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PubMed1992

Pericranial healing and the temporalis myo-osseous flap in the rabbit model.

The purpose of this animal study was to determine the rate of revascularization of a temporalis myo-osseous (TMO) flap after pericranial elevation. In 24 rabbits, the right pericranium was raised in entirety through a bicoronal flap at the first operation. The pericranium was then reapproximated in situ. The pericranium was allowed to heal for 1 to 28 days before the second operation. At the second operation, through the same bicoronal flap, right and left temporalis myo-osseous flaps were raised. The left temporalis myo-osseous flap served as a control. Revascularization and viability of the temporalis myo-osseous flaps were studied by using technetium bone scans, india ink injection studies, and histologic study. Results demonstrated that 4 days following pericranial elevation, the temporalis myo-osseous flap is viable and revascularized by the pericranium. Immediate bone scanning and india ink injection showed patent pericranial circulation to the osseous portion of the temporalis myo-osseous flap at 4 days. Histologic study confirmed the viability of the temporalis myo-osseous flap. In conclusion, after pericranial elevation, pericranial healing and revascularization were complete at 4 days. This allowed a viable temporalis myo-osseous flap to be raised successfully at this time.

Animals↗
PubMed1997

The peritoneal free flap: an anatomic study.

In view of a possible clinical application of an isolated microvascular peritoneal flap, an anatomic study was performed in order to determine the peritoneal vascular territory of the deep inferior epigastric artery. For this, the deep inferior epigastric artery was injected unilaterally with Araldite in 30 embalmed cadavers and bilaterally with india ink in 15 fresh cadavers. In 70 percent of the embalmed cadavers, a constant pattern of three branches from the deep inferior epigastric artery could be identified. The peritoneal vascular supply is not derived solely from these three branches but also from multiple small branches sprouting directly from the main stem of the deep inferior epigastric artery and from segmental and muscular branches. Therefore, classification of peritoneal branches arising from the deep inferior epigastric artery seems to be of little clinical importance. In all cases, the india ink injected in the deep inferior epigastric artery colored a similar territory of the parietal peritoneum. Considering the magnitude of the peritoneal vascularization by the deep inferior epigastric artery, implementation of an isolated free or pedicled peritoneal flap seems to be possible. Such a microvascular peritoneal flap vascularized by the deep inferior epigastric artery may be used, for example, for reconstruction of mucosal defects in the head and neck region.

Epigastric Arteries↗
PubMed1997

Arterial fasciocutaneous vascular territories of the lower leg.

The purpose of this study was to analyze the fasciocutaneous arterial circulation of the lower extremity to provide a quantitative guide to design reliable fasciocutaneous flaps. Thirty-one fresh cadaver limbs were studied using the techniques of dissection of latex injected specimens, selective ink injections, and barium latex radiographs. Fasciocutaneous perforator locations were recorded according to fascial septum of origin and distance relative to bony landmarks between the knee and the ankle. Selective ink injections of the trifurcation vessels identified four anterior tibial, three peroneal, and three posterior tibial fasciocutaneous territories. Although perforator site locations were randomly distributed along the trifurcation vessel within any vascular territory, the separate cutaneous regions that make up the fasciocutaneous territories occur in predictable locations with a measurable standard deviation. The transverse section radiographs confirmed the transverse dimensions of the vascular territories. Additionally, the summation of any two vascular territories calculated from the anatomical data conforms to the clinically observed 2.5:1 to 3:1 length-to-width ratios for fasciocutaneous flap viability as reported by Ponten and by Barclay et al. This study provides a quantitative anatomical framework using primary fasciocutaneous vascular territories to design potentially reliable fasciocutaneous flaps in the lower extremity.

Fascia↗
PubMed1997

A model for chronic nerve root compression studies. Presentation of a porcine model for controlled, slow-onset compression with analyses of anatomic aspects, compression onset rate, and morphologic and neurophysiologic effects.

STUDY DESIGN: Compression onset rate, anatomic aspects, and morphologic and neurophysiologic effects in spinal nerve roots were studied in a nerve root compression model in pigs. OBJECTIVES: To analyze the compression onset rate by measuring the gradual reduction of the inner diameter of the constrictor, the motor nerve conduction velocity by electromyography, the morphologic changes by light microscopy, and the gross and vascular anatomy by dissection and ink injections, respectively, in a model for experimental chronic nerve root compression. SUMMARY OF BACKGROUND DATA: Chronic nerve root compression is recognized to be related to back pain syndromes, including sciatica. Various aspects of morphologic and physiologic changes have been studied previously in models for acute compression and chronic nerve root irritation, but a controlled, graded chronic nerve root compression model has not been described. METHODS: An ameroid constrictor was applied around a spinal nerve root just cranial to the dorsal root ganglion. The inner diameter of this constrictor gradually becomes reduced. After 1 week or 4 weeks, electromyographic measurements were performed, and tissue samples were harvested for histologic analyses. The gross and vascular anatomy of the pigs' spinal nerve roots were studied by dissection and ink injections. RESULTS: There was a statistically significant decrease in the nerve conduction velocity in compressed compared with noncompressed spinal nerve roots after 1 week and after 4 weeks. The ameroid constrictors induced nerve fiber damage, endoneural hyperemia, bleeding, and inflammation at the compression zone. There was often a severe reduction in the number of myelinated fibers after 4 weeks. CONCLUSION: A model for controlled, chronic, partial nerve root injury using a gradual compression-onset constrictor is presented. This model allows for induction of a controlled graded chronic nerve root injury and can be used for research on basic pathophysiologic mechanisms and on the effects of various interventions on nerve root injury development.

Animals↗
PubMed2002

The vascular pattern of the human dorsal root ganglion and its probable bearing on a compartment syndrome.

STUDY DESIGN: A descriptive anatomic investigation of the vasculature of the dorsal root ganglions. OBJECTIVES: To determine whether the blood supply of the various spinal ganglions is sufficiently consistent to derive a "generic" description and illustration that would be applicable to all spinal levels, and to ascertain whether this vascular pattern is inherently predisposed to the development of a closed compartment syndrome. SUMMARY OF BACKGROUND DATA: The few previous descriptions of spinal ganglionic vasculature do not include photographic evidence showing uniformity in the arterial distribution plan at all ganglionic levels. The venous drainage, although verbally reconstructed from microscopic sections, lacks any indication of its probable role in the etiology of a compartment syndrome. METHODS: Three perinatal cadavers received latex/India ink injections, and their removed radiculomedullary systems were cleared, transilluminated, and macroscopically photographed. Paravertebral sections were grossly removed from the spines of two adult anatomic cadavers and received retrograde venous injections of a fine suspension of barium sulfate. The intervertebral foraminal tissues were then dissected from the bone, and radiographs of them were made. For comparative reference, a nerve root/ganglion complex of a rabbit was arterially injected with a more dilute preparation of the latex/India ink suspension. RESULTS: Macroscopic photographs of perinatal dorsal root ganglions showed that the pattern of the intraganglionic arterial distribution was sufficiently consistent to allow a graphic rendering and labeling of a "generic" ganglion. The series of incomplete retrograde venous injections adequately indicated the pressure labile location of a periganglionic venous plexus. CONCLUSIONS: The common development, structure, and function of the human dorsal root ganglions have resulted in the evolution of a uniform nutritional vascular pattern that can be conceptualized in a single visual image. Its plan of a primarily internal arterialization with a superficial venous drainage renders it vulnerable to the ischemic conditions consequent on external pressures and/or internal edematous swelling. This vascular arrangement may contribute to a propensity for the ganglion to develop a compartment syndrome when subjected to compression by periforaminal degenerative or neoplastic space-occupying lesions.

Adult↗
PubMed2006

Epithelial migration of the postmyringoplasty tympanic membrane.

OBJECTIVE: To determine whether the postmyringoplasty tympanic membrane (TM) behaves in a similar way compared with a healthy nonoperated eardrum by calculating and comparing the epithelial migration rate and pattern. STUDY DESIGN: Prospective, nonrandomized case-control study. Each subject who has undergone a myringoplasty and with healthy contralateral TM as control is included in this study. SETTING: Otolaryngology outpatient clinic. PATIENTS: A total of 23 subjects with 46 ears have been selected from otolaryngology outpatient clinic of the University Malaya Medical Center after excluding two subjects from this study. The study was conducted from June 2003 to September 2004. INTERVENTION: All the selected subjects' ears were examined under a microscope. Methylene blue ink dot was applied to the epithelium of the TM at the immediate vicinity of the umbo; then, weekly follow-up was performed until the migratory pattern and the rate of epithelial migration was noted. MAIN OUTCOME MEASURE: Direction and rate of epithelial migration in both study and control group were measured and compared. RESULTS: Epithelial migration did occur in postgrafted TM, from the umbo region toward the ear canal wall in a lateral radiating manner. Most of the patients (19 of 23 patients [82.6%]) showed nearly similar migratory pattern (same direction) in both the control and the study ear. None of the 46 ears showed any evidence of epithelial migration in the anteroinferior direction of TM. The rate of migration was slower in the beginning, especially near the umbo region (almost no movement at the first 1 or 2 weeks after ink dot application). For both groups, the rate of migration increased when approaching the periphery or annulus region. There was not much difference in the mean daily migratory rate between the control and the study ear (93.4 microm/d and 91.7 microm/d, respectively). The mean migratory rate was highest in subtotal perforation (97.2 microm/d) and lowest in small perforations (< 20%; 83.6 microm/d). CONCLUSION: There was no difference in epithelial migration rate and pattern between the postmyringoplasty TM and the healthy TM.

Adolescent↗
PubMed2005

Current perceptions regarding surgical margin status after breast-conserving therapy: results of a survey.

OBJECTIVE: The surgical margin status after breast-conserving surgery is considered the strongest predictor for local failure. The purpose of this study is to survey how radiation oncologists in North America (NA) and Europe define negative or close surgical margins after lumpectomy and to determine the factors that govern the decision to recommend reexcision based on the margins status. METHODS: A questionnaire was sent to active members of the European Society of Therapeutic Radiation Oncology and the American Society for Therapeutic Radiology and Oncology who had completed training in radiation oncology. Respondents were asked whether they would characterize margins to be negative or close for a variety of scenarios. A second survey was sent to 500 randomly selected radiation oncologists in the United States to assess when a reexcision would be recommended based on surgical margins. RESULTS: A total of 702 responses were obtained from NA and 431 from Europe to the initial survey. An additional 130 responses were obtained from the United States to the second survey regarding reexcision recommendations. Nearly 46% of the North American respondents required only that there be "no tumor cells on the ink" to deem a margin negative (National Surgical Adjuvant Breast and Bowel Project definition). A total of 7.4% and 21.8% required no tumor cells seen at <1 mm and <2 mm, respectively. The corresponding numbers from European respondents were 27.6%, 11.2%, and 8.8%, respectively (P <0.001). Europeans more frequently required a larger distance (>5 mm) between tumor cells and the inked edges before considering a margin to be negative. CONCLUSION: This study revealed significant variation in the perception of negative and close margins among radiation oncologists in NA and Europe. Given these findings, a universal definition of negative margins and consistent recommendations for reexcision are needed.

Attitude of Health Personnel↗
PubMed1987

Assessment of spatial distribution of growth in the elongation zone of grass leaf blades.

Knowledge about the spatial distribution of growth is essential for understanding the leaf growth process. In grasses the elongation zone is located at the base of the leaf blade and is enclosed by sheaths of older leaves. Assessment of spatial growth distribution, therefore, necessitates use of a destructive method. We used a fine needle to make holes through bases of tillers at the location of the leaf elongation zone of tall fescue (Festuca arundinacea Schreb.), then measured the displacement of the holes after a 6 or 24 h interval. Needle holes caused a 22 to 41% decrease in daily leaf elongation so experiments were conducted to investigate if the spatial distribution of growth in the elongation zone was altered. Leaf elongation rate was reduced similarly when needle holes were made within or above the zone where cell elongation occurs. Distribution of elongation within the zone was the same when estimated by displacement of needle holes or ink marks placed on the epidermis of the elongation zone after surrounding tissue had been removed. Making holes at different locations within the elongation zone did not differentially affect the relative contribution of the damaged or undamaged parts to leaf elongation. These findings demonstrate that needle holes or ink marks in paired leaves can be used to estimate the relative distribution of growth in the elongation zone of undamaged tall fescue leaf blades.

Journal Article↗
PubMedDate not supplied

Interactive tensor field design and visualization on surfaces.

Designing tensor fields in the plane and on surfaces is a necessary task in many graphics applications, such as painterly rendering, pen-and-ink sketching of smooth surfaces, and anisotropic remeshing. In this article, we present an interactive design system that allows a user to create a wide variety of symmetric tensor fields over 3D surfaces either from scratch or by modifying a meaningful input tensor field such as the curvature tensor. Our system converts each user specification into a basis tensor field and combines them with the input field to make an initial tensor field. However, such a field often contains unwanted degenerate points which cannot always be eliminated due to topological constraints of the underlying surface. To reduce the artifacts caused by these degenerate points, our system allows the user to move a degenerate point or to cancel a pair of degenerate points that have opposite tensor indices. These operations provide control over the number and location of the degenerate points in the field. We observe that a tensor field can be locally converted into a vector field so that there is a one-to-one correspondence between the set of degenerate points in the tensor field and the set of singularities in the vector field. This conversion allows us to effectively perform degenerate point pair cancellation and movement by using similar operations for vector fields. In addition, we adapt the image-based flow visualization technique to tensor fields, therefore allowing interactive display of tensor fields on surfaces. We demonstrate the capabilities of our tensor field design system with painterly rendering, pen-and-ink sketching of surfaces, and anisotropic remeshing.

Algorithms↗
PubMed2002

Constrained formation of object representations.

Viewers were presented with a rapid sequence of very brief stimulus pairs, each of which consisted of a pictured object followed by a related or unrelated word. The form of relatedness between the picture and word was manipulated across experiments (identical concept, associated concept, ink color of the picture). Recognition memory for the pictures was affected not only by whether or not paired items were conceptually identical or semantically related, but also by whether or not the words named an irrelevant feature, ink color. These results show that sequential items are integrated on the basis of similarity at whatever level is available, so that the stability of the memory representation of one or both items is increased. We propose that a common mechanism may underlie integration, priming, and selective attention.

Association Learning↗
PubMed2004

Light microscopy and computer three-dimensional reconstruction of the blood capillaries of the enamel organ of rat molar tooth germs.

We performed a light microscope and a computer three-dimensional reconstruction study of serial sections of the molar enamel organ of 3- and 5-day-old rats perfused with Indian ink through the arterial system. The tooth germs were fixed in Bouin's solution, embedded in paraffin, sectioned and stained with haematoxylin and eosin. For the three-dimensional reconstruction, light micrographs of the serial sections were digitized, and aligned using the serial EM Align software downloaded from http://synapses.bu.edu/tools/. After alignment, the boundaries of the India-ink-filled blood vessels were manually traced with a mouse using the software IGL trace (version 1.26b), also downloaded from the above website. After tracing, a three-dimensional representation of the blood vessel contours was generated in a VRML format and visualized with the help of the software Cortona Web3D viewer (version 4.0) downloaded from http://www.parallelgraphics.com/products/cortona/. Our results showed that in regions where ameloblasts are polarized the capillaries are arranged in three distinct levels: (1) penetrating and leaving capillaries in relation to the outer enamel epithelium; (2) capillaries crossing and branching inside the stellate reticulum; and (3) capillaries branching and anastomosing profusely within the stratum intermedium, thereby forming an extensive capillary plexus intimately associated with the cells of the stratum intermedium. The existence of a conspicuous capillary plexus intermingled with cells of the stratum intermedium, as shown in our results, suggests that some molecules produced by cells of the stratum intermedium could be released into the capillary plexus and thereafter carried to the dental follicle.

Animals↗
PubMed2003

Marking patch tests sites: description of a practical, clean, durable and inexpensive method.

We propose an original method to mark patch test sites. A transparency is applied on the patient's back, surrounding patch tests, immediately after their application. Marks are performed with an erasable pen on test sites and on permanent skin lesions like naevi. For further readings performed after patch test removal, the transparency applied on the patient's back permits the attribution of a positive reaction. This method is clean (without ink marks on skin and without contamination of clothing), more comfortable for the patient (allowing back washing immediately after the 1st reading, never inducing allergy to inks), durable (permitting delayed readings after several days or weeks) and inexpensive (because transparencies can be reused).

Dermatitis, Allergic Contact↗
PubMedDate not supplied

Defining negative margins in DCIS patients treated with breast conservation therapy: The University of Chicago experience.

Management of ductal carcinoma in situ (DCIS) has been evolving and the majority of women are now being treated with breast-conserving surgery and radiation therapy (i.e. breast conservation therapy [BCT]). Controversies still exist regarding the histologic features and margin status that are associated with local recurrence. The goal of this study was to review our institution's experience in patients diagnosed with DCIS and treated with BCT to determine pathologic features that can predict local recurrence, with particular emphasis on the final surgical margin status. We analyzed 103 consecutive patients with DCIS who were treated with BCT between 1986 and 2000. The slides were reviewed to determine the final margin status, type of DCIS, size of DCIS, nuclear grade, presence of necrosis and calcification, and volume of excised specimen. Margins were considered positive when DCIS touched or was transected at an inked margin. Negative margins were further categorized as close (less than 1 mm), 1--5 mm, and more than 5 mm. The size of the DCIS was determined based on either the maximal dimension on a slide or from the number of consecutive slides containing DCIS. Morphology and immunohistochemical profiles of the recurrent DCIS cases were compared with original DCIS. All patients were treated uniformly with external beam radiation therapy to the entire breast (median dose 46 Gy) with a boost to the tumor bed (median dose 14 Gy). The median follow-up was 63 months (range 7--191 months). The actuarial 5-year local control rate was 89%. The median time to local recurrence was 55 months. There were 13 local recurrences, of which 9 recurred as pure DCIS and 4 as invasive ductal carcinomas. Univariate analysis showed a significant association with local recurrence for positive margin (p=0.008), high nuclear grade (p=0.02), and young age at diagnosis (p=0.03). If margins were negative, the 5-year local control was 93%, as compared to 69% if margins were positive. A multivariate analysis showed that early age at diagnosis, positive margin status, and high nuclear grade were independently associated with local recurrence. The morphology and immunohistochemical stains of all nine recurrent DCIS were similar to those of the original DCIS. Breast conservation can be achieved with excellent local control by obtaining microscopically negative margins as strictly defined by DCIS not touching the inked surgical margins, and postoperative radiation that includes boost therapy to the tumor bed.

Adult↗
PubMed1994

Efficacy of silver nitrate pencils in the treatment of common warts.

Common warts are an important issue, especially in children and young adults. The methods proposed for common wart treatment are painful, destructive, and require special experience and devices. The chemical cauterization effect of silver nitrate on warts is noted in the classical pharmaceutical textbooks. In order to test this effect for this purpose, a placebo controlled clinical study was performed on 70 patients. On 35 randomly selected patients, a silver nitrate stick was used, and, on the other 35 patients, black ink was used. At the end of this study, of the patients treated with the silver nitrate stick, 15 (43%) were completely healed, 9 (26%) were partially healed, and 11 (31%) were not healed. Of the patients treated with black ink, 4 (11%) were completely healed, 5 (14%) were partially healed, and 26 (74%) patients were not affected. Thus the silver nitrate stick affected the warts of 24 (69%) patients. Therefore, this method appears to be an effective, economic, and easily applicable treatment for common warts.

Administration, Topical↗
PubMed2004

Assessment of the biological performance of the needle-free injector INJEX using the isolated porcine forelimb.

BACKGROUND: The development and utilization of novel needle-free injection devices in order to minimize needle stick injuries make increasing demands for suitable assay systems, which reflect the physiological situation in humans as close as possible. OBJECTIVES: It was therefore the goal of the present study to test the biological performance of a needle-free injector (INJEX) by the use of porcine skin as a model with a high predictive value for the feasibility in humans because of its close similarity to human skin. METHODS: In order to use porcine skin in the context of the underlying tissues, the isolated porcine forelimb was chosen as an assay model for use with the INJEX injector. Ink or the fluorescent dye fluorescein-isothiocyanate was injected and the penetration depth was determined metrically and dye distribution histologically. To assess the resorption of heparin, needle injection was compared with needle-free injection in a perfused limb model. RESULTS: Increasing amounts of ink increasingly penetrated into subcutaneous tissue layers in a cone-shaped manner mainly following lead structures. Penetration was hampered by skin thickness and by the deep muscle fascia, which served as a penetration barrier. Resorption of heparin was similar irrespective of injection by the use of a needle or the INJEX device. CONCLUSIONS: The isolated porcine forelimb serves as a versatile tool for the assessment of the biological performance of needle-free injection devices such as INJEX. Further studies are necessary to correlate the model for drug delivery in humans.

Absorption↗
PubMed1995

Pityrosporum (Malassezia) folliculitis in Saudi Arabia--diagnosis and therapeutic trials.

Pityrosporum folliculitis (PF) is frequently misdiagnosed as acne vulgaris, resulting in unnecessary and prolonged treatment. Sixty-two patients with PF seen in the Dermatology Clinic, King Gahad Hofuf Hospital, Saudi Arabia were evaluated clinically. The diagnosis was confirmed by routine histology with haemotoxylineosin staining and Periodic acid-Schiff staining. Scrapings of the lesions, especially the molluscum-like papules, were mounted in KOH/Parker blue ink and examined under the microscope. Patients, divided into three groups as follows, were given treatment for 4 weeks: (1) 20 were treated with ketoconazole, 200 mg orally in addition to ketoconazole shampoo 2% daily; (2) 20 were given only ketoconazole, 200 mg daily, orally; (3) 12 used econazole nitrate 1% solution applied twice daily; and (4) 10 used miconazole nitrate 2% cream twice daily. Cases in groups 3 and 4, who did not respond, were given the same treatment as for group 1. All who responded were kept on ketoconazole shampoo 2% twice weekly. PF was commoner in young adult females as the female to male ratio was 2:1 while the mean age was 21.5 years. The most common site involved was the trunk (95%) in the form of papules, pustules and molluscoid lesions. The latter type of lesion yielded the highest number of spores using KOH/Parker blue ink. Biopsy was positive in 87% of the patients but is usually not necessary. Combined topical and systemic ketoconazole produced clearance of the lesions in 20 patients (100%), while systemic therapy only resulted in 75% clearance (15 of 20). Topical econazole and miconazole failed in 20 of 22 (90%) and improved only two patients. There were no significant side-effects from the drugs.

Acne Vulgaris↗
PubMed1994

In vitro evaluation of apical leakage of root canal fillings after in situ obturation with thermoplasticized and laterally condensed gutta percha.

The purpose of this investigation was to compare in vitro the sealing ability of root canal treatments performed in situ with injected thermoplasticized gutta-percha compared with the lateral condensation technique. Thirty single-rooted teeth with straight canals, due for extraction for orthodontic or periodontal reasons were prepared using a step-back technique. Teeth were then divided randomly into groups A and B and obturated, respectively, with the sectional injection thermoplasticized gutta-percha and the lateral condensation techniques. Teeth were extracted after 15 days, immersed in India ink for 3 days and the leakage was determined using area-metric analysis. The roots were ground transversally and the root portion was incrementaly removed with successive steps of 250 microns each, up to the coronal level of the dye penetration. Every section was photographed with a stereoscopic microscope under the same magnification (x20); 310 photographs were taken. Measurements of the area covered by ink as well as the whole cross-sectional area of the canal were made from each photograph with the aid of a digitizing computer. The measurements were subjected to non-parametric statistical analysis. No significant difference was found (P > 0.05). The technique of lateral condensation proved to be better in the first five sections (1.2 mm from the foramen) while thermoplasticized gutta-percha technique was superior in the rest of the root canal.

Adolescent↗
PubMed1994

The effect of storage time on the degree of dye leakage of root-end filling materials.

The quality of the apical seal obtained by root-end filling materials generally has been assessed by the extent of dye penetration. Most investigators have allowed the root-end filling materials to set before they are tested. The purpose of this study was to evaluate if storage time had any influence on the amount of dye leakage. Seventy extracted single rooted human anterior teeth were used. After removal of their anatomical crowns, the roots were instrumented, filled with gutta-percha and Roth's sealer, and subjected to test conditions. The roots were coated with two coats of nail polish. Root ends were resected and class I cavities prepared. Sixty class I root-end cavities were filled with with either amalgam, super EBA, or IRM (20 each). Ten roots in each group were placed immediately into India ink for 48 h, the other 10 were stored for 24 h before placement into the ink. Ten roots (five per group) were used as positive (gutta-percha, no sealer) and negative (bone wax) controls. The roots were then demineralized, linear dye leakage was measured, and the data were evaluated statistically using ANOVA. Super EBA, and IRM showed significantly less dye leakage than amalgam (P < 0.0001). No significant difference existed between Super EBA and IRM. Storage time had no significant influence on the amount of dye leakage observed in this study.

Analysis of Variance↗