Evolution of MHC class II SLA-DRB1 locus in the Croatian wild boar (Sus scrofa) implies duplication and weak signals of positive selection.

The wild boar is an ancestor of the domestic pig and an important game species with the widest geographical range of all ungulates. Although a large amount of data are available on major histocompatibility complex (MHC) variability in domestic pigs, only a few studies have been performed on wild boars. Due to their crucial role in appropriate immune responses and extreme polymorphism, MHC genes represent some of the best candidates for studying the processes of adaptive evolution. Here, we present the results on the variability and evolution of the entire MHC class II SLA-DRB1 locus exon 2 in 133 wild boars from Croatia. Using direct sequencing and cloning methods, we identified 20 SLA-DRB1 alleles, including eight new variants, with notable divergence. In some individuals, we documented functional locus duplication, and SLA-DRB1*04:10 was identified as the allele involved in the duplication. The expression of a duplicated locus was confirmed by cloning and sequencing cDNA-derived amplicons. Based on individual genotypes, we were able to assume that alleles SLA-DRB1*04:10 and SLA-DRB1*06:07 are linked as an allelic combination that co-evolves as a two-locus haplotype. Our investigation of evolutionary processes at the SLA-DRB1 locus confirmed the role of intralocus recombination in generating allelic variability, whereas tests of positive selection based on the dN/dS (non-synonymous/synonymous substitution rate ratio) test revealed atypically weak and ambiguous signals.

Novel Y-chromosome short tandem repeats in Sus scrofa and their variation in European wild boar and domestic pig populations.

Y-chromosome markers are important tools for studying male-specific gene flow within and between populations, hybridization patterns and kinship. However, their use in non-human mammals is often hampered by the lack of Y-specific polymorphic markers. We identified new male-specific short tandem repeats (STRs) in Sus scrofa using the available genome sequence. We selected four polymorphic loci (5-10 alleles per locus), falling in one duplicated and two single-copy regions. A total of 32 haplotypes were found by screening 211 individuals from eight wild boar populations across Europe and five domestic pig populations. European wild boar were characterized by significantly higher levels of haplotype diversity compared to European domestic pigs (HD  = 0.904 ± 0.011 and HD  = 0.491 ± 0.077 respectively). Relationships among STR haplotypes were investigated by combining them with single nucleotide polymorphisms at two linked genes (AMELY and UTY) in a network analysis. A differentiation between wild and domestic populations was observed (FST  = 0.229), with commercial breeds sharing no Y haplotype with the sampled wild boar. Similarly, a certain degree of geographic differentiation was observed across Europe, with a number of local private haplotypes and high diversity in northern populations. The described Y-chromosome markers can be useful to track male inheritance and gene flow in wild and domestic populations, promising to provide insights into evolutionary and population genetics in Sus scrofa.

Revisiting the phylogeography and demography of European badgers (Meles meles) based on broad sampling, multiple markers and simulations.

Although the phylogeography of European mammals has been extensively investigated since the 1990s, many studies were limited in terms of sampling distribution, the number of molecular markers used and the analytical techniques employed, frequently leading to incomplete postglacial recolonisation scenarios. The broad-scale genetic structure of the European badger (Meles meles) is of interest as it may result from historic restriction to glacial refugia and/or recent anthropogenic impact. However, previous studies were based mostly on samples from western Europe, making it difficult to draw robust conclusions about the location of refugia, patterns of postglacial expansion and recent demography. In the present study, continent-wide sampling and analyses with multiple markers provided evidence for two glacial refugia (Iberia and southeast Europe) that contributed to the genetic variation observed in badgers in Europe today. Approximate Bayesian computation provided support for a colonisation of Scandinavia from both Iberian and southeastern refugia. In the whole of Europe, we observed a decline in genetic diversity with increasing latitude, suggesting that the reduced diversity in the peripheral populations resulted from a postglacial expansion processes. Although MSVAR v.1.3 also provided evidence for recent genetic bottlenecks in some of these peripheral populations, the simulations performed to estimate the method's power to correctly infer the past demography of our empirical populations suggested that the timing and severity of bottlenecks could not be established with certainty. We urge caution against trying to relate demographic declines inferred using MSVAR with particular historic or climatological events.

Mitochondrial DNA and Y-chromosome diversity in East Adriatic sheep.

Variation in mitochondrial DNA (mtDNA) and Y-chromosome haplotypes was analysed in nine domestic sheep breeds (159 rams) and 21 mouflon (Ovis musimon) sampled in the East Adriatic. Mitochondrial DNA analyses revealed a high frequency of type B haplotypes, predominantly in European breeds, and a very low frequency of type A haplotypes, which are more frequent in some Asian breeds. Mitochondrial haplotype Hmt-3 was the most frequent (26.4%), and 37.1%, 20.8% and 7.6% of rams had haplotypes one, two and three mutations remote from Hmt-3 respectively. In contrast, Y-chromosome analyses revealed extraordinary paternal allelic richness: HY-6, 89.3%; HY-8, 5.0%; HY-18, 3.1%; HY-7, 1.3%; and HY-5, 1.3%. In fact, the number of haplotypes observed is comparable to the number found in Turkish breeds and greater than the number found in European breeds so far. Haplotype HY-18 (A-oY1/135-SRYM18), identified here for the first time, provides a link between the haplotype HY-12 (A-oY1/139-SRYM18) found in a few rams in Turkey and haplotype HY-9 (A-oY1/131-SRYM18) found in one ram in Ethiopia. All mouflons had type B mtDNA haplotypes, including the private haplotype (Hmt-55), and all were paternally monomorphic for haplotype HY-6. Our data support a quite homogeneous maternal origin of East Adriatic sheep, which is a characteristic of European breeds. At the same time, the high number of haplotypes found was surprising and intriguing, and it begs for further analysis. Simultaneous analysis of mtDNA and Y-chromosome information allowed us to detect a large discrepancy between maternal and paternal lineages in some populations. This is most likely the result of breeder efforts to 'upgrade' local populations using rams with different paternal origins.

Experimental hematotympanum--aspects to the tympanosclerosis development.

The etiology of tympanosclerosis is very complex and has not been entirely clarified. In order to find out if blood could be a provoking factor and cause tympanosclerosis, the authors injected the blood into the ears of 32 guinea pigs. The animals were sacrificed at weekly intervals up to one month and the histological analysis of the temporal bones was performed. Their experiment has shown that mucosal changes, similar to those seen in the early stage of tympanosclerosis, can appear in a certain number of cases. As a result, they recommend the aspiration of blood from the middle ear cavity in the cases of middle ear trauma with hematotympanum. They recommend the same procedure for operations of the middle ear, which should be performed with adequate hemostasis to prevent possible tympanosclerosis development.

Tympanoplasty after war blast lesions of the eardrum: retrospective study.

AIM: To establish whether hearing loss after eardrum blast injury could be recovered by tympanoplasty performed immediately after injury and what material is the most suitable for eardrum closure. METHODS: Tympanoplasty was performed in 119 (a total of 181 injuries) out of 651 patients examined for blast injury of the ear between 1991 and 2000. The study included a total of 106 patients who underwent tympanoplasty: 51 patients with unilateral and 55 with bilateral blast eardrum rupture (a total of 161 injuries). Three different materials were used for eardrum rupture closure: temporal fascia in 81, perichondrium in 61, and heterograft in 19 cases. Injuries were divided in 4 groups, according to the time elapsed between the injury and tympanoplasty (0-20, 21-60, 61-180, and 181 days and more). Otomicroscopic finding, audiometry, and tympanometry were used for definitive evaluation of tympanoplasty outcome. RESULTS: Eardrum rupture was successfully closed with temporal fascia in 91%, perichondrium in 92%, and heterograft in 89% of the cases (p=0.429). There were no statistically significant differences in either values of postoperative air- bone gap (p=0.210) or in eardrum perforation closure rate (p=0.951) with respect to the time period between the injury and tympanoplasty. Also, there was no correlation between the postoperative air-bone gap and the number of days elapsed between the rupture and tympanoplasty (r=-0.037, p=0.641). CONCLUSION: Small ruptures of the eardrum should be left to heal spontaneously. The patients with subtotal and total rupture and rupture that did not heal spontaneously in three months should undergo tympanoplasty. Temporal fascia, perichondrium from tragus, and heterograft are equally acceptable materials for eardrum closure after blast injury.

Vasodilator and vitamins in therapy of sensorineural hearing loss following war-related blast injury: retrospective study.

AIM: To establish whether a better recovery of the sensorineural hearing loss can be achieved by systemic administration of a vasodilator and vitamins in a saline infusion immediately after a blast injury. METHODS: Retrospective analysis included 134 patients with pure sensorineural hearing loss after blast injury (a total of 192 injuries), who were treated between 1991 and 1995. A group of 82 patients (119 injuries) was treated with 250 mL infusion of saline containing vitamins and pentoxifylin, whereas the other group (52 patients or 73 injuries) did not receive any therapy. RESULTS: The patients who received infusion therapy with vitamins and pentoxifylin showed no better hearing recovery than the patients who did not receive any therapy. There was no statistically significant difference between the groups in the average values of the sensorineural hearing recovery (p=0.315). CONCLUSION: Therapy with vitamins and pentoxifylin does not improve recovery of sensorineural hearing loss caused by blast injuries of the internal ear.

Changes of intratympanic pressure in secretory otitis media.

A thick, gluey effusion was observed to out of the middle ear after myringotomy only in some cases of secretory otitis media. Cytologic analysis of intratympanic transudate in 321 cases of secretory otitis media in children showed the cellular infiltration to result in increased viscosity of the middle ear content. It caused gradual change in the middle ear pressure, from the initially negative to positive pressure in the advanced stages of the disease.

Hyperbaric oxygen as a therapy of Bell's palsy.

The purpose of this study was to compare the therapeutic effects of hyperbaric oxygen (HBO2) to the effects of prednisone treatment in 79 subjects with Bell's palsy. Patients were randomly assigned either to the HBO2-treated group (n = 42) or to the prednisone-treated group (n = 37). The HBO2 group was exposed to 2.8 atm abs of 100% oxygen for 60 min, twice a day, 5 days a week and was given a placebo orally. The prednisone group was exposed to 2.8 atm abs of 7% O2 (equivalent to 21% O2 in air at normal pressure) following the same schedule as the HBO2 group; prednisone was given orally (total of 450 mg in 8 days). Subjects from both groups were treated in the hyperbaric chamber for up to 30 sessions or to complete recovery, and were followed up for 9 mo. At the end of the follow-up period, 95.2% of subjects treated with HBO2, and 75.7% of subjects treated with prednisone recovered completely. The average time to complete the recovery in the HBO2 group was 22 days as opposed to 34.4 days in the control group (P < 0.001). In the HBO2-treated group, at the beginning, the altered nerve excitability test (NET) was abnormal in five subjects; three of them had normal NET by the end of the follow-up period. In the prednisone group the NET was abnormal in nine subjects at the beginning and they had not recovered by the end of the follow-up (P < 0.05). Our results suggest that HBO2 is more effective than prednisone in treatment of Bell's palsy.

Arachidonic acid metabolites and sinonasal polyposis. I. Possible prognostic value.

PURPOSE: The etiology of sinonasal polyps is sometimes obscure. This study was undertaken to evaluate the potential role of arachidonic acid metabolites (AAm) on recurrent polyposis. MATERIALS AND METHODS: Tissue production of prostaglandin E2 (PGE2), 6-keto-prostaglandin F1-alpha (PGI2), thromboxane A2 (TxA2), and leukotriene C4 (LTC4) by nasal mucosa was determined by radioimmunoassay in 27 patients with sinonasal polyposis (SNp) and in 10 volunteers. RESULTS: The group of patients with SNp with the evidence of recurrences in postoperative period (Group 1) showed significantly lower PGE2 concentrations than group of patients with SNp recurrences (Group 2). The differences in concentrations of PGI2 in mentioned groups were insignificant. In comparison with other groups, a group of patients who underwent surgery several times for SNp (Group 4) had a higher mean TxA2 concentration. The LTC4 concentrations were the highest in groups of patients where SNp recurrences were observed. When the incidence of polyposis recurrences (within 18 months after surgery) was correlated with the level of LTC4 production at the time of surgery, the rate of recurrence was significantly higher in patients with increased LTC4 level than in those with normal LTC4 levels. CONCLUSIONS: LTC4 might have a prognostic value. The possible role of AAm in occurrence of SNp is apparent and suggests possible role for medical intervention.

Effects of sulphur dioxide and smoke on the incidence of secretory otitis media.

The study deals with the effects of sulphur dioxide and smoke as air pollutants on the incidence of secretory otitis media. Although various atmospheric irritants, such as sulphur dioxide and smoke, have been reported to increase proneness to infection and allergy stabilization, and to cause Eustachian tube obstruction and development of secretory otitis media by way of infection and allergy reaction, such statements appear to be challenged by the results obtained in this study. No correlation was found between the number of children hospitalized for secretory otitis media, and the mean and maximum monthly concentrations of sulphur dioxide and smoke in the air. Although there has been a steady decrease in sulphur dioxide and smoke concentrations over the past decade, the number of cases of secretory otitis media has not only failed to follow that decrease but has actually increased.

Effects of sulfur dioxide and smoke on the incidence of laryngotracheitis (croup).

During the 1981-1990 period, 652 children, 70% boys and 30% girls, were hospitalized at the Zagreb University ENT Department for laryngotracheitis (croup). Most of the children were hospitalized at the age of 2, and 90% of all children were under the age of 7. The highest number of cases of laryngotracheitis were admitted during autumn, especially in October, and the lowest in July. No correlation was found between the mean and peak monthly values of sulfur dioxide and smoke in the air in Zagreb and the number of children hospitalized for laryngotracheitis.

Effect of climatic elements on the frequency of secretory otitis media.

A relationship between the number of children hospitalized for secretory otitis media and climatic elements was observed during a 10-year period (1981-1990). The highest correlation was obtained between the number of secretory otitis media cases and the number of sunny hours in a month, followed by that between the number of secretory otitis media cases and the mean monthly temperature and relative air humidity. The largest number of children were hospitalized for secretory otitis media toward the end of winter, and the lowest toward the end of summer (in August and September).

Immunohistochemistry of glomus jugulare and tympanicum tumors.

Glomus jugulare and tympanicum tumors differ from other paragangliomas, especially from aorto-sympathetic, visceral-autonomic and adrenal medulla tumors. They differ not only in their localization, but also in their histologic and clinical pictures. As differentiated from other paragangliomas, these tumors show no argyrophilia. Besides the difference in histochemistry, the glomus jugulare and tympanicum tumors also differ from other paragangliomas in the intensity of their immunohistochemical reaction to particular markers. As differentiated from other paragangliomas that react well to chromogranin with antisera, the chief cells of these tumors are poorly represented with chromogranin as a marker. As distinguished from chromogranin, however, synaptophysin produced a positive reaction, i. e., moderate to maximal reactivity, in all cases under study. Helper cells of these tumors, and likewise other paragangliomas, are well represented by antisera to S-100 protein and glial fibrillary acid protein.

Surgical treatment of Menière's disease--two techniques compared.

Two techniques for surgical treatment of Menière's disease, endolymphatic sac decompression according to Shambough and decompression on the stapes footplate, i. e., platino decompression according to Martin, were compared. A series of 81 patients were hospitalized for treatment, and 20 surgical procedures were performed: platino decompression in eight cases, and endolymphatic sac decompression in 12. None of the eight patients had relapse of the disease after platino decompression, whereas the endolymphatic sac operation was followed by relapse in 6 of the 12 patients. Considering the postoperative values of hearing loss, the considerably lower incidence of relapse, and the technically simple procedure of platino decompression, this technique is recommended as a method of choice for the surgical treatment of Menière's disease.

[Granuloma of the oval window after stapedectomy]. / Granulom fenestre ovalis nakon stapedotomije.

A case report is presented of a female patient who underwent stapedotomy with bad functional results Reoperation revealed a dislocated prosthesis and there was no successful hearing. Besides conduction deafness perceptive deafness occurred as well. The second reoperation disclosed reparative granuloma around the prosthesis, while histological analysis showed foreign-body granuloma. Foreign-body granuloma by stapedectomy is extremely rare and it was the first case in our 810 surgically treated ears.

[Secretory otitis media in childhood: epidemiologic and etio-pathogenic aspects, diagnosis and therapy]. / Sekretorni otitis u djecjoj dobi: epidemioloski i etiolosko-patogenetski aspekt, dijagnostika i lijecenje.

Secretory otitis media is still poorly understood. In spite of extensive investigations, there are nevertheless many unsolved problems, from etiology to therapy. This paper reviews the current knowledge on epidemiology, etiology, pathogenesis, diagnosis and therapy of secretory otitis media. The possibilities of both conservative and surgical treatments as well as still ununiform view-points on a surgical procedure are presented. In conclusion, on the basis of abundant literature and our experience, a treatment approach to secretory otitis media in children is proposed.

[Classification of secretory otitis based on cytological analysis of middle ear transudate]. / Tipizacija sekretornog otitisa na temelju citoloske analize transudata iz srednjeg uha.

On the basis of the number of cell elements in the cytologic smear of the transudate from the middle ear, secretory otitis media was divided into four groups providing indication for therapeutic procedures. The first group is characterized by a mass of mucus and very scarce cell elements, suggesting a relatively short duration of illness and the healing by conservative therapy and adenoidectomy. The second group contained somewhat more cell elements in the cytologic smear, suggesting a longer duration of illness and requiring the use of a ventilation tubule in the therapeutic procedure. The third group showed a considerable cell infiltration and the appearance of individual collagenic fibres scattered in disorder in the smear. The transudate is dense, sticky, and dark. Its aspiration from the cavum is not easy. The finding suggests a long duration of illness, requiring a more active approach of the otosurgeon. The fourth group is characterized by a very scarce but dense aspirate from the middle ear. The cytologic smear shows an intense cell infiltration and a mass of collagenic fibres, which speaks for a very long duration of illness and for the necessity, in view of the formation of connective tissue in the middle ear, of an explorative tympanotomy coupled with adhesiolysis and the use of the ventilatory tubule.

[Immunologic studies of secretory otitis]. / Immunologische Untersuchungen der sekretorischen Otitis.

Evaluation of total serum proteins and RIST immunoelectrophoresis of immunoglobulins IgG, IgA and IgM were carried out on a group of 35 children of both sexes suffering from secretory otitis. The transudate aspirated from the middle ear was examined only by immunoelectrophoresis. The values for albumin and gammaglobulins were decreased for all the patients. The values for RIST were significantly lower than those where an allergic cause was present. The values for immunoglobulins in transudate were also significantly lower than normal.