Treating vaginitis.

Vaginitis resulting from bacterial, fungal, or protozoal infections can be associated with altered vaginal discharge, odor, pruritus, vulvovaginal irritation, dysuria, or dyspareunia, depending on the type of infection. Bacterial vaginosis, which is primarily characterized by a malodorous discharge, is common in women with multiple sex partners and is caused by the overgrowth of several facultative and anaerobic bacterial species. Vulvovaginal candidiasis is characterized by pruritus and a cottage cheese-like discharge. Vaginal trichomoniasis, a sexually transmitted disease caused by an anaerobic protozoan parasite, is associated with a copious yellow or green, sometimes frothy, discharge. Differential diagnosis of these infections requires a thorough history, vulvovaginal examination, and simple laboratory tests, including microscopy of the vaginal discharge. The information obtained from this workup should enable an accurate diagnosis. Topical or oral metronidazole is the treatment of choice for bacterial vaginosis; terconazole, or other antifungals, for vulvovaginal candidiasis; and oral metronidazole for trichomoniasis.

The changes seen in balloon-injured porcine femoral arteries following sympathectomy.

On the basis that vasoconstriction may contribute to restenosis following angioplasty, the influence of lumbar sympathectomy on the morphometry of femoral arteries after balloon injury was examined in a pig model. Twenty-six juvenile pigs underwent balloon de-endothelialization of the right femoral artery followed by an open bilateral lumbar sympathectomy (n = 14) or a sham sympathectomy (n = 12). Four weeks later flow was measured in femoral arteries. Animals were then killed and the femoral arteries were perfusion-fixed and harvested. Sympathectomy resulted in a significant (P = 0.04) increase in flow in both the injured (right) and uninjured (left) femoral arteries. Sympathectomy did not inhibit intimal thickening following balloon injury: median (interquartile range) intimal area was 0.4 mm2 (0.3-0.9) in the sympathectomy group versus 0.5 mm2 (0.4-0.9) in the sham group. Sympathectomy did, however, result in a significant (P = 0.02) increase in the lumen area: 1.1 mm2 (0.8-1.8) versus 0.7 mm2 (0.6-0.9). Sympathectomy may reduce vasospasm following angioplasty with the potential for clinical benefit.

Vascular endothelial growth factor (VEGF) expression during arterial repair in the pig.

OBJECTIVES: Vascular endothelial growth factor (VEGF) is reported to be a potent and specific mitogen for endothelial cells (EC) and an inducer of angiogenesis in vivo. Originally called vascular permeability factor (VPF), VEGF also increases permeability of microvessels to circulating macromolecules. The aim of this study was to examine whether the VEGF gene was expressed in porcine arteries following denudation of EC. DESIGN: Experimental animal model with mechanical injury to large arteries. METHODS: The right iliac artery of juvenile pigs was de-endothelialised using an inflated balloon catheter. At a number of time-points after injury, these arteries were harvested together with uninjured contralateral arteries. Sections of arteries were used for RNA analysis by Northern blots and for protein localisation studies by immunohistochemistry. RESULTS: Two VEGF transcripts (2.0 kb, 4.5 kb) were markedly elevated in pig arteries soon after injury. Newly synthesised VEGF protein was located in smooth muscle cells (SMC) throughout the media of injured arteries. CONCLUSIONS: The elevated expression of VEGF by SMC in denuded porcine arteries is evidence that this cytokine plays a role in the injury response of large arteries. Since several biological activities have been identified for VEGF, the function of this cytokine in the arterial repair process remains to be determined.

Morphometric changes seen in balloon-injured porcine iliac arteries: the influence of sympathectomy on intimal hyperplasia and remodelling.

OBJECTIVES: To assess the influence of lumbar sympathectomy on intimal thickening and arterial remodelling following balloon de-endothelialisation. DESIGN: Experimental animal model with control and treated (sympathectomy) groups. METHODS: Unilateral common iliac artery de-endothelialisation was performed in 36 male pigs using a 5F balloon catheter introduced via the profunda femoris artery. Bilateral lumbar sympathectomies were performed in 18 animals. Both iliac arteries were perfusion-fixed and harvested 4 weeks later. Arterial morphometry was assessed using computer image analysis. RESULTS: Area measurements are expressed as median (interquartile range) in mm2. Balloon injury resulted in significant intimal thickening but no loss of lumen due to compensatory enlargement of the injured artery. Sympathectomy resulted in significant lumen enlargement (4.8 (2.6-6.3) vs. 1.9 (1.7-2.9)) in balloon-injured arteries. Although intimal thickening was reduced (0.9 (0.6-1.7) vs. 1.5 (0.9-2.0)), this was not statistically significant. CONCLUSIONS: Sympathectomy increases lumen area 4 weeks after balloon injury to porcine iliac arteries. This effect is due to a combination of reduced arterial wall thickening and increased arterial size.

Monocyte chemoattractant protein-1 gene expression in injured pig artery coincides with early appearance of infiltrating monocyte/macrophages.

Monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) are potent chemokines which attract circulating monocytes and neutrophils respectively to inflamed tissues. JE/MCP-1 gene expression has been previously studied in rabbit aortae after endothelial denudation and the rapid appearance of this transcript was thought to precede emigration of phagocytes. We now report MCP-1 gene expression following de-endothelialization of iliac arteries in the pig, a species which can develop spontaneous atherosclerosis. Using Northern blot analysis, we demonstrated that MCP-1 mRNA was rapidly induced in pig arteries at 2 h and continued to increase to reach a maximum at 8 h before returning to low levels at 16-24 h after injury. The increase seen for MCP-1 mRNA at 8 h was also observed for IL-8 mRNA but was not apparent for growth-related gene expressions, urokinase-type plasminogen activator (u-PA), and plasminogen activator inhibitor-1 (PAI-1). Since smooth muscle cells, endothelial cells, and phagocytes are all capable of expressing MCP-1, we examined pig arteries for immunostaining using a monoclonal antibody to human MCP-1 (5D3-F7). At 8 h after injury, the predominant cell type staining positive for MCP-1 was the monocyte/macrophage. Staining was also observed in occasional scattered neutrophils, but MCP-1 protein could not be detected in smooth muscle cells or on extracellular matrix within the sensitivity constraints posed by our methodology. Our results are consistent with invading monocyte/macrophages having a major input into the production of this chemokine in the arterial wall following injury. The fact that MCP-1 expression accompanied monocyte/macrophage presence in damaged artery, rather than preceding it, is suggestive that continued MCP-1 expression is required for functions other than chemoattraction.

Overexpression of p53 protein is an independent prognostic indicator in human endometrial carcinoma.

The important role of the p53 gene in tumour progression and cellular response to DNA damage has prompted investigation of the clinical significance of alterations to this gene. We examined both p53 overexpression and mutation of the gene in endometrial carcinoma in order to evaluate the prognostic significance of these changes. Of 122 endometrial carcinomas, 33 (27%) showed overexpression of p53 in the nucleus and 66 (54%) in the cytoplasm. Mutation in the p53 gene was found in 16 (13%) cases but showed no significant association with patient survival. Nuclear p53 overexpression was associated with poor survival (48% vs 80% alive in negative tumours 5 years post operatively, P < 0.001). In contrast, cytoplasmic p53 overexpression was associated with better survival (85% vs 55%, P < 0.001). When patients were separated into prognostic subgroups according to established clinical markers, these associations remained significant within most subgroups examined. In multivariate analysis adjusted for surgical stage, histological grade and type and vascular invasion, both nuclear p53 overexpression [hazard ratio 4.9 (95% CI 1.3-17.6). P = 0.016] and cytoplasmic overexpression [0.25 (0.06-0.98), P = 0.047] were independent prognostic factors. Immunohistochemical assessment of p53 overexpression in the nucleus and cytoplasm could provide useful prognostic information for the management of patients with endometrial cancer.

Vitamin D3 exacerbates intimal hyperplasia in balloon-injured arteries.

Although a high dose of vitamin D3 is known to cause arterial calcification and degeneration, its effect on intimal hyperplasia has never been studied. The aim of this study was to examine the influence of a moderate supplement of vitamin D3 on intimal hyperplasia in the balloon-injured rat carotid artery. Forty-four rats had balloon injury to the carotid artery; 22 were given oral vitamin D3 supplementation (0.25 microgram kg-1 day-1). Animals were killed at 4 weeks and the carotid arteries were perfusion fixed and assessed morphometrically by means of computerized image analysis of transverse sections. Mean (s.e.m.) intimal area was significantly greater in the vitamin D3-treated animals than in controls: 0.92(0.05) versus 0.71(0.07) mm2 (P = 0.02). The area of the media of both injured and uninjured arteries was not influenced by vitamin D3 administration. A small dose of vitamin D3 resulted in significant exacerbation of intimal hyperplasia in this rat carotid artery model and raises the question of the role of dietary vitamin D3 in restenosis following vascular intervention.

Expression of transforming growth factor-beta1 (TGF-beta1) and urokinase-type plasminogen activator (u-PA) genes during arterial repair in the pig.

OBJECTIVE: The transition from quiescence to proliferation in vitro is accompanied by early expression of proliferation-associated genes encoding products including cytokines and enzymes. We aimed to investigate TGF-beta1, u-PA and PAI-1 gene expressions in relation to proliferation and extracellular matrix (ECM) protein gene expressions in porcine arteries following injury. METHODS: Right iliac arteries of juvenile pigs were de-endothelialised and harvested at fixed times after injury. RNA was then extracted and analysed by Northern blot analysis. RNA transcripts in thickened neointima of arteries were examined by in situ hybridisation using digoxygenin-labelled cDNA probes. RESULTS: TGF-beta1, u-PA and PAI-1 transcripts were rapidly elevated (2-8h) and preceded a peak in histone mRNA at 24h after arterial injury. A second prolonged rise in TGF-beta1 mRNA at 4d coincided with elevated ECM protein gene expression. TGF-beta1 gene expression was detected in neointimal cells lining the arterial lumen at 4wk after injury. CONCLUSIONS: The timings of increases in TGF-beta1, u-PA and PAI-1 mRNAs in injured arteries are consistent with contributions to processes prior to proliferation. The observation of a second protracted elevation in TGF-beta1 expression is supportive of an additional role in stimulation of ECM protein synthesis. Functional specialisation exists within the thickened intima of arteries late in repair.

The role of vitamin D3 in the aetiology of abdominal aortic aneurysms.

Abdominal aortic aneurysms (AAAs) have traditionally been attributed to atherosclerosis, although there is increasing epidemiological, biochemical and genetic evidence that aneurysmal arterial disease is different from occlusive atherosclerosis. One of the most consistent biochemical findings in the aneurysmal aorta is a significant reduction in elastin protein; the cause, for this remains unclear. There is in vitro evidence that vitamin D3 (1,25 dihydrocholecalciferol) inhibits the production of elastin by smooth muscle cells. On the basis of this observation and the possibility that some subjects may be exposed to excess vitamin D3, the hypothesis that vitamin D3 may be a previously unrecognized aetiological factor in the pathogenesis of AAA is developed.

Gene expression of transforming growth factor-beta 1 and its type II receptor in giant cell tumors of bone. Possible involvement in osteoclast-like cell migration.

Giant cell tumor of bone (GCT) is a relatively rare skeletal neoplasm characterized by multinuclear giant cells (osteoclast-like cells) scattered in a mass of mononuclear cells. The currently favored hypothesis for the origin of cells within GCT is that the multinuclear giant cells are reactive osteoclasts, whereas the truly neoplastic cells are the major component of the mononuclear population. However, the pathological significance and the precise relationship of tumor cells and osteoclast-like cells in GCT have not been fully established. In this study, we evaluated two GCTs for the presence of transforming growth factor-beta 1 (TGF-beta 1) and TGF-beta type II receptor gene transcripts and attempted to establish a possible role for TGF-beta 1 in the interaction between tumor cells and osteoclast-like cells. By using in situ hybridization and Northern blot analysis, we have demonstrated that TGF-beta 1 mRNA transcript is consistently detected in both tumor mononuclear cells and osteoclast-like cells, whereas TGF-beta type II receptor gene transcript is only present in osteoclast-like cells. Moreover, isolated rat osteoclasts were tested for their ability to migrate in response to GCT-conditioned medium (GCTCM) in an in vitro chemotactic assay. Our results showed that GCTCM stimulates the migration of osteoclasts in a dose-dependent manner. Interestingly, only osteoclasts containing less than three nuclei can migrate through 12-mu pore filters. Addition of monoclonal antibody against TGF-beta significantly reduced but did not abolish the chemotactic activity of GCTCM. Moreover, TGF-beta type II receptor mRNA has been demonstrated in the normal rat osteoclasts and may be involved in the chemotactic action of TGF-beta 1. We concluded that TGF-beta 1, possibly in concert with other cytokines, is involved in the recruitment of osteoclast-like cells in GCT by acting in an autocrine or paracrine fashion.

Prognostic significance of pS2 mRNA in breast cancer.

The oestrogen-inducible pS2 protein has previously been associated with good prognosis for breast cancer patients. In 1987-1988 a series of 145 primary breast cancers were examined for pS2 mRNA using northern blots. On recent examination of mortality data, we were unable to find any association between tumour pS2 positivity and patient survival. One patient in 6 died within 5 years of surgery, regardless of pS2 status. In the oestrogen receptor positive/progesterone receptor positive tumour subgroup of patients, we found no evidence of increased survival for pS2-positive tumours. These results do not support use of pS2 as an indicator of increased survival in an average breast cancer patient population.

Detection of pS2 messenger RNA in gynecological cancers.

Estrogen-inducible pS2 mRNA was previously detected in human cancer cell lines the growth of which was sensitive to estrogen. In the present study, the expression of the pS2 gene was analyzed in 111 gynecological carcinomas. The pS2 message was detected in greatest abundance in 6 primary carcinomas of the ovary (6 of 29), 4 of these being mucinous cystadenocarcinomas. A secondary carcinoma of the ovary, and another of the omentum (1 of 4), also contained detectable levels of pS2 mRNA. Weak pS2 mRNA signals were occasionally observed in endometrial (2 of 55) and cervical carcinomas (2 of 33) as well. There was a poor correlation between estrogen receptor and pS2 mRNA in ovarian carcinomas.

Hormone-sensitive gene expression in breast tumours.

The expression of genes specifically induced by estrogens (pS2), prolactin (PIP) or progestins (Pg8) was measured in primary breast tumours. A highly augmented pS2 gene expression was evident in 55% of estrogen receptor (ER)+, progesterone receptor (PR)+ tumours but was absent in ER- PR- tumours. There was no clear cut correlation between augmented levels of PIP and Pg8 mRNAs in tumours and ER and PR status. Tumours from premenopausal patients were more likely to contain high levels of Pg8 mRNA (P less than 0.038), whereas tumours from postmenopausal patients tended to have augmented levels of PIP mRNA (P less than 0.053).

Prolidase deficiency: a patient without hydroxyproline-containing iminodipeptides in urine.

A 17-year-old girl was shown to have prolidase deficiency on the basis of the presence of large amounts of proline-containing dipeptides in urine and an almost complete absence of prolidase in plasma and erythrocytes. Unlike most earlier cases of this genetic defect our patient did not excrete hydroxyproline-containing dipeptides in her urine.

3-Hydroxy-3-methylglutaryl-coenzyme a lyase deficiency: a review.

Children with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency (HMG-CoA-LD; McKusick 24645), have inherited two areas of metabolic weakness. Firstly, they are unable to metabolize fully the carbon skeleton of leucine, and secondly, they cannot make ketone bodies in response to prolonged fasting. In the first year of life infants with HMG-CoA-LD run a high risk of developing severe hypoglycaemia which can lead to death if prompt intervention does not occur. The metabolic crisis develops when the infant is first introduced to dietary protein soon after birth, or later, when a reduced intake of glucose, often during a viral infection, results in a drain on the infant's circulating glucose levels. However, where diets are adequately adjusted to limit protein and fat intake, the metabolic handicaps of individuals with HMG-CoA-LD are not exposed and they are virtually symptomless. As children with HMG-CoA-LD grow older the incidence of hypoglycaemic attacks diminishes and they usually develop normally. This article reviews literature on cases of HMG-CoA-LD and interprets data on altered metabolism in these children.

Simultaneous assessment of fetal lung maturity and sebaceous gland function by gas chromatography.

Lipid profiles were analysed by gas chromatography of underivatised extracts from pellets and supernatant fractions of amniotic fluid (AFP and AFS). Peaks, identified as diglycerides resulting from on column decomposition of phosphoglycerides at high temperature, were obvious in gas chromatograms of AFS. The C32 diglyceride, mainly derived from dipalmityl lecithin, was not a prominent peak in AFS from pregnancies from which babies subsequently developed respiratory distress syndrome. There was evidence of a moderate increase of squalene in AFS and AFP for 10 pregnancies of 36-38 weeks gestation resulting in the birth of small for gestational age babies. The increase was not obvious for 21 out of 22 control pregnancies delivering appropriate for gestational age babies. This demonstrated that we had an assessment of two fetal functions in one chromatographic run, the ratio of C32/C34 diglycerides providing a measure of lung maturity and squalene providing an index of proximity to term or of the presence of intrauterine growth retardation.

Prenatal diagnosis of X-linked ichthyosis.

Prenatal diagnosis of X-linked ichthyosis in a case of steroid sulfatase deficiency was made at 16 weeks by the demonstration of (1) high levels of dehydroepiandrosterone sulfate in amniotic fluid; (2) gross deficiency of steroid sulfatase activity in cultured amniotic fluid cells; (3) very low estriol concentrations in maternal blood and urine; (4) increased maternal plasma dehydroepiandrosterone sulfate; and (5) a characteristic maternal urinary steroid profile with greatly increased levels of 16 alpha-hydroxydehydroepiandrosterone. The latter method is particularly useful since it requires no invasive procedures for the patient and is very specific.