Fast and furious: successful survival and resumption of meiosis in immature human oocytes vitrified and warmed using a short protocol.

RESEARCH QUESTION: Can immature oocytes vitrified and warmed using a short protocol survive and resume meiosis? DESIGN: This study examined modifications of oocyte vitrification and warming protocols that reduce the length of exposure to vitrification and warming solutions. In total, 561 germinal vesicles and 218 metaphase I oocytes that were immature at oocyte retrieval were vitrified at room temperature for 2 min. Warming was performed at 37°C for 2 min. Resumption of meiotic activity was evaluated after 24 and 48 h of culture. Two different commercially available vitrification and warming kits were used for comparison. RESULTS: Ninety-five percent of germinal vesicles survived, with no difference observed between the kits. The survival of metaphase I oocytes was, on average, 95.4% and did not differ significantly between the kits. Of the 533 germinal vesicles that survived, 491 converted to metaphase I oocytes (92.1%). After culture for 48 h, 54.4% converted to metaphase II oocytes. In addition, of the 208 metaphase I oocytes that survived warming, 84.1% converted to metaphase II oocytes after 24 h of culture. These maturation rates were similar to those of non-vitrified oocytes. CONCLUSIONS: Vitrification and warming of oocytes at different nuclear maturation stages can be performed with 2 min of exposure to hypertonic solution and 2 min of exposure to hypotonic solution, respectively. This approach reduces exposure of the oocytes to room temperature during dehydration and rehydration. Warming in 0.5M sucrose helps to maintain and support the potential of oocytes to resume nuclear meiotic activity, and conversion from germinal vesicles to metaphase I and metaphase II oocytes.

Oral Health Birth Cohort Studies: Achievements, Challenges, and Potential.

Birth cohorts are those among observational studies that provide understanding of the natural history and causality of diseases since early in life. Discussions during an International Association for Dental Research symposium in London, United Kingdom, in 2018, followed by a workshop in Bangkok, Thailand, in 2019, concluded that there are few birth cohort studies that consider oral health and that a broader discussion on similarities and differences among those studies would be valuable. This article aims to 1) bring together available long-term data of oral health birth cohort studies from the low, middle, and high-income countries worldwide and 2) describe similarities and differences among these studies. This work comprises 15 studies from all 5 continents. The most studied dental conditions and exposures are identified; findings are summarized; and methodological differences and similarities among studies are presented. Methodological strengths and weaknesses are also highlighted. Findings are summarized in 1) the negative impact of detrimental socioeconomic status on oral health changes over time, 2) the role of unfavorable patterns of dental visiting on oral health, 3) associations between general and oral health, 4) nutritional and dietary effects on oral health, and 5) intergenerational influences on oral health. Dental caries and dental visiting patterns have been recorded in all studies. Sources of fluoride exposure have been documented in most of the more recent studies. Despite some methodological differences in the way that the exposures and outcomes were measured, some findings are consistent. Predictive models have been used with caries risk tools, periodontitis occurrence, and permanent dentition orthodontic treatment need. The next steps of the group's work are as follows: 1) establishing a consortium of oral health birth cohort studies, 2) conducting a scoping review, 3) exploring opportunities for pooled data analyses to answer pressing research questions, and 4) promoting and enabling the development of the next generation of oral health researchers.

Potential impact of lingual retainers on oral health: comparison between conventional twistflex retainers and CAD/CAM fabricated nitinol retainers : A clinical in vitro and in vivo investigation.

PURPOSE: To evaluate the impact of a novel computer-fabricated lingual nitinol retainer compared to a conventional lingual flexible spiral wire twistflex retainer on oral health. METHODS: The study was based on a retrospective controlled clinical study with pilot character, an in vitro investigation of material-dependent biofilm formation and an analysis of biofilm formation after intraoral incubation. Sixty-one patients with completed fixed orthodontic treatment and retention phase for at least 6 months with twistflex retainers (group 1, n = 31) or computer-aided design/computer-aided manufacturing (CAD/CAM) nitinol retainers (group 2, n = 30) were included and examined regarding plaque index (PI), gingival index (GI), probing depths, bleeding on probing (BOP) and marginal recessions (MR). Material-dependent biofilm formation of twistflex, untreated nitinol and electropolished nitinol wire samples were assessed (1) in vitro: using optical density (OD) measurement of 10 samples of each and (2) in vivo: using histomorphometric analysis of 18 samples of each. RESULTS: Patients treated with nitinol retainers had significant better oral health indices (PI1 = 1.29 ± 0.06, PI2 = 0.94 ± 0.06; GI1 = 0.71 ± 0.05, GI2 = 0.56 ± 0.04; BOP1 = 0.11 ± 0.01, BOP2 = 0.08 ± 0.01; PD1 = 1.79 ± 0.03 mm, PD2 = 1.59 ± 0.04 mm) except for MR (0.08 ± 0.03 mm versus 0.08 ± 0.02 mm) compared to twistflex retainers. After 24 h intraoral incubation nitinol retainers demonstrated significant less biofilm formation compared to twistflex retainers. In the in vitro investigation the temporary significant differences between the groups were compensated in the end. CONCLUSIONS: Based on the results it can be assumed that nitinol-made CAD/CAM developed lingual retainers have a positive effect on oral health.

Evaluation of a regional German interdisciplinary oral health programme for children from birth to 5 years of age.

OBJECTIVES: The aim of this prospective birth cohort study was to evaluate the effect of the interdisciplinary oral health programme (OHP) for early childhood caries (ECC) in 5-year-old German children. MATERIAL AND METHODS: All parents of newborns (n = 1162; born 2009/2010) were visited after birth by the communal newborn visiting service of Jena, Thuringia, and advised on general and dental health. In the first year of life, children were invited to attend a dental examination in the Jena University Hospital. Participating children were included in a caries-risk-related recall system with continuous oral care over 5 years. The caries-risk assessment tool of the AAPD was used to determine the likelihood of carious lesion development and to categorize the children at low, moderate or high risk for caries. High-risk children received fluoride varnish. Families (n = 563) who gave their approval for final examination after 5 years were invited again and examined by a blinded clinician. Dental caries was scored using WHO diagnostic criteria expanded to d1-level without radiography. Children were allocated to prevention (PG) and control group (CG) and matched on the basis of age, sex, ethnicity and socio-economic status (SES). RESULTS: Two hundred and eighty-nine children (mean age 5.2 ± 0.7 years; 46.7 % female) were examined. Children in the PG (n = 174) showed significantly lower caries prevalence and experience (10.9 %, 0.2 ± 0.7 d3-4mft) than children in the CG (57.4 %, 2.9 ± 3.8 d3-4mft). Multivariate analysis found that low SES, early start of tooth brushing, supervision/regular second brush by parent, regular dental visits and duration of breast-/bottle-feeding >1 year were significantly related to d3-4mft. CONCLUSIONS: The OHP was an effective approach for preventing early childhood caries in preschool children. CLINICAL RELEVANCE: A programme consisting of early maternal counselling, establishment of a dental home, and inclusion of the children in a caries-risk-related recall system with continuous dental care and fluoride varnish application can prevent ECC. TRIAL REGISTRATION: German Clinical Trials Register DRKS00003438, https://drks-neu.uniklinik-freiburg.de/drks_web/navigate.do?navigationId=trial.HTML&TRIAL_ID=DRKS00003438.

Effect of a thin-neck pacifier on primary dentition: a randomized controlled trial.

OBJECTIVES: To evaluate changes in occlusal characteristics in the primary dentition that occur after introducing a thin-neck pacifier (TNP) to children with previously diagnosed pacifier-associated anterior open bite (AOB) and increased overjet. SETTING AND SAMPLE POPULATION: Department of Preventive and Pediatric Dentistry, Jena University Hospital, Germany. Subjects were 86 children (mean age 20.3 months) with a pacifier-associated open bite or overjet ≥2 mm. MATERIAL & METHODS: Randomized controlled trial. Subjects were randomly assigned: group I (n = 28), intervention group using a TNP; group II (n = 30), control group, using a conventional or physiological pacifier; and group III (n = 28), intervention group, Gold standard, weaned off pacifier. Participants were re-examined after 3, 6, 9 and 12 months by an operator, blinded for the treatment. RESULTS: After 12 months data for 63 children (mean age 33.1 months) were analyzed (I: n = 24; II: n = 22; III: n = 17). There was a significant difference between the groups regarding mean overjet (group I: 2.7 ± 0.5 mm, group II: 3.2 ± 0.7 mm, group III: 2.4 ± 0.5 mm, Kruskal-Wallis, p = 0.002) and AOB (group I: -1.2 ± 0.3 mm, group II: -2.2 ± 0.3 mm, group III: -0.8 ± 0.8 mm, Kruskal-Wallis, p < 0.001). The differences between group I and II regarding increased overjet (3.1 ± 0.2 mm vs. 3.6 ± 0.3 mm, Mann-Whitney, p < 0.001) and extent of AOB (-1.2 ± 0.3 mm vs. -2.2 ± 0.3 mm, Mann-Whitney, p < 0.001) were statistically significant. CONCLUSION: Use of TNP resulted in better clinical measurements for in overjet and overbite compared with the continuing use of conventional or physiological pacifiers.

Evaluation of an oral health promotion program at the time of birth on dental caries in 5-year-old children in Vorarlberg, Austria.

OBJECTIVES: The aim of this study was to evaluate the effect of an early oral health promotion program (OHPP) on oral health of 5-year-old children in Austria by providing new mothers with dental health counseling. METHODS: Qualified dental health educators visited all mothers at time after birth in regional hospitals in Vorarlberg and counseled them regarding the oral health of their children as a one-off intervention. Mothers were given comprehensive oral hygiene instructions for their children and themselves (daily use of fluoride toothpaste, restoration of own caries lesions), which included practical tooth brush training and dietary counseling by the use of brief motivational interviewing and anticipatory guidance approaches. After 5 years, a case-cohort study was conducted to evaluate the program with annual dental check-ups of 471 children and a parent survey. Dental caries was scored using WHO diagnostic criteria at dentine level without radiography. To compare oral health parameters (dmfs/dmft, care index), children were allocated to intervention and control group and matched on basis of age, sex, ethnicity, and socio-economic status (SES) and analyzed statistically. RESULTS: Children whose mothers participated in the OHPP showed significantly lower caries prevalence and experience (33.2%; 3.2 ± 7.4 d3-4 mfs/1.5 ± 2.5 d3-4 mft) than children whose mothers did not participate (42.6%; 5.2 ± 6.4 d3-4 mfs/2.4 ± 4.1 d3-4 mft). Bivariate analysis showed that ethnicity, SES, age at which toothbrushing started, supervision of toothbrushing, use of fluoride salt, and use of fluoride toothpaste were significantly related to d3-4 mft. CONCLUSIONS: The OHPP was an effective community-based preventive approach for improving oral health in preschool children.

Gabapentin in the treatment of post-dural puncture headache: a case series.

Gabapentin has been reported to be useful in the management of epilepsy, neuropathic pain and post-dural puncture headache. Seventeen obstetric cases are presented in which gabapentin was used either as a primary therapy for the management of severe headache following a diagnosed dural puncture or as an analgesic adjunct in patients with dural puncture headache unresponsive to epidural blood patch. In our patients, analgesic therapy was essential to enable care of their newborns as other management options had failed or were unavailable. In nine patients we observed an excellent result with reduction of headache severity within 24 hours. Gabapentin appears potentially beneficial in the management of patients with post-dural puncture headaches.

Prophylactic endovascular placement of internal iliac occlusion balloon catheters in parturients with placenta accreta: a retrospective case series.

BACKGROUND: Endovascular occlusion balloon catheters can be placed preoperatively in internal iliac vessels of patients perceived to be at risk of major obstetric haemorrhage during caesarean section. Their safety and efficacy remains undefined, and we report our experience of 14 patients over four years. METHODS: We undertook a chart review of all patients who had undergone prophylactic internal iliac balloon catheters before caesarean section in our institution. RESULTS: Balloon catheters were placed in 14 and inflated in 11 (78.6%) patients. Five of the 14 patients (35.7%) underwent emergency balloon catheter placement before unscheduled caesarean section. Surgeons reported that balloon inflation provided favourable surgical conditions in six of 11 cases (54.5%), no improvement in four and was not required in one due to lack of pathology. Within the balloon-inflated group, nine patients underwent a hysterectomy: two electively, the remaining seven because of perioperative confirmation of placenta accreta or for control of bleeding. One patient suffered massive haemorrhage leading to three perioperative hypovolaemic cardiac arrests. Four patients required intervention to avoid complications related to balloon catheters: three minor and one related to catheter displacement and prolonged resuscitation. CONCLUSION: Internal iliac balloon catheters can be inserted electively or in an emergency in patients at risk of major obstetric haemorrhage. Although useful in some, they are not universally effective; patients are still at risk of significant blood loss and at high risk of requiring a hysterectomy. In our experience, catheters can be placed electively or in an emergency but have been associated with adverse outcomes. These lessons have been important learning points in perioperative management.

Prevalence of short-segment Barrett's epithelium.

BACKGROUND/AIMS: The incidence of adenocarcinoma of the oesophagus has increased. Its major risk factor is Barrett's epithelium of which the sine qua non is microscopically diagnosed intestinal metaplasia. Short segment Barrett's epithelium may often be overlooked during routine endoscopy. In routine biopsies taken from normal-appearing mucosa of the distal oesophagus, the reported rates of short segment Barrett's epithelium in the distal oesophagus reached 36%. We compared these rates with the results obtained in a community hospital in Israel. METHODS: Consecutive patients undergoing oesophagogastroduodenoscopy were enrolled. Biopsy specimens taken from cardia, oesophagogastric junction and 2 cm above the oesophagogastric junction were stained with haematoxylin & eosin and Alcian blue. RESULTS: There were 112 study patients (mean age +/- SD 48. 9+/-18.3 years, 51.8% males). Nine (8.04%) patients had intestinal metaplasia (according to specimen from 2 cm above oesophagogastric junction), and symptoms of gastro-oesophageal reflux were found in only four (44.4%) of them. Of these nine patients, six (6.66%) had normal-appearing mucosa and three (3.33%) had macroscopic Barrett's epithelium. Alcian blue staining revealed two patients with intestinal metaplasia that haematoxylin & eosin staining had missed. CONCLUSION: We found an 8% prevalence of intestinal metaplasia compared to 18-36% reported in the literature. We also determined that the added advantage of routine biopsy was 5.4%.

[Cytokeratin and p53 expression of odontogenic cysts]. / Zytokeratin- und p53-Expression odontogener Zysten.

The histopathologic diagnosis of odontogenic cysts is based mainly on the morphological nature of the epithelial lining of cysts and their origin. We used the international histologic classification set up by the World Health Organisation in 1992. The aim of this study was to investigate the differentiation of various types of cyst using an immunohistochemical technique rather than by conventional morphological assessment. A standard immunocytochemical method (APAAP method), applying anticytokeratin monoclonal antibodies and a p53 antibody, was used for the diagnosis of odontogenic cysts. A total of 57 jaw cysts were diagnosed according to clinical, radiological and pathological criteria as radicular cysts (20), dentigerous cysts (20) and keratocysts (17). The results proved that cyst type can be distinguished by the pattern of staining using the monoclonal antibodies CK7, CK19, CK20 for cytokeratins and the clone DO-7 for the p53 protein. Staining with the monoclonal antibodies CK7 and CK20 did not distinguish type. CK19 was not detected in keratocysts and p53 was only expressed in keratocysts. This may prove to be diagnostically useful for the more precise distinction between different cyst types.

Comparison of various Helicobacter pylori detection methods: serology, histology and bacteriology.

The current gold standard test for diagnosis of Helicobacter pylori involves histological staining and/or urease testing of antral biopsy specimens. However, these methods are invasive, and alternative non-invasive methods, i.e. the urease breath test and serological tests, are available. The test for H. pylori-specific serum immunoglobulin G (IgG) is now available commercially. The aim of this study was to compare the gold standard tests for diagnosis of H. pylori to the non-invasive method of detecting IgG antibody in the serum. Two hundred and twenty-five (225) subjects were tested for H. pylori by histological staining, urease testing, direct microscopy of antral biopsy specimens and quantification of serum IgG antibody. The population examined was divided into 2 groups--a group of 52 patients with no gastrointestinal symptoms and a group of 173 patients with dyspepsia. Out of 173 dyspeptic patients, 22 (12.7%) were false-positive to H. pylori. Out of 52 non-dyspeptic subjects, 30 (57.7%) were false-positive (p < 0.0001). The sensitivity and specificity were 91.6% and 51.7%, respectively. The specificity and positive predictive value increased by approximately 30% when the subjects examined were in the younger age group (< 30 years), while the sensitivity and the negative predictive value did not change significantly. This study indicates that serological testing is not recommended for diagnosis nor is it recommended for follow-up treatment, especially among the older age group (> 30 years).

[The buried gastrostomy bumper syndrome].

The buried gastrostomy bumper syndrome is one of the rare complications of PEG (percutaneous endoscopic gastrostomy) insertion. It develops when there is a combination of a rigid bumper and a tension build-up between internal and external bumpers. This condition is manifested by complete occlusion of the internal opening of the gastrostomy by mucosa, making it impossible to feed the patient. We report a case in which the PEG was inserted a year prior to the appearance of this rare complication. It was embedded beneath the gastric mucosa and we had difficulty in removing it to insert a new PEG. The bumpers are anchor-like attachments to each end of the gastrostomy, which keep it stationary. The rigid bumper is an integral part of the gastrostomy. However, a "soft" bumper has been developed, but its costliness has restricted its use. In our case the gastrostomy was removed with the aid of the cutting wire of a sphincterotome in light contact with the external tissue.

The effect of the Gulf War on upper-gastrointestinal bleeding.

Upper-gastrointestinal bleeding may be related to tension or fear, which are commonly aggravating factors in digestive diseases. A survey was made of patients living in and around Hadera (located in Israel's central coastal region) during the Gulf War (January 18 through February 28, 1991) who suffered from upper-gastrointestinal bleeding; they were compared with patients seen during 1990 and 1992-1993 at the same time of the year. We found no appreciable difference in the rate of upper-gastrointestinal bleeding during the analogous periods covered in the survey.

The role of Helicobacter pylori in children with recurrent abdominal pain.

OBJECTIVES: Our major goals in this study were to determine the prevalence of Helicobacter pylori among Israeli children with recurrent abdominal pain and to establish whether a link exists between eradication of Helicobacter pylori and the recovery from abdominal pain. The alternative target was to examine whether the serological test can replace endoscopy in children. METHODS: Upper gastrointestinal endoscopy was performed in 80 children with recurrent abdominal pain. During endoscopy, antral biopsies were taken and sent for histological and bacteriological examination. RESULTS: The prevalence of H. pylori, which was indicated by Gram stain and urease test, was 54%. The positive cases were treated with bismuth combined with amoxycillin and metronidazole. Two months after completion of the treatment, 34 patients were reexamined. Twenty-nine of them (85%) became symptom free, and five (15%) remained symptomatic. These five children were retreated, and they also became symptom free. Eight months after completion of treatment, all 34 patients were reexamined and found to be asymptomatic. CONCLUSIONS: We found a high prevalence of H. pylori (54%) among Israeli children with recurrent abdominal pain; we also found that symptoms were effectively and significantly reduced by the eradication of H. pylori. The endoscopic examination cannot be replaced by serological test.

Cardiorespiratory effects of premedication for children.

Cardiovascular and respiratory effects of pediatric preanesthetic premedication have received only minimal attention, probably because most children tolerate such drugs without apparent ill effect. In children with congenital heart disease or other serious illness, there is often reluctance to use premedication. We sought to determine whether different premedication regimens produced significant cardiorespiratory effect. A randomized prospective study of the cardiovascular and respiratory effects of different oral, nasal, and rectal premedication regimens was conducted. Fifty-eight young children (average age 2.7 yr) were studied. Oral meperidine (3 mg/kg) with pentobarbital (4 mg/kg) decreased heart rate, mean arterial pressure, cardiac index, respiratory rate, and oxygen saturation. Stroke volume was maintained. Nasal ketamine (5 mg/kg) with midazolam (0.2 mg/kg) produced no significant cardiovascular or respiratory effects. Rectal methohexital (30 mg/kg) increased heart rate with a coincident decrease in stroke volume but had no other positive or negative cardiac or respiratory effect. This information documents disparate cardiorespiratory effects of different preanesthetic medications in normal children.

Studies of manometric abnormalities of the rectoanal region during defecation in constipated and soiling children: modification through biofeedback therapy.

Anorectal manometry was performed on 12 normal children and 18 patients suffering from constipation and soiling. In both groups, the results of the rectoanal inhibitory reflex and the squeezing anal pressure were similar. The pressure recording in all the normal children showed that the anal canal relaxed during defecation. Fourteen (78%) constipated children closed the anal canal while straining by contracting the anal sphincter. This paradoxical contraction appears to be the cause of chronic constipation. Twelve children with paradoxical anal closure were treated by biofeedback therapy. The results show that all these children were successfully conditioned to relax their anal sphincter during defecation. This therapy improved their bowel habits and relieved them from constipation and soiling. It is proposed that the paradoxical anal closure itself is the result of a self-conditioning process. In this process, the patient learns to paradoxically contract the external anal sphincter in response to the urge and the act of defecation. Biofeedback therapy seems to be the appropriate treatment in such cases.

Chediak-Higashi syndrome. Expression of the cytoplasmic defect by in vitro cultures of bone marrow progenitors.

Studies on proliferation and differentiation of granulocyte-monocyte progenitor cells in Chediak-Higashi syndrome (CHS) were done on a 1-month-old patient, using the soft-agar bone marrow culture technique. The number of granulocyte-macrophage colony-forming cells (GM-CFC) was markedly increased, but with a normal distribution into granulocyte, macrophage, or mixed colonies. Morphologic, cytochemical, and ultrastructural studies showed that 70% of the colonies consisted of cells with giant lysosomes typical of CHS, and in the remaining 30% abnormal cells were not detected. The supply of granulocyte-macrophage colony-stimulating factor (GM-CSF) by the patient's peripheral blood leukocytes was markedly decreased. Inhibition of normal in vitro granulopoiesis by the patient's lymphocytes or serum was not demonstrated. It appears that granulocyte progenitors in CHS proliferate normally, or even in excess, probably in response to intramedullary destruction of granulocytes. The majority of the progenitors are intrinsically defective and give rise to colonies that contain the abnormality. In others the defects are unidentifiable, probably due to the immaturity of the specific fusion process of the cytoplasmic granules. The abnormal leukocytes in CHS are also defective in their capacity to provide GM-CSF, and this may account in part to the overt neutropenia. These studies demonstrate that the basic cytoplasmic abnormalities of the granulocytes and monocytes in CHS are embedded in the granulocytic-monocytic committed stem cell.