter release.

SV2 proteins are abundant synaptic vesicle proteins expressed in two major (SV2A and SV2B) and one minor isoform (SV2C) that resemble transporter proteins. We now show that SV2B knockout mice are phenotypically normal while SV2A- and SV2A/SV2B double knockout mice exhibit severe seizures and die postnatally. In electrophysiological recordings from cultured hippocampal neurons, SV2A- or SV2B-deficient cells exhibited no detectable abnormalities. Neurons lacking both SV2 isoforms, however, experienced sustained increases in Ca2+-dependent synaptic transmission when two or more action potentials were triggered in succession. These increases could be reversed by EGTA-AM. Our data suggest that without SV2 proteins, presynaptic Ca2+ accumulation during consecutive action potentials causes abnormal increases in neurotransmitter release that destabilize synaptic circuits and induce epilepsy.

Functional properties of multiple synaptotagmins in brain.

At least four forms of synaptotagmin are expressed in neurons. Of these, synaptotagmin I has an essential function in mediating Ca(2+)-triggered neurotransmitter release at hippocampal synapses, but the functional implications of multiple synaptotagmins are unknown. Synaptotagmins I-III exhibit a strikingly differential distribution between synapses, with most neurons coexpressing either synaptotagmins I or II with III. Synaptotagmin IV is present uniformly throughout the brain at low levels. Synaptotagmins III and IV are both coexpressed with synaptotagmin I in hippocampal synapses, suggesting that these synaptotagmins are not functionally redundant. The first C2 domains of synaptotagmins I-III exhibit similar Ca2+ affinities in phospholipid-binding assays, whereas that of synaptotagmin IV is unable to bind Ca2+. All synaptotagmins tested bind the clathrin-adaptor protein AP-2 with high affinity. Our results suggest that different synaptotagmins serve distinct but overlapping functions in neuronal membrane traffic, with synaptotagmins I and II representing alternative Ca2+ sensors in exocytosis and all synaptotagmins functioning as AP-2 receptors in endocytosis.

Synaptic targeting of rabphilin-3A, a synaptic vesicle Ca2+/phospholipid-binding protein, depends on rab3A/3C.

rab3A, a low molecular weight GTP-binding protein of synaptic vesicles with a putative function in synaptic vesicle docking, interacts in a GTP-dependent manner with rabphilin-3A, a peripheral membrane protein that binds Ca2+ and phospholipids. We now show that rabphilin-3A is an evolutionarily conserved synaptic vesicle protein that is attached to synaptic vesicle membranes via its N terminus and exhibits a heterogeneous distribution among synapses. In rab3A-deficient mice, rabphilin-3A is decreased in synapses belonging to neurons that primarily express rab3A and accumulates in the perikarya of these neurons. In contrast, neurons expressing significant levels of rab3C still contain normal levels of rabphilin-3A in a synaptic pattern, and rabphilin-3A binds rab3C in vitro. These results suggest that analogous to the membrane recruitment of raf by ras, rab3A and rab3C may function in recruiting rabphilin-3A to the synaptic vesicle membrane in a GTP-dependent manner.

Neuropilin-2 is required in vivo for selective axon guidance responses to secreted semaphorins.

Neuropilins are receptors for class 3 secreted semaphorins, most of which can function as potent repulsive axon guidance cues. We have generated mice with a targeted deletion in the neuropilin-2 (Npn-2) locus. Many Npn-2 mutant mice are viable into adulthood, allowing us to assess the role of Npn-2 in axon guidance events throughout neural development. Npn-2 is required for the organization and fasciculation of several cranial nerves and spinal nerves. In addition, several major fiber tracts in the brains of adult mutant mice are either severely disorganized or missing. Our results show that Npn-2 is a selective receptor for class 3 semaphorins in vivo and that Npn-1 and Npn-2 are required for development of an overlapping but distinct set of CNS and PNS projections.

Rabphilin knock-out mice reveal that rabphilin is not required for rab3 function in regulating neurotransmitter release.

Rab3A and rab3C are GTP-binding proteins of synaptic vesicles that regulate vesicle exocytosis. Rabphilin is a candidate rab3 effector at the synapse because it binds to rab3s in a GTP-dependent manner, it is co-localized with rab3s on synaptic vesicles, and it dissociates with rab3s from the vesicles during exocytosis. Rabphilin contains two C(2) domains, which could function as Ca(2+) sensors in exocytosis and is phosphorylated as a function of stimulation. However, it is unknown what essential function, if any, rabphilin performs. One controversial question regards the respective roles of rab3s and rabphilin in localizing each other to synaptic vesicles: although rabphilin is mislocalized in rab3A knock-out mice, purified synaptic vesicles were shown to require rabphilin for binding of rab3A but not rab3A for binding of rabphilin. To test whether rabphilin is involved in localizing rab3s to synaptic vesicles and to explore the functions of rabphilin in regulating exocytosis, we have now analyzed knock-out mice for rabphilin. Mice that lack rabphilin are viable and fertile without obvious physiological impairments. In rabphilin-deficient mice, rab3A is targeted to synaptic vesicles normally, whereas in rab3A-deficient mice, rabphilin transport to synapses is impaired. These results show that rabphilin binds to vesicles via rab3s, consistent with an effector function of rabphilin for a synaptic rab3-signal. Surprisingly, however, no abnormalities in synaptic transmission or plasticity were observed in rabphilin-deficient mice; synaptic properties that are impaired in rab3A knock-out mice were unchanged in rabphilin knock-out mice. Our data thus demonstrate that rabphilin is endowed with the properties of a rab3 effector but is not essential for the regulatory functions of rab3 in synaptic transmission.

Identification of new SNPs in native South American populations by resequencing the Y chromosome.

The Y-chromosomal genetic landscape of South America is relatively homogenous. The majority of native Amerindian people are assigned to haplogroup Q and only a small percentage belongs to haplogroup C. With the aim of further differentiating the major Q lineages and thus obtaining new insights into the population history of South America, two individuals, both belonging to the sub-haplogroup Q-M3, were analyzed with next-generation sequencing. Several new candidate SNPs were evaluated and four were confirmed to be new, haplogroup Q-specific, and variable. One of the new SNPs, named MG2, identifies a new sub-haplogroup downstream of Q-M3; the other three (MG11, MG13, MG15) are upstream of Q-M3 but downstream of M242, and describe branches at the same phylogenetic positions as previously known SNPs in the samples tested. These four SNPs were typed in 100 individuals belonging to haplogroup Q.

Role of alpha-synuclein in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced parkinsonism in mice.

In humans, mutations in the alpha-synuclein gene or exposure to the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) produce Parkinson's disease with loss of dopaminergic neurons and depletion of nigrostriatal dopamine. alpha-Synuclein is a vertebrate-specific component of presynaptic nerve terminals that may function in modulating synaptic transmission. To test whether MPTP toxicity involves alpha-synuclein, we generated alpha-synuclein-deficient mice by homologous recombination, and analyzed the effect of deleting alpha-synuclein on MPTP toxicity using these knockout mice. In addition, we examined commercially available mice that contain a spontaneous loss of the alpha-synuclein gene. As described previously, deletion of alpha-synuclein had no significant effects on brain structure or composition. In particular, the levels of synaptic proteins were not altered, and the concentrations of dopamine, dopamine metabolites, and dopaminergic proteins were unchanged. Upon acute MPTP challenge, alpha-synuclein knockout mice were partly protected from chronic depletion of nigrostriatal dopamine when compared with littermates of the same genetic background, whereas mice carrying the spontaneous deletion of the alpha-synuclein gene exhibited no protection. Furthermore, alpha-synuclein knockout mice but not the mice with the alpha-synuclein gene deletion were slightly more sensitive to methamphetamine than littermate control mice. These results demonstrate that alpha-synuclein is not obligatorily coupled to MPTP sensitivity, but can influence MPTP toxicity on some genetic backgrounds, and illustrate the need for extensive controls in studies aimed at describing the effects of mouse knockouts on MPTP sensitivity.

Presynaptic opioid receptors modulating acetylcholine release in the hippocampus of the rabbit.

Slices of the rabbit hippocampus were preincubated with 3H-choline, rinsed and superfused continuously. The release of 3H-acetylcholine in these slices, evoked by electrical field stimulation, was strongly reduced by the preferential kappa-agonists ethylketocyclazocine, dynorphin A (1-13) and dynorphin A (1-17). Dynorphin A (1-9) and (-)MR 2034 [(-)5,9-dimethyl-2'-OH-2-tetrahydrofurfuryl-6, 7-benzomorphan] were less potent, the (+)enantiomer of (-)MR 2034 was ineffective. Whereas the mu-agonist DAGO (D-Ala2-Gly-ol5-enkephalin) showed significant depressant effects, two other mu-agonists morphine and morphiceptine, as well as the delta-agonists DADLE (D-Ala2-D-Leu5-enkephalin) and Leu-enkephalin were much less inhibitory. The preferential mu-antagonist (-)naloxone as well as (-)MR 2266 [(-)N-(3-furylmethyl)-alpha-noretazocine], a preferential kappa-antagonist, did not increase acetylcholine release when given alone, but antagonized the effect of ethylketocyclazocine; (-)MR 2266 (Ke: 1.6 nmol/l) was about 4 times more potent than (-)naloxone (Ke: 6.3 nmol/l). The inhibitory effects of DAGO and DADLE were abolished by (-)MR 2266 (0.1 mumol/l) but not by the delta-antagonist ICI 174864 (N,N-diallyl-Tyr-Aib-Phe-Leu-OH, 0.3 mumol/l). It is concluded that the release of acetylcholine in the hippocampus of the rabbit is inhibited at the level of the axon terminals via kappa-receptors; in addition, mu-receptors may be present. An inhibitory tone of endogenous opioid peptides on hippocampal acetylcholine release could not be demonstrated. Experiments on rat hippocampal slices showed that in this species mu- rather than kappa-receptors may modulate acetylcholine release.

Pain in the posterior aspect of the ankle in dancers. Differential diagnosis and operative treatment.

A retrospective review was performed of the results of operative treatment of stenosing tenosynovitis of the flexor hallucis longus tendon or posterior impingement syndrome, or both, in thirty-seven dancers (forty-one operations). The average duration of follow-up was seven years (range, two to thirteen years). The results were assessed with use of a questionnaire for all patients, and a clinical evaluation was performed for twenty-one patients (twenty-two ankles). Twenty-six operations were performed for tendinitis and posterior impingement; nine, for isolated tendinitis; and six, for isolated posterior impingement syndrome. A medial incision was used in thirty-three procedures; a lateral incision, in six; an anterior and a medial incision, in one; and a lateral and a medial incision, in one. Thirty ankles had a good or excellent result; six, a fair result; and four, a poor result. (The result of the second procedure on an ankle that was operated on twice was not included.) The result was good or excellent for twenty-eight of the thirty-four ankles in professional dancers, compared with only two of the six ankles in amateur dancers.

The lymphocyte-epithelial cell ratio in tubal mucosa of patients with cervical carcinoma.

The lymphocytes in the luminal subepithelial basement membrane of the uterine tube from patients with cervical carcinoma were counted, and the results compared with those obtained from women of the same age but without cervical carcinoma (cf. Geppert et al. 1977) to determine whether a relationship exists between emigration of lymphocytes into the tubal epithelium and the clinical diagnosis of cervical carcinoma. The comparison indicated that the number of lymphocytes emigrating into the tubal epithelium during the secretory and the proliferative phase in women with cervical carcinoma is significantly lower than that in healthy women from the control group. The differences during the secretory phase were particularly pronounced. The lymphocyte count in patients with cervical carcinoma was only one third that of the healthy women in the control group. Assuming that an immunologic defect is determinative for the development of carcinoma, a decline in the number of lymphocytes emigrating into the tubal epithelium can be interpreted as the reflexion of an immunologic deficiency. It is possible that a disturbance in the lymphocytes epithelial cell ratio in this stage of life could indicate a disposition for those tumors the incidence of which is high in this age group (e.g., squamous cell carcinoma of the cervix). Further study is necessary to establish whether these cells be classified as B or T lymphocytes.

Endometrial cancer associated with polycystic ovaries in young women.

Two women (17 and 36 years old) are presented, who complained about abnormal vaginal bleeding. The rare diagnosis of an adenocarcinoma of the corpus uteri with concomitant polycystic ovaries (Stein-Leventhal-Syndrome) was made.

[Influence of histopathological factors on dynamic MR mammography]. / Einfluss histopathologischer Faktoren auf die dynamische MR-Tomographie der Mamma.

AIM: To assess the histopathological background of enhancement mechanisms in dynamic MR mammography studies. METHODS: The dynamic MR mammography (MRM) examinations were done with a 1.5 T MR imager (Magnetom Vision, Siemens) using a double breast coil and a coronal FLASH-3D sequence. Enhancement data were acquired during 9 minutes post contrast medium injection (Gd-DTPA 0.2 mmol/kg). Acquisition time was 87 sec/slab. Early enhancement at the first post contrast measurement (E1) and slope of wash-out (SE2-L) were calculated. In immunohistology, proliferation was assessed by the monoclonal antibody Ki 67, capillaries were stained by a CD 31 antibody. Of a total of 48 operated patients, 58 lesions and 46 surrounding tissues were evaluated. RESULTS: Cellularity, capillary density and proliferation showed statistically significant correlations with E1 (p < 0.01). In multiple regression analysis, E1 was significantly associated only with high cellularity (p = 0.002) and the combination of high cellularity and high microvessel density (p = 0.002); a negative slope of wash out was significantly associated only with malignant histology (p = 0.027). CONCLUSIONS: Our findings indicate a direct influence of cellularity and microvessel density on early enhancement. The expression of the proliferation marker Ki 67 was not an independent predictor for contrast enhancement.

Pseudolymphoma of the nipple.

Pseudolymphoma of the breast is a rare benign disorder. To avoid over-treatment or misdiagnosis of a malignancy of the breast, exact histological diagnosis is important. We report on a 69-year-old patient, who presented with symptoms of mastitis.

Serial transplants of DMBA-induced mammary tumors in fischer rats as model system for human breast cancer: V. Myoepithelial-mesenchymal conversion during passaging as possible cause for modulation of pineal-tumor interaction.

An elevation of melatonin secretion parallel to an enhanced production of macrophage-derived biopterin was observed in female F344 Fischer rats bearing passage 2 serial transplants derived from a malignant mammary tumor induced by 7,12-dimethylbenz[a]anthracene (DMBA). As opposed to that both parameters were depressed at passage 12. These results indicate the presence of divergent immunoneuroendocrine interactions during different phases of tumor growth. Since these biochemical events must have their common origin in changes taking place within these tumor transplants the current histopathological study was initiated. The primary tumor used for serial transplantation was a moderately differentiated adenocarcinoma of the mammary gland showing cytokeratin-positive epithelial components located in the inner epithelial tubule layer. In addition, bland-looking round or elongated actin-positive myoepithelial cells were detected which apart from epithelial cells are known to constitute the main cellular components of the mammary ductal system which resemble smooth muscle cells both morphologically and functionally. The tumor of passage 1 showed glandular tubules, lined by an inner epithelial layer, and many nests of clear, bland-looking actin-positive myoepithelial cells lying around tubules as well as in the stroma between actin-negative epithelial elements. The tumor of passage 2 used for transplantation consisted of a chaotic mixture of epithelial carcinomatous cells, forming a few irregular small tubules or solid nests, and, predominantly, of elongated plump or spindle-shaped, "myoid" atypical myoepithelial cells with a strong actin-positive reaction and some of these cells showed a focal vimentin expression. The tumor was characterized as a carcinosarcoma. At passage 12 epithelial cells were not identified. The tumor displayed features of a pleomorphic sarcoma consisting mainly of giant cells with bizarre nuclei being cytokeratin- and desmin-negative, weakly vimentin-positive but strongly actin-positive. These results indicate that DMBA-induced mammary tumor cells in female F344 Fischer rats undergo dramatic morphological changes during serial transplantation characterized by a total loss of malignant epithelial (carcinomatous) cells and the emergence and subsequent predominance of malignant (sarcomatous) mesenchymal cells. It appears that these sarcomatous cells develop out of myoepithelial cells since atypical myoepithelial cells with a strong actin-positive reaction showed a focal vimentin expression at passage 2 indicating myofibroblastic differentiation as part of mesenchymal transition. The loss of epithelial cell elements as well as a parallel transition of myoepithelial to mesenchymal cell elements during passaging could lead to a lack of immunological recognition of these tumor transplants and to depression of melatonin. Possible mechanisms involved in these phenomena as well as the relevance of these findings for a better understanding of the role of melatonin in human mammary cancer are discussed.

Histologic changes in dog lymph nodes after endolymphatic application of bleomycin oil suspension.

Bleomycin oil suspension (Oil-Bleo) was injected endolymphatically in a dog experiment. The drug remained in lymph nodes over many weeks in detectable concentrations. The histologic changes in these lymph nodes were examined and compared with 3 controls (aqueous Bleomycin, Lipiodol, and no treatment). Granulomatous reaction, microabscesses, and small necrotic foci were observed. After a month, fibrosis was detectable in all lymph nodes. The damage to microscopic lymph node structures after Oil-Bleo was moderate. These findings suggest that Oil-Bleo may be useful in local treatment of lymph node metastases.

Retroperitoneal lymph nodal visualization using 30% Guajazulen blue (chromolymphography).

Guajazulen, a dark blue lipid-soluble dye was used to stain retroperitoneal lymph nodes before lymphadenectomy. Initially, a 30% solution of Guajazulen (Merck, Darmstadt, FRG) in iodinated oil (Lipiodol, Byk-Gulden, Konstant, FRG) was injected endolymphatically in the hindlimbs of pigs (n = 10) and dogs (n = 8) to better visualize retroperitoneal lymph nodes. Because of encouraging results, 11 patients with testicular cancer undergoing staging or therapeutic laparotomy underwent endolymphatic injection of this dye solution preoperatively. The excellent staining of retroperitoneal lymph nodes facilitated nodal selectivity during lymphadenectomy.

Painful os peroneum syndrome: a spectrum of conditions responsible for plantar lateral foot pain.

Plantar lateral foot pain may be caused by various entities and the painful os peroneum syndrome (a term coined by the authors) should be included in the differential diagnosis. Painful os peroneum syndrome results from a spectrum of conditions that includes one or more of the following: (1) an acute os peroneum fracture or a diastasis of a multipartite os peroneum, either of which may result in a discontinuity of the peroneus longus tendon; (2) chronic (healing or healed) os peroneum fracture or diastasis of a multipartite os peroneum with callus formation, either of which results in a stenosing peroneus longus tenosynovitis; (3) attrition or partial rupture of the peroneus longus tendon, proximal or distal to the os peroneum; (4) frank rupture of the peroneus longus tendon with discontinuity proximal or distal to the os peroneum; and/or (5) the presence of a gigantic peroneal tubercle on the lateral aspect of the calcaneus which entraps the peroneus longus tendon and/or the os peroneum during tendon excursion. Familiarity with the various clinical and radiographic findings and the spectrum of conditions represented by the painful os peroneum syndrome can prevent prolonged undiagnosed plantar lateral foot pain. Clinical diagnosis of the painful os peroneum syndrome can be facilitated by the single stance heel rise and varus inversion stress test as well as by resisted plantarflexion of the first ray, which can localize tenderness along the distal course of the peroneus longus tendon at the cuboid tunnel.(ABSTRACT TRUNCATED AT 250 WORDS)

Gross, histological, and microvascular anatomy and biomechanical testing of the spring ligament complex.

In recent years there has been an increased interest in the treatment of acquired pes planus. The breakdown of the medial longitudinal arch is most often seen at the talonaviculocalcaneal articulation. This suggests a relationship between the ligamentous complex at this articulation and acquired pes planus. This study was undertaken to gain a better understanding of the gross, histologic, and microvascular anatomy, as well as the biomechanics of the ligamentous structures surrounding the talonaviculocalcaneal articulation. Cadaver dissections of 38 fresh-frozen feet were performed. Detailed descriptions of the gross anatomy of the superomedial calcaneonavicular ligament, inferior calcaneonavicular ligament, and the superficial deltoid ligament were recorded. Their relationships to the posterior tibialis tendon and to the bones of the talonaviculocalcaneal articulation are described. The histology and microvascularity of these structures were also studied. Preliminary biomechanical testing was performed. It was found there are two definitive anatomic structures that are commonly called the spring ligament: the superomedial calcaneonavicular ligament (SMCN) and the inferior calcaneonavicular ligament (ICN). The SMCN ligament was found to have histologic properties that suggest significant load bearing. The histology of the ICN ligament suggests a pure tensile load function. The deltoid ligament and the posterior tibialis tendon had direct attachments to the SMCN ligament in all specimens. An articular facet composed of fibrocartilage was found in each SMCN ligament specimen. The microvascular structures showed an avascular articular facet present in the ligament. The biomechanical testing showed that the SMCN ligament and ICN ligament had strength similar to ankle ligaments. This study suggests this "spring ligament complex" has more of a "sling" function for the talar head. It is hoped that the better understanding of this region will add to our understanding of the etiology of pes planus and possible treatment alternatives.