Adenosine triphosphatases as histochemical markers for the cell of origin in experimental mammary carcinoma.

Different adenosine triphosphatase (ATPase) activities were detected at an ultrastructural level in order to differentiate epithelial and myoepithelial cells in normal and neoplastic mouse mammary tissues. Mg2+ dependent and Na+-K+-dependent ATPase activities were studied in: BALB/c mouse mammary gland; a BALB/c carcinoma from a transplantable D2 hyperplastic nodule; a stable cell line, MCF-8, derived from the BALB/c carcinoma; and a BALB/c scirrhous-like carcinoma induced by MCF-8 cell inoculation. Mg2+-dependent ATPase was detected in the plasma membranes of the normal mouse mammary epithelial cells, the epithelial component of the BALB/c carcinoma, the MCF-8 cells in culture, and the atypical epithelial component of the scirrhous-like carcinoma. Na+-K+-dependent and Mg2+-dependent ATPase were localized in the plasma membranes of the myoepithelial cells of the normal mammary gland and the BALB/c carcinoma. The results from these histochemical studies established that the cell of origin in both the BALB/c carcinoma and the scirrhous-like carcinoma was the mammary epithelial rather than the myoepithelial cells. Furthermore, these results indicated that the MCF-8 cell line was derived from the epithelial component of the primary BALB/c carcinoma. These conclusions, which were based on histochemical study, were supported by the presence of intracisternal type A viral particles in the epithelial cells of the primary BALB/c carcinoma, the MCF-8 cells in culture, and the epithelial cells of the scirrhous-like carcinoma. Thus, the enzymatic markers were specific for cell type and remained unchanged by the process of cell transformation.

Cyclic nucleotide modulation of herpes simplex virus latency and reactivation.

Clinical observations and experimental studies suggested that the relative proportions of ganglionic neuronal intracellular cyclic adenosine monophosphate (c-AMP) and cyclic guanosine monophosphate (c-GMP) concentrations may influence the state or activity of herpes simplex viral DNA in its relationship with the host cell DNA. We studied the effects of putative modulators of intracellular cyclic nucleotide levels on herpes simplex virus (HSV) reactivation from latency in murine trigeminal ganglion cells. We also investigated the effects of these same mediators on the c-GMP and/or c-AMP concentrations in HSV-latently infected trigeminal ganglion cells and in acyclovir-suppressed, HSV-infected neuroblastoma cells. Cholera toxin and theophylline increased c-AMP levels (2-fold and 5-fold at 1 min and 30 sec, respectively for cholera toxin and 2-fold and 1.5-fold at 1 min and 30 sec for theophylline) and enhanced the rapidity of HSV reactivation from latency (P less than 0.005). Exogenous dibutyryl c-AMP also stimulated viral reactivation (P less than 0.005). Carbamylcholine increased c-GMP levels (7-fold and 6-fold at 15 sec and 30 sec, respectively), produced no significant change in c-AMP levels, and delayed HSV reactivation from latency (P less than 0.005). None of these mediators had a demonstrable effect on HSV replication.

Histology and immunohistology of Igh-1-restricted herpes simplex keratitis in BALB/c congenic mice.

In order to characterize the local ocular immunologic milieu of Igh-1-restricted herpes simplex keratitis (HSK), we investigated histologic and immunohistologic correlates of disease over a 21-day time course. Clinically observable keratitis began 10 days postinoculation in susceptible C.AL-20 (Igh-1d) and moderately susceptible BALB/c (Igh-1a) mice, whereas HSV-1-resistant C.B-17 (Igh-1b) mice rarely developed disease. Igh-1-restricted histologic differences were observed by day 11 postinoculation; C.AL-20 and BALB/c mice showed augmented recruitment of neutrophils and mononuclear cells in conjunctival, limbal, and corneal tissues compared to C.B-17 mice. On immunohistologic study, Lyt-1 to Lyt-2 cell ratios by day 11 postinoculation were 7:1, 2:1, and 1:8 in corneas from C.AL-20, BALB/c, and C.B-17 mice, respectively. Macrophages and neutrophils were absent in corneas from C.B-17 mice at this time, but could be found in large numbers in the corneas of susceptible mouse strains through day 21. These data demonstrate a strong relationship between Igh-1 phenotype and inflammatory cell recruitment in response to corneal infection with HSV-1, and support a role for T cell subpopulations in mediating Igh-1-restricted HSK.

Ganglionic permissivity to HSV-1 replication and acyclovir-induced latency: an in vitro model.

HSV-1 is known to establish latent infections in murine trigeminal ganglia. In order to study the genetic influence of the host on permissivity to HSV-1 replication and latency in mouse trigeminal ganglia, an in vitro culture system was developed. Ganglia from HSV sensitive A/J and resistant C57BL/6J mice were harvested and inoculated in vitro with either MP or KOS strains of HSV-1. Infected ganglia were placed in culture for 7 days with or without 150 micrograms/ml acyclovir. Ganglia were then homogenized and assayed for infectious particles or frozen whole for standard immunostaining. A/J ganglia without acyclovir consistently demonstrated a three-fold higher viral replication compared to C57BL/6J ganglia without acyclovir for both MP and KOS. Indirect immunofluorescent staining of actively infected ganglia using rabbit anti-HSV antiserum showed increased staining in A/J ganglionic axons compared to C57BL/6J ganglionic axons. Ganglia from both murine strains showed total suppression of viral replication with acyclovir treatment. Immunostaining for latent viral protein with mouse monoclonal anti-ICP-4 (VP 175) was positive for both A/J and C57BL/6J strains following acyclovir treatment. Desuppression of acyclovir-treated whole ganglia resulted in 54% spontaneous HSV reactivation. These results suggest that: (1) HSV-1 can establish an active infection in whole murine trigeminal ganglia in vitro; (2) HSV-1 replicates to a greater extent in A/J ganglia compared to C57BL/6 ganglia; and (3) in vitro HSV-1 ganglionic infection in both murine strains is equally suppressed to a latent state by acyclovir treatment.

Immunogenetic influence of Igh-1 phenotype on experimental herpes simplex virus type-1 corneal infection.

Patterns of herpes simplex virus type-1 (HSV-1) infection were studied in BALB/c congenic, Igh-1 disparate murine strains to establish the influence of Igh-1 phenotype on the development of keratopathy, trigeminal ganglionic latency and keratocyte permissivity. Eighty-two percent of C.AL-20 (Igh-1d) mice, 40% of BALB/cByJ (Igh-1a) mice and 12% of the C.B-17 (Igh-1b) mice developed herpes simplex keratitis (HSK) following corneal challenge with 2.5 X 10(4) PFU HSV-1 strain KOS. While disease frequency was directly proportional to HSV-1 challenge dose, relative resistance and susceptibility patterns in the congenic mice were constant and highly significant. F1 progeny from C.AL-20 X C.B-17 matings demonstrated the HSK pattern of the C.B-17 parent suggesting that Igh-1 linked resistance to HSK is dominantly inherited. Equivalent trigeminal ganglionic latency was established following ocular HSV-1 inoculation in the three congenic Igh-1 disparate murine strains. Cultured keratocytes from the three Igh-1 disparate murine strains demonstrated equivalent in vitro permissivity to HSV-1 replication. These data illustrate a strong correlation between Igh-1 phenotype and the development of a HSK in congenic mice. The susceptibility/resistance to HSK in these mice is unrelated to trigeminal ganglionic latency or keratocyte permissivity.

Topical nonsteroidal agents and corneal wound healing.

The effects on corneal wound healing of two topical nonsteroidal anti-inflammatory agents, flurbiprofen sodium (0.03%) and diclofenac sodium (0.1%), and the topical corticosteroid, prednisolone sodium phosphate (1%), were evaluated in masked, controlled rabbit studies. Healing of epithelial scrape wounds was significantly retarded in all three treatment groups for the first 3 days after wounding. There was no difference in the epithelial healing rate between the two nonsteroidal or corticosteroid treatment groups. Clinical grading of epithelial quality, conjunctival hyperemia, keratitis, stromal edema, and corneal haze were similar in all groups. There was a significant early decrease in the iritis score in the diclofenac treatment group. The strength of 2-mm central penetrating corneal trephination wounds and the collagen content of these wounds were similar in all groups. Both the topical nonsteroidal anti-inflammatory agents and the corticosteroid used in the preparations and dosages investigated in this study decreased early epithelialization of scrape wounds but had no apparent effect on corneal stromal healing. No toxic effects of the various drugs were found.

Scanning electron microscopy of conjunctival surfaces in patients with ocular cicatricial pemphigoid.

The conjunctival surfaces of ten patients with active, ocular cicatricial pemphigoid, three patients with drug-controlled ocular cicatricial pemphigoid, and six patients with normal conjunctivas were studied using scanning electron microscopy. A homogeneous granular sheet of amorphous mucin-like material was observed covering extensive areas of the conjunctiva in eight of ten patients with active ocular cicatricial pemphigoid. This sheet of amorphous material was absent on drug-controlled ocular cicatricial pemphigoid and normal conjunctival specimens. Our study demonstrates that patients with active ocular cicatricial pemphigoid possess ocular surface mucus that appears thicker and more continuous than normal ocular mucus when observed with scanning electron microscopy. This observation is in agreement with clinical observations of thick mucus strands in the inferior fornix of patients with active ocular cicatricial pemphigoid.

Effect of phototherapy light, sodium bisulfite, and pH on vitamin stability in total parenteral nutrition admixtures.

The three sections of this study extend previous research into losses of vitamins A, C, E, thiamin, riboflavin, and folic acid from total parenteral nutrition (TPN) admixtures. First, phototherapy light on TPN admixtures containing one of four amino acid solutions was studied. Experimental conditions included presence or absence of Intralipid iv fat emulsion, plastic bag or glass bottle storage container, and storage time of up to 48 hrs. The second phase studied stability of the same vitamins (except vitamin E) for 48 hrs in admixtures containing the amino acid solution which has no bisulfite, in glass bottles; with or without Intralipid; and with added sodium bisulfite (final concentrations of 0, 1, 2, 3, 4, 5 and 10 mEq/liter). Third, vitamin C and thiamin levels were measured in admixtures containing the amino acid solution with no bisulfite, without Intralipid, stored in glass bottles with various bisulfite concentrations (0, 1, 2, or 3 mEq/liter) and three pH levels (5.5, 6.5, and 6.75 pH). Exposure of TPN admixtures to phototherapy light caused losses of vitamins A, C, and riboflavin. Intralipid inclusion significantly reduced losses of vitamin A and riboflavin, but did not appear to affect vitamin C levels. The smallest vitamin C losses were noted in admixtures containing amino acid solutions A or B. Phototherapy light did not affect thiamin levels. Bisulfite had no affect on vitamin C, riboflavin, or folic acid levels. Vitamin A levels were maintained with bisulfite concentrations less than 3 mEq/liter. At 3 mEq/liter bisulfite, admixtures with Intralipid showed 50% loss of vitamin A.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Intralipid, amino acids, container, temperature, and duration of storage on vitamin stability in total parenteral nutrition admixtures.

This study was designed to determine the stability of certain vitamins added to total parenteral nutrition (TPN) admixtures with or without Intralipid iv fat emulsion and with each of four amino acid solutions stored in either glass bottles or plastic bags at either ambient room (25 degrees C) or refrigerator (5 degrees C) temperature for a 48-hr period. Riboflavin and folacin were not affected by the experimental conditions. The presence of Intralipid resulted in higher levels of vitamin E due to Intralipid's inherent vitamin E content; no other experimental conditions affected vitamin E. Thiamin levels decreased in admixtures containing the amino acid solution C and stored at 25 degrees C. Vitamin A levels were lower in admixtures stored in plastic but were maintained in admixtures containing Intralipid and stored in glass bottles at either temperature. Vitamin C levels were maintained in admixtures stored at 5 degrees C for all experimental conditions. The greatest vitamin C losses occurred in admixtures containing amino acid solutions C or D stored in plastic bags, or containing D stored in glass bottles at 25 degrees C.

Quantitation of immunoglobulins in mouse tears using enzyme linked immunosorbent assay (ELISA).

Aqueous tears were collected by glass capillary tubes at weekly intervals from 6-9 week old, Swiss-Webster mice. Mouse IgA, IgG1, IgG2a and IgM were quantitated in the tears using sandwich type enzyme linked immunosorbent assays (ELISAs). IgE was quantitated using an indirect two-step ELISA. The results established IgA as the predominant antibody in mouse tears at a level of 110 micrograms/ml. This was determined using a mouse IgA standard which contained both polymeric and monomeric IgA. IgG1, IgG2a and IgM were detected in the tears at low levels, 13 ng/ml, 21 ng/ml and 442 ng/ml, respectively. IgE was not detectable in mouse tears with a lower limit of detection equal to 10 ng/ml. These studies establish the relative levels of IgA, IgG1, IgG2a and IgM in the tears of normal adult mice.

SDS-gradient polyacrylamide gel electrophoresis of individual ocular mucus samples from patients with normal and diseased conjunctiva.

Individual mucus samples were collected from normal individuals and from patients with ocular cicatricial pemphigoid (CP), Stevens-Johnson syndrome (SJS), and various types of conjunctival inflammation (rosacea, meibomianitis, atopy, keratoconjunctivitis sicca, etc.). The mucus samples were dissolved in sample buffer containing 8M urea, 2% SDS and 5% 2-mercaptoethanol and were electrophoresed on gradient 2-16% polyacrylamide gels. Four glycoproteins with molecular weights greater than 200,000 daltons were consistently observed in both individuals with normal conjunctiva and patients with CP, SJS, and other diseases exhibiting conjunctival inflammation. The amounts of each glycoprotein appeared to vary from one individual to another; however, the presence or absence of specific glycoproteins could not be correlated with the different ocular diseases. The techniques described for mucus analysis offer advantages over previously published techniques since improved resolution of the mucous glycoproteins can be achieved by electrophoresis on 2-16% gradient gels, and individual samples can be analyzed. Our results suggest that substantial amounts of ocular mucous glycoprotein are present in the eyes of patients with CP and SJS, diseases which have been previously described as mucin-deficient dry eye syndromes.

Immunomodulation of experimental murine herpes simplex keratitis: II. Glycoprotein D protection.

Subcutaneous immunization with purified HSV-1 glycoprotein D (gD) protects susceptible A/J mice against keratitis and encephalitis following corneal HSV-1 challenge. Mice were immunized with gD, in complete Freund's adjuvant, 3.0 micrograms/mouse followed by two booster doses of 1.5 micrograms/mouse at weeks 2 and 4. Control groups of A/J mice were injected with either complete Freund's adjuvant (unimmunized controls) or live HSV-1 MP strain (immunized controls). All mice were challenged ocularly at week 5 with HSV-1, F strain (6.5 x 10(3) PFU) after corneal scarification. None of the 16 animals immunized with gD developed stromal keratitis; only 3 out of 12 animals immunized with live HSV-1 developed a stromal keratitis; 13 out of 16 CFA primed unimmunized mice developed severe stromal keratitis within 14 days post corneal challenge, and 3 out of 16 control CFA primed animals died within 16 days post corneal challenge. At the time of sacrifice (3 weeks post corneal challenge), the ipsilateral trigeminal ganglia were removed and assayed for latent HSV-1 using cocultivation on Vero cell monolayers. The results of these experiments indicate that immunization with gD produces protection against latent ganglionic infection in 56% of the immunized animals, and provides protection against keratitis and death following HSV corneal challenge.

Immunomodulation of experimental murine herpes simplex keratitis: I. UV-HSV protection.

A/J mice were immunized subcutaneously with ultraviolet light (UV) inactivated herpes simplex virus type-1, MP strain (HSV-MP). Control A/J mice were immunized subcutaneously either with media (unimmunized controls) or with live HSV-MP (immunized controls). Immunized and control mice were challenged ocularly with either MP or mP strain HSV-1 after corneal scarification and were followed for 3 weeks post corneal challenge. The mice were observed during this time period for signs of herpes simplex keratitis (HSK), lid lesions and encephalitis. At the time of sacrifice, the ipsilateral trigeminal ganglia were removed and assayed for latent HSV-1 using cocultivation on Vero cell monolayers. The results of these studies demonstrated that immunization with UV inactivated HSV (UV-HSV) gave the same protection against keratitis and encephalitis as immunization with live virus. Furthermore, the cocultivation assays indicated that immunization with either live HSV-1 or UV inactivated HSV-1 protected against the establishment of latency.

Survey of student clinical practice. Implications for educational programs.

The purpose of this survey was to analyze the content of physical therapy students' clinical education programs. We developed and used patient and student record forms that elicited information on the age and sex of the patient groups, the types of pathological conditions, the number and types of treatments administered, the levels of assessments, the levels of supervision of students by clinical instructors, the time spent with patients, and the specialized educational activities at each facility. One hundred seventy-six students participated in the three-year survey; each student completed 600 hours of study for a total of 105,600 hours. The results of this study revealed that musculoskeletal disorders (47.26%) were reported most frequently; fractures of the lower extremities constituted 10.5% of those disorders. Therapeutic exercise was the most frequently used treatment procedure (57.3%). The data obtained through this survey may be useful for evaluating curricula and developing improved practice-specific education programs.

Does fortune favour the bald? Psychological correlates of hair loss in males.

In order to investigate the relationship between male hair loss and psychological distress, 182 men were recruited who had a wide range of ages and hair loss varying from none to severe. Care was taken to ensure that hair loss and age were uncorrelated in the sample. Multiple regression was used to predict possible consequences of baldness, controlling for age, and examining the interaction between baldness and age to see if consequences were especially severe in cases of premature baldness. Increasing degrees of hair loss were associated with loss of self-esteem, depression, introversion, neuroticism and feeling unattractive. These effects were more marked for young men in the case of self-esteem, introversion and feeling unattractive.

Handedness, anxiety and sex differences.

Anxiety scores of 35 male and 35 female students (28 right-handers, 23 left-handers and 19 mixed-handers) were assessed in relation to the handedness classification of each individual. Females were found to show higher anxiety scores than were males; but in contradiction to previous findings, left-handers and mixed-handers were not found to be more anxious than right-handers. Extremes of handedness, however, were closely related to anxiety scores, with the strongly right-handed and the strongly left-handed both showing higher anxiety scores than those whose hand preferences were less extreme.

[T-cell-induced prevention of HSV-1 keratitis by immunization with the synthetic peptide of glycoprotein D]. / T-Zell-vermittelter Schutz gegen HSV-1-Keratitis durch Immunisierung mit synthetischem Peptid des Glykoprotein D.

C.AL-20 mice susceptible to herpes simplex virus (HSV) were protected against HSV keratitis (HSK) and encephalitis by subcutaneous immunization with synthetic peptide corresponding to the N-terminal amino acid residues 5-23 of HSV glycoprotein D, which is a dominant immunogen of HSV-1. Protection against HSV was related to a potent humoral anti-HSV response. FACScan analysis revealed that CD4+V beta 8(1.2)+ cells in the spleen were markedly decreased 2 days after HSV challenge, and CD8+ cells were increased. Numerous CD4+ and V beta 8(1.2)+ cells were found in the corneal tissue from HSV-infected sham-immunized mice; no such cells were seen in gD(5-23) immunized mice. No cytotoxic cells were detected in the corneas or spleens of gD(5-23) immunized mice, and these mice had decreased DTH responses. Protection against HSV through immunization with gD(5-23) involves humoral and cellular immune mechanisms. CD4+V beta 8(1.2)+ maybe critical in mediating the pathology of HSK. CD8+ cells may be protective by non-cytotoxic mechanisms. Our results suggest that gD peptides may be potent as vaccines against HSV.

Children as patients: a communications process study in family practice.

To determine how family physicians divide their attention between children and parents, 115 videotaped pediatric encounters of children (aged 4 to 14 years) and parents with family physicians in a family medicine center were analyzed. It was learned that physicians tended to involve children actively in the diagnostic stages of interviews but much less so in discussions of treating or dealing with their problems. Physicians did not alter this pattern as they advanced in training. Older children were more likely to receive direct communication from physicians in all phases of office encounters. Though the family physicians in this sample had more direct communication with children than reported in previous research, it is argued that greater involvement of children in all phases of pediatric visits is warranted. A developmental framework indicates that children gradually evolve their attitudes, concepts, and behaviors around illness and health. Family physicians and family medicine programs are in an excellent position to explore and utilize the practice of actively incorporating a child in a medical interview.