bluuebunny/crossref_metadata_2025 huggingface dataset:
Residual heat deposition in dental enamel during IR laser ablation at 2.79, 2.94, 9.6, and 10.6 μm
Authors: Fried Daniel , Ragadio Jerome , Champion Allen
Published: August 2002

Background and Objective

The principal factor limiting the rate of laser ablation of dental hard tissue is the risk of excessive heat accumulation in the tooth. Excessive heat deposition or accumulation may result in unacceptable damage to the pulp. The objective of this study was to measure the residual heat deposition during the laser ablation of dental enamel at those IR laser wavelengths well suited for the removal of dental caries. Optimal laser ablation systems minimize the residual heat deposition in the tooth by efficiently transferring the deposited laser energy to kinetic and internal energy of ejected tissue components.

Study Design/Materials and Methods

The residual heat deposition in dental enamel was measured at laser wavelengths of 2.79, 2.94, 9.6, and 10.6 μm and pulse widths of 150 nsec −150 μsec using bovine block “calorimeters.” Water droplets were applied to the surface before ablation with 150 μsec Er:YAG laser pulses to determine the influence of an optically thick water layer on reducing heat deposition.

Results

The residual heat was at a minimum for fluences well above the ablation threshold where measured values ranged from 25–70% depending on pulse duration and wavelength for the systems investigated. The lowest values of the residual heat were measured for short (< 20 μs) CO2 laser pulses at 9.6 μm and for Q‐switched erbium laser pulses at 2.79 and 2.94 μm. Droplets of water applied to the surface before ablation significantly reduced the residual heat deposition during ablation with 150 μsec Er:YAG laser pulses.

Conclusions

Residual heat deposition can be markedly reduced by using CO2 laser pulses of less than 20 μsec duration and shorter Q‐switched Er:YAG and Er:YSGG laser pulses for enamel ablation. Lasers Surg. Med. 29:221–229, 2001. © 2001 Wiley‐Liss, Inc.
Treatment of rhinophyma with Er:YAG Laser
Authors: Orenstein Arie , Haik Josef , Tamir Jeremy , Winkler Eyal , Frand Jacob , Zilinsky Isaac , Kaplan Haim
Published: August 2002

Background and Objective

Rhinophyma is a benign, disfiguring disorder of the nose which presents the end stage of acne rosacea, and can also occasionally result in nasal airway obstruction. We describe the first series of patients treated with Erbium laser.

Study Design/Materials and Methods

Severe‐to‐moderate rhinophyma in six patients were treated at our institution between 1995 and 1996, using the Erbium:YAG (Erb:YAG) laser.

Results

All patients achieved marked cosmetic improvement, with no complications. Post‐operative healing time was seven to fourteen days‐significantly shorter than similar other modalities.

Conclusions

The Erbium laser provides very accurate tissue ablation and allows the sculpturing of the hypertrophied areas, offering good cosmetic results, with a very short healing period as shown in our study. Lasers Surg. Med. 29:230–235, 2001. © 2001 Wiley‐Liss, Inc.
Gene expression profile in BALB/c‐3T3 cells transformed with beryllium sulfate
Authors: Joseph Pius , Muchnok Timothy , Ong Tong‐man
Published: August 2002

Differential gene expression was studied to understand the potential molecular mechanism responsible for cell transformation and tumorigenesis induced by beryllium. Cell lines were derived from tumors developed in nude mice injected subcutaneously with BALB/c‐3T3 cells morphologically transformed with beryllium sulfate. Using the Atlas mouse 1.2 cDNA expression microarray, the expression profiles of 1176 genes, belonging to several different functional categories, were studied in the tumor cells as well as in the nontransformed control cells. Expression of 18 genes belonging to two functional groups was found to be consistently and reproducibly different (at least twofold) in the tumor cells compared with the control cells. The functional groups and the differentially expressed genes are as follows: The cancer‐related genes (nine genes) were the ets‐related transcription factor activated by ras, colony‐stimulating factor, A‐myb, sky, cot1, c‐fos, c‐jun, c‐myc, and R‐ras proto‐oncogenes. The DNA synthesis, repair, and recombination genes (nine genes) were the DNA replication licensing factor MCM4, the DNA replication licensing factor MCM5, the DNA mismatch repair gene PMS2, the DNA excision repair gene, the DNA mismatch repair gene MSH2, the ultraviolet excision repair gene Rad23 DNA ligase 1, Rad51, and Rad52. The differential gene expression profile was confirmed with reverse transcription–polymerase chain reaction using primers specific for the differentially expressed genes. In general, expression of the cancer‐related genes was upregulated, while expression of genes involved in DNA synthesis, repair, and recombination was downregulated in the tumor cells compared with the control cells. Using c‐fos and c‐jun, two of the differentially expressed genes, as model genes, we have found that in the nontransformed BALB/c‐3T3 cells, the beryllium‐induced transcriptional activation of these genes was dependent on pathways of protein kinase C and mitogen‐activated protein kinase and independent of reactive oxygen species. These results indicate that beryllium‐induced cell transformation and tumorigenesis are accompanied by and are possibly a product of alterations in expression of genes related to cancer and to DNA synthesis, repair, and recombination. © 2001 Wiley‐Liss, Inc.
Electrophoretic separations in PMMA capillaries with uniform and discontinuous buffers
Authors: Caslavska Jitka , Thormann Wolfgang
Published: August 2002

The basic phenomena occurring within polymethylmethacrylate (PMMA, Plexiglass) capillaries under the conditions of capillary zone electrophoresis (CZE), isotachophoresis (CITP), and isoelectric focusing (CIEF) have been explored in our laboratory and are compared to those characteristic for untreated fused‐silica (FS) capillaries. The origin of charge at the inner wall of the PMMA capillary is different compared to that of FS, this having a significant impact on the magnitude of the electroosmotic flow. While both column materials are characterized with a negative surface charge, electroosmosis in PMMA is considerably smaller over the entire pH range. Furthermore, electroosmosis in PMMA and FS is shown to follow different dependencies on the ionic strength (I). Using a set of experimentally determined values, the electroosmotic mobility (μEO) of a PMMA capillary is best described using the relationship μEO=a+b/√I. For a FS capillary, μEO follows the expression μEO=a+b log(I). A dynamic capillary electrophoresis simulation model comprising the determined pH and ionic strength dependent wall titration data as input for calculation of electroosmosis is shown to provide CZE and CITP electropherograms that qualitatively agree well with those obtained experimentally. In contrast to FS and glass, electroosmosis at any pH in PMMA is too weak to perform bidirectional (i.e., simultaneous cationic and anionic) CZE and CITP analyses with a detector placed toward the cathodic capillary end. Furthermore, the same is true for the performance of CIEF with electroosmotic zone displacement. Imposed flow is demonstrated to provide the required net buffer flow. © 2001 John Wiley & Sons, Inc. J Micro Sep 13: 69–83, 2001
Focal pathological startle following pontine infarction
Authors: Watson Shaun R.D. , Colebatch James G.
Published: August 2002

A 36‐year‐old male developed an acute right‐sided weakness due to left‐sided pontine infarction. Two months later, he first noticed sudden right elbow flexion in response to a loud unexpected noise. Detailed electrophysiological assessment was performed. A large, short‐latency (median 39 msec), synchronous electromyographic discharge occurred in the right biceps brachii electrodes following a 50‐msec, 120‐dB 1‐kHz tone burst, with habituation only with very short (30‐second) interstimulus intervals. Less synchronous activity at longer latencies was present both in a number of right‐sided arm muscles at rest and on the clinically unaffected side during a tonic voluntary contraction. We discuss possible underlying mechanisms and our reasons for considering this a focally enhanced startle response. Our report broadens the range of expression of acquired startle disorders. © 2001 Movement Disorder Society.
Placebo‐associated improvements in motor function: Comparison of subjective and objective sections of the UPDRS in early Parkinson's disease
Authors: Goetz Christopher G. , Leurgans Sue , Raman Rema
Published: August 2002

The Unified Parkinson's Disease Rating Scale (UPDRS) is primarily composed of an investigator‐derived objective rating of motor function and a patient‐derived assessment of activities of daily living (ADL). Using a stringent definition of placebo effect, we examined the frequency, temporal development, and stability of improvements during placebo treatment over 6 months in a large placebo‐controlled trial of deprenyl and tocopherol in early Parkinson's disease (DATATOP). One hundred ninety‐nine subjects received placebo treatment in the randomized, multicenter, placebo‐controlled DATATOP study. We compared the baseline UPDRS motor section scores with follow‐up scores at 4, 13, and 26 weeks. Placebo‐associated improvement was defined as an improvement over baseline score in motor UPDRS of at least 50% or a change in at least two motor items at any one visit by two or more points. Seventeen percent of the 185 subjects who qualified for analysis met the placebo response criteria. The group prevalence of response was steady (7% to 10%) at any one visit without a marked predominance of an early study effect. Older subjects with more motor impairment at baseline were most likely to show a placebo‐associated improvement. ADL scores were low throughout the study, and ADL improvements did not identify the subjects with objectively defined placebo‐associated improvement. Prominent improvements in investigator‐derived objective measures of Parkinson's disease motor impairment occur during clinical trials, including one that was not aimed at showing improved short‐term efficacy. Although the notion of placebo effect often implies patient‐based perceptions, we found subjective changes to be infrequent in placebo‐treated patients, suggesting that either: (1) the placebo effect was rater‐driven; (2) the ADL questionnaire is insensitive to transient but objectively demonstrable motor changes; or (3) that the objective changes, albeit major, are within the realm of natural variation in the UPDRS motor scale from visit to visit. © 2002 Movement Disorder Society.
Do risk factors for Alzheimer's disease predict dementia in Parkinson's disease? An exploratory study
Authors: Levy Gilberto , Tang Ming‐Xin , Cote Lucien J. , Louis Elan D. , Alfaro Brenda , Mejia Helen , Stern Yaakov , Marder Karen
Published: August 2002

The extent to which concomitant Alzheimer's disease (AD) is etiologically related to the development of dementia in Parkinson's disease (PD) remains controversial. We explored the association of four risk factors associated with AD, including head injury, smoking, hypertension, and diabetes mellitus, with incident dementia in PD. A cohort of 180 nondemented PD patients from the Washington Heights community in northern Manhattan, New York, completed a risk factor questionnaire at baseline and was followed annually with neurological and neuropsychological evaluations. The association of baseline variables with incident dementia was analyzed by using Cox proportional hazards models. All analyses controlled for age at baseline, gender, years of education, duration of PD, and total Unified Parkinson's Disease Rating Scale (UPDRS) motor score at baseline. Of 180 patients (mean age, 71.0 ± 10.3 years), 52 (29%) became demented during a mean follow‐up period of 3.6 ± 2.2 years. Head injury risk ratio ([RR] 0.9; 95% confidence interval [CI], 0.4–2.2; P = 0.9), hypertension (RR, 0.7; 95% CI, 0.4–1.4, P = 0.3), and diabetes mellitus (RR, 0.8; 95% CI, 0.3–2.3; P = 0.7) were not significantly associated with incident dementia in the Cox models. Patients who reported having ever smoked were at increased risk for the development of dementia compared with nonsmokers (RR, 2.0; 95% CI, 1.0–3.9; P = 0.05). Current smoking was significantly associated with incident dementia (RR, 4.5; 95% CI, 1.2–16.4; P = 0.02), whereas past smoking approached significance (RR, 1.9; 95% CI, 0.9–3.7; P = 0.07). Although an inverse association between smoking and PD has been reported in several studies, our study showed a positive association between smoking and dementia in the setting of PD. The association of smoking with incident dementia in PD deserves further study. © 2002 Movement Disorder Society.
Nocturnal body core temperature falls in Parkinson's disease but not in multiple‐system atrophy
Authors: Pierangeli G. , Provini F. , Maltoni P. , Barletta G. , Contin M. , Lugaresi E. , Montagna P. , Cortelli P.
Published: August 2002

OBJECTIVE:

To evaluate whether the circadian rhythm of body core temperature (CRT°) can differentiate Multiple‐System Atrophy (MSA) from Idiopathic Parkinson's disease (IPD).

METHODS:

We evaluated 14 patients with probable MSA, seven with IPD, and eight controls. After a preliminary evaluation of cardiovascular autonomic function, rectal temperature and sleep‐wake cycle were monitored continuously for 48 hours in a temperature‐controlled room, at constant bed rest with controlled food intake and fixed light‐dark schedule.

RESULTS:

MSA patients showed cardiovascular autonomic sympathetic and parasympathetic failure. IPD had normal cardiovascular autonomic function. A 24‐hour rhythm of body core temperature (BcT°) was present in all subjects. IPD had CRT° comparable to controls. In MSA the mesor was higher and mean BcT° of each hour was significantly higher from 11 p.m. to 7 a.m. The analysis of mean BcT° during the different sleep phases showed significantly higher values during both NREM (1–2, 3–4) and REM sleep stages in MSA.

CONCLUSIONS:

The physiological nocturnal fall of BcT° is blunted in MSA patients mainly because BcT° did not decrease during sleep. This CRT° pattern is not justified by differences in sleep structure and may reflect an impairment of central sympathetic nervous system function. © 2001 Movement Disorder Society.
Quantitative assessment of daytime motor activity provides a responsive measure of functional decline in patients with Huntington's disease
Authors: van Vugt J.P.P. , Siesling S. , Piet K.K.E. , Zwinderman A.H. , Middelkoop H.A.M. , van Hilten J.J. , Roos R.A.C.
Published: August 2002

Voluntary motor impairment is a functionally important aspect of Huntington's disease (HD). Therefore, quantitative assessment of disturbed voluntary movement might be important in follow‐up. We investigated the relation between quantitatively assessed daytime motor activity and symptom severity in HD and evaluated whether assessment of daytime motor activity is a responsive measure in the follow‐up of patients.

Sixty‐four consecutive HD patients and 67 age‐ and sex‐matched healthy controls were studied. Daytime motor activity was recorded using a wrist‐worn activity monitor that counts all movements during a period of five consecutive days. Patients were rated clinically for voluntary motor impairment, dyskinesias, posture & gait, depression, cognitive impairment and functional capacity. Follow‐up was available from 40 patients (mean follow‐up 2.0 years) and 29 controls (mean follow‐up 5.9 years).

Despite chorea, patients had less daytime motor activity than controls (P < 0.005). This hypokinesia correlated with impaired voluntary movements (r = 0.37; P < 0.01), disturbed posture & gait (r = 0.38; P < 0.005) and especially with reduced functional capacity (r = 0.51; P < 0.0005). During follow‐up, hypokinesia remained unchanged in clinically stable patients, but became worse in those whose functional disability progressed (P < 0.005).

Hypokinesia seems a core symptom of HD which is related to functional capacity. Actimetric assessment of hypokinesia is responsive to disease progression and can be used as an objective tool for follow‐up. © 2001 Movement Disorder Society.
Penicillamine‐induced lethal status dystonicus in a patient with Wilson's disease
Authors: Svetel Marina , Šternić Nadežda , Pejović Sanja , Kostić Vladimir S.
Published: August 2002

A 37‐year‐old man with Wilson's disease is described, in whom the introduction of penicillamine therapy was followed after 3.5 weeks by the development of the status dystonicus with a fatal outcome.
Acute paroxysmal dystonia induced by fluoxetine
Authors: Domínguez‐Morán José A. , Callejo Juan M. , Fernández‐Ruiz Luis C. , Martínez‐Castrillo Juan C.
Published: August 2002

We report a case of a 32‐year‐old woman who developed paroxysmal episodes of right hemidystonia 2 days after taking fluoxetine. The attacks subsided 2 days after fluoxetine was withdrawn and did not recur afterwards. To our knowledge, this is the first report of paroxysmal dystonia induced by fluoxetine. © 2001 Movement Disorder Society.
Botulinum toxin and short‐term electrical stimulation in the treatment of equinus in cerebral palsy
Authors: Detrembleur C. , Lejeune T.M. , Renders A. , Van den Bergh P.Y.K.
Published: August 2002

Intramuscular botulinum toxin type A (BT‐A) has been shown to reduce spasticity and to improve gait in children with cerebral palsy. To determine whether the efficacy of BT‐A may be enhanced by electrical stimulation, as suggested in focal dystonia or in adult spastic patients, 12 children with dynamic foot equinus deformity were randomly assigned to two groups in a blinded, clinically controlled trial. Intramuscular BT‐A into calf muscles was followed by adjuvant electrical stimulation in Group A (n = 6) but not in Group B (n = 6). Clinical assessment and instrumented gait analysis were performed before and 1, 3, and 6 months after treatment. The combined treatment of BT‐A and electrical stimulation was not superior to BT‐A alone. For all patients, improvement of the clinical and gait variables occurred at 1 and 3 months after BT‐A injection. © 2001 Movement Disorder Society.
Natural products as probes of cell biology: 20 years of didemnin research
Authors: Vera Matthew D. , Joullié Madeleine M.
Published: August 2002

The discovery of the didemnin family of marine depsipeptides launched an exciting and intriguing chapter in natural product chemistry. The unusual structure of the didemnin congeners has led to several total syntheses by research groups from around the world. The impressive in vitro and in vivo biological activities of the didemnins resulted in the first human clinical trials in the U.S. of a marine natural product against cancer, and additional clinical trials of a second‐generation didemnin, dehydrodidemnin B (aplidine), are underway. As we mark the 20‐year anniversary of the discovery of the didemnins, this class of natural products continues to stimulate active research in fields ranging from synthetic and medicinal chemistry to clinical oncology and cell biology. While some progress was made in dissecting the molecular mechanism of action and in establishing structure‐activity relationships, there are still more questions than answers. This review covers the recent didemnin literature, highlighting the work directed towards understanding how this group of natural products interact with fundamental processes such as cell proliferation, protein biosynthesis, and apoptosis. The didemnin field illustrates how natural product chemistry may be used as a critical tool for the study of cell biology. © 2002 John Wiley & Sons, Inc. Med Res Rev, 22, No. 2, 102–145, 2002; DOI 10.1002/med.10003
Conditions for the parameter dyadics of lossy bianisotropic media
Authors: Lindell I. V. , Dahl F. M.
Published: August 2002

Conditions for the parameter dyadics are derived for bianisotropic media to be lossy (power absorbing). The conditions, which can be expressed easily in terms of the 6×6 matrix of four parameter dyadics as being Hermitian and negative definite, are brought to the level of individual parameter dyadics as four separate dyadic conditions. The theory is applied to three particular medium classes: bi‐isotropic media, uniaxial bianisotropic media, and gyrotropic bianisotropic media as a set of inequalities for the scalar parameters. The conditions are needed for judging the physical realizability of parameter values of new composite media. © 2001 John Wiley & Sons, Inc. Microwave Opt Technol Lett 29: 175–178, 2001.
Pharmacokinetic longitudinal studies of antibiotics administered via a permanent intraosseous device in micropigs
Authors: Chastagner Pascal , Lozniewski Alain , Lascombes Pierre , Barberi‐Heyob Muriel , Merthes Paul Michel , Merlin Jean‐Louis
Published: August 2002

Background

In critically ill children, intraosseous (IO) administration of a medicine provides an alternate vascular access route when IV access is not readily available. We investigated the short and long‐term efficacy and safety of a totally intraosseously implantable device.

Procedure

This study was undertaken in micropigs (i) to compare serum levels achieved by equal bolus dosages of two antibiotics (amikacin and vancomycin) administered through an intratibial needle, an intraosseous implantable device and central IV routes to determine whether standard parenteral dosing guidelines for antibiotics may be applied without modification for IO injection, and (ii) to show the efficiency of the implantable device over a prolonged period, as a permanent access to intraosseous space.

Results

The area under the plasma concentration time curves were similar for IV, intratibial and through the intraosseous implantable device, for intermittent administrations of amikacin and vancomycin, over a period of 6 months. However, vancomycin did not reach therapeutic levels via both IO routes. We did not find any alteration of amikacin and vancomycin pharmacokinetics over a period of 6 months using the implantable device. No complication occurred.

Conclusions

Long‐term administration of antibiotics through a totally implantable intraosseous device is feasible and safe in micropigs. Although the procedure seems promising, additional studies of the continuous infusion of chemotherapeutic agents, blood products and antimicrobial solutions are needed prior to use in humans. Med. Pediatr. Oncol. 36:635–640, 2001. © 2001 Wiley‐Liss, Inc.
Complete assignment of 1H and 13C NMR data for nine protopanaxatriol glycosides
Authors: Teng Rongwei , Li Haizhou , Chen Jiangtao , Wang Dezu , He Yineng , Yang Chongren
Published: August 2002

Nine protopanaxatriol glycosides isolated from mild acid hydrolysis products of crude root saponins of Panax notoginseng were identified as 20(R)‐ginsenoside‐Rh1, 20(S)‐ginsenoside‐Rh1, ginsenoside‐Rg1, ‐Re and ‐Rg2, notoginsenoside‐R2 and ‐R1, a mixture of 25‐hydroxy‐20(S)‐ginsenoside‐Rh1 and its C‐20 (R) epimer, ginsenoside‐Rh4. The complete assignments of the 1H and 13C NMR chemical shifts for these glycosides were obtained by means of 2D NMR techniques, including 1H–1H COSY, ROESY, HMQC, HMBC and HMQC‐TOCSY spectra. The glycosylation shift effect of protopanaxatriol and the differences in chemical shifts between 20(R)‐ and 20(S)‐protopanaxatriol isomers are also discussed. Except for ginsenoside‐Re and ‐Rg2, complete NMR assignments of the other seven glycosides are reported for the first time. Copyright © 2002 John Wiley & Sons, Ltd.
Structure elucidation and antibacterial activity of a new coumarinolignoid from Daphne gnidium L.
Authors: Cottiglia F. , Bonsignore L. , Loy G. , Garau D. , Floris C. , Casu M.
Published: August 2002

Phytochemical investigation of the dichloromethane extract from the leaves of Daphne gnidium L. afforded 2‐(4‐hydroxy‐3,5‐dimethoxyphenyl)‐3‐hydroxymethyl‐2,3‐dihydro‐1,4,5‐trioxaphenanthren‐6‐one, and its structure was determined by a combination of spectral methods (1D and 2D 1H, 13C NMR and HRMALDI and EI mass spectrometry). The new compound was found to possess moderate antibacterial activity against Staphylococcus aureus and Bacillus cereus. However, it showed no cytotoxic activity against KB cells. Copyright © 2002 John Wiley & Sons, Ltd.
27Al and 7Li spin‐echo and spin‐echo double resonance NMR applications to the study of lithium depletion and surface oxidation in Al–Li alloy
Authors: Ba Yong , Ripmeester John A.
Published: August 2002

The physical properties and the near‐surface microstructures of Al–Li alloys are known to be affected by thermo‐mechanical processes during alloy manufacture. The research reported here explored 27Al and 7Li spin‐echo and spin‐echo double resonance NMR to follow the structural changes that accompany lithium depletion and oxidation processes at the alloy surface. The NMR analysis generally agrees with conclusions reached previously using other analytical methods. The major oxide compound generated is Li2CO3 and the minor oxide compound is LiAlO2, which is formed at higher temperatures. LiAlO2 is shown to be formed in spatially confined clusters on the surface of the Li‐depleted, Al‐rich near‐surface layer. Copyright © 2001 John Wiley & Sons, Ltd.
Molecular structures of organotin tropolonato complexes with tin coordination numbers 5–7, seen by single‐crystal x‐ray diffraction and solid‐ and solution‐state 119Sn NMR
Authors: Camacho‐Camacho Carlos , Contreras Rosalinda , Nöth Heinrich , Bechmann Matthias , Sebald Angelika , Milius Wolfgang , Wrackmeyer Bernd
Published: August 2002

Organotin tropolonato complexes of the type R3Sn(trop) 1 [R = Bu (b), Ph (c)], R2Sn(trop)2 2 [R = Me (a), Bu (b), Ph (c)] and RSn(trop)3 3 [R = Bu (b), Ph (c)] were studied by single‐crystal X‐ray diffraction (1c, 2a,b, 3b), 13C and 119Sn MAS NMR and 119Sn NMR in solution. The tin coordination numbers from 5 to 7, as determined for the solid state, are retained in solution. The different distorted octahedral surroundings (cis‐like arrangement of the methyl groups in 2a; trans‐like arrangement of the butyl groups and weak intermolecular association in 2b) of the tin atoms in solid compounds 2 are reflected both by the 119Sn chemical shift anisotropies and the magnitude of the coupling constants |1J(117/119Sn,13C)|. The molecular structures of 2a and 2b become more similar to each other in solution, as indicated by differences in the isotropic 119Sn chemical shifts in solution and in the solid state, and even more obvious by changes in the magnitude of the corresponding coupling constants |1J(117/119Sn,13C)|. Copyright © 2001 John Wiley & Sons, Ltd.
Inhibition of apoptosis in vitellogenic ovarian follicles of rainbow trout (Oncorhynchus mykiss) by salmon gonadotropin, epidermal growth factor, and 17β‐estradiol
Authors: Wood Antony W. , Van Der Kraak Glen
Published: August 2002

We have examined the ability of selected hormones and growth factors to suppress the spontaneous onset on apoptotic DNA fragmentation in isolated vitellogenic rainbow trout ovarian follicles cultured in serum‐free conditions. Primary culture of isolated follicles for 24 hr in serum‐free conditions resulted in a 3–5‐fold increase in the amount of fragmented DNA as compared to non‐cultured controls, measured by radioactive 3′end‐labeling. Culture in medium containing salmon gonadotropin (SG‐G100; 1, 5 μg/ml) suppressed the spontaneous onset of DNA fragmentation in dose‐dependent fashion. Culture with 1 ng/ml 17β‐estradiol, or 100 ng/ml epidermal growth factor also suppressed the spontaneous onset of apoptosis, whereas culture with higher concentrations of 17β‐estradiol (10 and 100 ng/ml), insulin‐like growth factor I (IGF‐I; 1, 10, and 100 ng/ml), or 8‐bromo‐cAMP (0.1, 1, and 5 mM) was ineffective in suppressing apoptosis. Apoptosis was confirmed as the mode of cell death through positive identification of nuclear morphological characteristics associated with apoptosis, and positive staining for fragmented DNA using in situ end‐labeling (TUNEL); apoptotic cells identified in situ were almost exclusively localized to the thecal/epithelial region of the follicle. In summary, this study shows that vitellogenic ovarian follicles are susceptible to apoptosis and that both endocrine and locally‐derived growth factors may play a role as cell survival factors by preventing apoptosis. The study also suggests that rainbow trout differ markedly from mammals both in terms of the cell types susceptible to apoptosis and the responsiveness to specific growth factors in terms of inhibiting apoptosis. Mol. Reprod. Dev. 61:511–518, 2002. © 2002 Wiley‐Liss, Inc.
On the regulation of Crisp‐1 mRNA expression and protein secretion by luminal factors presented in vivo by microperfusion of the rat proximal caput epididymidis
Authors: Turner T.T. , Bomgardner D.
Published: August 2002

Synthesis and secretion of certain epididymal proteins are regulated by lumicrine factors from the testis or from upstream regions of the excurrent ducts. Cysteine‐rich secreted protein‐1 (Crisp‐1) is a major androgen regulated protein in the epididymal lumen fluid of the rat and other species. Previous research has demonstrated that disturbance of the luminal microenvironment through obstruction of the tract reduces Crisp‐1 synthesis and secretion. The present study was undertaken to determine the influence of the luminal microenvironment on rat proximal caput epididymal Crisp‐1 secretion into lumen fluid and on Crisp‐1 gene expression in the same tubules. Western blot analysis demonstrated that Crisp‐1 protein concentrations were reduced from control levels by perfusion with artificial caput fluid containing no testicular factors and were not increased by perfusion with fluids containing rete testis fluid proteins. Crisp‐1gene expression was also reduced by perfusion with artificial caput fluid and not increased by perfusion with rete testis fluid proteins. Perfusion with artificial caput fluid containing 5α‐dihydrotestosterone did increase one Crisp‐1 transcript. This study demonstrates that intraluminal testicular proteins are not important co‐regulators with androgens of Crisp‐1gene expression or resulting Crisp‐1 secretion into the rat proximal caput tubule lumen in vivo. Mol. Reprod. Dev. 61:437–444, 2002. © 2002 Wiley‐Liss, Inc.
Transgenic pig expressing the enhanced green fluorescent protein produced by nuclear transfer using colchicine‐treated fibroblasts as donor cells
Authors: Lai Liangxue , Park Kwang‐Wook , Cheong Hee‐Tae , Kühholzer Birgit , Samuel Melissa , Bonk Aaron , Im Gi‐Sun , Rieke August , Day Billy N. , Murphy Clifton N. , Carter David B. , Prather Randall S.
Published: August 2002

Fetal‐derived fibroblast cells were transduced with replication defective vectors containing the enhanced green fluorescent protein (EGFP). The transgenic cells were treated with colchicine, which theoretically would synchronize the cells into G2/M stage, and then used as donor nuclei for nuclear transfer. The donor cells were transferred into the perivitalline space of enucleated in vitro matured porcine oocytes, and fused and activated with electrical pulses. A total of 8.3% and 28.6% of reconstructed oocytes showed nuclear envelope breakdown and premature chromosome condensation 0.5 and 2 hr after activation, respectively. Percentage of pronuclear formation was 62.5, 12 hr after activation. Most (91.4%) of the 1‐cell embryos with pronuclei did not extrude a polar body. Most (77.2%) embryos on day 5 were diploid. Within 2 hr after fusion, strong fluorescence was detectable in most reconstructed oocytes (92.3%). The fluorescence in all NT embryos became weak 15 hr after fusion and disappeared when culture to 48 hr. But from day 3, cleaved embryos at the 2‐ to 4‐cell stage started to express EGFP again. On day 7, 85.8% of cleaved embryos expressed EGFP. A total of 9.4% of reconstructed embryos developed to blastocyst stage and 71.5% of the blastoctysts expressed EGFP. After 200 reconstructed 1‐cell stage embryos were transferred into four surrogate gilts, three recipients were found to be pregnant. One of them maintained to term and delivered a healthy transgenic piglet expressing EGFP. Our data suggest that the combination of transduction of somatic cells by a replication defective vector with the nuclear transfer of colchicine‐treated donors is an alternative to produce transgenic pigs. Furthermore, the tissues expressing EGFP from descendents of this pig may be very useful in future studies using pigs that require genetically marked cells. Mol. Reprod. Dev. 62: 300–306, 2002. © 2002 Wiley‐Liss, Inc.
Cloning and sequencing of cDNA encoding for a novel human testis‐specific contraceptive vaccinogen: Role in immunocontraception
Authors: Naz Rajesh K. , Zhu Xiaolong , Kadam Arjun L.
Published: August 2002

Sperm‐specific antigens are attractive candidates for the development of a contraceptive vaccine. Using the subtractive cDNA hybridization technology, the present study was conducted to obtain a human sperm‐specific antigen. The 32P‐labeled single stranded cDNA of human testis, subtracted with poly(A)+ RNA of human peripheral white blood cells, was used to screen the human testis cDNA‐ZAP II library. The putative positive clones were further screened for binding with the solubilized human oocyte zona pellucida preparation (HZP). After screening 107 colonies, one positive clone, designated contraceptive vaccinogen (CV), was obtained. It had an insert of ∼1.3 kb, that was cloned and sequenced. The sense strand was identified by using the in vitro transcription and translation procedures, and the full‐length sequence was obtained by using the 5′ rapid amplification of 5′ ‐cDNA ends (5′‐RACE) procedure. The full‐length CV cDNA has an ORF of 312 amino acids (aa) with the first ATG Met start codon at nucleotide (nt) 35 and the stop codon TAA, at nt 959. The translated protein has a calculated molecular mass of 35.3 kD and four potential N‐linked glycosylation and several phosphorylation sites. Hydropathy plot generated from the deduced aa sequence showed it to be a membrane‐anchored peptide. Extensive computer search in the database did not find any homology of existing sequences with CV both for nt and aa. Northern blot analysis indicated the human testis‐specific expression of CV antigen. The coding region of CV cDNA was subcloned into pET22b(+) vector and expressed. The expressed recombinant (r)CV protein had a molecular size of ∼44 kD, and it specifically reacted with the ZP3 component of HZP. Rabbit rCV antibodies recognized the rCV, and a cognate antigen of ∼64 kD in the human sperm extract. The antibodies showed binding with the live and methanol‐fixed human sperm, and significantly (P < 0.001) inhibited human sperm penetration of zona‐free hamster oocytes, as well as human sperm binding to human oocyte zona pellucida. These findings indicate that the testis/sperm‐ specific CV antigen has a role in human sperm function and may find clinical applications in the contra ceptive vaccine development and in the specific diagnosis and treatment of male infertility. Mol. Reprod. Dev. 60: 116–127, 2001. © 2001 Wiley‐Liss, Inc.
Expression of retinol‐binding protein messenger RNA and retinoic acid receptors in preattachment bovine embryos
Authors: Mohan M. , Malayer J.R. , Geisert R.D. , Morgan G.L.
Published: August 2002

In cattle, retinoic acid (RA) has been indirectly associated with developmental potential of the embryo. RA is transported by retinol‐binding protein (RBP) and actions of RA are mediated by several subtypes of nuclear retinoic acid receptors (RAR). Bovine embryos, produced in vitro from oocytes harvested from ovaries collected at a local abattoir, were frozen in liquid nitrogen at the oocyte, 2‐, 4‐, 8‐, 16 to 20‐cell, morula, blastocyst, and hatched blastocyst stages. Employing reverse transcription polymerase chain reaction (RT‐PCR) we investigated mRNA expression for RBP, RARα, RARβ, RARγ, and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH). Total RNA was extracted from 25 pooled embryos at each stage and RT‐PCR analysis was repeated thrice. GAPDH transcript was detected in all stages. Transcripts for RBP, RARα, and RARγ were also detected in all stages from the oocyte through to the hatched blastocyst. Expression of RARβ was not detected at any stage. Whole‐mount immunohistochemistry was performed with intact and hatched blastocysts using polyclonal antibodies against RARα and RARγ2 to investigate if these embryonic mRNAs were translated to the mature protein. Strong immunostaining was observed for both RARα and RARγ2 in the inner cell mass and trophectoderm of intact and hatched blastocysts. Expression of mRNA for RBP, RARα, RARγ, and of the RARα and RARγ2 receptor proteins in the bovine embryo suggests that RA is likely to directly regulate gene expression during preimplantation development in that species. Mol. Reprod. Dev. 60: 289–296, 2001. © 2001 Wiley‐Liss, Inc.
Solubilized zona pellucida proteins and progesterone induce calcium influx and the acrosome reaction in capacitated dog spermatozoa
Authors: Brewis I.A. , Morton I.E. , Moore H.D.M. , England G.C.W.
Published: August 2002

Spermatozoa from the sperm‐rich fractions of the semen of 6 beagle dogs were capacitated and the effect of both zona pellucida (ZP) proteins and progesterone on calcium flux and the acrosome reaction measured. Sperm calcium flux was determined using the dual wavelength calcium probe indo‐1/AM (6 μM) in a flow cytometric assay (one ejaculate from each dog examined; n = 6). No calcium flux was observed in the negative control treatments (RPMI medium or DMSO). Both heat‐solubilized bitch ZP proteins and progesterone caused a similar response characterized by a gradual but marked influx of calcium ions which was sustained over 2 min. Acrosomal status was assessed by indirect immunofluorescence using a specific monoclonal antibody following 1 hr incubation for each treatment (four ejaculates from each dog examined; n = 24). The level of acrosomal exocytosis was very high for samples treated with ZP proteins (70.3 ± 2.1%) and progesterone (84.6 ± 1.5%) and was significantly different from the respective controls (P < 0.001). Interestingly the patterns of calcium flux in response to both ZP proteins and progesterone were in contrast to the situation in other species studied to date raising the possibility that the mechanism for triggering the acrosome reaction may be different in dog spermatozoa. In addition the high degree of progesterone‐induced acrosomal exocytosis compared to other species raises the probability that the majority of dog spermatozoa are already undergoing the acrosome reaction before they reach the egg ZP. Mol. Reprod. Dev. 60: 491–497, 2001. © 2001 Wiley‐Liss, Inc.
Acrocentric centromere organization within the chromocenter of the human sperm nucleus
Authors: Gurevitch Merav , Amiel Aliza , Ben‐Zion Moshe , Fejgin Moshe , Bartoov Benjamin
Published: August 2002

It has recently been reported that in human sperm cells, the centromeres are clustered in a chromocenter in the interior region of the nucleus. The aim of the present study was to determine the intra‐chromocenter organization of the five centromeres of the acrocentric chromosomes responsible for the biosynthesis of rRNA. The acrocentric centromeres were labeled by fluorescence in situ hybridization (FISH) after mild decondensation of the sperm nuclei to preserve the tail structure. The tail was used as a topographical marker for the orientation of the nucleus. The following results were obtained: (a) the association among the five centromeres was higher than expected from random distribution; (b) all the centromeres observed were randomly located within the chromocenter, occupying about 87% of the total area of the internal nucleus; (c) a major subpopulation of centromeres was located in a preferred area occupying 8.3% of the total nuclear area, with a peak 0.6 μm on the lateral axis and 1.0 μm on the apical side of the longitudinal axis; and (d) The dispersion of the centromeres was not influenced by the degree of the nuclear decondensation. We conclude that in human sperm nuclei, the acrocentric centromeres are organized within a nonlocalized structural element in the chromocenter. The chromocenter can range from an expanded size of 87% of the whole nucleus to a preferred size of 8.3% independent of the degree of nuclear decondensation. These findings have important implications for nuclear function (rRNA) that is not directly related to sperm cell function or early embryo development. Mol. Reprod. Dev. 60: 507–516, 2001. © 2001 Wiley‐Liss, Inc.
Apoptosis in the preimplantation mouse embryo: Effect of strain difference and in vitro culture
Authors: Kamjoo Marziel , Brison Daniel R. , Kimber Susan J.
Published: August 2002

Cell death by apoptosis occurs predominantly in the inner cell mass (ICM) of the blastocyst, the cell population which carries the germ line and gives rise to the foetus. The frequency of apoptosis in blastocysts varies widely within outbred species such as human and cow. We have addressed the basis of this variation by examining the relative influence of strain difference and in vitro culture conditions on apoptosis, using embryos from two different strains of mice (MF1 and C57BL6/CBA) in two different culture media (M16 and kSOM). In both strains and all crosses apoptosis was first detected by nuclear fragmentation or TUNEL [Terminal deoxynucleotidyl transferase mediated d‐UTP nick end‐labelling] labelling at the early blastocyst stage. This was true for embryos which had developed in vivo, and in vitro in both M16 and kSOM. The apoptotic index in blastocysts was found to be significantly different between both media and strain (P < 0.0001). Blastocysts from MF1 × MF1 at equivalent stages had an apoptotic index of 32.4% in M16 and 20.3% in kSOM. Blastocysts from C57BL6/CBA × C57BL6/CBA had an apoptotic index of 19.3% in M16 and 14.4% in kSOM. When embryos of similar cell number were compared, a significantly greater apoptotic index was found for cultured MF1 × MF1 embryos with a cell number between 40 and 59 compared to similar directly flushed C57BL6/CBA embryos (P = 0.001), and MF1 embryos (P < 0.0005). MF1 × MF1 embryos and C57BL6/CBA × MF1 embryos of 60–79 cells had a greater apoptotic index in M16 than kSOM (P < 0.0005) but the difference between media was not significant for C57BL6/CBA × C57BL6/CBA. When strain was compared MF1 × MF1 embryos of 60–79 cells had a significantly greater apoptotic index than C57BL6/CBA × MF1 in both media (P < 0.0005 M16; P = 0.002 kSOM) and than C57BL6/CBA × C57BL6/CBA in M16 (P = 0.019). Our data suggest that genetic make‐up and the chemical composition of simple medium are equally important in determining the level of apoptosis. Mol. Reprod. Dev. 61: 67–77, 2002. © 2002 Wiley‐Liss, Inc.
Ultrastructural studies on developing follicles of the spotted ray Torpedo marmorata
Authors: Prisco Marina , Loredana Ricchiari , Piero Andreuccetti
Published: August 2002

Light and ultrastructural investigations on sub‐adult and adult sexually mature females, demonstrates that in Torpedo marmorata folliculogenesis starts in the early embryo and that the two ovaries in the adult contain developing follicles of various sizes and morphology. Initially, the follicle is constituted by a small oocyte, surrounded by a single layer of squamous follicle cells. The organization is completed by a basal lamina and, more externally, by a theca, that at this stage is composed by a network of collagen fibers. As the oocyte growth goes on, during previtellogenesis and vitellogenesis, the organization of the basal lamina and of the oocyte nucleus does not change significantly. The basal lamina, infact, remains acellular and constituted by fibrils intermingled in an amorphous matrix; the nucleus always shows an extended network of chromatin due to the lampbrush chromosomes, and one or two large nucleoli. By contrast, the granulosa (or follicular epithelium), the ooplasm, and the theca cells significantly change. The granulosa shows the most relevant modifications becoming multi‐layered and polymorphic for the progressive appearance of intermediate and pyriform‐like cells, located respectively next to the vitelline envelope, or spanning the whole granulosa. The appearance of intermediate cells follows that of intercellular bridges between small follicle cells and the oocyte so that one can postulate that, as in other vertebrates, small cells differentiate into intermediate, and then pyriform‐like cells, once they have fused their plasma membrane with that of the oocyte. Regarding the ooplasm, one can observe as in previtellogenic follicles, it is characterized by the presence of intermediate vacuoles containing glycogen, while in vitellogenic follicles by an increasing number of yolk globules. The theca also undergoes significant changes: initially, it is constituted by a network of collagen fibers, but later, an outermost theca esterna containing cuboidal cells and an interna, with flattened cells, can be recognized. The role of the different constituents of the ovarian follicle in the oocyte growth is discussed. Mol. Reprod. Dev. 61: 78–86, 2002. © 2002 Wiley‐Liss, Inc.
Transition to adulthood: Supporting young adults to access social, employment, and civic pursuits
Authors: Hughes Carolyn
Published: August 2002

“Transition support” is a concept new to the field of secondary transition. This paper describes the development of a model of transition support as derived from the empirical literature (Hughes and Carter. [2000] The Transition Handbook. Baltimore, MD: Paul H. Brookes). The components of the Transition Support Model are both empirically based and field‐tested among practitioners. The over 500 transition support strategies that compose the model have been used by practitioners in the field to improve educational and postschool outcomes for high school students. These strategies are grouped according to two main goals (i.e., “developing support in the environment” and “increasing students' competence”) and corresponding components. Literature supporting the Transition Support Model is described as well as the use of the model in the transition planning process. Suggestions are made for application of transition support in high schools and the need for future research. MRDD Research Reviews 7:84–90, 2001. © 2001 Wiley‐Liss, Inc.
Glutamate signaling and the fetal alcohol syndrome
Authors: Olney J.W. , Wozniak D.F. , Jevtovic‐Todorovic V. , Ikonomidou Chrysanthy
Published: August 2002

It has been known for three decades that ethanol, the most widely abused drug in the world, has deleterious effects on the developing human brain, but progress has been slow in developing animal models that are optimal for studying this problem, and the underlying mechanisms have remained elusive. Recently, we have shown that during the synaptogenesis period, also known as the brain growth spurt period, ethanol has the potential to trigger widespread neuronal suicide (apoptosis), deleting many millions of neurons from the in vivo mammalian brain. It appears that ethanol triggers apoptotic neurodegeneration by a dual mechanism (blockade of NMDA glutamate receptors and excessive activation of GABAA receptors), in that ethanol has both NMDA antagonist and GABAmimetic properties; we have shown that other drugs which have either of these properties trigger apoptotic neurodegeneration in the developing brain. The brain growth spurt period in humans spans the last trimester of pregnancy and the first several years after birth. Thus, our findings provide a likely explanation for the reduced brain mass and neurobehavioral disturbances associated with the human fetal alcohol syndrome. Furthermore, since NMDA antagonist and GABAmimetic drugs are sometimes abused by pregnant women and also are used as anticonvulsants, sedatives, or anesthetics in pediatric medicine, our findings suggest the possibility that exposure of the developing brain to these various drugs either pre or postnatally could contribute to mental disability syndromes that have heretofore been attributed to unknown causes. In addition, the observation that ethanol and related drugs trigger massive neuronal apoptosis in the developing brain provides an unprecedented opportunity to study both neuropathological aspects and molecular mechanisms of apoptotic neurodegeneration in the in vivo mammalian brain. MRDD Research Reviews 2001;7:267–275. © 2001 Wiley‐Liss, Inc.