Micro-autologous Fat Transplantation (MAFT) for Forehead Volumizing and Contouring.

BACKGROUND: Frontal fullness in Asians is often considered to indicate one's public popularity and leadership skills. Numerous materials and techniques have been applied clinically to recontour or volumize the frontal area, with variable results. The micro-autologous fat transplantation (MAFT) technique proposed by Lin et al. (2nd academic congress of Taiwan Cosmetic Association Taipei, Taiwan) in 2007 has demonstrated its feasibility in facial rejuvenation. In the present study, we used an innovative instrument to apply the MAFT technique to frontal augmentation with fat grafting and reported the results. METHODS: MAFT was performed on 178 patients (167 female, 11 male) during a 5-year period starting in January 2010. Fat was harvested by liposuction, processed and refined by centrifugation at 1200×g for 3 min. The purified fat was micro-transplanted for frontal contouring with the assistance of an instrument, the MAFT-GUN. The patients were followed up regularly, and photographs were taken for comparison. RESULTS: On average, the MAFT procedure took 52 min to complete. The average amount of delivered fat was 10.2 mL. The follow-up period was 34 months on average. No complications, including neurovascular injury, skin necrosis, abscess, nodulation, calcification or irregularity, were noted. A patient-rated satisfaction 5-point Likert scale demonstrated that 83.1% of all patients had favorable results (48.3% were satisfied, and 34.8% were very satisfied). CONCLUSION: The concept and technique of MAFT has changed fat grafting from an operation with unpredictable clinical results to an easy, reliable and consistent procedure. Furthermore, the use of a precisely controlled instrument enabled surgeons to perform highly accurate micro-fat grafting. In comparison with other strategies for volume restoration, the MAFT procedure demonstrated high patient satisfaction with the long-term results. Therefore, the use of MAFT as an alternative approach to forehead contouring and volumizing was addressed. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

For t 2 DNA vaccine prevents Forcipomyia taiwana (biting midge) allergy in a mouse model.

BACKGROUND: Forcipomyia taiwana (biting midge) is the most prevalent allergenic biting insect in Taiwan, and 60% of the exposed subjects develop allergic reactions. Subjects with insect allergy frequently limit their outdoor activities to avoid the annoyingly intense itchy allergic reactions, leading to significant worsening of their quality of life. Allergen-specific immunotherapy is the only known therapy that provides long-term host immune tolerance to the allergen, but is time-consuming and cumbersome. This study tested whether the For t 2 DNA vaccine can prevent allergic symptoms in For t 2-sensitized mice. MATERIALS AND METHODS: Two consecutive shots of For t 2 DNA vaccine were given to mice with a 7-day interval before sensitization with recombinant For t 2 proteins, using the two-step sensitization protocol reported previously. RESULTS: The For t 2 DNA vaccine at 50 µg prevented the production of For t 2-specific IgE (P < 0.05), as well as midge allergen-challenge-induced scratch bouts, midge allergen-induced IL-13 and IL-4 production from splenocytes, and inflammatory cell infiltrations in the lesions 48 h after intradermal challenge. CONCLUSIONS: This study is the first to demonstrate that DNA vaccine encoding midge allergen is effective in preventing allergic skin inflammation induced by biting midge. Immunotherapy using For t 2 DNA vaccine can protect mice from being sensitized by midge allergen and may be a promising treatment for biting midge allergy in the future.

Asp68His mutation in the A1 domain of human factor V causes impaired secretion and ineffective translocation.

Congenital factor V (FV) deficiency is a rare inherited disorder. We determined the mechanism of a missense mutation, Asp68His, in the A1 domain of the FV protein, is associated with severe FV deficiency. We characterized the mutant FV-Asp68His protein using in vitro expression studies by using specific secretion and degradation pathway inhibitors and analysed the intracellular translocation of the mutant protein by immunofluorescence staining. The Asp68His mutation caused very low levels of FV protein in the conditioned media, with normal specific FV activity. Similar mRNA degradation rates between FV-wild-type (wt) and FV-Asp68His mRNA showed that the Asp68His mutation does not affect FV expression at the transcriptional level. A specific secretion pathway inhibitor, brefeldin A, was used to demonstrate that the lower efficiency of transport to the outside of the cell for FV-Asp68His mutant protein compared with that of the FV-wt protein. Furthermore, we showed that the Asp68His mutation resulted in increased intracellular degradation through a MG132-mediated proteasomal degradation pathway. In the transfected cell lysates, FV-wt protein had multiple posttranslational modified forms, but the FV-Asp68His protein was not completely glycosylated. We further observed that the FV-Asp68His protein was retrieved in the endoplasmic reticulum only and did not undergo transport to the Golgi apparatus, leading to impaired secretion. These results strongly suggest that the Asp68His mutation may result in intracellular defective trafficking and enhanced degradation, and impaired secretion of FV protein.

An analysis of the implant-supported overdenture in the edentulous mandible.

This investigation examined the cumulative survival rate of the implant-supported overdenture using two types of attachments in patients treated at Show Chwan Memorial Hospital Implant Center from 1992 to 2006. Fifty-one patients (30 men and 21 women) were treated with mandibular implant-supported overdentures. Attachment systems used were the Hader bar with bilateral, cast ERA attachments (Group A, 31 patients with 15 men and 16 women, 134 implants) and the Hader bar with bilateral, distal extension cantilevers (Group B, 20 patients with 15 men and 5 women, 85 implants). Two hundred and four implants remained at the end of the follow-up period. Among failed implants, 10 implants were in Group A (failure rate: 10/134 = 7·5%), whereas five implants were in Group B (failure rate: 5/85 = 5·9%). Sixty-six point seven per cent (10/15) of failed implants were placed in the distal anterior mandible, and 33·3% (5/15) were placed in the middle anterior mandible. Survival was also examined with respect to condition of the opposing arch. Patients wearing a maxillary removable partial denture had the highest implant failure rate (5/51 = 9·8%), whereas the failure rate of the maxillary complete denture group was only 5·7%. The most frequent need for maintenance was wear over patrix component of ERA or Hader clip (n = 56). Eight patients experienced connector fracture between ERA and Hader bar, and one experienced distal extension cantilever fracture. The implant-supported overdenture can be an effective and reliable alternative to the conventional complete mandibular denture. Fewer prosthetic complications were seen in overdentures retained with distal extension cantilever attachments.

Functional characterisation of a complex mutation in the α(1,4)galactosyltransferase gene in Taiwanese individuals with p phenotype.

BACKGROUND AND OBJECTIVE: Individuals with p phenotype lack P1, P(k) and P antigens on red blood cells, presumably as a result of deficiency in the enzyme α(1,4)galactosyltransferase (A4GALT). The aim of this study was to explore the molecular background of a Taiwanese family with p phenotype. MATERIALS AND METHODS: Blood samples from two p siblings and seven family members were investigated. The coding region of the A4GALT gene was analysed by polymerase chain reaction and direct sequencing. The wild- and mutant-complementary DNAs (cDNAs) of A4GALT were cloned into an expression vector and transfected to Chinese hamster ovary (CHO) cells. P(k) expression on the transfected cells was analysed by flow cytometry and the activities of A4GALT were measured by high-performance liquid chromatography. RESULTS: The two individuals with p phenotype were homozygous for the complex mutation, which was caused by a combined deletion and insertion between nt 418 and 428. No expression of P(k) and no enzyme activity were observed in cells transfected with the mutant construct. CONCLUSION: The first case of p phenotype in Taiwan was caused by a non-functional allele resulting from a homozygous complex mutation of A4GALT gene.

In vitro organization of dissociated rat cardiac cells into beating three-dimensional structures.

When isolated beating ventricular heart cells from newborn rats were grown in tissue culture on untreated polystyrene surfaces, they showed a striking tendency to grow focally in three dimensions from the single layer cell sheets which were formed early in growth. During this process, they frequently formed miniature spherical heart-like masses, which continued to beat and grow in size. These often were somewhat lobulated in appearance, and grew up to 2 mm in diameter. Histological sections of such structures sometimes revealed evidence of appreciable orientation of the cells to each other, in fiber-like units. Electron microscope sections of such mini-hearts showed structures resembling intercalated discs between myocardial cells. The precise factors which induced the cardiac cells to apparently organize into these heart-like structures are not presently known.

Placental localization of human pregnancy--associated plasma proteins.

Frozen sections of human placenta were examined for the presence of four human pregnancy proteins, pregnancy-associated plasma proteins A and C (PAPP-A and PAPP-C), human chorionic somatomammotropin (hCS), and pregnancy zone protein (PZP), by the indirect immunofluorescence technique. Monospecific rabbit antiserums to PAPP-A, PAPP-C and hCS all stained the trophoblast cytoplasm equivalently in a continuous layer, usggesting that the same trophoblast cells synthesize all three pregnancy proteins. In contrast, PZP was localized in blood vessel walls, parenchymal structures within the villous, as well as in the trophoblast cytoplasm. Its distribution in the latter was relatively inhomogeneous, tending to be more intense on the basement membrane side.

Quantitative analysis of pregnancy-associated plasma proteins in human placenta.

By immunochemical methods and simultaneous measurements of several normal plasma proteins, human placenta was shown to contain elevated quantities of four pregnancy-associated plasma proteins (PAPP's). In the order of increasing amounts, PAPP-A, PAPP-C, PAPP-B, and human chorionic somatomammotropin (PAPP-D) all were present in placenta extracts in quantities greater than could be expected on the basis of their content in maternal blood. In sharp contrast, the placental content of pregnancy zone protein could be entirely accounted for by the maternal plasma present in the placenta. All of the PAPP's appeared to be readily extractible from placental tissue with buffered saline, the large bulk of them being solubilized in the first extraction procedure. However, absorption studies indicated that appreciable quantities of the PAPP's were still present in the insoluble placental residue after 12 sequential extractions with saline. The chorioamniotic membranes were not significantly enriched in any of the PAPP's. Immunochemical analysis of unwashed placental tissue extracts for the PAPP's IgA, and IgM (maternal blood derived), as well as albumin and transferrin (maternal and fetal blood derived), permitted calculations to be made of the amount of blood and PAPP's in placenta. On the basis of these data, it was roughly estimated that a 400-g placenta (wet weight) would occupy 312 ml in volume, and would contain 144 ml of blood. Of this blood, 36 ml would be derived from the mother.

Measurement of pregnancy-associated plasma proteins during human gestation.

Studies of four pregnancy-associated plasma proteins (PAPP's) were made on 206 plasma samples obtained from 175 pregnant women between 12 and 44 wk of gestation. The concentration of three PAPP's (A, C, and D) were assayed by quantitative crossed immunoelectrophoresis. They showed a gradual but small rate of increase during the 2nd trimester, which became more rapid in the 3rd trimester. PAPP-A continued to rise steadily during this period, while PAPP-C and PAPP-D (recently identified as human placental lactogen) tended to reach a plateau. Although PAPP-B could not be quantitated because of technical problems, it was detected in over 50% of the samples from the last 2 mo of gestation, but was almost never seen in those obtained during the 12th-28th wk of gestation. Various parameters were analyzed to determine their possible correlation with the PAPP levels during the last month of gestation. The race and age of the mother showed no influence on any of the PAPP's, while parity, sex of fetus, and infant birth weights appeared to affect the plasma concentration of some of the PAPP's. In the two instances studied, mothers of twins showed abnormally high PAPP levels.

Maternal age and birth rank of women with breast cancer.

Data from a large international case-control study of breast cancer suggested that women born to young mothers had a 25% lower risk of breast cancer. The association was not secondary to a tendency for these women themselves to have had children at early ages. The data provided no indication of a meaningful association between breast cancer risk and birth rank. Confounding was controlled by stratification according to a summary confounder score.

Malignant neoplasms among residents of a blackfoot disease-endemic area in Taiwan: high-arsenic artesian well water and cancers.

The objective of this study is to elucidate the association between high-arsenic artesian well water and cancers in endemic area of blackfoot disease, a unique peripheral vascular disease related to continuous arsenic exposure. As compared with the general population in Taiwan, both the standardized mortality ratio (SMR) and cumulative mortality rate were significantly high in blackfoot disease-endemic areas for cancers of bladder, kidney, skin, lung, liver, and colon. The SMRs for cancers of bladder, kidney, skin, lung, liver, and colon were 1100, 772, 534, 320, 170, and 160, respectively, for males, and 2009, 1119, 652, 413, 229, and 168, respectively, for females. A dose-response relationship was observed between SMRs of the cancers and blackfoot disease prevalence rate of the villages and townships in the endemic areas. SMRs of cancers were greater in villages where only artesian wells were used as the drinking water source than in villages using both artesian and shallow wells, and even greater than in villages using shallow wells only.

Tetrachlorodibenzo-p-dioxin exposure alters radial arm maze performance and hippocampal morphology in female AhR mice.

Perinatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been reported to alter spatial learning in rats tested on a radial arm maze (RAM). TCDD is believed to exert most of its effects through binding to the aryl hydrocarbon receptor (AhR). To determine whether the AhR mediates TCDD-induced alterations in spatial learning, we tested male and female AhR-knockout (AhR-/-), heterozygous (AhR+/-) and wild-type (AhR+/+) mice on the RAM. AhR+/- male and female mice were time mated, and treated dams were dosed with 5 microg TCDD/kg body weight on day 13 of gestation. When offspring reached adulthood, male and female AhR+/+, AhR+/- and AhR-/- mice from TCDD-exposed and unexposed litters were tested on the eight-arm RAM. After testing, we examined hippocampal morphology as visualized by the Timm's silver sulfide stain. TCDD-exposed female AhR+/- mice made more errors than their respective controls on the RAM and exhibited a decrease in the size of the intra- and infrapyramidal mossy fiber (IIP-MF) field of the hippocampus. None of the other TCDD-exposed groups differed from their respective control groups with regard to maze performance or hippocampal morphology. The reduction of IIP-MF field indicates a possible morphological basis for the learning deficit that was observed in the female AhR+/- mice. It is hypothesized that the effect of TCDD exposure is AhR dependent and that TCDD may alter GABAergic activity in the hippocampus of female mice during development.

Biological effects of sex hormone-binding globulin on androgen-induced proliferation and androgen metabolism in LNCaP prostate cells.

The effects of purified human sex hormone-binding globulin (SHBG) on androgen-sensitive cell proliferation were examined using a human prostatic cell line (LNCaP-FGC). Cells were grown for 5 days in medium supplemented with 10% charcoal-dextran-stripped human serum (10% CDHuS) and various concentrations of 5 alpha-dihydrotestosterone (DHT). In 10% CDHuS, without SHBG, the proliferative response of these cells to androgens was typically biphasic. At low androgen concentrations, cell yields were increased in a dose-dependent manner, reaching maximal levels at 0.3 nM DHT. However, at high androgen concentrations, cell proliferation was inhibited. Addition of purified human SHBG to the medium reduced the effectiveness of DHT on both phases of the proliferative response in a dose-dependent manner. These effects of SHBG appeared to be due primarily to the high affinity binding of DHT by SHBG. Proliferative responses induced by the synthetic androgen methyltrienolone (R1881), which binds poorly to SHBG, were not affected by added SHBG. Furthermore, analysis of the protein binding of DHT revealed that cell proliferation correlated best with the concentration of DHT not bound to SHBG. The presence of SHBG in the medium also altered the uptake and metabolism of DHT. LNCaP-FGC cells rapidly metabolized DHT to a polar glucuronidase-sensitive conjugate of DHT. In 10% CDHuS, LNCaP-FGC cells conjugated virtually all of the added DHT during the 5-day experiment. However, in medium containing SHBG, the SHBG-bound DHT remained unconjugated; more than 90% of the DHT initially bound to SHBG was present in the medium at the end of the experiment as unconjugated DHT. Uptake of radiolabeled DHT by cells was also inhibited by SHBG. In summary, these experiments provide evidence that 1) SHBG-bound DHT is not a signal for DHT-induced cell proliferation and 2) SHBG inhibits the uptake and metabolism of DHT by LNCaP-FGC cells.

An enzyme-linked immunoassay for circulating immune complexes using solid phased goat Clq.

An ELISA procedure was developed for measuring circulating immune complexes (IC), using solid phased goat Clq. The use of purified Clq from this species significantly diminished the background uptake of the enzyme-labeled goat antibodies used in the assay, in comparison with Clq isolated from human, guinea pig or rabbit serum. The test specimen results are reported as micrograms equivalent (microgram eq)/ml of heat aggregated human immunoglobulin G (HAIgG), and are based on standard curves developed with this latter reagent for each assay. The 2 World Health Organization (WHO) reference preparations for immune complex determinations (HAIgG and a human tetanus antitoxin-toxoid immune complex) were assayed by this ELISA procedure, and the results obtained were in very good agreement with the WHO established values. All the reagents in the ELISA, including the lyophilized HAIgG standard and the solid phased Clq are stable for more than one year at 4 degrees C. The range of accurate quantitation in test serum is 2-500 micrograms/ml, when using a 1:100 specimen dilution. The total incubation is less than 2 h, with no preliminary preparation of test specimens. The average concentration of IC reactivity in 126 healthy adults was 6 micrograms eq/ml, and the normal upper limit was determined to be 12 micrograms eq/ml.

Computed tomography findings of bony regeneration after radiotherapy for nasopharyngeal carcinoma with skull base destruction: implications for local control.

PURPOSE: To evaluate the response of bony destruction (BD) of the skull base following radiotherapy in nasopharyngeal carcinoma (NPC) and investigate the implications of bony regeneration (BR) on local control and its related factors. METHODS AND MATERIALS: Ninety patients with NPC with skull base destruction clearly demonstrated on computed tomography (CT) were reviewed. These patients have completed the prescribed treatment and received regular CT follow-up. A total of 338 sets of CT images of the head and neck were reviewed. The tumor response and the appearance of BR in the previous destructive part of the skull base were recorded and analyzed. The tumor response was divided into complete, partial, or no response. BR was defined as recalcification or sclerotic change with partial or complete healing in the previous osteolytic bony defect. Local failure was confirmed either by pathological or merely by imaging studies showing progression of tumor in consecutive radiological pictures. RESULTS: The distribution of specific sites of bony destruction (BD) in these patients included the sphenoid bone (68%), paracavernous sinus area (48%), petrous apex (47%), clivus (44%), pterygoid plates (20%), and others (7%). The CT showed 57 patients (63%) had BR. All were observed within 1 year after treatment. Sixty-two patients (69%) had complete tumor response after treatment. Analyzed by logistic regression method, tumor response after treatment was found to have a statistically significant correlation with BR (p = 0.0004). Most BR (55/57) was demonstrated in patients with complete tumor response. The 3-year actuarial local control rate was 54 % in these patients. The local control was quite different in the comparison of patients with BR versus those with persistent BD (77% vs. 21%, p < 0.0001). Multivariate analysis showed that patients with complete tumor response or with BR on imaging had statistically better local control than those without either of the two findings (p < 0.05). CONCLUSION: Appearance of BR at previous destructive skull base following radiotherapy for NPC patients could be clearly demonstrated on CT. Bony regeneration significantly correlated with treatment response and local control. Although the underlying significance of BR was unknown, to predict the outcome after treatment, the appearance of BR shown on CT may imply the complete eradication of tumor in this area.

Dynamics of tumor cell clonogen repopulation in a murine sarcoma treated with cyclophosphamide.

Experiments were performed to establish the extent and kinetics of tumor cell repopulation in a murine sarcoma, designated SA-NH, treated with cyclophosphamide (CY). Mice bearing 8-mm leg tumors were treated with 200 mg/kg CY which caused a transient tumor regression. Changes in the absolute clonogen content of tumors was determined by the change in TCD50 values (50% tumor control) obtained under hypoxic conditions of local tumor irradiation at different times after CY treatment until tumors regrew to the pretreatment size. For comparison, hypoxic TCD50 values were determined during the growth of tumors not treated with CY. CY greatly depleted tumors of clonogenic cells as manifested by the reduction in the control TCD50 value of 64.5 Gy to 32.8 Gy 1 day after CY treatment. The reduced TCD50 value remained unchanged for 2 weeks after treatment with CY, at which time the TCD50 began to rapidly increase, continuing until the end of the observation period of 21 days when tumors reached the pretreatment size. In contrast, there was a constant but slower increase in TCD50 values during the growth of tumors not treated with CY. The daily increase in TCD50 was more than twice as high in CY-treated than in CY-untreated tumors: 4.5 Gy/day versus 2.1 Gy/day. This implies that the rate of clonogen production in CY-treated tumors was twice as high as that of unperturbed tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

Abnormalities of sexual development in male rats with in utero and lactational exposure to the antiandrogenic plasticizer Di(2-ethylhexyl) phthalate.

Several members of the phthalate ester family have antiandrogenic properties, yet little is known about how exposure to these ubiquitous environmental contaminants early in development may affect sexual development. We conducted experiments to determine effects of in utero and lactational exposure to the most prevalent phthalate ester, di(2-ethylhexyl) phthalate (DEHP), on male reproductive system development and sexual behavior. Sprague-Dawley rats were dosed with corn oil or DEHP (0, 375, 750, or 1,500 mg/kg/day, per os) from gestation day 3 through postnatal day (PND) 21. Dose-related effects on male offspring included reduced anogenital distance, areola and nipple retention, undescended testes, and permanently incomplete preputial separation. Testis, epididymis, glans penis, ventral prostate, dorsolateral prostate, anterior prostate, and seminal vesicle weights were reduced at PND 21, 63, and/or 105-112. Additional dose-related effects included a high incidence of anterior prostate agenesis, a lower incidence of partial or complete ventral prostate agenesis, occasional dorsolateral prostate and seminal vesicle agenesis, reduced sperm counts, and testicular, epididymal, and penile malformations. Many DEHP-exposed males were sexually inactive in the presence of receptive control females, but sexual inactivity did not correlate with abnormal male reproductive organs. These results suggest that in utero and lactational DEHP exposure also inhibited sexually dimorphic central nervous system development. No major abnormalities were found in any of eight control litters, but DEHP caused severe male reproductive system toxicity in five of eight litters at 375 mg/kg/day, seven of eight litters at 750 mg/kg/day, and five of five litters at 1,500 mg/kg/day. These results demonstrate that the male reproductive system is far more sensitive to DEHP early in development than when animals are exposed as juveniles or adults. The effects of DEHP on male reproductive organs and sexual behaviors and the lack of significant effects on time to vaginal opening and first estrus in their littermates demonstrate that DEHP (and/or its metabolites) affects development of the male reproductive system primarily by acting as an antiandrogen. The pattern of effects of in utero and lactational DEHP exposure differed from patterns caused by other phthalate esters, and the preponderance of anterior prostate agenesis appears to be unique among all chemicals. These results suggest that DEHP acts partly by mechanisms distinct from those of other antiandrogens.

Expression of novel genes linked to the androgen-induced, proliferative shutoff in prostate cancer cells.

Androgens control cell numbers in the prostate through three separate pathways: (a) inhibition of cell death, (b) induction of cell proliferation (Step-1) and (c) inhibition of cell proliferation (Step-2, proliferative shutoff). The mechanisms underlying these phenomena are incompletely understood. The human prostate carcinoma LNCaP variants express these pathways as follows: LNCaP-FGC express both steps, LNCaP-LNO expresses Step-2, LNCaP-TAC expresses Step-1, and LNCaP-TJA cells express neither step. These cells facilitated the search for mediators of the androgen-induced proliferative shutoff pathway. Androgen exposure for 24 h or longer induced an irreversible proliferative shutoff in LNCaP-FGC cells. The Wang and Brown approach for identifying differentially expressed mRNAs was used to search for mediators of Step-2. Ten unique inserts were identified and from those ten, three genes were further studied. The basal expression of these genes in shutoff-negative variants was not affected by androgen exposure. They were induced by androgens in shutoff-positive LNCaP variants and the androgen receptor-transfected, shutoff-positive, MCF7-AR1 cells. These genes were induced only in the range of androgen concentrations that elicited the shutoff response. Time course analysis showed that their induction precedes the commitment point by 12-18 h. In addition, they were expressed in the normal prostate during proliferative shutoff. These features suggest that the candidate genes have a role in the regulation cascade for proliferative shutoff.

Increased incidence of cytomegalovirus but not Chlamydia pneumoniae in atherosclerotic lesions of arteries of lower extremities from patients with diabetes mellitus undergoing amputation.

AIMS: To evaluate the association between cytomegalovirus (CMV) or Chlamydia pneumoniae infection and the development of accelerated atherosclerotic lesions in patients with diabetes who are known to have an impaired immune response to infection and a high incidence of atherosclerosis. METHODS: Two hundred arterial samples from patients with diabetes who had undergone surgical amputation for gangrenous lower limbs were selected to assess the presence of CMV or C pneumoniae nucleic acid by means of the polymerase chain reaction. RESULTS: CMV nucleic acid sequences were detected in 64 of 200 (32%) samples and C pneumoniae in seven of 200 (3.5%) arterial samples with severe atherosclerosis. Of those positive for C pneumoniae, six were also positive for CMV. CONCLUSION: The significantly higher incidence of CMV nucleic acid sequences in the arterial samples of patients with diabetes supports the hypothesis that this organism is involved in the pathogenesis of atherosclerosis in patients with diabetic mellitus. It is possible that the potential role of different infectious agents in the pathogenesis of atherosclerosis might rely on their biological properties and their infectivity in hosts with varying immunological status.

Physiological role of the aryl hydrocarbon receptor in mouse ovary development.

The aryl hydrocarbon receptor (AhR) regulates the toxicity of environmental contaminants such as 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD). As the physiological role of the AhR in the ovary is unknown, the purpose of this study was to test the hypothesis that the AhR regulates the appearance and numbers of ovarian follicles. Ovaries were harvested from AhR-deficient (AhRKO) and wild-type mice on gestational day 18 (GD 18) and postnatal days (PND) 2-3, 8, 32-35, and 53. Complete serial sections of ovaries were evaluated histologically for the presence of germ cells and follicles. On GD 18, there was no difference in the number of germ cells per ovary between AhRKO and wild-type fetuses. However, by PND 2-3, AhRKO mice had significantly more fully formed primordial follicles (AhRKO = 38,440 +/- 3632 versus wild-type = 21,120 +/- 2688) and fewer single germ cells than wild-type mice (AhRKO = 12,696 +/- 1192 vs. wild-type = 18,160 +/- 720). On PND 8 and 32-35, there was no difference in the number of follicles between AhRKO and wild-type mice but by PND 53, AhRKO mice had significantly fewer antral follicles than wild-type (AhRKO = 3416 +/- 480 vs. wild-type = 6776 +/- 1024). Taken together, these results suggest that the AhR may play a role in the formation of primordial follicles and the regulation of antral follicle numbers.