Simple differences in the protein-membrane attachment mechanism have functional consequences for surface mechanics.

This paper describes two methods for propagating coupled membrane and embedded particle dynamics with ensembles that are valid to second order in the deformation of the membrane. Proteins and functional lipids associate with cellular membranes, and their attachments influence membrane physical and dynamical properties. Therefore, it is necessary to accurately model the coupled dynamics of the membrane and any associated material of interest. We have developed two methods for coupling membrane and particle dynamics that differ in the binding mechanism of the particle to the surface. The "on-surface" mechanism should be used for particles that slide along the membrane; this description leads to an effective reduction in the membrane surface tension. The "in-surface" mechanism treats the particles as tightly bound to the lipidic binding sites; the method avoids double counting lateral entropy of implicitly modeled lipids. We emphasize the differences between these two mechanisms, when it is appropriate to use them, and how the methods differ from previously used dynamic methods.

Emerging outbreaks associated with equine coronavirus in adult horses.

The purpose of this study was to describe clinical, hematological and fecal PCR results from 161 horses involved in outbreaks associated with ECoV. The outbreaks happened at four separate boarding facilities between November 2011 and April 2012 in the States of CA, TX, WI and MA. Following the molecular detection of ECoV in the feces from the initial index cases, the remaining herdmates were closely observed for the development of clinical signs. Fecal samples were collected from sick and healthy horses for the PCR detection of ECoV. All four outbreaks involved primarily adult horses. Fifty-nine horses developed clinical signs with 12-16 sick horses per outbreak. The main clinical signs reported were anorexia, lethargy and fever. Four horses from 3 different outbreaks were euthanized or died due to rapid progression of clinical signs. The cause of death could not be determined with necropsy evaluation in 2 horses, while septicemia secondary to gastrointestinal translocation was suspected in 2 horses. Blood work was available from 10 horses with clinical disease and common hematological abnormalities were leucopenia due to neutropenia and/or lymphopenia. Feces were available for ECoV testing by real-time PCR from 44 and 96 sick and healthy horses, respectively. 38/44 (86%) horses with abnormal clinical signs tested PCR positive for ECoV, while 89/96 (93%) healthy horses tested PCR negative for ECoV. The overall agreement between clinical status and PCR detection of ECoV was 91%. The study results suggest that ECoV is associated with self-limiting clinical and hematological abnormalities in adult horses.

Mechanisms underlying digenean-snail specificity: role of miracidial attachment and host plasma factors.

Digenetic trematodes usually show a high degree of specificity for their molluscan intermediate hosts. A panel of 4 digenean species (Echinostoma paraensei, E. trivolvis, Schistosoma mansoni, and Schistosomatium douthitti) and 5 snail species (Biomphalaria glabrata, Helisoma trivolvis, Lymnaea stagnalis, Stagnicola elodes, and Helix aspersa representing 3 gastropod families) was used to assess the relative contributions of miracidial behavior, host plasma osmolality, and host plasma factors in dictating specificity. Additional experiments were undertaken with a fifth digenean, Echinoparyphium sp. Expected patterns of compatibility were first confirmed; each parasite species produced patent infections in its known snail host, but not in the other snail species. One exception was S. douthitti, which unexpectedly did not infect L. stagnalis. As judged by direct observation and by noting their disappearance after exposure to snails, miracidia were generally less likely to attach to or penetrate incompatible than compatible hosts. However, over half of the miracidia of each parasite species attached to or attempted penetration of both compatible and incompatible hosts, suggesting that under the experimental conditions used, miracidial host location and attachment behaviors were not of overriding importance in dictating observed patterns of specificity. For each digenean species, the percentage of larvae that became immobile, rounded, showed tegumental damage, or died over a 6-hr interval in plasma of the various snails was assessed. In no case was plasma from a compatible host harmful to sporocysts or rediae. In contrast, in 8 of 16 (50%) incompatible combinations, snail plasma had a significant negative effect on sporocyst condition. In 4 of 12 (33%) incompatible combinations, plasma had a significant negative effect on rediae. In 9 of 10 combinations tested, lymnaeid plasma was toxic for the parasites of planorbid snails and in 2 of 4 combinations, planorbid plasma was toxic for the parasites of lymnaeid snails. Toxicity was not attributable to differences in plasma osmolality between snail species. The ability of plasma from incompatible snails to affect viability of both sporocysts and rediae was surprisingly strong, suggesting that humoral factors play a greater role in dictating patterns of digenean-snail specificity than previously appreciated.

A comparative study of mechanisms underlying digenean-snail specificity: in vitro interactions between hemocytes and digenean larvae.

A panel of 4 digenetic trematode species (Echinostoma paraensei, E. trivolvis, Schistosoma mansoni, and Schistosomatium douthitti) and 5 snail species (Biomphalaria glabrata, Helisoma trivolvis, Lymnaea stagnalis, Stagnicola elodes, and Helix aspersa) was examined to determine if known patterns of host specificity could be explained by the tendency of digenean larvae to be bound by snail hemocytes, or by the ability of larvae to influence the spreading behavior of hemocytes. In short-term (1 hr) in vitro adherence assays, there was no overall pattern to suggest that sporocysts were more likely to be bound by hemocytes from incompatible than compatible snails. Compared with the other parasites, sporocysts of E. paraensei were less likely to be bound by hemocytes from any of the snail species tested. All rediae examined, including those of another species Echinoparyphium sp., were also remarkably refractory to binding by hemocytes from any of the snails. Of all the larvae examined, only sporocysts and young daughter rediae of E. paraensei caused hemocytes to round up in their presence. This was true for hemocytes from the compatible species B. glabrata and the incompatible lymnaeid species S. elodes and L. stagnalis. The patterns of host specificity shown by this particular panel of parasites and snails were not predicted by either the extent of hemocyte adherence to digenean larvae or by the ability of larvae to affect hemocyte spreading behavior. The results of this study suggest that a role for hemocytes, although likely, may require different assays, possibly of a more prolonged nature, for its detection. Also, different parasite species (notably E. paraensei) and intramolluscan stages have distinctive interactions with host hemocytes, suggesting that the determinants of specificity vary with the host-parasite combination, and with the parasite life cycle stage.

Computerized medical record in a private neurotology practice.

OBJECTIVE: To develop a computerized medical record for a private neurotology practice using a relational database with templates. BACKGROUND: Computers have been used in the medicine many years for billing, scheduling appointments, and word processing. Neurotology represents a relatively narrow field of medicine, which is particularly suited to a computerized record. A computerized medical record should allow for better data storage and retrieval as well as better documentation of the patient care given. METHODS: Using a commercially available, record-keeping software in a local area computer network. templates were developed for several common neurotologic problems. A template for a neurotologic examination was also developed. These were adapted from templates developed by a group otolaryngology practice in a neighboring city. Data entry templates for vestibular and audiologic tests were also developed. Implementation of the computerized record was accomplished in phases, allowing elimination of the paper record. RESULTS: A fully computerized medical record has been developed and maintained in a private neurotology practice over the past year. Patient response to the computerized record system has been favorable. Office personnel have adapted well to a paperless record, and referring physicians have welcomed the documents generated by the computer. CONCLUSION: Computerizing the medical records of a neurotology practice is possible and allows for improved documentation, ongoing assessment of treatment results, and improved efficiency among office staff.

Cytologic findings in the sarcomatoid variant of large cell anaplastic (Ki-1) lymphoma. A case report.

Anaplastic large cell (Ki-1) lymphomas are a recently described subtype of non-Hodgkin's lymphoma. A single case of the sarcomatoid variant has recently been reported. Below we report the fine needle aspiration findings in an additional case of the sarcomatoid variant of Ki-1 lymphoma. The diagnosis was made only in retrospect with the aid of immunostains because the cytologic findings were more in keeping with a sarcoma. Striking vascular structures in the smears suggested angiosarcoma or liposarcoma. Additional features were pleomorphic cells with eccentric nuclei and abundant, tapering cytoplasm; spindle cells; multinucleation; cytoplasmic vacuolization; and prominent nucleoli. The smear pattern was composed of dispersed cells with some cohesive groups within an acute inflammatory background with a virtual absence of lymphoglandular bodies. These findings, atypical of lymphoma, broaden the spectrum of possible cytologic findings in lymphoid malignancies and highlight the possible utility of immunostaining. The distinction of lymphoma from sarcoma or carcinoma is of therapeutic importance.

The conservative surgical treatment of endometriosis: evaluation of pregnancy success with respect to the extent of disease as categorized using contemporary classification systems.

A homogeneous group of 214 infertile women with endometriosis treated at the Johns Hopkins Hospital from 1960 to 1979 received conservative surgery as the sole therapeutic modality. Among this group, 115 patients (54%) conceived following surgery; of these conceptions, 109 resulted in a living child. Among 49 patients with secondary infertility, the spontaneous abortion rate was reduced from 49% to 20% after conservative surgery (P less than or equal to 0.01). Three contemporary classification systems were utilized to categorize patients according to the sites and amount of endometriosis at the time of conservative surgery. Those systems suggested by Buttram (Fertil Steril 30:240, 1978) and by Kistner and coauthors (Fertil Steril 28:108, 1977) revealed differences among fecundability rates among the different categories (P less than or equal to 0.01); however, the system suggested by The American Fertility Society (AFS) (Fertil Steril 32:633, 1979) revealed significant differences only if categories were combined (mild plus moderate versus severe plus extensive, P less than or equal to 0.05). Nevertheless, the AFS system revealed that pregnancy success was significantly reduced if an ovarian endometrioma was greater than 3 cm or had ruptured (P less than or equal to 0.01).

Gestational outcome of clomiphene-related conceptions.

The experience of the gynecologic endocrinology and infertility clinic at The Johns Hopkins Hospital has been subjected to a nonconcurrent prospective analysis in an attempt to evaluate the gestational fate of clomiphene-related conceptions (study series, n = 86). This latter series was contrasted with a series of pregnancies following bilateral ovarian wedge resection (BOWR) (n = 51) in a comparative analysis of gestational outcome event rates. Post-therapy follow-up was available for varying time spans of up to 15 years. A 12.8% twinning rate constituted the single most important complication of clomiphene therapy, resulting in measurable increments in perinatal morbidity and mortality rates. The observation of a 26.5% spontaneous abortion rate would seem to suggest that clomiphene-related conceptions are at little or more risk for spontaneous abortion than would have been expected from the infertile population under discussion. A 3.1% incidence of post-clomiphene birth defects was not increased as compared with commonly quoted rates for the population at large. The corresponding incidence rates of twinning, spontaneous abortion, and birth defects for the BOWR series were 0%, 21.6%, and 0%, respectively.

Stimulation of follicular growth with "pure" FSH in patients with anovulation and elevated LH levels.

Ovarian follicular growth was stimulated in 4 patients with clomiphene resistant polycystic ovary (PCO)-like disease, with "pure" human pituitary FSH during 10 cycles. In 8 cycles additional hCG was given to induce ovulation. Serum LH and FSH and plasma estradiol and progesterone were determined daily. From the response patterns of steroidogenesis, four functional stages of follicular development can be distinguished, each subsequent stage being characterized by an increasing spontaneous estradiol production and a decreasing capacity to produce extra estradiol in response to stimulation with additional hCG. It is concluded that the unpredictable response to gonadotropin stimulation of patients with PCO-like disease is due to the varying state of development of the follicles at the start of the stimulation. The suggestion is made, based on the different responses in 3 cycles of the same patient (EW), that the lack of positive estrogen feedback in patients with PCO-like disease may be a hyposensitivity to estrogens rather than an absolute insensitivity.

Danazol as a luteolytic agent.

Danazol, a synthetic 2,3-isoxazol derivative of 17 alpha-ethinyltestosterone, was administered to healthy nonpregnant volunteers to determine wheter a luteolytic effect could be detected by observation of cycle length, duration of the luteal rise, and luteal steroidogenesis. Danazol administration resulted in a decreased duration of the luteal rise and a decrease in progesterone output in three of four subjects, but no decrease in total cycle length. The administration of human chorionic gonadotropin during danazol administration increased progesterone output. Therefore, danazol would be unlikely to be effective as a luteolytic contraceptive agent.

Studies of pathophysiology in primary amenorrhea.

Patients with primary amenorrhea underwent stimulation with luteinizing hormone releasing hormone (LRH) before and after clomiphene administration to determine the maturational status of the hypothalamic-pituitary-gonadal axis, and to assess the development of feedback control mechanisms. Four patients with clinically recognizable conditions served as models. One patient had panhypopituitarism with undetectable baseline FSH and LH values, no response to LRH or clomiphene, and no detectable feedback control. One patient with a "specific" hypothalamic hypogonadotropism had low baseline gonadotropins with a normal response to LRH and no positive or negative feedback mechanisms reflected by no change in LRH response after clomiphene. A patient with delayed puberty due to low bodyweight had low FSH and LH baseline values and an "immature" response to clomiphene, characterized by lowered baseline gonadotropins and an increased peak LH following LRH stimulation. The fourth patient, with an adolescent polycystic ovarian syndrome, had a high baseline LH, normal FSH, hyperresponse to LRH, and a "mature feedback control" as evidenced by an increase in baseline FSH and LH, and a decrease in the LRH response while on clomiphene. The remaining patients showed inappropriate responses. They are regarded as having defects rather than dysfunctions, and the possible abnormalities are discussed in relation to the clinical findings.

Progestational activity of danazol in the human female subject.

A progestational action of Danazol has not been reported in the human subject. Danazol administration for 1 month suppressed serum FSH and estradiol values, and the midcycle LH surge; ovulation promptly resumed after treatment. Administration to patients with polycystic ovarian disease resulted in increased LH values, elevated basal temperature, secretory findings in the endometrial biopsy, and menses. Ovarian biopsy did not document corpus luteum formation, and plasma progesterone did not rise. Administration of Danazol to an ovariectomized patients pretreated with estrogen resulted in an atypical endometrial secretory change. Clinical responses to Danazol may in part be explained by a progestational-like effect of the drug.

Effect of clomiphene citrate on gonadotropin responses to LRH administration in secondary amenorrhea and oligomenorrhea.

Clomiphene citrate was administered to 17 patients with either secondary amenorrhea or oligomenorrhea to study its effect on hypothalamic-pituitary response. Measurement of pulsatile gonadotropin output was accomplished utilizing samples collected every 20 minutes for 6 to 8 hours before and after clomiphene administration. Response to 100 mug synthetic luteinizing hormone releasing hormone (LRH) was measured at the end of the sampling period. Patients with polycystic ovarian disease who ovulated showed increased baseline FSH and LH, decreased peak LH, and a decrease in the percentage increase over baseline for both LH and FSH. On patient wil clinical anorexia nervosa responded to clomiphene as an estrogen, with lowered baseline LH and FSH values. Patients with resolving anorexia responded to clomiphene as an antiestrogen, with increased baseline gonadotropins, and decreased gonadotropin peak values. A patient with a surgically treated chromophobe adenoma showed no change in parameters measured before and after clomiphene administration. Baseline and pulsatile LH output appear to reflect the tonic output of gonadotropin as affected by estrogen acting in a negative feedback system, and the peak response to LRH administration, the response to the positive feedback of estrogen. Thus, LRH stimulation in clomipheneresistent patients may be of value in diagnosing the site and degree of the defect and aid in improving therapy.

Investigation of Danazol as a contraceptive agent.

PIP: An attempt to determine the efficacy of Danazol in producing an inadequate luteal phase either by inhibiting follicular development or by producing an insufficient luteinzing hormone (LH) surge at the time of ovulation is reported. Danazol is the 2,3-isoxasol derivative of 17alpha-ethinyl testosterone. Subjects were 17 healthy women treated during the first 7 days of their menstrual cycles. Daily blood samples were analyzed for plasma estradiol, progesterone, and serum follicle stimulating hormone (FSH) and LH. A group received Danazol 400 mg/day for 7 days. Another group was given 800 mg and a 3rd group 1600 mg/day for 7 days. The follicular span, from onset of menses to LH surge, was lengthened and ovulation delayed for at last 1 week, in all but 2 cases. Although delayed, the LH surge was normal, and a normal progesterone output occurred during the luteal phase. The luteal span was 12 days or more which was considered normal. Inhibition of gonadotropin output was noted during Danazol administration but prompt recovery followed. Serum chemistries showed no significant abnormalities. Hematologic and urinary parameters were normal. It is thought that FSH was inhibited more than expected resulting in the delayed ovulation with effects similar to those when estrogen is given during the proliferative phase. However, an inadequate luteal phase did not follow the inhibition of FSH-stimulted follicular development.^ieng

Pulsatile gonadotropin output and response to luteinizing hormone releasing hormone (LRH) in primary amenorrhea.

The pulsatile gonadotropin output and resonse to luteinizing hormone releasing hormone (LRH) administration was evaluated in 23 patients with primary amenorrhea. Pulsatile LH output and associated FSH pulses were readily observed in patients with hypergonadotropism. Responses in patients with congenital adrenal hyperplasia varied with the adequacy of the prior therapeutic management, which in turn apparently determined the maturational status of the individual. A patient with Kallmann's syndrome had both FSH and LH pulsatile activity. Patients thought to have simple delayed puberty had normal baseline gonadotropin values, and regular LH pulsations, but no correlation of FSH and LH pulsatile activity. Correlated analysis of LRH response and pulsatile gonadotropin activity may allow differentiation of individuals showing a normal progression of maturation from those with maturational dysfunction.

Pulsatile gonadotropin output in menstrual dysfunction.

Patterns of gonadotorpin output were studied in normal individuals and in patients with menstrual dysfunction by radioimmunoassay measurement of LH and FSH output in samples taken every 20 minutes for 6-8 hours, and following administration of synthetic luteinizing hormone releasing hormone (LRH). Follicular phase LH pulses occurred every 1-2 hours, whereas those in the luteal phase occurred less frequently and with higher amplitude. FSH output was irregular, and had no correlation with LH dynamics. In anorixia nervosa, pulsatile LH activity was minimal, and the response to LRH variable, correlating somewhat with the clinical status of the patient. Pulsatile LH activity was observed in patients with postpill and postpartum amenorrhea, and also in one individual with a probable prolactin-producing pituitary tumor. Patients with polycystic ovarian disease had obvious LH pulsatile activity, with a greater amplitude and frequency than seen in the luteal phase, and a decrease in percentage increment, suggesting some difference in the hypothalamic-pituitary control mechanisms under these conditions. The character and pattern of the LH pulsatile activity does vary with different forms of menstrual dysfunction and may be predictive of the LRH response when considered in relation to the LH baseline values. Pulsatile LH activity, analyzed in conjunction with response patterns following LRH stimulation, may reflect the degree of hypothalamic dysfunction.