The homeobox gene mirror links EGF signalling to embryonic dorso-ventral axis formation through notch activation.

Recent studies in vertebrates and Drosophila melanogaster have revealed that Fringe-mediated activation of the Notch pathway has a role in patterning cell layers during organogenesis. In these processes, a homeobox-containing transcription factor is responsible for spatially regulating fringe (fng) expression and thus directing activation of the Notch pathway along the fng expression border. Here we show that this may be a general mechanism for patterning epithelial cell layers. At three stages in Drosophila oogenesis, mirror (mirr) and fng have complementary expression patterns in the follicle-cell epithelial layer, and at all three stages loss of mirr enlarges, and ectopic expression of mirr restricts, fng expression, with consequences for follicle-cell patterning. These morphological changes are similar to those caused by Notch mutations. Ectopic expression of mirr in the posterior follicle cells induces a stripe of rhomboid (rho) expression and represses pipe (pip), a gene with a role in the establishment of the dorsal-ventral axis, at a distance. Ectopic Notch activation has a similar long-range effect on pip. Our results suggest that Mirror and Notch induce secretion of diffusible morphogens and we have identified TGF-beta (encoded by dpp) as such a molecule in germarium. We also found that mirr expression in dorsal follicle cells is induced by the EGF-receptor (EGFR) pathway and that mirr then represses pip expression in all but the ventral follicle cells, connecting EGFR activation in the dorsal follicle cells to repression of pip in the dorsal and lateral follicle cells. Our results suggest that the differentiation of ventral follicle cells is not a direct consequence of germline signalling, but depends on long-range signals from dorsal follicle cells, and provide a link between early and late events in Drosophila embryonic dorsal-ventral axis formation.

Stress response after race and endurance training sessions and competitions in Arabian horses.

Assuring a high level of animal welfare is a critical aspect of contemporary animal husbandry. Equine athletes begin their careers at a very young age when they are still developing and they are both physically and mentally immature. Lack of scientific knowledge of the stress related to horse racing impedes the development of optimal training programs to attain equilibrium between the best sport results and optimal welfare. This study aimed to determine the influence of the intensity and type of physical activity on peripheral blood cortisol concentration. Thirty untrained Arabians, 9 endurance and 21 race horses were enrolled in this longitudinal study. Blood samples were analysed every 3-weeks in 4 training sessions and 2 races in racing horses and monthly after 2 training sessions and 1 competition in endurance horses. Cortisol concentration was measured at rest and 30 min. after physical effort. Racing horses were divided into two groups of the best and the worst performers. Cortisol concentration increased significantly after training and competition, however both in racing and endurance horses the increase was more intensive after competition. In the racing horses, cortisol concentration tended to gradually increase after the subsequent trainings during the racing season, however the starting gate did not appear to exert any impact on cortisol concentration. The best performing race horses appeared to have a lesser increase in cortisol concentration after the race than the worst performing horses, however the cortisol concentration after the race was not significantly different between these two groups of horses. This study suggests that an optimal training program can induce a stress response which is likely not to have any harmful impact on an athlete's welfare. The more intense effort associated with competition events, both in race and endurance horses, results in a greater stress response, indicating that these kinds of events should be limited to assure animal welfare. Finally, better performance horses adapt more readily to physical activity as they experience less increase of serum cortisol concentration after the race.

The value of 68 Gallium-DOTATATE PET/CT in sinonasal neuroendocrine tumor management: A case series.

BACKGROUND: 68 Gallium-DOTATATE (68 Ga-DOTATATE) is a somatostatin analog used as a PET tracer to successfully identify neuroendocrine tumors (NETs). Due to the rarity of sinonasal NETs, there are few recommendations for 68 Ga-DOTATATE imaging in these patients. METHODS: We discussed the impact of 68 Ga-DOTATATE imaging on the management of six sinonasal NET cases and reviewed existing literature. RESULTS: 68 Ga-DOTATATE PET/CT revealed an unknown primary in one case and identified metastatic disease in a primary sinonasal small cell neuroendocrine carcinoma (SNEC) patient missed on conventional imaging. In two esthesioneuroblastoma (ENB) patients, 68 Ga-DOTATATE detected abnormal radiotracer uptake not present on 18F-FDG PET/CT and identified a patient for treatment with 177 Lu-DOTATATE. CONCLUSIONS: This is the one of the first few reports, and the largest series to our knowledge, demonstrating the utility of 68 Ga-DOTATATE imaging for primary sinonasal SNEC and ENB. Further study is required to determine its role in sinonasal NET management.

Virtual Reality as Distraction Analgesia and Anxiolysis for Pediatric Otolaryngology Procedures.

OBJECTIVES/HYPOTHESIS: While virtual reality (VR) has been used as analgesia and anxiolysis for invasive procedures, no literature exists on the use of VR in the pediatric otolaryngology setting. The purpose of this study was to determine the efficacy of VR in reducing pain and anxiety for pediatric otolaryngology patients. STUDY DESIGN: Randomized controlled trial. METHODS: A total of 53 patients aged 7-17 undergoing in-office nasal endoscopies were included. Patients were randomized to receive VR or standard of care. Procedural pain, anxiety, and satisfaction scores were recorded from patients and caregivers. The physician filled out a childhood emotional manifestation scale (CEMS). RESULTS: Patients in VR group reported a significant decrease in pain (0.80 ± 1.06 vs. 2.26 ± 2.38, P = .018) and anxiety (9.50 ± 12.48 vs. 38.48 ± 29.83, P = .0002) and increase in procedural satisfaction (6.40 ± 0.77 vs. 4.74 ± 1.74, P = .0002) compared to patients in control group. CEMS scores were significantly reduced in VR group (5.15 ± 0.46 vs. 9.64 ± 5.66, P = .0001) and caregiver anxiety levels were significantly reduced in VR group (11.50 ± 17.67 vs. 27.39 ± 30.48, P = .041) compared to control group. There were no reported side effects. Procedural time did not significantly differ between groups. CONCLUSIONS: For pediatric otolaryngology patients undergoing in-office nasal endoscopies, VR is a safe and effective form of distraction analgesia and anxiolysis, significantly reducing pain and increasing procedural satisfaction for patients. In addition, VR significantly reduces anxiety for both patients and caregivers without disrupting procedural efficiency and workflow. LEVEL OF EVIDENCE: 2. Laryngoscope, 131:E1714-E1721, 2021.

Nuclear factors that mediate intrathymic signals are developmentally regulated.

Thymocytes mature through several stages of development, defined by cell surface markers such as CD3, CD4, and CD8, in response to environmental cues. Signal transduction resulting from lymphocyte-stromal cell interactions is likely to activate inducible transcription factors which in turn govern stage-specific gene expression. In this report we show that inducible transcription factors such as AP-1 and NF-AT are constitutively nuclear, in response to intrathymic signals, in freshly isolated thymocytes at all stages of maturation. In CD4+CD8+ double positive (DP), but not in the more immature CD4-CD8- double negative (DN) thymocytes, constant stimulus from the thymic environment is required to maintain nuclear AP-1. Thus, disruption of the thymus and incubation of thymocytes at 37 degrees C downregulates DNA binding by nuclear factors AP-1 and NF-AT. Similar treatment of thymocytes has previously been shown to downregulate CD3 zeta chain phosphorylation and increase T cell receptor CD3 expression on DP thymocytes, which is a feature of repertoire selection. Since mature T cells maintain inducible nuclear factors in an inactive form until an encounter with antigen, we propose that downregulation of nuclear DNA binding proteins may reflect another feature of this stage of T cell maturation.

Infiltrative ophthalmopathy and primary hypothyroidism: a rare clinical manifestation of a common disease.

The association of infiltrative ophthalmopathy with primary hypothyroidism is uncommon. We describe two such cases manifesting at different times during the course of primary hypothyroidism. The successful outcome of the present cases suggests that the timely addition of levothyroxine treatment alone or in combination with steroids is useful and effective in the management of hypothyroid Graves' ophthalmopathy.

Identification and validation of two peptide markers for the recognition of Clostridioides difficile MLST-1 and MLST-11 by MALDI-MS.

OBJECTIVES: Clostridioides difficile infection (CDI) has become the main cause of nosocomial infective diarrhoea. To survey and control the spread of different C. difficile strains, there is a need for suitable rapid tests. The aim of this study was to identify peptide/protein markers for the rapid recognition of C. difficile strains by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). METHODS: We analysed 44 well-characterized strains, belonging to eight different multi-locus sequence types (MLST), using ultrahigh-resolution Fourier transform ion cyclotron resonance (FTICR) MS. The amino acid sequence of two peptide markers specific for MLST-1 and MLST-11 strains was elucidated by MALDI-TOF-MS/MS. The investigation of 2689 C. difficile genomes allowed the determination of the sensitivity and specificity of these markers. C18-solid-phased extraction was used to enrich the MLST-1 marker. RESULTS: Two peptide markers (m/z 4927.81 and m/z 5001.84) were identified and characterized for MLST-1 and MLST-11 strains, respectively. The MLST-1 marker was found in 786 genomes of which three did not belong to MLST-1. The MLST-11 marker was found in 319 genomes, of which 14 did not belong to MLST-11. Importantly, all MLST-1 and MLST-11 genomes were positive for their respective marker. Furthermore, a peptide marker (m/z 5015.86) specific for MLST-15 was found in 59 genomes. We translated our findings into a fast and simple method that allowed the unambiguous identification of the MLST-1 marker on a MALDI-TOF-MS platform. CONCLUSIONS: MALDI-FTICR MS-based peptide profiling resulted in the identification of peptide markers for C. difficile MLST-1 and MLST-11.

Human milk fortification and gastric emptying in the preterm neonate.

Fortification of expressed breast milk (EBM) is widely recommended for preterm feeding. Fortification of EBM results in increased caloric density and osmolarity, both of which may retard gastric emptying. As gastric emptying is a major determinant of feed tolerance, we investigated the effect of fortification (with Lactodex HMF) of EBM on gastric emptying in preterm neonates. The half gastric emptying time was measured using real time ultrasonography in 25 consecutive preterm neonates first on EBM alone, then on EBM + Lactodex HMF. Each baby served as its own control. The students t-test was used for statistical analysis. The mean gestation age was 34.48 +/- 0.77 weeks. The mean birth weight was 1.92 +/- 0.14 kg. The mean half gastric emptying time at an age of 5.4 +/- 0.86 days on EBM was 24.00 +/- 5.00 min and 24.40 +/- 5.06 min on EBM + human milk fortifier (HMF). The same at 2nd assessment (15.2 +/- 1.79 days), with EBM was 22.80 +/- 4.58 min vs. 23.60 +/- 4.89 min when given EBM + HMF. These differences were not statistically significant. Fortification of EBM with Lactodex HMF does not affect the gastric emptying in preterm neonates and therefore is unlikely to affect feed tolerance in them.

Metabolic failure precedes intracranial pressure rises in traumatic brain injury: a microdialysis study.

BACKGROUND: Cerebral microdialysis (MD) is able to detect markers of tissue damage and cerebral ischaemia and can be used to monitor the biochemical changes subsequent to head injury. In this prospective, observational study we analysed the correlation between microdialysis markers of metabolic impairment and intracranial pressure (ICP) and investigated whether changes in biomarker concentration precede rises in ICP. METHODS: MD and ICP monitoring was carried out in twenty-five patients with severe TBI in Neurointensive care. MD samples were analysed hourly for lactate:pyruvate (LP) ratio, glutamate and glycerol. Abnormal values of microdialysis variables in presence of normal ICP were used to calculate the risk of intracranial hypertension developing within the next 3 h. FINDINGS: An LP ratio >25 and glycerol >100 micromol/L, but not glutamate >12 micromol/L, were associated with significantly higher risk of imminent intracranial hypertension (odds ratio: 9.8, CI 5.8-16.1; 2.2, CI 1.6-3.8; 1.7, CI 0.6-3, respectively). An abnormal LP ratio could predict an ICP rise above normal levels in 89% of cases, whereas glycerol and glutamate had a poorer predictive value. CONCLUSIONS: Changes in the compound concentrations in microdialysate are a useful tool to describe molecular events triggered by TBI. These changes can occur before the onset of intracranial hypertension, suggesting that biochemical impairment can be present before low cerebral perfusion pressure is detectable. This early warning could be exploited to expand the window for therapeutic intervention.

Free phenytoin concentration measurement in brain extracellular fluid: a pilot study.

This article investigates the relationship between brain extracellular fluid free phenytoin concentration and plasma free phenytoin concentration in adults with acute brain injury. Daily cerebral microdialysate free phenytoin concentration was measured in eight adults with acute brain injury and compared with simultaneous measurement of plasma free phenytoin concentration. The group data revealed no significant correlation between microdialysate and plasma free phenytoin concentration (r = 0.34, p = 0.41). However, in two patients, with a sufficient number of samples for intra-individual analysis, there was a significant correlation between microdialysate and plasma free phenytoin concentration (r = 0.92, p < 0.001 and r = 0.88, p < 0.01). In vitro microdialysis relative recovery for phenytoin was 2.1%. In the context of acute brain injury, measurement of free plasma phenytoin concentration may not provide an accurate reflection of regional brain extracellular fluid free phenytoin concentration and may have limitations with respect to achieving reproducible brain extracellular fluid free phenytoin concentrations. This has implications for dosing regimens relying on plasma phenytoin levels.

Axonal pathology in subarachnoid and intracerebral hemorrhage.

Electrically active axons degenerate in the presence of nitric oxide (NO) in vitro. High CSF NO concentrations have been observed in patients with hemorrhagic brain injury such as subarachnoid hemorrhage (SAH) and intracerebral hemorrhage (ICH). This study investigated the evidence for axonal injury in SAH and ICH and related this to CSF NO levels. In this study, neurofilament phosphoforms (NfH(SMI34), NfH(SMI35), NfH(SMI38), NfH(SMI310)), surrogate markers for axonal injury, and NO metabolites (nitrate, nitrite = NOx) were measured by ELISA in cerebrospinal fluid (CSF) from patients with SAH and ICH and from a group of controls. Injury severity was classified using the Glasgow Coma Scale, and survival was used as the outcome measure. Compared to the control group, a higher proportion of patients with SAH and ICH had elevated NfH(SMI34) levels from day 0 to day 6 (p < 0.001), elevated NfH(SMI35) levels from day 1 to 6 (p < 0.001), and elevated NfH(SMI310) levels at day 0, 1, 4, and 6 (p < 0.001). The NOx levels were higher in the SAH and ICH patients than in the controls (p < 0.05) and distinguished the non-survivors from the survivors (p < 0.05). No direct correlation was found for NOx with any of the NfH phosphoforms. This study provides evidence for primary and secondary axonal injury in patients with SAH and ICH, with non-survivors also having higher NOx levels. CSF NfH phosphoforms might emerge as a putative surrogate marker for monitoring the development for secondary axonal degeneration in neurocritical care and guiding targeted neuroprotective strategies.

Dermatoglyphic variation among the Limboo of Sikkim, India.

Variations in finger and palmar dermatoglyphic patterns are investigated among the Limboo (18-60 years, 150 males and 150 females), a little known population of Sikkim. The results for Limboo were compared with other North-East Indian populations. The most commonly occurring pattern was loop (males: 64.33%; females: 75.00%) followed by whorl (males: 31.00%; females: 21.33%) and finally arch (males: 4.67%; females: 3.66%). There were no significant differences between sexes in pattern types. The overall values of pattern intensity (P.I.I.), Dankmeijer's (D.I.) and Furuhata's (F.I.) indices were 14.08, 12.60 and 96.06 respectively. The P.I.I. was within the range for East Asian populations of North-East India. The D.I. was similar to those reported for Rajbanshi, Kalita, Rabha and Newar populations, while F.I. was higher than in other populations of Eastern Himalaya and North-East India. The most frequently occurring mainline formulae in all palm prints (left and right combined) were 7.5'.5.- followed by 9.7.5.- and finally 11.9.7.- (p>0.05) and these were similar to the reported values for other North-eastern populations of India. The mean values of total finger ridge count (TFRC) and absolute finger ridge count (AFRC) were greater among males (138.03; s=42.26 and 198.78; s=77.4) than females (137.91; s=44.15 and 194.47; s=86.71). The a-b ridge count was greater among females than males. Sex differences in AFRC and a-b ridge count were both statistically significant (p<0.05). The mean TFRC values were within ranges for populations of North-East India. Cluster analysis based on P.I.I., D.I. and F.I. shows affinity of the Limbo to some of the populations of Assam and North-East India.

Identification, sequence and developmental expression of invertebrate flotillins from Drosophila melanogaster.

Caveolae are vesicular organelles that represent a sub-compartment of the plasma membrane. Caveolins (Cav-1, -2 and -3) and flotillins (FLO-1 and FLO-2 [also known as epidermal surface antigens (ESAs)] are two families of mammalian caveolae-associated integral membrane proteins. Although a caveolin gene family has recently been described in the invertebrate Caenorhabditis elegans, it remains unknown as to whether flotillin homologues exist in invertebrates. Here, we report the identification, cDNA sequence and embryonic expression pattern of the first invertebrate flotillin, i.e. flotillin from Drosophila melanogaster (FLODm). FLODm is most closely related to mammalian flotillin-1. Remarkably, the invertebrate FLODm protein behaves like mammalian flotillins and is targeted to the caveolae-enriched membrane fraction after transient expression in mammalian cells. Localization of the FLODm message in D. melanogaster embryos reveals that expression of FLODm is confined primarily to the developing nervous system. This is consistent with our previous observation that mammalian flotillin-1 mRNA and protein is expressed abundantly in brain tissue. Interestingly, the FLODm gene is localized to chromosomal region 52 B1-B2. In addition, we find that at least two flotillin-related genes are expressed in D. melanogaster. Our current results provide a starting point and systematic basis for dissecting the role of flotillin in caveolae and neuronal development using Drosophila as a genetic system.

High intracellular calcium concentrations in transformed lymphoblasts from subjects with bipolar I disorder.

OBJECTIVE: Higher basal concentrations of intracellular calcium Ca2+ in platelets and lymphocytes from subjects with bipolar affective disorder than in unipolar depressed and healthy subjects implicate abnormal intracellular Ca2+ signaling in bipolar disorder. The purpose of this study was to clarify whether these intracellular Ca2+ abnormalities are trait related. METHOD: Basal Ca2+ concentration was measured by using ratiometric fluorescence assay with fura-2 for T lymphocytes and Epstein-Barr-virus-immortalized B lymphoblasts from physically healthy subjects with DSM-IV diagnoses of bipolar mood disorder (bipolar I, N = 28; bipolar II, N = 11) or major depressive disorder (N = 14), mixed psychiatric patients with non-mood disorders (N = 14), and health subjects (N = 20). Phytohemagglutinin-stimulated (10 micrograms/ml) intracellular Ca2+ levels were also determined in T lymphocytes. RESULTS: The basal Ca2+ concentration was significantly higher in the B lymphoblasts from patients with bipolar I disorder, but not bipolar II disorder or major depression, than in healthy subjects or psychiatric patients with nonmood disorders. There was a significant interaction between gender and diagnosis in the effect on basal Ca2+ levels in T lymphocytes. Contrasts of diagnoses within gender revealed significantly higher basal Ca2+ concentrations in T lymphocytes in male bipolar I patients, but not mean with bipolar II disorder or major depression, than in healthy male comparison subjects. Phytohemagglutinin-stimulated Ca2+ concentrations did not differ among groups, but the percent differences from basal Ca2+ levels were lower in bipolar I and depressed patients than in healthy subjects. CONCLUSIONS: These findings support the occurrence of abnormal calcium homeostasis in bipolar disorder and suggest that trait-dependent factors account, at least partly, for the higher basal lymphocyte Ca2+ concentration in bipolar I subjects.

A prospective study of the rate of falls before and after cataract surgery.

BACKGROUND: There has been considerable interest in the development of intervention programmes aimed at reducing the risk of falls. The primary objective was to ascertain whether cataract surgery reduced the risk of falls in elderly patients with age related cataract. METHODS: 97 patients scheduled for cataract surgery were enrolled in this prospective clinical study. The patients were assessed for established risk factors for falls preoperatively and postoperatively. Patients were issued with a diary to record any falls and phoned at 2 monthly intervals during the 6 month preoperative and postoperative periods. RESULTS: Of the 84 patients who completed the study, 31 recorded falls during the preoperative period (37%). This group showed a statistically significant reduction in the number of fallers in the postoperative period (n = 6, p<0.001) CONCLUSION: These results suggest that cataract surgery is an effective intervention to reduce the risk of falls in elderly patients with cataract related visual impairment.

Congophilicity (Congo red affinity) of different beta2-microglobulin conformations characterized by dye affinity capillary electrophoresis.

The amyloidogenic protein beta-microglobulin was characterized by affinity capillary electrophoresis (CE). CE could separate conformational variants of beta2-microglobulin and with the amyloid-specific dye Congo red as a buffer additive it was possible to measure different Congo red-affinities of native and abnormally folded beta2-microglobulin. We find that native beta2-microglobulin has an intermediate affinity for Congo red at pH 7.3 and that binding involves electrostatic interactions. The conformational variant of beta2-microglobulin that appears in acetonitrile solutions binds Congo red more strongly. Affinity CE using Congo red as a buffer additive is a new, simple, fast, and quantitative micromethod for the characterization of soluble conformational intermediates of amyloidogenic proteins.

Subcutaneous cysticercosis involving the eyelid: sonographic diagnosis.

A 25-year-old man and a 14-year-old boy presented with neurocutaneous cysticercosis involving the eyelid. Both patients had hundreds of scattered subcutaneous cysticerci. They were arranged in clusters over the sternocleidomastoid muscle in the neck. Such clustering of cysticerci is highly suggestive of central nervous system (CNS) involvement, as both the sternocleidomastoid muscle and the CNS are supplied by the carotid artery and cysticerci travel via the hematogenous route. We used ultrasonography to diagnose subcutaneous cysticercosis, which showed characteristic low reflective cysts and high reflective scolices inside. Although subcutaneous cysticerci are inconsequential, their verification is important in the diagnosis of more severe CNS involvement. They may be confused with other painless swellings such as lymphadenopathies, neurofibromas, and epidermoid cysts.

Clinical profile in gelatinous bone marrow transformation.

OBJECTIVE: To study the clinical spectrum associated with gelatinous bone marrow transformation (GMT). METHODS: All subjects whose bone marrow aspiration showed pink purple material on Leishman stain underwent a detail history, clinical examination and investigation (biochemical/microbiological/radiological). Additionally, in each subject the smear was stained with special stains of Periodic Acid Schiff and Alcian blue. RESULTS: Out of total 1498 marrows, 65 showed evidence of GMT. All of these had anaemia. The associated clinical spectra of diseases noticed were: Infection (31 cases), Nutritional deficiency (5 cases), Haematological disorders (Aplastic/toxic depression) (17 cases), Malignancies (3 cases), and Miscellaneous (9 cases). CONCLUSION: Based on the heterogenecity of associated clinical disorders, GMT indicates severe illness and not a particular disease. GMT may be a result of bioregulatory process (which presently needs further prospective studies) that are activated in different pathologic conditions but resulting in similar lesion in the bone marrow and so till then it may be concluded that GMT is a symptom of bone marrow.