The Foulage Test: Proof of Concept of a Novel Stepping Test Using a Stabilometer.

Objective In this article, we aimed to describe the Foulage test (FT) and investigate the test-retest reliability of parameters recorded during stepping execution in healthy adults. Materials and methods This was a single-center prospective cohort study conducted at an outpatient clinic. It included five healthy male participants [mean age ± standard deviation (SD): 27 ± 5.4 years]. The FT was performed first with the participants' eyes open and again with their eyes closed. If the heel height was not within 2-6 cm, the participant was asked to restart. The FT value and variance of steps were automatically calculated. To verify the influence of heel height, measurements were taken at different heel heights. We also evaluated the Romberg ratio (calculated from the parameters with eyes open and closed) and defined it as the dynamic Romberg ratio. Correlations between parameters were also assessed. Results The parameters' FT value (front-back width of the band of locus shape) and variance of steps plateaued under stable conditions within a heel height of 2-6 cm. FT values and variance of steps were strongly correlated. The dynamic Romberg ratios by FT value and by the variance of steps were also strongly correlated. Conclusions The FT is a dynamic and reproducible equilibrium function test that can quantify agitation with the eyes open or closed in general outpatient clinics, and it may be employed as a clinically useful method for the observation of clinical courses in patients with vestibular disorders.

Intercalation: Constructing Nanolaminated Reduced Graphene Oxide/Silica Ceramics for Lightweight and Mechanically Reliable Electromagnetic Interference Shielding Applications.

There is a critical need to develop lightweight and mechanically reliable materials for electromagnetic interference (EMI) shielding applications in the harsh environment. In this study, we propose a low-density (∼2.2 g/cm3) reduced graphene oxide (rGO)/silica ceramic with multilayer rGO sheets parallelly aligned inside the silica matrix through a new intercalation strategy. The parallel rGO sheets lead to outstanding EMI shielding effectiveness values of 29-33 dB in the X-band, owing to the interlayer multiple reflections of the electromagnetic wave. Meanwhile, the parallel rGO sheets elevate the flexural strength by 110-130% and improve the fracture toughness by 100-130% compared with the monolithic silica by capturing and deflecting the propagating cracks. The nanolaminated structure constructed by the intercalation approach can effectively break the trade-off between mechanical properties and EMI shielding performances in the graphene/ceramic composites, thus opening up new opportunities in the lightweight and mechanically reliable EMI applications.

Identification and characterization of a cell division-regulating kinase AKB1 (associated kinase of Trypanosoma brucei 14-3-3) through proteomics study of the Tb14-3-3 binding proteins.

We used a proteomics approach to identify the binding partners of Trypanosoma brucei 14-3-3 (Tb14-3-3) which led to the identification of a novel kinase, AKB1. The binding between these two proteins was mediated by an amphipathic groove structure in Tb14-3-3 and 1-438 amino acid sequence of AKB1. Recombinant AKB1 but not its ATP-binding-deficient mutant (DFG to NFG) possessed an auto-phosphorylation activity as well as a kinase activity towards a peptide substrate in vitro. However, the autophosphorylation was not required for the binding of AKB1 to Tb14-3-3. Interestingly, the kinase activity of AKB1 was inhibited by calcium, and the kinase was found to utilize GTP, and dATP in addition to ATP as phospho-donors. AKB1 formed homodimers through a leucine-zipper structure. Either knockdown of AKB1 or overexpression of AKB1, but not kinase-dead AKB1 mutant, deregulated cytokinesis and cell division, suggesting that kinase activity of AKB1 is crucial for its function. Furthermore, we showed that AKB1 exists in a detergent insoluble fraction. Laser confocal microscopy revealed that the majority of AKB1 is co-localized with α-tubulin. Taken together, these findings suggest that AKB1 might regulate cytokinesis and cell division by phosphorylating cytoskeleton-associated proteins.

Trypanosoma brucei 14-3-3I and II proteins predominantly form a heterodimer structure that acts as a potent cell cycle regulator in vivo.

Hetero- and homodimerization of 14-3-3 proteins demonstrate distinctive functions in mammals and plants. Trypanosoma brucei 14-3-3I and II (Tb14-3-3I and II) play pivotal roles in motility, cytokinesis and the cell cycle; however, the significance and the mechanism of Tb14-3-3 dimerization are remained to be elucidated. We found that ectopically expressed epitope-tagged Tb14-3-3I and II proteins formed hetero- and homodimers with endogenous Tb14-3-3I and II proteins. However, we also found the ability to form hetero- or homodimers between Tb14-3-3I and II proteins was clearly affected by the sequence and location of the epitope tag used. We found a blue native polyacrylamide gel electrophoresis system followed by western blotting may distinguish monomer from dimer structure, and stable from unstable conformation of Tb14-3-3. Combined with co-immunoprecipitation results, we revealed that Tb14-3-3 proteins mainly existed as heterodimeric form. Furthermore, co-overexpression of Tb14-3-3I and II proteins in T. brucei induced aberrant numbers of organelles in cells, but overexpression of either isoform alone rarely produced such morphology. These results suggest that heterodimers play more significant roles than homodimers not only in the maintenance of steady-state levels of the 14-3-3 proteins but also in the regulation of cytokinesis.

Aerial dispersal of blood-contaminated aerosols during dental procedures.

OBJECTIVES: Dental procedures with high-speed instruments produce large amounts of aerosols. The present study aimed to clarify whether blood-contaminated aerosols were existent and floating in air during dental procedures and to evaluate the effect of an extraoral evacuator system. METHOD AND MATERIALS: An extraoral evacuator system with a test filter was used for sample collection at distances from 50 cm (n = 102) and 100 cm (n =124) behind the patient. A leucomalachite green presumptive test was performed on the test filter. One or more positive reactions on the test filter was considered a positive result, and the positive ratio in each procedure was compared. To assess the effect of the extraoral evacuator, an additional extraoral evacuator was introduced, and the positive ratio during third molar extractions at 100 cm behind the patient was compared (n = 55). RESULTS: At 50 cm from the mouth of the patient during third molar surgery, full-crown preparation, inlay cavity (Black Class II) preparation, and scaling with an ultrasonic scaler, positive results were obtained in 92% (12/13), 70% (21/30), 35% (9/26), and 33% (11/33) of cases, respectively. At a distance of 100 cm, positive ratios sustained 90% (35/39), 48% (15/31), 29% (6/21), and 12% (4/33), respectively. When the second extraoral evacuator was utilized, the positive ratio, at a distance of 100 cm, decreased significantly from 90% (35/39) to 60% (33/55) (P = .0015, chi-square test). CONCLUSION: This study showed that blood-contaminated aerosols can be suspended in air, even in general dental settings, and that extraoral evacuators are useful for reducing contaminated aerosols.

Inferior alveolar nerve canal and branches detected with dental cone beam computed tomography in lower third molar region.

PURPOSE: To evaluate the course of the inferior alveolar nerve and its branches, the detectable branches were investigated with dental cone beam computed tomography (CBCT). MATERIALS AND METHODS: Patients in whom the lower third molar (M3) and inferior alveolar nerve canal showed overlapping in the initial panoramic image were included. One hundred twelve impacted lower M3s were extracted after examination with dental CBCT. The detection ratio, the course of the branches, and their relation with the M3 were retrospectively investigated. RESULTS: One hundred fifty-five branches were observed in 106 cases (94.6%, 106/112) around the M3. Most branches coursed under the M3 (55.5%, 86/155), and 85 branches (54.8%, 85/155) were in contact with the M3. The inferior alveolar nerve canal and branch(es) were mostly in contact with the M3 (57.5%, 61/106). CONCLUSIONS: Dental CBCT can detect most tubular structures representing branches in the impacted lower M3 region.

Radiotherapy for glottic T1N0 carcinoma with slight hypofractionation and standard overall treatment time: importance of overall treatment time.

OBJECTIVE: We retrospectively investigated treatment outcomes in patients with glottic T1 carcinoma treated with 65 Gy in 26 fractions four times a week and discuss the importance of the overall treatment time. METHODS: Two hundred one patients with glottic T1 carcinoma were evaluated. Sixty-five Gray in 26 fractions were delivered for 200 patients, whereas 1 patient received 62.5 Gy in 25 fractions. We delivered radiotherapy once daily four times a week in this period, for a weekly dose of 10 Gy. Weekdays except Wednesday were treatment days. RESULTS: The overall survival rate was 96.8 ± 1.3% (standard error) at 3 years and 90.8 ± 2.2% at 5 years. The local control rate was 91.9 ± 2.0% at 3 years and 89.8 ± 2.3% at 5 years. In patients with an overall treatment time equal to or longer than 47 days, the local control rate was 82.6 ± 6.0% at both 3 and 5 years. In the patients with overall treatment time equal to or less than 46 days, the local control rate was 94.6 ± 1.9% at 3 years and 91.8 ± 2.4% at 5 years. There was a significant difference between these two groups (P = 0.0349). A severe late radiation reaction occurred in one patient. He experienced severe laryngeal edema that required tracheotomy at 6 months after the completion of radiotherapy. The tracheotomy was closed at 14 months after completion of radiotherapy. CONCLUSIONS: Overall treatment time seems to be an important factor for a good local control rate for glottic T1N0 carcinoma even when treated with slight hypofractionation.

Long-term outcomes of fractionated stereotactic radiotherapy for intracranial skull base benign meningiomas in single institution.

OBJECTIVE: To investigate the outcome of linac-based fractionated stereotactic radiotherapy over the last 10 years for intracranial skull base benign meningiomas in patients who were inoperable, who had residual tumors with some components of high mitotic index after surgery and who experienced relapse of the tumor. METHODS: Twenty-seven patients with intracranial skull base benign meningiomas treated with fractionated stereotactic radiotherapy were retrospectively reviewed. Twenty-seven cases were diagnosed as benign meningiomas on pathological (17 cases) or radiological (10 cases) examination. The median follow-up time was 90 months after initial treatment and 63 months after fractionated stereotactic radiotherapy. The median biological equivalent dose calculated using an α/ß ratio of 2.0 Gy was 82.0 Gy (range, 60-106 Gy). RESULTS: The 5-year overall survival was 95.7 (95% confidence interval: 87.3-100)% after initial treatment and 96.2 (88.8-100)% after fractionated stereotactic radiotherapy. The 5-year overall survival and local control rate of patients who received fractionated stereotactic radiotherapy alone were both 100%. The 5-year progression-free survival and local control rate after fractionated stereotactic radiotherapy were all 100% with a tumor volume of <9.1 cc and 68.2 (37.2-99.2) and 75.8 (45.2-100)% for the tumors 9.1 cc, respectively. The difference was significant in progression-free survival (P = 0.022) and local control rate (P = 0.044). The local control rate was significantly worse in patients who received fractionated stereotactic radiotherapy for relapsed tumors (P = 0.01). No late radiation damage was observed in the follow-up period. CONCLUSIONS: The long-term outcome suggests that fractionated stereotactic radiotherapy is a safe and effective treatment for intracranial skull base benign meningioma, especially for those who have tumors <9.1 cc or would receive fractionated stereotactic radiotherapy with or without surgery as the initial treatment.

Bone formation without lamina dura in the middle-aged and elderly: possible dependence on enamel.

Bone formation below the crown of mandibular horizontal incompletely impacted third molar is frequently seen in the middle-aged and elderly. The phenomenon shows lamina dura loss without radiolucency and we hypothesized the participation of mature enamel without any influence on the environmental oral status. In order to investigate the characteristics of the phenomenon based on the presence/absence of the lamina dura and radiolucency below the crown, we studied the relationship between 58 men and 43 women with a lamina dura without radiolucency, 12 men and 8 women without a lamina dura with radiolucency, 34 men and 16 women without a lamina dura without radiolucency, and the status of teeth in the ipsilateral mandible. Subjects without a lamina dura without radiolucency were significantly older than those with a lamina dura without radiolucency in both men (P < 0.0001) and women (P <0.01), indicating different chronological causes. Men without lamina dura with radiolucency showed significantly more tooth loss than those with a lamina dura without radiolucency (P < 0.00001) and those without a lamina dura without radiolucency (P < 0.0001), indicating the influence of poor oral health. Thus, the phenomenon without a lamina dura without radiolucency may show the clinical importance of bone formation in the elderly.

A comparison of the postnatal development of muscle-spindle and periodontal-ligament neurons in the mesencephalic trigeminal nucleus of the rat.

The trigeminal mesencephalic nucleus (Vmes) is known to include primary afferent neurons of jaw muscle spindles (MS neurons) and periodontal ligament receptors (PL neurons). The aim of this study was to clarify the postnatal development of Vmes neurons by comparing MS neurons with PL neurons using horseradish peroxidase labeling. We measured somal diameter and somal shape of MS and PL neurons in rats from postnatal day (P)7 to P70. No significant changes were seen between postnatal day P7 and P70 in somal diameter or somal shape of MS neurons. Conversely, PL neurons showed a larger somal diameter at P7 than at P14, and in terms of somal profile, multipolar neurons comprised 0% at P7, but 4.8% at P14 and 16.9% at P70. These findings suggest that PL neurons develop with the eruption of teeth, taking into account the fact that tooth eruption occurs from around P14 in rats. Conversely, the lack of postnatal changes in MS neurons is due to the fact that these neurons have been active since the embryonic period, as swallowing starts in utero.

Phosphorylation-dependent protein interaction with Trypanosoma brucei 14-3-3 proteins that display atypical target recognition.

BACKGROUND: The 14-3-3 proteins are structurally conserved throughout eukaryotes and participate in protein kinase signaling. All 14-3-3 proteins are known to bind to evolutionally conserved phosphoserine-containing motifs (modes 1 and/or 2) with high affinity. In Trypanosoma brucei, 14-3-3I and II play pivotal roles in motility, cytokinesis and the cell cycle. However, none of the T. brucei 14-3-3 binding proteins have previously been documented. METHODOLOGY/PRINCIPAL FINDINGS: Initially we showed that T. brucei 14-3-3 proteins exhibit far lower affinity to those peptides containing RSxpSxP (mode 1) and RxY/FxpSxP (mode 2) (where x is any amino acid residue and pS is phosphoserine) than human 14-3-3 proteins, demonstrating the atypical target recognition by T. brucei 14-3-3 proteins. We found that the putative T. brucei protein phosphatase 2C (PP2c) binds to T. brucei 14-3-3 proteins utilizing its mode 3 motif (-pS/pTx(1-2)-COOH, where x is not Pro). We constructed eight chimeric PP2c proteins replacing its authentic mode 3 motif with potential mode 3 sequences found in Trypanosoma brucei genome database, and tested their binding. As a result, T. brucei 14-3-3 proteins interacted with three out of eight chimeric proteins including two with high affinity. Importantly, T. brucei 14-3-3 proteins co-immunoprecipitated with an uncharacterized full-length protein containing identified high-affinity mode 3 motif, suggesting that both proteins form a complex in vivo. In addition, a synthetic peptide derived from this mode 3 motif binds to T. brucei 14-3-3 proteins with high affinity. CONCLUSION/SIGNIFICANCE: Because of the atypical target recognition of T. brucei 14-3-3 proteins, no 14-3-3-binding proteins have been successfully identified in T. brucei until now whereas over 200 human 14-3-3-binding proteins have been identified. This report describes the first discovery of the T. brucei 14-3-3-binding proteins and their binding motifs. The high-affinity phosphopeptide will be a powerful tool to identify novel T. brucei 14-3-3-binding proteins.

Occupational safety among dental health-care workers.

Compared to other health-care workers, dental health-care workers come in close contact with patients and use a variety of sharp and high-speed rotating instruments. It is important to understand the characteristics of the occupational accidents that occur. We reviewed incident reports from April 1, 2005, to March 31, 2010, at Matsumoto Dental University Hospital. In addition, questionnaires dealing with identification of occupational safety issues, especially splash exposures, were conducted for dentists, dental hygienists, and nurses. Thirty-two occupational injuries were reported during the study period, including 23 sharp instrument injuries (71.9%), 6 splash exposures (18.8%), and 3 others. Of the six splash exposures, only two cases involved potential contamination with blood or other potentially infectious patient material. Of the 66 workers who experienced sharps injuries, 20 workers (30.3%, 20/66) reported them to the hospital work safety team. The questionnaire revealed high incident of splash exposures and conjunctiva exposures: 87.9% (51/58) and 60.3% (35/58) in dentists and 88.6% (39/44) and 61.4% (27/44) in dental hygienists. The compliance rate for routine use of protective eyewear was 60.3% (35/58) for dentists and 34.1% (15/44) for hygienists. Of the presented informational items included in the questionnaire, those that strongly persuaded respondents to use protective eyewear were 'splatters from the patient's mouth contain blood' (90%, 99/110) and 'dental operations at our clinic are performed based only on a questionnaire without serious examinations for HBV, HCV, and HIV' (71.8%, 79/110). The reason of low compliance of protective eyewear among dentists might relate to fine dental procedures. Appropriate information is important for the motive of wearing personal protective equipment, and an early educational program may have a potential to increase compliance with the use of that equipment.

Acute inflammation in horizontal incompletely impacted third molar with radiolucency in the elderly.

Although radiolucency has been shown as a risk of infection, the poorly understood effects of aging on radiolucency correlate with acute pericoronitis, which has a high risk of infection extending any complications. We reviewed the records of 346 consecutive patients aged more than 41 years to evaluate whether pericoronal radiolucency below the crown in mandibular horizontal incompletely impacted third molars is related to acute inflammation. The frequency of acute inflammation in teeth with pericoronal radiolucency below the crown was similar to that in teeth without; however, the odds ratio of acute inflammation exhibited in women aged more than 61 years compared to women aged 41-50 years was 9.77 (95% confidence interval [CI]: 1.67-57.29; P < <0.05), and in women aged more than 61 years compared to women aged 51-60 years was 26.25 (95% CI: 2.94-234.38; P < 0.01). The odds ratio of severe acute inflammation exhibited in men aged more than 61 years compared to men aged 41-50 years was 16.67 (95% CI: 1.76-158.27; P < 0.01). These odds ratios indicate an association of acute pericoronitis, including the severe forms of acute inflammation that result from pericoronitis, with pericoronal radiolucency below the crown in the elderly.

Bone resorption in incompletely impacted mandibular third molars and acute pericoronitis.

Acute pericoronitis (AP) arises frequently in incompletely impacted mandibular third molars, but it remains unknown whether bone resorption in aging is associated with acute inflammation of the third molar. We conducted an experiment to compare the ratio of bone resorption to root length in the distal surface of the second molar (A), the proximal surface (B), and distal surface (C) in mesio-angular, incompletely impacted third molars in 27 young and 58 older adults with AP and 77 young and 79 older adults without a history of AP. Bone resorption in A, B, and C in older adults with AP demonstrated a significantly higher ratio when compared to those without AP, whereas there was no difference between those with and without AP in young adults except for B in women. However, there were no differences between bone resorption in B with AP in young and older women, and between bone resorption in C with AP in young and older adults. These indicate that AP and bone resorption are associated with incompletely impacted mandibular third molars in older adults.

Radiolucency below the crown of mandibular horizontal incompletely impacted third molars and acute inflammation in men with diabetes.

Although mandibular third molar has a high risk of infection extending any complications, the influence of diabetes on radiolucency and acute inflammation in pericoronitis remains unclear. The present study was to evaluate whether radiolucency below the crown is related to acute inflammation in mandibular horizontal incompletely impacted third molars and to review the records of 140 men more than 45 years with and without diabetes. The odds ratio of exhibiting acute inflammation was 3.38 (95% CI: 1.13-10.16, p < 0.05) and that of exhibiting severe acute inflammation was 15.38 (95% CI: 3.56-66.49, p < 0.0001), indicating an association of acute pericoronitis in diabetes. The frequency of radiolucency below the crown and below the root in diabetics was similar to that in nondiabetics. However, the odds ratio of exhibiting both radiolucency below the crown and acute inflammation under the diabetic condition was 4.85 (95% CI: 1.60-14.73, p < 0.01), whereas that of diabetics showing both radiolucency below the root and acute inflammation was 0.46 (95% CI: 0.06-3.74, p = 0.74). Radiolucency below the crown and acute inflammation were associated with diabetes, but that below root and acute inflammation were not associated with diabetes, indicating that the region below the crown carries susceptibility to acute pericoronitis, whereas the periodontium shows a protective effect against acute pericoronitis.

Acute inflammation at a mandibular solitary horizontal incompletely impacted molar.

Acute inflammation is frequently seen in the elderly around incompletely impacted molars located apart from molars or premolars. To identify the factors causing acute inflammation in the solitary molars without second molars or without second and first molars, ages of patients and rates of acute inflammation in 75 horizontal incompletely impacted mandibular molars in contact or not in contact with molars in subjects 41 years old or older were studied using orthopantomographs. Acute inflammation was seen in nine third molars out of 48 third molars in contact with second molars (18.8%), whereas acute inflammation was seen in 11 molars out of 19 solitary molars without second molars or without first and second molars (57.9%) (p < 0.01). The mean age of 48 subjects with third molars in contact with the second molar was 50.42 +/- 7.62 years, and the mean age of 19 subjects with isolated molars was 65.16 +/- 10.41 years (p < 0.0001). These indicate that a solitary horizontal incompletely impacted molar leads more frequently to acute inflammation along with aging due to possible bone resorption resulting from teeth loss.

Postnatal changes in 5HT and NK1 receptors in rat trigeminal motor nucleus and surroundings.

Our previous study showed that developmental changes to serotonin and substance P coexist in the trigeminal motor nucleus (Vmo), dorsolateral subnucleus (Vmo.dl), ventromedial subnucleus (Vmo.vm) and the area within 300 microm surrounding Vmo (SVmo). This occurred in rats from embryonic day (E)19 to postnatal day (P)70, with density of these terminals peaking at P7 in these three areas. The present study examined postnatal development of serotonergic 1A receptor (5HT1A receptor) and substance P receptor (NK1 receptor) expression in Vmo and SVmo in rats from E19 to P70. No significant changes in percentages of 5HT1A and NK1 receptor-expressing cells were seen between E19 and P70 by immunohistochemical study or in situ hybridization. In a real-time PCR study, quantities of 5HT1A and NK1 receptor expression peaked at P7, indicating that expression of these receptors was maximized in each neuronal cell body in the Vmo and SVmo at P7. This result corresponds with postnatal changes in serotonin/substance P-coexisting terminals found in our previous study. Furthermore, 5HT1A and NK1 receptors displayed very similar patterns of expression, which may support the hypothesis that potentiation of serotonin and substance P are involved in excitability regulating trigeminal motor functions, including mastication and breathing.

NK1 receptor activation by geniohyoid primary afferents modulates parasympathetic postganglionic neuronal excitability in the rat.

Previous studies have indicated that the geniohyoid (GH) muscle receives innervation via both the hypoglossal nerve (CNXII) and the ansa cervicalis. Our recent studies revealed that the efferent root that contributes to the ansa cervicalis is a parasympathetic pathway and contains postganglionic cell bodies. Afferent axons from the GH muscle also travel via the ansa cervicalis, and afferent cell bodies are located in spinal ganglia. The present study attempts to locate the central terminations of these afferents. From the peripheral cut end of the ansa cervicalis, we recorded afferent discharges that coincided with inspiration and these were elicited by stretch of the GH muscle. After cutting CNXII proximal to its union with the ansa cervicalis, we applied horseradish peroxidase to the branch of CNXII that innervates the GH muscle. This procedure labeled cells ipsilaterally in the C2 spinal ganglia but not in the brainstem or upper spinal cord. Substance P-reactive terminals in the peripheral CNXII trunk were in apparent contact with vasoactive intestinal peptide-reactive cell bodies. Addition of the NK1 receptor agonist SP(NK1) excited parasympathetic postganglionic neurons and the specific NK1 receptor antagonist GR82334 blocked these effects in vitro. These results suggest that GH primary afferents synapse on parasympathetic postganglionic neurons in the CNXII trunk and that activation of SP(NK1) receptors modulates activity in these neurons.

Colocalization of serotonin and substance P in the postnatal rat trigeminal motor nucleus and its surroundings.

Trigeminal motoneurons are involved in a variety of oral motor activities, including mastication and breathing, which must be adapted to postnatal environmental change. Serotonin has both an excitatory and an inhibitory effect on trigeminal motor function, whereas substance P has mainly an excitatory effect. In the present study, we measured the density of serotonin- and substance P-immunoreactive nerve terminals in the trigeminal motor nucleus and the area 300 microm surrounding it in rats from embryonic day 19 to postnatal day 70. The density of these terminals gradually increased from embryonic day 19 to postnatal day 7 and decreased thereafter. The density was greatest in the ventromedial subnucleus of the trigeminal motor nucleus at embryonic day 19 and postnatal day 0 and in the area 300 microm surrounding trigeminal motor nucleus at postnatal day 4 and older. Two-color fluorescence immunohistochemistry was used to identify nerve processes immunoreactive for both substance P and serotonin. Approximately, 90% of serotonergic terminals also contained substance P at all ages examined, which suggests that the physiological function of terminals in which these neurotransmitters are colocalized is similar throughout development.

The 14-3-3 proteins of Trypanosoma brucei function in motility, cytokinesis, and cell cycle.

The cDNAs for two isoforms (I and II) of the 14-3-3 proteins have been cloned and functionally characterized in Trypanosoma brucei. The amino acid sequences of isoforms I and II have 47 and 50% identity to the human tau isoform, respectively, with important conserved features including a potential amphipathic groove for the binding of phosphoserine/phosphothreonine-containing motifs and a nuclear export signal-like domain. Both isoforms are abundantly expressed at approximately equal levels (1-2 x 10(6) molecules/cell) and localized mainly in the cytoplasm. Knockdown by induction of double-stranded RNA of isoform I and/or II in both bloodstream and procyclic forms resulted first in a reduction of cell motility and then significant reduction in cell growth rates and morphological changes; the changes include aberrant numbers of organelles and abnormal shapes and sizes that mimic phenotypes produced by various cytokinesis inhibitors. Morphological and fluorescence-activated cell sorting analysis of the cell cycle suggested that isoforms I and II might play important roles in nuclear (G2-M transition) and cell (M-G1 transition) division. These findings indicate that the 14-3-3 proteins play important roles in cell motility, cytokinesis, and the cell cycle.