Differential expression of tissue-specific adhesion molecules on human circulating antibody-forming cells after systemic, enteric, and nasal immunizations. A molecular basis for the compartmentalization of effector B cell responses.

Expression of the adhesion molecules CD44, L-selectin (CD62L), and integrin alpha 4 beta 7 by antibody-secreting cells (ASC) was examined in human volunteers after oral, rectal, intranasal, or systemic immunization with cholera toxin B subunit. Almost all blood ASC, irrespective of immunization route, isotype (IgG and IgA), and immunogen, expressed CD44. On the other hand, marked differences were observed between systemically and intestinally induced ASC with respect to expression of integrin alpha 4 beta 7 and L-selectin, adhesion molecules conferring tissue specificity for mucosal tissues and peripheral lymph nodes, respectively. Thus, most ASC induced at systemic sites expressed L-selectin, whereas only a smaller proportion of ASC expressed alpha 4 beta 7. In contrast, virtually all IgA- and even IgG-ASC detected after peroral and rectal immunizations expressed alpha 4 beta 7, with only a minor fraction of these cells expressing L-selectin. Circulating ASC induced by intranasal immunization displayed a more promiscuous pattern of adhesion molecules, with a large majority of ASC coexpressing L-selectin and alpha 4 beta 7. These results demonstrate that circulating ASC induced by mucosal and systemic immunization express different sets of adhesion molecules. Furthermore, these findings provide for the first time evidence for differential expression of adhesion molecules on circulating ASC originating from different mucosal sites. Collectively, these results may explain the anatomical division of mucosal and systemic immune responses in humans as well as the compartmentalization of mucosal immune responses initiated in the upper vs. the lower aerodigestive tract.

Recovery from orbital floor fractures: a prospective study of patients' and doctors' experiences.

This study aimed at investigating the extent to which remaining symptoms and signs troubled patients in the year after suffering from zygomatico-orbital fractures, and whether there was any discrepancy between patients' and doctors' opinions as to the presence of symptoms and signs. Over the course of 1 year, 46 patients were included. Symptoms and clinical findings were registered in a 'doctor's protocol', and patients described self-reported symptoms and signs using a visual analogue scales (VAS) in a questionnaire administered 5 times during the year after injury. The VAS proved to be a useful instrument for evaluating patient discomfort and indicating differences between patients' and doctors' opinions regarding the presence of symptoms and signs. Agreement between the two was good regarding the presence of objective and measurable signs, such as facial asymmetry and diplopia. When it came to sensibility and mouth-opening ability, however, discrepancies were evident. It is desirable that reliable methods for measuring sensibility and evaluating mouth opening are included in follow-up routines. This would increase our knowledge of the course of healing, prognosis and possibilities for the prevention and active treatment of these problems.

A microradiographic investigation of cancellous bone healing after irradiation and hyperbaric oxygenation: a rabbit study.

PURPOSE: To analyze the effect of irradiation on cancellous bone healing at different times after irradiation and to study if hyperbaric oxygen therapy (HBO) would affect the bone healing capacity, when delivered directly after irradiation. METHODS AND MATERIALS: Rabbits were given a single dose of 15 Gy (60)Co radiation to one hind leg, the other hind leg serving as control. A standardized defect through the femoral metaphysis of the rabbits was created by a trephine drill biopsy at different times after irradiation. New bone formation in the defect was evaluated by a new biopsy through the previous defect after a healing time of 8 weeks. The mineral contents of the biopsies were analyzed by microradiography and microdensitometry. RESULTS: There was a large variation in the bone-forming capacity expressed as bone mineral content between the animals. No statistically significant differences could be detected regarding the effect of irradiation, HBO, or delayed surgery. Qualitative histology revealed more pronounced inflammation, fibrosis, and bone resorption in the irradiated bone. CONCLUSIONS: No definite conclusions can be drawn from the results of this study, however it might be hypothesized that cancellous bone recovers faster than cortical bone from radiation trauma.

A comparison of ATP-degrading enzyme activities in rat incisor odontoblasts.

In active odontoblasts from the rat incisor, used as a model system for biologic calcification, two distinguishable enzyme activities capable of degrading adenosine monophosphate (ATP) exist. Once can be inhibited ny 1-tetramisole, (+/-)-2,3,5,6,-tetrahydro-6-phenylimidazo (2.1B) THIAZOLE HYDROCHLORIDE (Levamisol) and (+/-)-6(m-bromophenyl)-5.6-dehydroimidazo (2.1-b) thiazole oxalate (R823) and is probably identical with nonspecific alkaline phosphatase (EC 3.1.3.1). The activity of the other enzyme, named Ca2+-ATPase, is dependent on the presence of Ca2+ or Mg2+ and is activated by these ions. The pH optimum of Ca2+-ATPase is 9.8. The Ca2+-ATPase is unaffected by Levamisole, R 8231, ouabain, ruthenium red, Na+ and K+ ions. Maximal activity was found against ATP, whereas adenosine diphosphate, guanosine triphosphate, inosine triphosphate and adensoine monophosphate were hydrolysed at lower rate. It may be speculated that the Ca2+-ATPase is concerned with the transmembranous transport of Ca2+ ions to the mineralization front.

Ultrastructural localization of alkaline phosphatases in rat incisor odontoblasts.

The localization of alkaline phosphatases in dentinogenically active rat incisor odontoblasts was studied by means of subcellular fractionation and electron microscopical histochemistry. Subcellular fractionation revealed the predominant phosphatase activity to be present in the microsome fraction and to a lesser extent in the mitochondrial fraction. Adenosine triphosphate degrading enzyme activity was determined in the presence or absence of (+/-)-6(m-bromophenyl)-5, 6-dihydroimidazo(le) (2,1-b) thiazole oxalate (R 8231). Before the histochemical study, the effects on phosphatase activities by aldehyde fixation were studied by biochemical assay. A method of fixation for optimal preservation of phosphatase activity is presented. Phosphatase electron microscopic histochemistry was performed by using ATP as a substrate and with or without addition of the inhibitor R 82319 Precipitates were seen in the membranes of vesicles present in the odontoblast process and the Golgi region. When there were signs of insufficient fixation, precipitates were also seen in the outer membranes of mitochondria. No phosphatase activity was seen in the cell membrane. ATP degrading enzyme activities mediated by nonspecific alkaline phosphatase (APase) and Ca2+ -adenosine triphosphatase thus have the same morphological localization. This close association is consistent with earlier biochemical studies.

Cytological and bacteriological aspects of secretory otitis media.

Bacteriological and cytological examinations were performed on 105 middle ear secretions from 66 children with middle ear effusion (MEE) of more than 3 months' duration. The secretions were searched for granulocytes and the activity of these cells was judged by their capacity for random locomotion and their ability to reduce nitroblue tetrazolium. The functional characteristics of the granulocytes were compared with the bacteriological findings on cultures from MEE. Bacteria commonly regarded as pathogens in middle ear infections (Hemophilus influenzae, Branhamella catarrhalis or Streptococcus pneumoniae) were found in 25% of the secretions. Granulocytes with activity or lacking activity, virtually dead, were found in all secretions where these bacteria were isolated. In secretions where bacteria commonly regarded as commensals, mainly staphylococci, were isolated, about two thirds of the secretions showed phagocytes with or without activity. No relation between bacterial growth and the functional state of the granulocytes was observed. In contrast, no phagocytes were found in over 60% of MEE lacking bacterial growth. These findings suggest a role for bacteria in the development and maintenance of secretory otitis media.

Evaluation of an improved DNA probe for diagnosis of pertussis.

A Bordetella pertussis specific subclone, pRZ61, of a Bordetella genus-specific clone, pB23, was evaluated on nasopharyngeal aspirates of 179 patients with suspected pertussis. Hybridization was performed directly after spotting or after 1-3 days of preculture of the nylon membranes on solid culture medium. A direct comparison of the two probes was obtained by reprobing with the subclone the same membranes that had been hybridized with the parent probe. pRZ61 detected 50% of the serologically defined cases of pertussis, that is, had the same sensitivity as standard culture. Specificity as compared with serology was close to 100%. The increasing sensitivity and the corresponding decreasing specificity after preculture noted for pB23 was not seen with the subclone. The study showed that the improved probe represents a rapid diagnostic method in pertussis.

Effect of levamisole on bone resorption in cultured mouse calvaria.

The effects of levamisole (LT), dexamisole (DT), levo-p-bromotetramisole (LBT) and dextro-p-bromotetramisole (DBT) on bone were examined in an organ culture system using calvarial bones from newborn mice. LBT and DBT at concentrations 30 microM and greater and LT and DT at concentrations 100 microM and greater caused a dose-dependent, reversible inhibitory effect on mineral mobilization and matrix degradation. LBT, DBT (100 and 300 microM) as well as LT and DT (greater than or equal to 100 microM) reduced the spontaneous release of beta-glucuronidase without having any marked effect on the release of lactate dehydrogenase (LDH). LT and DT did not influence protein synthesis but LBT and LBT were inhibitory in concentrations at and above 100 microM. Mitotic activity, as assessed by incorporation of [3H]thymidine, was inhibited by LBT and DBT (0.1, 1 mM). LT and LBT caused a stereospecific inhibition of GPase, PPiase and ATPase. It is concluded that tetramisoles are potent, non-stereospecific inhibitors of bone resorption in vitro.

Changes of serum alkaline phosphatase following mandibular osteotomy in the rat.

Activity of alkaline phosphatase in serum (S-ALP) was characterized and compared with that of non-specific alkaline phosphatase (APase) of hard-tissue origin in the rat. The enzyme was characterized biochemically, and optimal incubation procedures were determined. S-ALP levels were determined before and after mandibular osteotomy combined with different degrees of periosteal reflections. Separation of S-ALP isoenzymes by isoelectric focusing revealed four major bands. A marked decrease of S-ALP activity was seen after osteotomy, and all isoenzymes were affected similarly. After treatment with hyperbaric oxygen, a smaller decrease in S-ALP was seen. S-ALP could be used as a marker for hard-tissue turn-over after surgical procedures.

Diagnostic value of clinical and bacteriological findings in pertussis.

Clinical and bacteriological findings in the diagnosis of pertussis were evaluated in 300 consecutive patients with parental or the patient's own suspicion of the disease. Serology was used as a reference method. Of the 285 (95%) patients fully sampled, 163 (57%) were diagnosed as having pertussis while the remaining 122 patients constituted the non-pertussis control group. The clinical and epidemiological data were collected at the first visit made on median day seven of illness. In this population of mainly unimmunised children, the highest predictive values were obtained for the physician's diagnosis of pertussis (100%) and for the physician's diagnosis of some other illness (93%). The only clinical symptom with a high predictive value for pertussis was the report of whoops (92%). Among epidemiological data the highest predictive value (90%) was obtained for reported household exposure in unimmunised children more than 1 year of age. Culture of Bordetella pertussis was found to have an overall 50% sensitivity. Isolation of other bacteria had no predictive value in the differential diagnosis of pertussis.

Acute traumatic tympanic membrane perforations. Cover or observe?

The influence of a paper prosthesis on healing in acute traumatic tympanic membrane perforations was studied in 60 patients randomly selected for a treatment group or a control group. The majority (42 [70%] ) were seen within two days. All patients were treated with oral antibiotics for ten days. The overall healing rate two months after the trauma was 94% (37 of 39 patients). No significant difference between the study group and the control patients was observed. Intermittent secretion was seen in six patients but did not influence the outcome. The perforations occurred in patients with normally sized mastoid air cell systems as seen on roentgenograms. We conclude that acute traumatic tympanic membrane perforations do not need to be treated routinely with paper patching.

Further evidence of an intravesicular Ca2+- pump in odontoblasts from rat incisors.

Intracellular vesicles containing alkaline phosphatases were isolated from isolated odontoblasts using several centrifugation techniques, gradient media and filtering procedures. With a combined centrifugation technique using 0.32 M sucrose layered on 1.23 M sucrose, a fraction containing alkaline phosphatases with a 36-40-fold increased specific activity was obtained. This fraction also revealed a high Ca2+-accumulating ability. The vesicle fraction was totally free from mitochondria but to some extent contaminated by lysosomes. Characteristics of Ca2+-uptake were obtained. The Ca2+-uptake was maximal at 37-40 degrees C whereas no Ca2+-accumulated at 4 degrees C. Temperatures above 40 degrees C strongly inhibited Ca2+-uptake. ATP was the most potent stimulator of Ca2+-uptake whereas ITP, GTP, CTP, ADP, PPi and AMP also promoted Ca2+-uptake. Cysteine, EDTA and Triton X-100 were inhibitory to Ca2+-uptake. A correlation between alkaline phosphatases in intracellular vesicles as well as their relation to extracellular matrix vesicles and to the mineralization process is suggested.

Influence of bivalent cations, phosphate and complexing substances on inorganic pyrophosphate in the microsomal fraction of isolated rat odontoblasts.

The influence of three diphosphonates, ethane-1-hydroxy 1.1. diphosphonate, methane-diphosphonate and dichloromethane diphosphonate, on inorganic pyrophosphatase (PPiase, E.C. 3.6.1.1.) was investigated. The inhibition by the diphosphonates appeared to be due to their complexation with a bivalent cation, probably Zn2+, which acts as a cofactor for the enzyme. The influence of calcium and inorganic phosphate (Pi) on the PPiase activity was studied in the presence and absence of several complexing substances, including the diphosphonates. Ca2+ alone inhibited the enzyme whereas, in the presence of EDTA, a Ca2+ stimulation of the PPiase was observed which was substrate-dependent and had an optimal activity at a Ca2+:PP4-i ratio of 1:1. There was a small inhibition of the PPiase activity by Pi which was not influenced by the substrate used.

Changes in alkaline-phosphatase isoenzymes of hard tissue origin during facial development in the rat.

Non-specific alkaline phosphatase (APase) activity was demonstrated histochemically in the hard-tissue-forming areas of the developing rat face from the 14th embryonic day. Isoenzyme distribution of APase was recorded by isoelectric focusing. Three APase isoenzyme bands with lower isoelectric points than adult forms were recorded. Inhibition studies with levamisole dexamisole, L-p-bromotetramisole, D-p-bromotetramisole, diphosphonates and urea showed the embryonic isoenzymes to have the same sensitivity as the adult forms except to urea. Heated to 56 degrees C, both embryonic and adult APase were rapidly inactivated. The differences between the embryonic and adult forms of APase may represent the expression of different gene loci or a difference in post-translational modification.

First and second branchial arch syndrome: aspects on the embryogenesis, elucidations, and rehabilitation using the osseointegration concept.

BACKGROUND: The osseointegration concept has dramatically changed the possibility of rehabilitating patients with craniofacial defects due to branchial arch syndromes. PURPOSE: This article describes some problems related to the investigative routines and rehabilitation of individuals with malformations of the first and second branchial arches of the craniofacial region. Animal model systems have increased the knowledge of basic embryonic processes that can explain the extent of the malformations. Though most clinical first and second branchial arch syndromes are likely to be caused by sporadic mutations, inherited syndromes occur and also teratogenically induced syndromes are known. Prenatal diagnosis ruling out heredity and exogenous influence seems possible in the future. The possibility of preventing and alleviating fulminant syndromes prenatally also could be conceivable in the future. PATIENTS AND METHODS: The rehabilitation process starts early after birth and should involve a team of specialists including clinical geneticists, pediatricians, audiologists, plastic surgeons, maxillofacial surgeons, otosurgeons, anaplastologists, speech pathologists, pedodontists, and orthodontists. With the development of the osseointegration concept in which craniofacial prostheses and hearing aids can be adapted on implants anchored in the craniofacial skeleton, a new field in the rehabilitation of these malformations has opened. RESULTS: Important aspects in the use of the osseointegration concept include determination of the lowest age for implant surgery, accessibility of adequate bone for implants, the growth of the craniofacial skeleton during childhood, and the possibility for the patient and his or her parents to care for the skin penetration. Adverse tissue reactions, durability of craniofacial prostheses, and the possibility of unknown adverse reactions to metal implants in the body over a long time are other aspects of concern. CONCLUSIONS: Patients with branchial arch syndromes benefit from a well-planned multidisciplinary rehabilitation process in which osseointegrated bone-anchored hearing aids and bone-anchored ear prostheses can be useful tools.

A histomorphometric and biomechanical study of the effect of delayed titanium implant placement in irradiated rabbit bone.

BACKGROUND: The time interval from irradiation to implant surgery has been considered an impact factor for implant integration in irradiated bone and the importance of a long interval between the radiation trauma and reconstructive bone surgery has been suggested. PURPOSE: The present study was undertaken to histomorphometrically and biomechanically analyze the effect of delayed implant placement on bone healing around titanium implants in irradiated bone. MATERIALS AND METHODS: Rabbits were given a single dose of 15 Gy Cobalt60 radiation to one hind leg, the other hind leg serving as a control. Titanium screws were inserted into the femur and tibia directly and at 12 weeks and 52 weeks after irradiation. The implants were evaluated after a healing time of 8 weeks. The torques necessary for removal of the implants were measured. Histomorphometry with respect to bone-metal contact and amount of bone surrounding the implants was performed. RESULTS: The biomechanical force necessary to unscrew the titanium implants in the irradiated bone was significantly increased after a resting period of 1 year between irradiation and implant placement, compared to direct implant placement. The histomorphometric evaluation showed an improvement of bone healing around the implants in the irradiated bone, both after a resting period of 12 weeks and after 1 year compared to direct implant placement. CONCLUSIONS: It is concluded that a relatively long interval between irradiation and reconstructive bone surgery will improve osseointegration of titanium implants in irradiated rabbit bone.

Retinoid-induced ear malformations.

Treatment of pregnant Sprague-Dawley rats with retinoic acid or etretinate pregnancy day 8.5 to 9.0 resulted in craniofacial defects in 100% of the embryos. A morphologic investigation of the malformations occurring in the ear was performed. Outer ears were missing, microtic, low-placed, and dorsally situated. External acoustic meatus was short or absent. Middle ear structures were delayed in differentiation, middle ear ossicle primordia were hypoplastic and malformed, the stapedial artery and facial nerve were hypoplastic, and their relation to the stapes was variable. In the inner ear, the otic capsule was thick, the cochlea had fewer turns and the semicircular ducts showed poor differentiation.

The bone-anchored hearing aid and bone-anchored epithesis for congenital ear malformations.

Surgery for correction of aural atresia was performed on 156 ears in 111 patients. A comparison was made between plastic surgery for auricular reconstruction and the bone-anchored epithesis. Another comparison was performed between hearing improvement after reconstructive surgery for meatal and middle ear atresia and the bone-anchored hearing aid. It was found that the bone-anchored epithesis is an excellent alternative to plastic reconstructive surgery of the auricle. The bone-anchored hearing aid could be considered as a strong alternative to surgery in patients with meatal and middle ear atresia--especially in patients with advanced malformations. Neither the bone-anchored epithesis nor the bone-anchored hearing aid excludes the patient from reconstructive surgery later in life.

Evaluation of implant losses and skin reactions around extraoral bone-anchored implants: A 0- to 8-year follow-up.

This is an 8-year follow-up of a group of 214 patients who underwent surgical insertion of titanium implants in the mastoid process for the retention of bone-anchored hearing aids and auricular prostheses. The skin reactions around the implants and the various factors dealing with implant loss were evaluated. The number of patients who never had any episode of adverse skin reactions during the 8-year period is 70% and is about the same as previously reported. The frequency and degree of adverse skin reactions were noted to be decreasing with time. The young age group had the highest incidence of adverse skin reactions, and this high frequency is consistent with results of earlier reports. None of the remaining group of patients (30%) who had 1 or more episodes of adverse skin reactions lost their implants because of this problem; most implant losses were primarily the result of loss of integration. The probability of losing an implant because of adverse skin reactions is quite low; however, these skin reactions, if left untreated, may eventually lead to implant loss or withdrawal.

Osseointegrated implants in children: experience from our first 100 patients.

This study was undertaken on 100 children aged 16 years or under intended for installation of osseointegrated implants. Of these, 76 had implants installed for bone-anchored hearing aids or prostheses. The main indication for implant installation was a bilateral ear malformation. Surgery was generally performed as a two-stage procedure with a healing time of 3 to 4 months in between. Available bone thickness was on average 2.5 mm, and lack of bone necessitated bone augmentation in 12 patients. Thirty-nine percent of implants were installed in contact with the dura, sigmoid sinus, or an air cell. Implant failures were 5.8% of 170 inserted fixtures. Adverse skin reactions appeared in 9.1% of patients over a 21-year follow-up period. Revision surgery was undertaken in 22% of patients because of appositional growth of the temporal bone. Of the 24 children considered but not found suitable for osseointegration surgery, plastic surgery was considered a better option, or growth of the temporal bone was awaited. It is concluded that the rate of implant failure is lower in children; the frequency of skin reactions is the same as in adults, but revision surgery is more common in young patients because of new bone formation. Our clinical experience supported by the data presented in this article suggests that the concept could be used with good functional and aesthetic results in children.